Serine hydroxymethyltransferase(SHMT),as an important enzyme involved in basic metabolism,plays an important role in plant cell metabolism,photorespiration and defense activities.To understand the bioinformatics function of the SHMT gene family in watermelon,explore its gene expression characteristics under abiotic stress,and provide a basis for the functional development of watermelon SHMT and the breeding of watermelon stress-resistance genes.Bioinformatics methods were used to identify SHMT family,and RT-qPCR was used to analyze the expression patterns of ClSHMTs in different tissues and abiotic stresses.The results showed that 8 ClSHMTs gene family members were identified in the whole genome of watermelon,which were unevenly distributed on 6 chromosomes and named ClSHMT1—ClSHMT8 in turn.There were some differences in the physical and chemical properties of each gene family member,such as the number of amino acids,molecular weight,isoelectric point.The protein contained 471—585 amino acids,with molecular weight of 51.87—65.00 ku and isoelectric point of 6.57—8.52,all of which were hydrophilic proteins.The subcellular localization prediction was mainly distributed on mitochondria.Gene structure and protein conserved motifs analysis showed that the ClSHMTs structure consisted of 4—15 exons and 3—14 introns,and all ClSHMTs contained conserved SHMT domains.Furtherly,phylogenetic analysis with 6 species such as cucumber and wheat showed that 50 SHMTs were divided into 3 sub-families,Group Ⅰ—Ⅲ.Promoter of ClSHMTs contained cis-acting elements related to light response,plant hormone response and stress response.The expression pattern analysis showed that 6 ClSHMTs were expressed in different tissues of watermelon,and the expression levels of ClSHMT1,ClSHMT4,ClSHMT5,ClSHMT8 in leaves were significantly higher than those in other tissues.Under low temperature,drought and salt stress,the expression abundance of ClSHMTs varies,but the expression was mainly up-regulated.In conclusion,this study systematically analyzed the SHMT gene family in watermelon,and will provide a reference for the further study of the biological functions of ClSHMTs.

The effects of sowing date on the yield and yield components of winter wheat,and the response characteristics of growth and development and plant type structure to accumulated temperature were studied in order to clarify the growth characteristics and reasonable plant type structure of wheat adapting to climate change.From autumn 2017 to summer 2019,field trials were conducted in Gaocheng,Hebei Province.Five sowing dates were set,September 25,October 5,October 15,October 25 and November 4.The results showed that the yield of wheat was higher sown from October 5 to 15 than other treatments,and the accumulated temperature before winter was 410-549 ℃.When accumulated temperature before winter was suitable,spike number was high and grain number per spike was moderate.When the accumulated temperature was as high as 733 ℃,the number of invalid tillers was more,the effective tiller rate was lower,and spike number was lower.When the accumulated temperature was insufficient to 279 ℃,the spike number and grain number per spike decreased.Accumulated temperature had a significant effect on growth and development indicators.Under the condition of high and stable yield,the individual index of wheat before winter was put forward:the number of main stem and tillers per plant was 2.3,the number of secondary roots was 2.5,the number of leaves of main stem was 4.1,and the spike differentiation of over winter was single edge stage.The accumulated temperature had an obvious effect on the plant structure of wheat.Delay with sowing date,the flag leaf became longer and wider,the leaf area increased,and the leaf from the top third to the top fifth became narrower and the leaf area decreased.Delay with sowing date,stem diameter of the base increased,the stem length of the top first to top second increased and length of the top third to top fifth decreased,and plant height decreased significantly.According to the equation of yield and accumulated temperature before winter,it was recommended that the suitable sowing time of wheat was October 8 to 14 under high and stable yield conditions,and the accumulated temperature range before winter was 433-541 ℃.Under late sowing conditions,the plant type structure was more reasonable,the growth and development of the plant were moderate before winter,the flag leaf was smaller and its stem length was shorter,and the leaf from the top third to the top fifth from the top was larger and their stem was slightly longer.

To select the suitable sorghum varieties for brewing in Qitai County,and provide reference for breeding and generalization of the new sorghum varieties in Xinjiang,20 new sorghum varieties were introduced from other provinces and their growth periods,agronomic traits and yields were compared to identify the highest and stable yield of each variety in 2020-2022.And the nutritional qualities such as grain starch,lysine,tannin,soluble total sugar,crude protein,crude ash and crude fiber were measured.The results showed that the growth period,agronomic traits,yields and nutritional composition were different between varieties and planting years.The average growth periods of 20 varieties were 84-146 days and plant heights ranged from 68.44 to 250.46 cm.The two species,Longza 18 and Longza 20,belonged to extremely early maturing and short varieties.The 13 short stem varieties with plant heights of less than 150 cm were identified as suitable for mechanized cultivation,such as Fengza 4,Liaonuo 11,Jiza 124,Jiza 127,Liaoza 37,Liaonian 3,Jiniang 3,Jinza 34,Jinnuo 3,Tongza 108,Jinza 109,Chiza 106 and Jinnuoliang 5.Hongyingzi was the only late-maturing high-stem variety.The yield ranged at 4 364.32-13 779.84 kg/ha,Jinza 109 had the highest yield while Shenza 5 had the lowest yield.There were differences in grain quality among different varieties;Liaoza 10 had the highest starch content of 759.93 g/kg while Jinza 34 had the lowest tannin content of 1.81 g/kg.In summary,based on the characteristics of growth period,agronomic traits,yield and quality,the tested short-stem and mid-mature variety Jinza 109 showed the highest yield,best stability and best comprehensive performance,which can be planted and popularized as the preferred variety suitable for mechanized cultivation in the Qitai area.

Temperature stress is one of the main nonbiological stresses that affect the quality and yield of spinach.Investigating the molecular response mechanism of spinach to temperature stress is crucial for spinach stress tolerance breeding.To provide a theoretical basis for the research of the mechanism of spinach resistant to cold stress and heat stress,this study used the cold-tolerant inbred line D3 and the heat-tolerant inbred line M10 of spinach as experimental materials and analyzed their transcriptomes and metabolomes under cold and heat stress to explore the transcriptional and metabolic mechanisms underlying spinach tolerance.Transcriptomic analysis showed that the pathways in which the DEGs were the most enriched in D3 and M10 were essentially the same under cold stress and heat stress.Metabolomics analysis showed that under cold stress,they were coenriched in the pyrimidine metabolism and lysine degradation in KEGG pathways.Under heat stress,these were mainly enriched in the tryptophan metabolism,toluene degradation,biosynthesis of various other secondary metabolites,and glycine,serine and threonine metabolism in KEGG pathways.The joint transcriptomic and metabolomic analysis indicated that through data analysis and gene annotation.SpADH(sov2g036390),SpSHMT(sov1g001130)and SpALDH-1(sov4g007150)were identified as the candidate genes for cold stress tolerance in spinach.SpALDH-1(sov4g007150),SpALDH-2(sov1g043320)and SpNPC(sov1g040610)were identified as candidate genes for heat stress in spinach.Among them,SpALDH-1(sov4g007150),which may be a regulatory gene for spinach stress tolerance,was significantly expressed under both cold and heat stress.

Tiller-related traits are important characteristics of wheat plant type,which determine plant structure and affect grain yield.In order to understand the inheritance and drought resistance of tiller-related traits in wheat under different water conditions,and to excavate the loci related to tiller-related traits,240 wheat varieties (lines) were selected as the subjects of this study,based on the phenotypic identification of tiller angle,effective tiller number and yield per unit area under normal irrigation (NI) and drought stress (DS) conditions,and the comprehensive evaluation of drought resistance,combined with 90K gene chip,genome-wide association study (GWAS) was to identify genetic loci for tiller-related traits and to screen for superior germplasm.The tiller angle,effective tiller number and yield per unit area showed significant difference,and the coefficient of variation ranged from 0.07 to 0.33.According to D-value,the drought resistance of Zhongyou 206 was the best.A total of 54 stable genetic loci significantly associated with tiller angle and other traits were detected, distributed on all chromosomes except 3D, 4D and 5D. Three identical stable loci were commonly detected under both treatments, located on chromosomes 2B, 4B, and 6B. Additionally, four pleiotropic loci were commonly detected in different traits, located on chromosomes 2B, 2D, and 5B.At the same time,the haplotype analysis of Ra_c491_902 (R²=5.45%—17.91%),which was significantly correlated with tiller angle on chromosome 2B,showed that there were three haplotypes:TA-Hap1,TA-Hap2 and TA-Hap3,the haplotypes (lines) containing TA-Hap1 were mainly derived from Huanghuai winter wheat regain.Five candidate genes related to tiller angle were screened by screening the stable genetic loci detected under different treatments.Gene annotation of the genes selected on Ra_c491_902 showed that the genes encoding cytochrome P450 family protein can be used as important genes such as regulating tillering angle,plant drought resistance and defense,to explore the association between genes and phenotype,and lay the foundation for the genetic improvement of tiller-related traits in wheat.

To clarify the regulatory effect of nitrogen(N)fertilizer reduction and postponing on the productivity of wheat-maize double cropping system in Huang-Huai-Hai Plain.The annual N fertilizer experiment of summer maize and winter wheat was established of four N application systems:annual N fertilizer application 400 kg/ha of traditional farmer treatment(F₄₀₀),10% reduction of annual N fertilizer(FN),20% reduction of annual N fertilizer(FH),and 30% reduction of annual N fertilizer(FL)from 2020 to 2023 at Jiyang Experimental Base of the Shandong Academy of Agricultural Sciences in Jinan.The grain yield,aboveground N accumulation characteristics,N use efficiency,and the nitrate residue after harvest in the 0—200 cm soil layer of wheat-maize double cropping system were tested,in order to provide the theoretical basis for further optimization of N fertilizer management in Huang-Huai-Hai Plain.The results indicated that N fertilizer postponing was optimized the grain yield of summer maize and winter wheat under the condition of N reduction,and the averaged across the three years,FL significantly increased by 9.2%—18.1%,13.5%—20.5%,and 11.1%—19.1%,respectively,compared with F₄₀₀ and FN.N fertilizer postponing improved the N accumulation rate,and promoted aboveground N accumulation at wheat-maize different growth stages,and the averaged across the three years,FL significantly increased plant N accumulation by 5.7%—12.3% and 5.0%—12.8% under silking and maturity,respectively,compared with F₄₀₀,FN,and FH,as well as 8.2%—17.2% in grain N accumulation.For winter wheat,FL and FH treatments were significantly higher than F₄₀₀ and FN at jointing,anthesis,and maturity,and the averaged across the three years,FL and FH significantly increased by 23.4%—28.1%,20.7%—26.3%,and 12.6%—20.8%,respectively,compared with F₄₀₀,FN and FH,at the same time the grain N accumulation under FL significantly increased by 16.4%,15.0% and 5.8%,respectively,compared with F₄₀₀ and FN.N fertilizer postponing optimized the N use efficiency of wheat-maize double crop system,the averaged across the three years,FL significantly increased N uptake efficiency by 4.8%—57.7% and 32.0%—72.4% of summer maize and winter wheat,respectively,compared with F₄₀₀,FN,and FH;and FL significantly increased N partial factor productivity by 68.8% and 40.4% in summer maize,respectively,compared with F₄₀₀ and FN,as well as by 38.4%—71.8% in winter wheat compared with F₄₀₀,FN,and FH.At harvest of summer maize and winter wheat,the soil nitrate residue was mainly enrichment in the 0—40 cm soil layer under four N application systems,the averaged across the three years,accounted for 40.0%,38.9%,44.9%,42.5% and 37.3%,36.9%,46.7%,38.3% of the 0—200 cm soil layer,respectively.In addition,the obvious accumulated effects in 0—200 cm soil layer nitrate residue under F₄₀₀ and FN treatments at harvest of summer maize and winter wheat,but there was the relative balance was achieved under FL and FH treatments.In conclusion,a 30% reduction of annual N fertilizer by N fertilizer postponing could optimize plant N accumulation characteristics and realized synergistic improve grain yield and N use efficiency.Therefore,FL treatment was an optimal N application system for realizing the collaborative target of high-yield,high-efficiency,and environment-friendly of wheat-maize double cropping system in Huang-Huai-Hai Plain.

Investigating the predominant genotypes of Bovine viral diarrhea virus(BVDV)infecting cattle in Tongliao,Inner Mongolia,to provide reference for the BVDV epidemiology and prevention and control.In the preliminary phase of the experiment,fecal samples from diarrheic calves were collected at a cattle farm in Tongliao,Inner Mongolia.These samples were tested using PCR to detect BVDV positivity.Positive fecal samples were then inoculated into madin-darby bovine kidney cells(MDBK)for isolation.The isolated strains were identified using RT-PCR and indirect immunofluorescence staining.Subsequently,the full-length genome of the isolates was sequenced,followed by genetic evolution analysis and genotype determination based on sequences of the 5'UTR,N^pro,and E2 genes.The results indicated that this experiment successfully isolated a strain of BVDV,designated as NM-21.Inoculation of NM-21 into MDBK did not induce cytopathic effects,indicating it was a non-cytopathic strain(NCP).Both RT-PCR and indirect immunofluorescence staining confirmed its positivity,with a virus titer of 10^-3 TCID₅₀/mL.Based on the full-length genomic sequence,and homology and genetic evolution analysis of the 5'UTR,N^pro,and E2 gene sequences,the isolate NM-21 showed the highest nucleotide homology with the BVDV-1c subtype strain NM2103(GenBank accession number ON337882.1)from Inner Mongolia,China.

Light-harvesting chlorophyll a/b binding proteins are important in plant photosynthesis and abiotic stress response.To study the characteristics of GhLhcb2A1 and its expression patterns and functions in low temperature and drought response in upland cotton,full-length CDS of GhLhcb2A1 gene was cloned from the leaf cDNA of Jimian 262 by PCR.Bioinformatic analysis was conducted to learn the basic characteristics of the gene.The expression patterns and functions in low temperature and drought response were evaluated by qRT-PCR and virus-induced gene silencing.It was shown that the length of GhLhcb2A1 CDS was 798 bp,encoding 265 amino acids.GhLhcb2A1 was highly expressed in leaves and was significantly up-regulated in leaves and roots under low temperature and drought treatment.Compared with the control,its expression maximized at 3 h under low temperature and drought in leaves with 17.42 and 30.03 folds increase respectively,whereas maximized at 6 h under low temperature and 12 h under drought in roots with 11.65 and 65.04 folds respectively.Subcellular localization assay verified that GhLhcb2A1 was expressed in the chloroplasts of cells.Compared with the control plants,GhLhcb2A1 silenced plants showed a more severe phenotype of water loss and dryness under both low temperatures and drought.The accumulated malondialdehyde content in the leaves of the silenced plants was significantly higher than that of the control,while the proline content and superoxide dismutase activity were significantly lower than those of the controls,suggesting that GhLhcb2A1 silenced plant reduced the resistance to low temperature and drought.The above results implied that this gene played a positive role in regulating low temperature and drought response.

In order to explore the difference in starch composition content and the change of rice digestion rate in rice lines. In this study, the contents of amylose (AC), total starch (TS), rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS)in 126 indica rice strains were analyzed by enzyme digestion method in vitro. And then detected the digestion rate of the cooked rice and estimated the glycemic index (eGI) of 18 rice lines with significant differences in AC,SDS and RS. The results indicated that the starch contents of these lines differ greatly, AC was 4.29%—25.58% with an average of 10.43%, TS was 71.69%—82.45% with an average of 77.73%, RDS was 43.31%—57.47% with an average of 50.07%, SDS was 18.96%—32.56% with an average of 25.26%, and RS was 0.59%—4.87%with an average of 2.39%. There was a certain correlation between eGI values and starch content in different rice lines. The eGI values of high AC were significantly lower than low AC, but it was also found that the eGI values of Diangu 2030 with low AC and Dianpan 3429 with low SDS were also low. The eGI values of high SDS were lower than low SDS, but there were also cases where the eGI values of low SDS were also low. The eGI value of high RS was significantly lower than low RS. The rice digestion rate was fast and the sugar release was the highest of all the rice stains within 30 min after the meal, which continued to decrease after 60 min. The eGI values of the lines with high AC, SDS or RS content were generally lower than those with low starch content. The grain digestion rates were different among the tested rice lines, and it suggested that the digestion rates of rice grain could be affected by other factors except the AC, SDS and RS. These results can offer valuable references for the development of low-GI rice varieties.

To provide theoretical basis and technical support for rational nitrogen fertilizer management in high-yield cultivation of hybrid wheat,this study aims to investigate the effects of nitrogen application rate on yield formation,dry matter accumulation and distribution,nitrogen absorption and utilization,as well as the nitrogen absorption ratio of wheat fertilizers in two-line hybrid wheat.From 2020 to 2021,three two-line hybrid wheat combinations and one conventional variety were used as materials.A split plot design was adopted,with nitrogen (labeled with ¹⁵N urea) treatment as the main zone and varieties as sub zones.Four nitrogen level experiments were set up at N0,N120,N180,and N240.The dry matter accumulation and distribution,plant nitrogen absorption and utilization,and grain yield of wheat organs were analyzed and measured under different treatments during the flowering and maturity stages.The results demonstrated that a highly significant effect of nitrogen application rate and combination (variety) on wheat yield and yield components.Compared with the conventional varieties Jingdong 17,the average yield of Jingmai 21 and BH9613 increased by 10.47% and 4.07% respectively,mainly due to their higher number of spikes and grains per ear.The application of nitrogen fertilizer significantly increased the number of spikes and grains per ear of wheat,but reduced the thousand grain weight.The application of nitrogen fertilizer to four varieties significantly increased the accumulation of dry matter in wheat during the flowering and maturity stages.The dry matter weight of various organs in wheat during the flowering stage was as follows:stem>leaf>spike,and during the maturity stage,it was as follows:grains>stem>spike-stalk+glume >leaf.The average values of nitrogen fertilizer agronomic utilization efficiency and nitrogen fertilizer partial productivity under different nitrogen application rates were as follows:Jingmai 21>BH9613>Jingdong 17>BH3606,which was consistent with the yield trend.The ¹⁵N atomic percentage of the four combinations (varieties),the nitrogen content from fertilizer,and the proportion of nitrogen from fertilizer all showed the following order:grains>straw,and they significantly increased with the increase of nitrogen application rate.Compared with Jingdong 17,the proportion of soil nitrogen in the three hybrid combinations (varieties) significantly increased,indicating that hybrid wheat had stronger tolerance to barrenness from the perspective of nitrogen utilization.After comprehensive consideration and analysis,the nitrogen application rate of N240 significantly increased wheat yield compared to other treatments,making it the optimal nitrogen fertilizer application rate for wheat cultivation.The comprehensive performance of the two hybrid combinations (varieties) of Jingmai 21 and BH9613 is better than the performance of the control Jingdong 17.

The aim was to elucidate the effects of different foliar fertilizer nitrogen application amounts on nitrogen accumulation,translocation,and utilization in maize.This experiment was conducted in 2021—2022 using a randomized block design,with maize variety Lihe 1 as the research object.No fertilization treatment (CK),conventional root fertilization treatment (CF),foliar nitrogen reduction treatment of 20% (LF1),foliar conventional nitrogen application treatment (LF2),and foliar nitrogen increase treatment of 20% (LF3) were set up to analyze the differences in nitrogen accumulation,translocation,and utilization of maize under different nitrogen fertilizer application amounts,no fertilization and conventional root fertilization.The results showed that the nitrogen accumulation in maize stems and leaves showed a trend of first increasing and then decreasing with the advancement of the growth stage,reaching the maximum value at the tasseling and silking stage.The nitrogen accumulation per plant gradually increased with the advancement of the growth stage and reached its maximum value at mature stage,the nitrogen accumulation per plant was highest in the LF2 treatment.The proportion of nitrogen distribution in leaves was highest before the silking stage;after the silking stage,the proportion of nitrogen distribution in grains gradually increased,reaching its peak at mature stage.The CK had the lowest proportion of nitrogen accumulation in grains,while the LF1 treatment had the highest proportion of nitrogen distribution in grains in 2021,and the LF3 treatment had the highest proportion of nitrogen distribution in grains in 2022.The nitrogen transport rate and the contribution rate of nitrogen transport to grains first increased and then decreased with the increase of nitrogen application amounts,the nitrogen harvest index and nitrogen utilization efficiency decreased with the increase of nitrogen application amounts;in 2021 and 2022,the nitrogen utilization efficiency of LF1 treatment,LF2 treatment,and LF3 treatment was higher than that of CF treatment,and the nitrogen utilization efficiency of LF1 treatment was the highest.The nitrogen absorption efficiency of foliar nitrogen application treatment for two years was higher than that of CF treatment.There were no significant differences in ear length,ear thickness,and ear row number among the treatments.CK had the longest bald tip length,and the row number and hundred grain weight of each nitrogen application treatment were higher than those of CK.CF treatment had the highest biological yield,while LF1 treatment had the highest grain yield and harvest index.The grain yield of each treatment was significantly higher than CK,and the harvest index decreased with increasing nitrogen application amounts.Therefore,maize can achieve better growth effects under LF1 foliar nitrogen application in the central and western regions of Inner Mongolia.

Soil nitrogen(N)cycling enzyme activity serves as a crucial indicator for characterizing soil fertility and N transformation.To investigate the effects of long-term application of fertilizers on the soil enzyme activities correlated with N cycling in rhizosphere soil of double-cropping rice fields in southern China,our project was based on a continue 37-year fertilization localization field experiment,including four fertilization treatments:without fertilizer as a control(CK),single fertilizer(MF),rice straw residue and mineral fertilizer(RF),and 30% organic manure and 70% mineral fertilizer(OM).The activities of N cycling enzymes in the rhizosphere soil were measured,and their correlation with soil chemical properties was analyzed.The results were as follows:compared to MF and CK treatments,OM and RF treatments significantly increased the contents of total N(TN),organic carbon(SOC),ammonium N($\mathrm{NH}_{4}^{+}-\mathrm{N}$),nitrate N($\mathrm{NO}_{3}^{-}-\mathrm{N}$)and microbial biomass N(SMBN)in rhizosphere soil,and also increased rice yield.The urease(Ure)and nitrite reductase(NiR)activities of rhizosphere soil in OM and RF treatments were significantly higher than those in MF and CK treatments.The RF treatment significantly increased rhizosphere soil hydroxylamine reductase(HyR)activities compared to the other three treatments,by 21.7%,13.0%,and 8.7%,respectively.This finding shown that OM treatment significantly increased protease(Pro),nitrogenase(Nit),nitrate reductase(NR)and nitrous oxide reductase(Nos)in rhizosphere soil compared to RF,MF and CK treatments.In comparison to MF treatment,OM treatments increased Pro,Nit,NR and Nos activities in rhizosphere soil by 20.0%,26.1%,426.1% and 26.7%,respectively.Nonetheless,the activity of nitric oxide reductase(Nor)on rhizosphere soil was considerably higher in the CK treatment than in MF,RF and OM treatments.Pearson correlation analysis revealed a substantial positive correlation between soil NR,NiR,Nit,Nos,Ure,Pro and soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN as well as rice yield.Soil Nor activity was observed to have a significantly negative connection with soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN and rice yield.The findings presented above showed that soil chemical properties and yield were substantially related to rhizosphere soil N cycling enzyme activities.Redundancy analysis(RDA)showed that the first order axis could explain 93.34% of the enzyme activity in rhizosphere soil and soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$,TN and SOC contents were the key factors affecting the pattern of rhizosphere soil enzyme activities.Therefore,the long-term application of organic materials such as organic manure and rice straw can enhance soil chemical and biological characteristics,stimulate soil N cycling enzyme activities,and effectively fertilize paddy soils by partially replacing chemical fertilizers.

To explore candidate genes for resistance of Aphis gossypii to the insecticide imidacloprid,the transcriptome data of the two strains were obtained and compared with imidacloprid indoor resistant and relatively sensitive lines of Aphis gossypii by using Illumina HiSeq 2500 high-throughput sequencing technology.The gene annotation was performed using the NCBI database,and bioinformatics analysis of the differential genes at the transcriptional level included GO function,KEGG metabolic pathway,and other analyses.The relative expression of eight candidate differentially expressed genes(CYP6a2,CYP6a13,CYP6k1,CYP6j1,CYP4c1,AChE2,CarE and ALP3)was detected using qRT-PCR technology,and the evolutionary relationships of resistance related genes were analyzed.After sequencing and sequence splicing,a total of 70 101 Unigenes were obtained,with an average length of 654.37 bp.29 131,27 861 and 2 993 Unigenes were annotated in NR,GO and KEGG databases,respectively.According to the NR annotation analysis of the differential genes of insecticide resistance and sensitivity strains,a total of 22 differential genes that may be related to insecticide resistance were found,including 9 detoxification enzyme genes(CYP6a13,CYP6k1,CYP6j1,CYP4c1 and ALP3 each,two CYP6a2,two CarE),8 cuticle protein genes(CP)and their precursors(CPP),target enzyme genes(AChE2),2 transcription factors(WRKY1,leucine zipper transcription factor-like protein 1 gene,LZTFL1),one pancreatic lipase-related protein 2 gene(PLRP2)and one multidrug resistance-associated protein gene(MRP).The results of qRT-PCR indicated that the expression levels of CYP6a2,CYP6a13,CYP6k1,CYP6j1,CarE and ALP3 genes in insecticide resistance strains were significantly higher than those in sensitive strains.The phylogenetic tree analysis of CYP,ALP,GST and CP genes obtained by NR annotation indicated that the genetic relationship between the Aphis gossypii and Aphis glycines and Acyrthosiphon pisum was relatively close.We found that 87.50% of the candidate differentially expressed genes in the two strains showed consistent changes in expression levels at the transcriptional and mRNA levels.

Soybean P34 protein mainly exists in soybean seeds,and its upstream promoter was likely to regulate the high expression of downstream genes in seeds.In order to further study the tissue expression pattern of soybean P34 protein gene and the regulatory activity of soybean P34 protein gene promoter,qRT-PCR was used to detect the expression of soybean P34 protein gene in soybean tissues.The 5'upstream sequence of soybean P34 protein gene(GmP34P)was cloned.The transcription initiation sites and cis-elements were analyzed by bioinformatics.The expression vector was constructed and the tobacco was transformed by Agrobacterium-mediated leaf disk method to detect GUS expression in transgenic tobacco.The results showed that the expression of P34 protein gene in soybean seeds was significantly higher than that in roots,stems,leaves and flowers.The length of GmP34P sequence obtained by cloning was 1 380 bp.Predictive analysis showed that the transcriptional start site of this sequence was base A at position 1 342,and the sequence contained a variety of cis-acting elements related to high seed expression,such as RY element,Skn-1 motif,2S seed protbanapa,etc.The plant expression vector pCAM-GmP34P containing GUS gene driven by GmP34P promoter was obtained.The positive transgenic plants were screened by hygromycin,PCR and RT-PCR.The results showed that GUS gene expression was extremely significant in transgenic tobacco seeds compared with other tissues by qRT-PCR with positive pCAM-GmP34P transgenic tobacco plants.GUS histochemical staining showed that the GmP34P promoter could regulate the high expression of downstream GUS gene in seeds.

As a key regulating factor in response to various abiotic stresses,plant heat shock transcription factor (Hsf) has a big family,and diverse structure,characteristics and functions.Hsf not only directly regulates Hsp and other relative gene expression and participates in the processes of response and adaption to various abiotic stresses,but also mediates many life activities regulation.Since the first Hsf was cloned from yeast in the 1980s,more and more Hsfs from other species have been identified and studied.In the previous reports,the identification of the Hsf family in plants was performed only in model species such as Arabidopsis and tomato.Furthermore,the studies is mainly focused on the HsfA subfamily,with few studies on the HsfB subfamily.And,the precise function of HsfC family is also largely unknown.With global climate change,the frequent occurrence of extremely high temperature events has seriously threatened the yield and quality of wheat,maize and other crops.To deal with the threat posed by heat stress,unraveling the mechanism of thermotolerance,identifying functional the targeted Hsfs and improving stress tolerance of crop through biotechnology methods is important.The number of Hsf family in field crops is various,the genome is complex,and the related research started lately compared with model species.To this end,our laboratory began to study the Hsf family of crops in 2009.Based on the latest genomic information,we confirmed the number of members,the modular structure and the spatio-temporal expression pattern of Hsf family.At the same time,with the help of transgenic wheat and mutant by genetic transformation and the CRISPR/Cas9 clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein mediated genome editing technology,several Hsfs were cloned and their regulatory functions of thermotolerance were identified,and some mechanism of thermotolerance was clarified.Our research not only enriched the theoretical basis of thermotolerance,but also provided new germplasm for biological breeding.At present,many studies have reported on functional identification and transcriptional regulation of Hsfs,however,evidences lack on which upstream component mediate Hsf's participation in regulation of thermotolerance,and the related mechanism is still unknown.Based on previous research results about wheat and maize Hsf families of lab,and many relative reports published in public,we reviewed the roles and mechanisms of plant Hsf in regulating process reported in recent years,aiming to promote research in illustrating the extensive and special roles and regulation network of plant Hsf family further,and dig useful genes and selective QTLs for biological breeding for plant thermotolerance.

In order to cultivate good quality rice,it utilized 139 rice germplasm resources from home and abroad as materials to analyze the total protein content of rice grains in 2022-2023,combined with 255 501 SNP markers obtained from whole-genome sequencing(depth of coverage of 10×),and performed genome-wide linkage analysis by using a general linear model to avoid the influence of false positives to select the genes with the highest thresholds for haplotype analysis.The genes related to the total protein content of rice grains were predicted based on the results of previous studies and gene function annotation,and the relative expression of the predicted genes was analyzed by Real-time Fluorescence PCR.The results showed that the total protein content of 139 rice seeds belonged to moderate variation,with coefficients of variation of 21.66% and 20.65%,respectively,which conformed to the normal distribution.Through genome-wide association analysis,a total of 55 significant SNPs were obtained in both environments,distributed on chromosomes 1,2,4,5,6,8,11,and 12,of which 16 consecutive and with upstream and downstream intervals of no more than 100 kb SNPs were distributed on chromosome 11.Further haplotype analysis of genes with strong correlation between the upstream and downstream intervals within 50 kb(±50 kb)of the loci of significant SNPs on chromosome 11 was conducted,combined with the results of functional annotation of the genes and the analysis of the relative expression of the seed grain at the irrigating stage,we preliminarily hypothesized that LOC_Os11g08460 was associated with the total protein content of the seed grain of rice,which encodes the Dnak/Hsp70s protein family.In conclusion,candidate gene prediction and haplotype analysis of total protein content of 139 rice germplasm resources using genome-wide association analysis can provide new genes for genetic improvement of rice quality and accelerate the process of rice improvement.

The aim of this study is to analyze the structure and biological function of ORF2 protein of Avian hepatitis E virus (aHEV)CaHEV-GDSZ01 strain,and the recombinant ORF2 protein was expressed in E.coli prokaryotic expression system and polyclonal antibodies against ORF2 protein were prepared to verify the immunogenicity and reactivity of the recombinant protein.The physicochemical properties,structure and function of ORF2 proteins of CaHEV-GDSZ01 strain were analyzed by bioinformatics software.The prokaryotic expression plasmids of pET-32a-ORF2-His and pET-32a-ORF2 were constructed by digesting and ligating pET-32a(+)vector with ORF2 gene of CaHEV GDSZ01 strain,and then transformed into DE3 competent cells for induction and expression,and the expression form was analyzed.The recombinant ORF2 protein was purified by gel cutting,its reactivity was verified by Western Blot test,and then rabbits were immunized to obtain rabbit anti-ORF2 protein polyclonal antibody.The polyclonal antibody was identified and analyzed by Western Blot,and the antibody titer was detected by indirect ELISA.The results of bioinformatics analysis showed that the ORF2 protein of CaHEV-GDSZ01 strain was hydrophilic and unstable,and the probability of the existence of signal peptide was as high as 93.59%.The secondary structure of the protein was mainly irregular curl and had the structural conditions for binding antibodies.And the overall prediction score of protective antigen of ORF2 protein showed that it had good immunogenicity.ORF2 gene prokaryotic expression plasmids was successfully constructed,and the target protein was successfully expressed.SDS-PAGE showed that the protein was expressed as insoluble protein,and the protein was proved to have good reactivity by Western Blot.The titer of the prepared polyclonal antibody was 1∶102 400,and it could specifically recognize immunogen ORF2 protein,non-immunogen ORF2-His protein and truncated expressed sORF2 protein.The physicochemical properties and some biological functions of ORF2 protein was compared and analyzed,the related proteins were expressed successfully,and the rabbit polyclonal antibody against ORF2 protein was prepared.

The tail type of sheep is a complex trait formed by the interaction of genetic and environmental factors.circRNA is closely related to lipogenesis.To investigate the effect of circular RNA(circRNA)on the tail fat deposition of sheep,transcriptome sequencing and differential expression analysis of sheep tail fat were performed.Candidate circRNA associated with sheep tail fat were screened,and the regulatory network diagram of circRNA-miRNA-mRNA associated with sheep tail fat deposition was constructed,the selected circRNA were located,and their functions were verified.The results showed that a total of 679 differentially expressed circRNA were detected in the transcripts of adipose tissue of two different tail types of sheep,of which 422 were up-regulated and 257 down-regulated.Moreover,GO and KEGG functional enrichment analysis was performed on differentially circRNA target genes,which involved many biological development processes such as DNA metabolism,anatomical structure development,catabolic process,autophagy,carbohydrate absorption process,cell proliferation and lipid metabolism related to fat deposition.Target gene enrichment was involved in cell growth and apoptosis,cell motility,transport and catabolism,signal transduction,transcription and translation,amino acid anabolism and other functions,suggesting that these circRNA may participate in the deposition process of sheep tail fat through the above pathways.The selected differential circRNA_0018 was localized by fluorescence in situ hybridization and verified in the precursor adipocytes.The results showed that circRNA_0018 was a true and stable cytoplasmic ring molecule,and functional verification of circRNA_0018 showed that it could promote adipocyte differentiation.circRNA_0018 may be involved in the process of fat deposition and lipid metabolism in sheep.

In order to investigate the expression pattern and biological function of lncRNA TCONS_00143126 in E.coli type mastitis of cows in depth.This study used cDNA from bovine mammary epithelial cells as a template,and confirmed the presence of lncRNA TCONS_00143126 using PCR cloning and sequencing techniques.Subcellular localization analysis of lncRNA was performed,and potential target miRNAs and genes were predicted.The potential mechanism of its action in bovine mastitis was explored through KEGG pathway enrichment analysis.In addition,LPS was used to induce bMECs to construct an in vitro model of bovine mastitis,and the expression of lncRNA TCONS_00143126 in LPS-induced bMECs at 6,12 and 24 h was detected by RT-qPCR.The results showed that lncRNA TCONS_00143126 was real,and its expression was significantly up-regulated in LPS-induced bMECs,and it was mainly distributed in the nucleus.The results of target gene prediction and KEGG enrichment analysis showed that lncRNA TCONS_00143126 might regulate inflammatory signaling pathways such as JAK-STAT,mTOR and MAPK by targeting miRNAs(bta-miR-133a,bta-miR-193a-5p and bta-miR-375,etc.)and target genes(IFNE,SLC2A10,MEX3B),and then play a role in the inflammation of bovine mammary epithelial cells.

Bioinformatics and expression patterns of Dicer-like(DCL),Argonaute(AGO)and RNA-dependent RNA polymerase(RDR)gene families in the whole genome of Solanum habrochaites were analyzed,so as to provide references for further study on the functions of DCL,AGO and RDR gene families in the response of S.habrochaites to abiotic and viral infection.Using Arabidopsis thaliana DCL,AGO and RDR genes as reference sequences,the genome of S.habrochaites LA1777 was searched by local perl language and software such as Pfam and SMART,and the members of ShDCL,ShAGO and ShRDR gene families were determined.Bioinformatics analysis of DCL,AGO and RDR family genes in S.habrochaites was carried out by means of ExPASy,GSDS 2.0,MEGA,Tbtools and SWISS-MODEL.According to abiotic stress treatment,Tomato chlorosis virus(ToCV)treatment and Real-time Fluorescence Quantitative PCR technology,the expression patterns of these genes were analyzed.Seven ShDCL,15 ShAGO and 6 ShRDR genes were identified from S.habrochaites,which were distributed on chromosome 5,7 and 6 respectively.The encoded proteins were similar in structure to DCL,AGO and RDR in other plants,and all of them contained conserved domains unique to this family.Phylogenetic analysis showed that these genes were divided into 4 subgroups,and there were high structural and functional similarities between S.habrochaites and S.lycopersicum.ShDCL2a,ShDCL2c,ShDCL3,ShDCL4,ShAGO1b,ShAGO3,ShAGO4b,ShAGO5,ShAGO7,ShAGO10a,ShAGO10b,ShRDR1,ShRDR2,ShRDR3a,ShRDR6a and ShRDR6b were significantly up-regulated after various abiotic stresses and ToCV infection.It is speculated that these genes play important roles in abiotic stress and virus infection.

The aim is to establish a rapid and accurate method for detecting Avian hepatitis E virus(aHEV)antibodies.The ORF2 protein of the CaHEV-GDSZ01 strain was expressed in an E.coli prokaryotic expression system and utilized as the coating antigen.The reaction conditions were optimized to determine the optimal working conditions,leading to the establishment of an indirect ELISA method for detecting aHEV chicken serum antibodies.The specificity,sensitivity and repeatability of the method were tested,and compared with Western Blot method,chicken clinical serum samples were preliminarily detected.The results showed that ORF2 protein was successfully expressed and purified,and Western Blot assay confirmed that ORF2 protein could specifically react with aHEV-positive chicken serum.The optimal reaction conditions for ELISA were as follows:ORF2 protein was coated with 200 ng per well at 4 ℃ overnight(12 h);blocked with 5% nonfat milk at 37 ℃ for 1-2 h;serum was diluted at 1∶1 600,incubated at room temperature for 1 h;rabbit anti-chicken IgY-HRP was diluted at 1∶5 000,incubated at 37 ℃ for 90 min;TMB was incubated at 37 ℃ for 20 min.The ELISA method could detect the positive sera of aHEV diluted 51 200 times,and did not cross-react with positive sera of other chicken-borne viruses such as NDV,AIV-H5,AIV-H9,ALV,REV,and MDV.The intra-batch and inter-batch coefficients of variation were both less than 10%,rate between the Western Blot method was 95.12%. Clinical serum samples from chicken farms in different areas of Guangdong were tested by this ELISA method, and the total positive rate reached 66.34%.In conclusion,this study successfully established a simple,effective,specific,sensitive and reproducible serological detection method for aHEV infection,which can provide technical support for the surveillance,prevention and control of aHEV infection at grassroots.

WRKY is a unique class of transcription factors in plants,which plays an important role in plant abiotic stress response,seed dormancy and germination,growth and development,etc.In order to reveal the function and underlying molecular mechanism of GmWRKY44 gene in soybean WRKY transcription factor family,bioinformatics analysis and biology function verification of soybean Williams 82 GmWRKY44 were performed.GmWRKY44 gene was 1 077 bp in length and encoded 358 amino acids;the results of structural prediction and evolutionary analysis showed that,the secondary structure was composed of 23.46% α-helix,4.75% β-fold,58.94% irregular coil and 12.85% extended chain,and the tertiary structure was unified with the secondary structure;It contained a conserved WRKY domain, the zinc finger structure was of the C₂H₂ type, and it belonged to the WRKY IIc subfamily; GmWRKY44 is a homologous gene of Arabidopsis thaliana AtWRKY71 with a similarity of 35.56%, and the two genes had similar gene structures. RT-qPCR analysis showed that GmWRKY44 responded to salt stress and its expression level first decreased and then increased.Under salt stress,the germination rate and root length of wild-type(Col-0)and GmWRKY44 overexpressing Arabidopsis lines were inhibited to a certain extent,but GmWRKY44 overexpressing lines were significantly better than Col-0.In addition,under salt stress,the growth inhibition of GmWRKY44 overexpressing lines was lower than that of Col-0.Physiological index analysis revealed that under salt stress,the overexpression lines of GmWRKY44 exhibited significantly higher activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)than Col-0,while the content of malondialdehyde(MDA)was significantly lower than Col-0.These data indicated that overexpression of GmWRKY44 could improve salt tolerance in transgenic Arabidopsis.

To study the structure and function of phosphotyrosine interaction domain 1 (PID1) gene in yak,and to explore its expression in various tissues.The CDS region of Sangsang yak PID1 gene was cloned using Sangsang yak adipose tissue cDNA as template,and the sequence was analyzed bioinformatically.Meanwhile,the relative expression level of PID1 gene was detected in seven tissues of yak namely heart,liver,spleen,lung,kidney,longissimus dorsi muscle and adipose tissue by Real-time Fluorescence Quantitative PCR(RT-qPCR).The results showed that the PID1 gene in yaks had a coding region length of 654 bp,which encoded 217 amino acids.Homology comparison showed that yak and wild yak were closely related,and the similarity reached 100%.The molecular weight of yak PID1 protein was about 24.84 ku and the theoretical isoelectric point was 6.30.According to the calculation results of instability coefficient,the instability of the protein was high (47.96),and it belonged to an unstable protein.The protein had one N-glycosylation site and 23 phosphorylation sites,with no signal peptide or transmembrane structure.The results of RT-qPCR showed that the expression of PID1 gene could be detected in all tissues,with the highest expression in the lung.The yak PID1 gene was cloned and its protein structure was analyzed.The expression of PID1 gene in yak tissue was also studied.Further study on the role of PID1 gene in yak fat deposition provided preliminary data.

To reveal the mechanism of drought resistance rice in booting stage,six single chromosome segments substitution lines (SSSLs) constructed from Oryza meridionalis and O.glumaepatula and their recipient parent Huajingxian 74 (HJX74) were used as experimental materials for potted drought treatment.Six biochemical indexes during 0,5,10 days of drought treatment and 5 days of rewatering and 11 agronomic traits after setting stage were measured,and the drought tolerance of 7 materials was comprehensivly evaluated by correlation analysis and principal component analysis.The result showed that under drought stress,there were extremely significant difference in agronomic traits among the 7 materials.There were extremely significant difference between M78-1 and HJX74 in relative panicle length,relative empty grain number and relative grain number per panicle,and there was extremely significant difference between M148 and HJX74 in relative dented grain number,there were extremely significant differences between M107 and HJX74 in relative panicle length and relative number of secondary branches; six drought-tolerant QTLs were identified at booting stage,including qRPL1-1,qRPL2-1,qRNSB2-1,qRNDG11-1,qRNEG1-1,qRGNP1-1,which were distributed on chromosomes 1,2,and 11.The superoxide dismutase (SOD) activity and peroxidase(POD) activity increased by 3.76%—18.20% and 31.88%—100.00% after 5 days of drought,while malondialdehyde (MDA)concentration decreased by 41.07%—81.65%.After 10 days of drought,SOD activity and POD activity decreased by 9.20%—48.53% and 44.74%—79.79%,while malondialdehyde (MDA)concentration was extremely significantly higher than that on the 5th day of drought.Osmoregulatory substances such as proline,soluble sugar and soluble protein continued to increase at the 5 d and 10 d drought treatment stages,and the biochemical indexes basically returned to normal level after 5 days of rewater.The comprehensive analysis revealed that the eigenvector and contribution rates of relative seed setting rate,POD activity and proline were the largest,indicating that these three indexes could better represent the drought tolerance of rice at booting stage.In conclusion,drought stress can affect agronomic traits and biochemical indexes of rice at booting stage,and rice can regulate its metabolic process in response to drought stress.

In order to explore new approaches for high-oil peanut breeding and establish a new method for directly developing high-oil peanut germplasm,this study employed in vitro mutagenesis breeding technology to create new high-oil peanut germplasm.Jihua 9 embryo leaflet was used as mutagenic test materials,Jihua 9 and Jihua 54 were used as control test materials,and bleomycin was used as mutagenic agent.The ovules were sterilized and placed in gradient mutagenesis medium and screened for semi-lethal concentrations of bleomycin.After somatic embryos germinated into seedlings,sterile peanut seedlings were used as rootstocks,and transplanted to the field.Bioinformatics analysis of two known regulated peanut fat synthesis genes WRI1 and experimental feasibility validation by the correlation of WRI1 gene expression in grain and crude fat content of mutagenic plants were conducted.The results were best when the bleomycin was 3 mg/L.The crude fat content of IM13-3 was higher than that of Jihua 9(CK₁,test variety control)and Jihua 54(CK₂,high oil variety control).Two WRI1 genes,WRI1X2 and WRI1X1,encoding 366 and 357 amino acids,respectively,were both unstable hydrophilic proteins. WRI1 gene expression and crude fat content were significantly positively associated in grain.Bleomycin was first used as a peanut vitro mutagenesis agent,and IM13-3 was obtained with a crude fat content of 56.64%.It further proves the authenticity of Jihua 9 high oil mutant and the feasibility of peanut in vitro mutagenesis method. The gene expression level of the high-oil mutant WRI1 was determined and was significantly different from the control varieties. Demonstrate the feasibility of breeding methods for in vitro mutagenesis of peanut.

In order to analyze the alterations in the expression profile of circular RNA(circRNA)in chicken liver cancer cells line(LMH)infected by fowl adenovirus serotype 4(FAdV-4),and the regulatory role of circRNA in the FAdV-4 infecting process,transcriptome sequencing was carried out on FAdV-4-infected LMH cells and uninfected ones.Enrichment analysis of GO functions and KEGG signaling pathways was executed for differentially expressed circRNAs,and five randomly selected circRNAs were verified by Real-time Fluorescent Quantitative PCR(qRT-PCR).The results demonstrated that the circRNAs in the infected and uninfected groups were distributed on the preponderance of chromosomes,and their lengths were mainly concentrated between 300 and 1 000 bp.Differential expression analysis identified 72 circRNAs,with 32 showing significantly upregulated expression levels and 40 presenting downregulated expression levels.GO functional analysis revealed that the genes from which the differential circRNAs originated were mainly enriched in processes such as cellular processes,metabolic processes,catalytic activity,and nucleic acid-binding transcription factor activity.KEGG pathway analysis indicated that the differentially expressed circRNAs were primarily enriched in the Notch signaling pathway,RNA degradation,and the MAPK signaling pathway.The qRT-PCR results showed that the expression levels of the five verified circRNAs were consistent with the sequencing results,further validating the reliability of the sequencing results.This study analyzed the expression profile of circRNAs in FAdV-4-infected LMH cells and screened out differentially expressed circRNAs,providing data support for exploring the functions of circRNAs during the FAdV-4 infection process and the interaction mechanism between the host and FAdV-4.

This study explored the relationship between cotton GhERF14 gene and Fusarium oxysporum pathogenicity,analyzed the molecular mechanism of F.oxysporum pathogenicity,and tentatively explored the response of cotton GhERF14 gene to Fusarium wilt disease and its regulatory effect on related resistance genes,to provides some theoretical basis for breeding new cotton cultivars resistant to wilt.Gene cloning and virus-induced gene silencing(VIGS)were used to construct the non-conserved domain interference vector pTRV2-GhERF14.Using Real-time fluorescence quantification(qRT-PCR)technology and VIGS technology,the expression characteristics of GhERF14 and downstream genes related to lignin,ethylene(ET),jasmonic acid(JA),salicylic acid(SA),antioxidant enzymes and disease progression-related protein(PR)were analyzed after F.oxysporum stress and hormone treatment,and the role of GhERF14 in the process of cotton disease resistance was analyzed.The results indicated that inhibition of GhERF14 gene expression could significantly reduce the synthesis of jasmonic acid(JA),salicylic acid(SA)and ethylene(ET)and the expression of genes related to the signaling pathway.After GhERF14 gene silencing by VIGS technology,cotton plants were more susceptible to Fusarium wilt.These results suggested that GhERF14 may play an important role in the pathogenesis and host-pathogen interaction of F.oxysporum.

The purpose of this study was to screen the candidate genes and key pathways related to traits at first laying through transcriptomic analysis of liver tissues on laying and non-laying Kangle yellow chickens,and to provide a theoretical basis for studying the molecular mechanisms of laying traits regulation in the liver of chickens.It also provides certain reference for the selection and breeding of Kangle yellow chickens.Three individuals of liver tissues at 154 days of age that had started laying (Group H) and had not started laying (Group L) were selected.Total RNA was extracted using the TRIzol method,and transcriptome sequencing was performed using the Illumina sequencing platform.Differentially expressed genes between the two groups were identified.The differentially expressed genes were subjected to functional enrichment and protein-protein interaction analysis.Nine candidate genes were randomly selected,and their expression levels were verified by qRT-PCR in the livers of the nine individuals.A total of 21 465 genes were detected to be expressed in the liver tissues,and a total of 227 differentially expressed genes were identified,among which 48 were up-regulated and 179 were down-regulated.The qRT-PCR validation results showed that the expression trends of nine genes in the two groups of individuals were basically consistent with the RNA-Seq results,and they showed a decreasing or increasing trend in the H,M(to be laid),and L three groups.Six candidate genes for the age at egg-laying trait were initially identified,namely VTG1,VTG2,VTG3,APOV1,RBP,and RNF186.The five crucial signaling pathways were fat digestion and absorption,cholesterol metabolism,ECM-receptor interaction,estrogen signaling pathway,D-glutamine and D-glutamate metabolism.Six genes and five key signaling pathways were preliminarily identified related to the traits at the first laying of Kangle yellow chicken.

Saline-alkali soils are an important reserve arable land resource in China,and are generally characterized by high inorganic salt content,lack of organic matter and low soil fertility,which seriously affects China's food security.Therefore,there is a need to improve the saline soil condition,enhance soil fertility,and provide a more reliable guarantee for China's food security.In China,straw is an important agricultural organic waste,the resource utilization of straw has become an important part of sustainable agricultural development.Among them,straw return to field has been widely promoted as a core measure.A large number of studies have shown that by returning straw to the field in a reasonable manner,the structure of soil can be effectively improved,the organic carbon content of soil can be increased,and the fertility of soil can be improved.At present,how straw resources can be efficiently utilized in saline-alkali soils so that the organic carbon of saline-alkuli soils can be improved is a major issue facing Chinese agriculture.Therefore,this paper summarizes the different ways and amounts of straw return to the field to investigate its effects on the organic carbon pool and components of saline-alkali soils.On this basis,the shortcomings of straw return technology are proposed and solutions are found.In practice,appropriate treatment measures are taken for further realizing the improvement of saline-alkali land by straw return to the field.In addition,conducting saline-alkali land research is not only an in-depth analysis of a specific soil environment,but also a complex systematic project involving the intersection of multiple disciplines.In this process,various factors,including but not limited to land management,hydrogeology,ecosystem,and agricultural economics,must be fully considered.Through such efforts,we will be able to gain a deeper understanding of the characteristics of saline-alkali soils and provide a scientific basis and technical support for the improvement and utilization of these valuable resources.

The hexameric Paf1 (RNA polymerase Ⅱ associated factor 1) complex is a crucial transcription regulator in eukaryotes.Paf1-regulated expression of specific genes in plants is closely related to diverse biological processes including growth,development,and stress responses.In order to get information on the responses of Paf1 to abiotic stresses in common wheat,homologous sequence searches were performed to identify all of the genes encoding each of the Paf1 subunits in the wheat genome.mCherry fusions of the wheat Paf1 subunit proteins were expressed in protoplasts and tobacco leaves for determination of protein subcellular localization by fluorescence microscopy.qRT-PCR assays were conducted to profile the expression of wheat Paf1 subunit genes in response to different abiotic stresses.The results showed that,in wheat,five of the Paf1 subunits,TaVIP3,TaVIP4,TaVIP5,TaVIP6,and TaPHP,were each encoded by one set of homeologous genes while the sixth subunit TaVIP2 was encoded by two sets.Plant VIP2 sequences had an N-terminal proline-rich region with variable length,and wheat TaVIP2 sequences had an additional glutamine-rich region.Protein subcellular localization assays revealed the nuclear localization of TaVIP2,TaVIP4,TaVIP5,and TaVIP6 proteins and the nuclear and cytoplasmic localization of TaVIP3 and TaPHP proteins.Gene expression analyses revealed similar tissue-dependent constitutive expression variations and similar stress-induced expression patterns of wheat Paf1 subunit genes.These genes coordinately responded to the stress of high temperature by expression upregulation and to the stresses of salt and drought by expression downregulation.Collectively,our results suggested the involvement of expression regulation of Paf1 subunit genes in the responses of wheat to abiotic stresses.

In order to deeply explore the diversity of mycoviruses in Ustilaginoidea virens,this study used an abnormal strain Uv263 of U.virens isolated from diseased rice samples collected from Hainan Province as experimental material to identify potential mycoviruses in this strain,and analyze the relationship between the genome organization and function of mycoviruses.The results showed that strain Uv263 was infected by a novel mycovirus named Ustilaginoidea virens RNA virus 7 (UvRV7).UvRV7 was a double stranded RNA virus with 5 082 bp in total length and 60.29% GC content.UvRV7 encoded two large open reading frames (ORF1 and ORF2),which encoded the coat protein (CP) and RNA-dependent RNA polymerase (RdRP),respectively.The BlastP comparison showed that the RdRP amino acid sequence of UvRV7 shared the highest similarity with that of Thelebolus microsporus totivirus 1,at 48.49%.The results of multiple alignment based on the amino acid sequence of UvRV7 RdRP showed that the RdRP sequence contained a total of eight conserved motifs,among which the most typical GDD motif in the RdRP conserved domain was identified in the Ⅵ motif.The phylogenetic analysis showed that UvRV7 was the most closely related to Thelebolus microsporus totivirus 1 and clustered with representative viruses of the genus Victorivirus in the family Totiviridae.The results of genome organization and evolutionary analyses both indicated that UvRV7 was a novel mycovirus in the genus Victorivirus.Transmission electron microscopy observations showed that UvRV7 formed a spherical viral particle of about 45 nm.Horizontal and vertical transmission experiments showed that UvRV7 could be efficiently transmitted vertically by conidia and efficiently transmitted horizontally between vegetatively compatible strains.Taken together,this study elucidated the genome organization and evolutionary relationships of the novel mycovirus UvRV7 in U.virens,and provided a potential biocontrol agent and theoretical basis for the biological control of rice false smut.

Anoctamin 5(ANO5)is a multichannel membrane protein localized in the sarcoplasmic and sarcoplasmic reticulum that primarily plays a role in myosin membrane repair and phospholipid scrambling,mutations in the ANO5 gene can lead to jaw hypoplasia as well as various myopathies.It aimed to investigate the association of SNPs in the ANO5 gene with fat deposition traits in sheep.A population of 1 005 healthy and clearly genealogical Hu sheep male lambs was selected for the study,and PCR amplification and KASPar typing techniques were used to detect the locus polymorphisms of the ANO5 gene in the experimental population and analyze the associations with fat deposition traits.The expression level of ANO5 gene in different tissues was analyzed by qPCR.The results showed that sheep ANO5 gene was widely expressed in a variety of tissues in Hu sheep,and the highest expression of ANO5 gene was found in heart tissue compared with other tissues.Three genotypes of CC,CT and TT with the g.58010 C>T polymorphic locus were detected in the 10th intron of the sheep ANO5 gene.Descriptive statistics showed that the perirenal fat weight was the most different and had the highest degree of variability compared with other fat weights.Correlation analysis showed that fat deposition related traits were positively correlated with growth and feed efficiency traits,and the results of association analyses showed that the polymorphic locus was significantly associated with perirenal fat weight and its related traits in the Hu sheep.Among them,the perirenal fat weight of individuals with CC genotype was significantly lower than that of individuals with TT genotype.In conclusion,the g.58010 C>T mutation locus of the ANO5 gene can be used as a candidate molecular marker for perirenal fat deposition traits in Hu sheep.

In order to study the effects of different fertilization treatments on the growth and development of aromatic rice,the present experiment was conducted with Qingxiangyou 19 xiang as the experimental material,and five fertilization treatments were designed,broadcasting compound fertilizer(T1),6 cm deep application of compound fertilizer(T2),broadcasting urea(T3),6 cm deep application of urea(T4),and no fertilizer applied treatment(T5),to explore the impacts of different fertilizer applications on the yield,quality,aroma,photosynthetic efficiency,and several other physiological properties of aromatic rice.The results of the experiment showed that the different fertilizer treatments had significant effects on the yield and quality of aromatic rice.The yields of aromatic rice were significantly higher in the deep-fertilization treatments(T2 and T4)than broadcasting-fertilization treatments(T1 and T3).In addition,the yield of aromatic rice was 19.61%,20.03%,39.57% and 32.28% higher than T5 treatment under T2 and T4 treatments in 2022 and 2023,respectively.In terms of net leaf photosynthetic rate,deep fertilization treatments significantly increased the net photosynthetic efficiency of aroma rice leaves by 25.69%,15.95%,17.83% and 11.28% under T2 and T4 treatments in 2022 and 2023,respectively,compared with T5 treatment.Moreover,2-acetyl-1-pyrroline(2-AP)content,2-AP synthesis-related precursor contents,and major enzyme activities were increased in aromatic rice under the deep fertilization treatments.Compared to the T5 treatment,the 2-AP content was significantly increased in the T2 treatments,reaching 161.31,180.17 μg/kg in 2022 and 2023,respectively.Furthermore,a significant increase in precursor content and major enzyme activities were also observed under deep fertilization treatments.The contents of proline,pyrrolidine-5-carboxylic acid and 1-pyrrolidine were increased by 9.90%,10.08%,4.38% and 8.13%,8.26%,6.06% under T2 treatment in 2022 and 2023,respectively.The activities of pyrrolidine-5-carboxylic acid synthetase and proline dehydrogenase activities were enhanced by 8.72%,27.79%,5.52% and 30.91% under the T2 treatment in 2022 and 2023,respectively.In conclusion,the deep fertilization treatment was able to significantly increase the yield,quality,net photosynthetic rate of leaves and promote the biosynthesis of 2-AP in aromatic rice.

The folding mode of Chinese cabbage leaf ball is the main character that determines the appearance shape,taste and stress resistance of commercial organs.In order to explore the internal molecular mechanism of the formation of the folding mode of Chinese cabbage,we cloned the full length sequence of the transcription factor BrPIF5 gene from overlaping and outward-curling Chinese cabbage as experimental materials,and conducted bioinformatics analysis,constructed the plant overexpression vector,and used Agrobacterium to mediate the transformation into tobacco to obtain positive transformation plants.The expression level of BrPIF5 gene in tobacco was detected by qRT-PCR.The results showed that the protein encoded by BrPIF5 gene was a hydrophilic protein with a continuous and complete open reading frame of 634 bp,containing 210 amino acids.The protein was composed of more α-helical structure and random curl,including an AP2/ERF domain.BrPIF5 protein and the other 9 gene family members contained a conserved motif 1,and the position was different from that of other gene family members,which was located in the front of the protein sequence.Phylogenetic tree showed that BrPIF5 gene had close evolutionary relationship with SoPIF15,BhPIF1,BoPIF4,AtPIF4 and BrPIF4 family members.The tobacco strain with overexpression of BrPIF5 was obtained by Agrobacterum-mediated genetic transformation,and the leaves of the tobacco positive transformation strain showed inward curling.qRT-PCR showed that the expression level of BrPIF5 gene in the overlaping Chinese cabbage was higher than that in the outward-curling Chinese cabbage,and the gene expression level in the positive tobacco plants was higher than that in the control.It was further proved that BrPIF5 gene controlled the inward curling of Chinese cabbage leaves,thus promoting the formation of leaf ball folding type.

The aim was to clarify the effects of different nitrogen application levels on soil organic nitrogen fractions and nitrogen use efficiency in maize fields in the central-western region of Inner Mongolia,so as to provide a reference for the scientific management of soil nitrogen and sustainable development of modern agriculture.Six nitrogen application levels were set up,N0(0 kg/ha),N8(120 kg/ha),N12(180 kg/ha),N16(240 kg/ha),N20(300 kg/ha),and N24(360 kg/ha).The dynamic changes with effects of nitrogen application on soil total nitrogen content,particulate organic nitrogen content,light fraction organic nitrogen content and heavy organic nitrogen content,as well as maize yield and nitrogen use efficiency were analyzed at different soil layers at pre-sowing and post-harvest.The results showed that soil total nitrogen,particulate organic nitrogen,light fraction organic nitrogen,and heavy fraction organic nitrogen content decreased with deepening of the soil layer at the same nitrogen application level;soil total nitrogen content at pre-sowing increased with the nitrogen application levels.Soil total nitrogen content in the N16,N20,and N24 treatments was significantly higher than that in the N0,N8,and N12 treatments at post-harvest.Soil particulate organic nitrogen content of N16 treatment was highest in the 0—10 cm,10—20 cm,and 20—40 cm soil layers at pre-sowing,with 0.14,0.13,and 0.09 g/kg,respectively.At post-harvest,N16 treatment had the highest content in the 10—20 cm,20—40 cm,and 40—60 cm soil layers,with 0.19,0.10,and 0.09 g/kg,respectively.The highest increase of soil light fraction organic nitrogen content of 37.27% was in the N16 treatment,and the highest increase of soil heavy fraction organic nitrogen content of 7.35% was in the N24 treatment,followed by the N16 treatment,at 6.84%.The N16 treatment had the highest maize biological yield of 31 443.50 kg/ha;the highest maize economic yield of 18 526.47 kg/ha;and the nitrogen use efficiency decreased with the increase in nitrogen fertilizer application levels,N16 treatment in the nitrogen harvest index was the highest, at 79.20%.In conclusion,the more suitable nitrogen fertilizer application level in the central-western region of Inner Mongolia should be maintained under 240 kg/ha,in order to achieve the best soil nitrogen management ang crop yield.

The purpose of this study is to screen candidate genes related to the body weight at 8 weeks age of Kangle yellow chicken based on the omics data,and provide the theoretical foundation for molecular marker-assisted breeding of growth traits in Kangle yellow chickens.It also provides key basic data for improving molecular breeding methods for high-quality broilers and accelerating,the progress of breed selection.To detect SNPs significantly associated with body weight traits at 8 weeks age of Kangle yellow chickens,the body weight was measured from 8 to 22 weeks of age of 434 Kangle yellow chickens.Genome-wide association study(GWAS)was performed using the gene chip technology.Genes in the candidate regions with 2 Mb windows surrounding each significant SNP were found for GO and KEGG function analysis.Combined with the data of transcriptomic sequencing and published literature,key candidate genes related to the body weight at 8 weeks age of Kangle yellow chicken were screened.Two potential SNPs significantly associated with target traits were detected,located on chromosome 2(131 485 613 bp)and chromosome 4(60 413 848 bp).A total of 118 candidate genes were screened near SNP sites.Gene function annotation analysis showed the most significant enrichment of biological processes was retinoic acid metabolism.The significant enrichment of 12 KEGG pathways was found,including fatty acid degradation,tyrosine metabolism and drug metabolism-cytochrome P450.OXR1, RSPO2,EIF3E,TRHR,BMPR1B, ADH1C, MTTP, LAMTOR3, PPP3CA and PDLIM3 were preliminarily identified as key candidate genes for body weight traits at 8 weeks age of Kangle yellow chickens.Two SNPs and 10 key candidate genes were preliminarily identified related to the body weight at 8 weeks age of Kangle yellow chicken.

This study explored the effects of intercropping and rotation on the growth,yield and quality of continuous cropping peanut,to provide theoretical basis for achieving high yield in peanut production.From 2022 to 2023,sweet potato-peanut rotation system(PSP)and maize-peanut intercropping and rotation system(PMP)were set up in the experimental farm of Henan University of Science and Technology on the basis of continuous cropping peanut for 2 years and 11 years respectively,with continuous cropping peanut as control(CCP1 and CCP2,respectively).The effects of PSP and PMP on photosynthetic characteristics,root characteristics,dry matter accumulation and distribution and yield of peanut were studied.The results showed that compared with CCP1,the leaf area index(LAI)of rotating peanut in PSP system(SRP)was significantly increased by 35.08%—53.68% and 24.32%—33.52% at pod-setting stage(PSS)and full pod maturity stage(PMS),respectively.The SPAD value at PSS and pod bulking stage(PBS)increased by 11.93%—18.55% and 5.95%—9.63%,respectively.Compared with CCP2,the LAI of rotating peanut in PMP system(MRP)increased by 46.81%—57.96% and 27.00%—61.78% at PSS and PMS,respectively.At PSS and PBS,compared with CCP2,the SPAD value of MRP and intercropping peanut(MIP)increased by 3.32%—3.69%,7.50%—8.64% and 5.47%—18.37%,15.73%—31.11%,respectively.At PSS and PBS,compared with CCP1,the net photosynthetic rate of SRP increased by 23.68%—41.31% and 26.52%—32.55%,and compared with CCP2,MRP increased by 12.77%—17.81% and 16.88%—62.07%,respectively.They both significantly improved the root length and root tip number,and promoted the dry matter accumulation and the distribution to pods during PMS,and the yields increased by 31.42%—47.36% and 54.12%—75.09%,respectively.Compared with CCP2,MIP reduced the LAI,net photosynthetic rate,root length,root tip number,as well as dry matter accumulation and yield of peanut under the influence of maize shading.At the same time,the content of peanut oleic acid and oleic acid-linoleic acid ratio was significantly increased after rotation.Among them,SRP increased by 1.63—1.65 percentage point and 6.59%—10.52%,respectively,compared with CCP1,and MRP increased by 1.95—2.82 percentage point and 9.75%—14.16% compared with CCP2,respectively.In summary,sweet potato-peanut rotation and maize-peanut rotation increased the peanut yield compared with continuous cropping peanut,the reason was that sweet potato-peanut and maize-peanut rotation promoted peanut root growth,delayed the leaf senescence,and increased photosynthetic rate,especially the photosynthetic rate during late growth period,which promoted the dry matter accumulation and distribution to seeds.Besides that,they could improve the quality of peanut to a certain extent.

To clarify the effects of different tillage methods on soil moisture dynamics changes and maize yield under the condition of straw return to field in the black soil area along the foothills of Daxing'anling,based on six consecutive years of tillage positioning experiments,this study analyzed the effects of seven types of tillage methods,namely,full-crushing and deep-tillage (SCD),full-crushing and shallow-tillage (SSS),full-crushing and deep-tillage (SCS),full-crushing and re-harrowing (SCR),full-crushing and rotary tillage (STR),full-crushing and no-tillage (NTS),and conventional tillage without returning straw to the field (CK)on soil moisture characteristics,water consumption,water use efficiency,and agronomic traits and yield of maize in the 0—60 cm soil layer at different growth stages of maize in each treatment.The results showed a bimodal pattern of soil quality moisture content in 2022 and 2023.0—10 cm soil layer soil quality moisture content was significantly higher than that of CK,and the NTS treatment had the highest soil quality moisture content in several periods.10—20 cm soil layer soil quality moisture content was lower than that of CK in the SSS and NTS treatments at the jointing stage,and soil quality moisture content was higher than that of CK in the 20—40 cm and 40—60 cm soil layers in 2022 and 2023 for all the treatments.In 2022 and 2023,maize plant height in all treatments except NTS treatment was significantly higher than CK at different growth stages.The SCD treatment was the tallest and the NTS treatment was the shortest at mature stage.Leaf area index (LAI) varied little among treatments at seedling stage of maize,STR treatment had the highest LAI after the jointing stage,and all treatments had significantly higher LAI than NTS treatment at the big trumpet stage.Dry matter accumulation in all treatments except for the SCS,NTS treatment was significantly higher than in CK,and dry matter accumulation was highest in the SCD treatment and lowest in the NTS treatment at mature stage.All tillage treatments increased maize yield and water use efficiency compared with CK,but the SCD treatment was significantly higher than CK.Comprehensive analysis of the indexes showed that the two tillage methods of full-crushing and deep-tillage and full-crushing and shallow-tillage were favorable to improve soil structure,maize yield and water use efficiency in the black soil area along the foothills of Daxing'anling.

In order to study the problem of strong winter/spring Brassica napus seed germination and flowering period under different winter sowing dates.Two strong winter rapeseeds and two spring rapeseeds provided by Gansu Agricultural University were used as materials.The experiment was carried out in the experimental field of Gansu Agricultural University from October 2022 to August 2023.The winter rapeseeds was carried out on October 11,2022.The winter/spring rapeseeds was sown every 20 days from December 10,2022,and the sowing ended on February 8,2023.The flowering period was recorded,and the germination seeds of winter rapeseed were sampled every 20 days to determine their physiological and biochemical characteristics and analyze the expression characteristics of vernalization genes(FLC,VRN2,FRI,FT).The results showed that the flowering period of winter/spring rape seeds was different by 22—34 days.The difference of flowering time between autumn sowing and spring sowing was 4—7 days.The flowering time of winter rapeseed in autumn sowing(October 11 th)was close to that of spring rapeseed under different winter sowing dates(December 10th,December 30th,January 19th,February 8th),and the flowering overlap time was as long as 15—20 days.With the delay of the sowing date,the relative expression levels of FLC,FRI and FT genes in germinating seeds of winter sowing were down-regulated.The relative expression of VRN2 gene was down-regulated in the early vernalization and up-regulated in the late vernalization.The activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and the contents of soluble protein(SP),gibberellin(GA3)and salicylic acid(SA)in germinating seeds were increased in the early vernalization,but those were decreased in the late vernalization.The contents of malondialdehyde(MDA)and abscisic acid(ABA)were increased in rapeseed germinating with the increase of vernalization time.

This study systematically investigates the effects of silicon-modified biochar (MSC) on the chemical properties of acidic soil,organic carbon and silicon fractions,and the growth of tomato plants.Silicon-modified biochar was prepared,with a focus on investigating its impacts on carbon and silicon chemical fractions,and the availability in acidic soils;tomato growth and soil microbial activity were also evaluated.The results showed that silicon-modified biochar significantly increased soil pH,cation exchange capacity,electrical conductivity,available phosphorus and potassium.MSC also raised the levels of water-soluble sodium and iron in the soil and enhanced the activities of hydrogen peroxidase and sucrase enzymes,thereby improving soil quality.Both biochar modification and unmodified biochar significantly increased the content of different carbon fractions in the soil.Compared with unmodified biochar,silicon-modified biochar significantly increased soil microbial biomass carbon(21.9%) and water-soluble organic carbon (898.3%).Furthermore,silicon-modified biochar significantly increased the contents of soil available silicon,water-soluble silicon,free silicon,active silicon,iron-manganese-bound silicon and amorphous silicon by 362.6%,158.9%,18.1%,34.9%,193.8%,and 74.1%,respectively.Meanwhile,the application of biochar promoted the growth of tomato plants and the absorption of silicon nutrients,with modified biochar showing more pronounced effects.The accumulation of plant dry matter,silicon content,and absorption rate increased by 82.0%,98.9%,and 261.5%,respectively.In summary,silicon-modified biochar significantly affected the carbon and silicon chemical forms and transformation in the soil,increased soil effectiveness and enzyme activity,thereby promoting nutrient absorption and growth of crops,demonstrating its good potential application in agricultural production.

Preliminary transcriptomic analysis identified ZmRAV1 as a candidate gene involved in maize's response to drought stress. To further investigate its function, this study cloned the ZmRAV1 gene, conducted bioinformatics analysis of its coding sequence, and overexpressed this gene in Arabidopsis thaliana. The function of ZmRAV1 was validated by assessing the phenotypes and physiological and biochemical indices of the transgenic Arabidopsis lines under drought conditions. The results showed that the ZmRAV1 gene had a total length of 1 176 bp and encoded 389 amino acids.It had the highest proportion of irregular coils in its secondary structure and was a hydrophilic protein that did not contain signal peptides and was non transmembrane.Subcellular localization indicated that the protein was located in the nucleus.ZmRAV1 exhibited high conservation across different species.Phylogenetic analysis indicated that ZmRAV1 shares the closest evolutionary relationship with its homolog in Miscanthus sinensis, showing a high degree of homology. After drought stress treatment,the root length of Arabidopsis thaliana lines overexpressing ZmRAV1 during germination was significantly higher than that of wild-type (WT)lines.In the seedling stage,WT showed withering or even death after drought stress,while the survival rate was lower than that of overexpressing lines.Moreover,the POD and SOD activities of ZmRAV1 overexpressing lines were higher than those of WT after drought treatment,indicating that overexpression of ZmRAV1 gene could enhance Arabidopsis thaliana's resistance to drought stress.

To investigate the function of lysophosphatidicacid acyltransferase 2(LPAT2)in Brassica napus,from which one copy(A07)of BnaLPAT2 was cloned by PCR.we constructed the overexpression vectors p35S∷BnaLPAT2-A07 and the seed-specific expression vector pNapin∷BnaLPAT2-A07,and by utilizing Agrobacterium-mediated genetic transformation method,obtained a total of 15 and 11 transgenic Brassica napus cv.Zhongshuang 6 respectively by PCR positive detection.Real-time Quantitative PCR(qRT-PCR)showed that the transcript levels of BnaLPAT2-A07 in most tissues of T₃ overexpressed rapeseed were higher than that of CK.However,in the seed-specific expression tissues of T₃ transgenic rapeseed,the BnaLPAT2-A07 genes were strongly expressed in the development and maturation stages of silique.Soxhlet extraction results showed that the oil content in the transgenic seeds driven by the 35S or Napin promoter accumulated 1.39 and 2.36 percentage point more oil than control seeds,respectively.The fatty acid components of transgenic rape were detected by gas chromatography.Compared with CK,the content of linolenic acid was increased by 3.13 and 1.47 percentage point,respectively.Taken together,the BnaLPAT2-A07 could promote seed oil synthesis,however,the specific selection function of BnaLPAT2-A07 for linolenic acid needs to be further verified.

Bone morphogenetic protein 6 gene(bmp6) plays an important role in regulating the formation and development of intermuscular bones in fish.In order to explore the sequence characteristics and expression characteristics of bmp6 gene in Carassius auratus var.Qihe,the coding sequence(CDS) of bmp6 gene was cloned and analyzed by bioinformatics.Through Quantitative Real-time PCR (qRT-PCR) detection,the expression differences of bmp6 gene in different tissues and developmental stages were identified.The results showed that the bmp6 gene of C.auratus var.Qihe was composed of two homologous genes,each with three alleles,namely bmp6a-1,bmp6a-2,bmp6a-3 and bmp6b-1,bmp6b-2,bmp6b-3,with full-length CDS of 1 245,1 257 bp,encoding 414 and 418 amino acids,respectively.Both bmp6a and bmp6b were unstable hydrophobic secretory proteins containing a signal peptide sequence and lacking a transmembrane structure.bmp6a was mainly distributed in mitochondria,nucleus and cytoplasm,while bmp6b was mainly distributed in nucleus and mitochondria.The secondary structure of bmp6a and bmp6b was mainly random coil,consistent with the results of tertiary structure prediction,and both contained one TGF-β-rel family conserved domain.The amino acid sequence homology analysis showed that the amino acid sequence of bmp6a had higher similarity with bmp6 of Megalobrama amblycephala,but lower similarity with mammals.The amino acid sequence of bmp6b had higher similarity with that of C.auratus and C.gibelio.Phylogenetic tree analysis showed that the bmp6a of C.auratus var.Qihe and the bmp6a of Cyprinus carpio were clustered in the same clade,while the bmp6b of C.auratus var.Qihe and the bmp6b of C.auratus and C.gibelio were clustered in the same clade.The results of qRT-PCR showed that bmp6a and bmp6b genes were expressed in brain,muscle,gill,spleen,intestine,ovary,liver and kidney tissues of C.auratus var.Qihe,but the expression levels were higher in liver,gill and kidney tissues.The expression patterns of bmp6a and bmp6b were similar in different developmental stages of juvenile C.auratus var.Qihe.The expression levels of bmp6a and bmp6b were the highest before the ossification of intermuscular spines,and then decreased with the development of intermuscular spines.To sum up,the CDS of bmp6 gene of C.auratus var.Qihe was obtained by cloning,and bioinformatics analysis and qRT-PCR detection were carried out,providing a reference for further creating a new strain of C.auratus var.Qihe without intermuscular bones.

In order to investigate the function of watermelon calcium-dependent protein kinase (CDPK) in grafted seedlings and abiotic stress environments, this study used RT-PCR technology to clone the ClCDPK(Cla97C01G019720) gene from watermelon grafted seedlings and performed bioinformatics analysis on it. Further designed specific primers with Kpn Ⅰ and Sal Ⅰ enzyme cleavage sites based on the ClCDPK sequence,conducted amplification and double enzyme cleavage, and connected with pCAMBIA1300 to successfully construct the expression vector pCAMBIA1300-35S-ClCDPK for the target gene.Using RT-qPCR technology, the gene expression levels of ClCDPK were measured in self rooted seedlings (ZG) and grafted seedlings (JJ) after being subjected to salt and drought stress, respectively.The results showed that the ORF of ClCDPK gene was 1 647 bp, encoding 548 amino acids. Its protein contained STKc_CAMK and FRQ1 functional domains, and was a hydrophilic protein. Subcellular localization prediction showed that the protein was located in the nucleus. Evolutionary tree analysis of ClCDPK with CDPK from six other plants revealed that it was closely related to CDPK from Cucurbitaceae melons and pumpkins, with protein sequence homology alignment exceeding 92.64%, indicating high homology.The RT-qPCR expression results showed that the expression level of ClCDPK in grafted seedlings was significantly higher than that in self rooted seedlings. With the duration of stress, the expression levels of ClCDPK in grafted and self rooted seedlings first increased and then decreased, and under the same stress treatment, the expression level of ClCDPK in grafted seedlings was higher than that in self rooted seedlings.This study indicated that ClCDPK responded positively to salt and drought stress, and the ability of grafted seedlings to resist stress was higher than that of self rooted seedlings. It is speculated that ClCDPK is one of the key factors in watermelon's response to grafting, thereby improving the salt and drought resistance of watermelon grafted seedlings.

To compare the effects of different nitrogen enhanced fertilizer on apple growth and to identify suitable nitrogen enhanced fertilizer and application methods for apple cultivation,this study examined the impact of four different types of nitrogen enhanced fertilizer on apple growth,yield,quality,and soil nitrogen supply capacity.The experiment consisted of seven treatments:no nitrogen fertilizer(CK),regular application of ordinary urea by farmers(U),coated urea mixed with ordinary urea 3∶7(CU1),basal application of coated urea and follow-up application of ordinary urea(CU2),loss-control urea(KSU),stabilizing urea(WDU),and humic acid stabilizing urea(FZU).A field experiment was conducted in Qixia City,Shandong Province,to analyze the effects of different fertilization treatments on the growth,yield,and quality of 6-year-old Golden Crown apple trees,as well as nitrogen utilization and soil nitrogen supply capacity.The results demonstrated that the application of diverse synergistic nitrogen fertilizers could markedly enhance spring and autumn growth,elevate leaf SPAD,and enhance apple yield and quality.Among these,the yield of the FZU treatment exhibited a significant increase of 8.89% in comparison to that of the control.The Vc content of the fruits of the CU1 treatment was found to be significantly increased by 66.73% in comparison to the U treatment.Furthermore,the CU1 treatment demonstrated the most pronounced impact on the improvement of growth in both spring and autumn.Nitrogen enhanced fertilizer had been demonstrated to significantly enhance nitrogen accumulation in new shoots and fruit nitrogen accumulation,nitrogen fertilizer agronomic efficiency,nitrogen fertilizer bias productivity and nitrogen utilization in the middle and late stages of apple fertility.Of these,the FZU treatment had been observed to exert the most pronounced effect.The nitrogen accumulation of new shoots in FZU treatment was significantly increased by 37.25%,15.91% and 37.85% compared with that in U treatment at bud differentiation,fruiting and ripening stages,respectively.The CU2 treatment was found to have the most beneficial effect on nitrogen utilization.Nitrogen enhanced fertilizer treatments demonstrated the capacity to significantly enhance the nitrate and ammonium content of soil.Among these treatments,the FZU treatment exhibited the most pronounced effect.The FZU treatment was observed to enhance soil urease activity during the flowering and bud differentiation stages.Additionally,the WDU and FZU treatments were found to significantly reduce the nitrification of soil ammonium and nitrogen loss.The {\mathrm{NO}}_3^{-}-N of the FZU treatment was concentrated at a depth of 20—60 cm soil,which was consistent with the distribution of apple roots in the soil and reduced the risk of {\mathrm{NO}}_3^{-}-N leaching.A comprehensive analysis demonstrated that the FZU treatment exhibited clear advantages in promoting apple growth,increasing apple yield and quality,and enhancing soil nutrient supply capacity.Consequently,the FZU treatment was identified as the optimal treatment.

To study the photosynthetic characteristics,fluorescence parameters and yield response to phosphorus in maize,and to clarify the optimal phosphorus application rate for maize under drip irrigation and water fertilization technology.Providing solid theoretical basis and technical support for high-yield and high-efficiency cultivation of maize in Ningxia region.The experiment was carried out at Pingjipu Farm,Yinchuan,Ningxia,from 2019 to 2020,with six phosphorus treatments in the order of 0(P0),60(P1),120(P2),180(P3),240(P4),and 300 kg/ha(P5).Analysis of the changing patterns of photosynthetic and fluorescence parameters of spring maize leaves and their correlation with yield under different phosphorus fertilizer treatments.In two years,during the big bell mouth stage,leaf area index (LAI) was increased by 4.21% to 12.78% and 4.68% to 15.60% for P3 compared to other treatments,respectively.Phosphorus fertilizer at 180 kg/ha was most effective in promoting leaf area index and photosynthetic potential(LAD) of maize.LAD was significantly increased by 14.42% under P3 treatment compared to no phosphorus fertilizer treatment during the full two year period.The photosynthetic characteristics of maize responded differently to the intensity of phosphorus application,and as the intensity of phosphorus application increased,the net photosynthetic rate (Pn) all reached the maximum value after the stamen pumping stage,and at the R1 stage of the 2 years,the Pn was significantly increased by 10.68% under the P3 treatment as compared to the no-phosphorus-fertilizer treatment.The coefficient of determination (R²) was 0.926 5,0.889 9,and 0.832 0,respectively.Phosphorus application increased the maize photosystem Ⅱ composite performance index (PI),which had its maximum peak at the R1 stage in 2 years,and PI increased by 1.12% to 8.50% and 8.47% to 15.40% under the P3 treatment compared with the other treatments,respectively.The maximum yield was obtained at 180 kg/ha of phosphorus application,which was 17.27% higher as compared to no phosphorus treatment.Based on the analysis of the yield fitting equation,it was shown that the maximum corn yield of 13 823.84 kg/ha was reached at 179.34 kg/ha of phosphorus applied.Pearson's correlation analysis showed that appropriate leaf area index significantly affected maize yield in the late stage,and the photosynthetic parameters all had highly significant effects on maize yield completion;principal component analysis showed that the P3 treatment had the highest composite score for the optimization effect on maize yield.Reasonable transportation of phosphorus fertilizer can effectively ensure higher SPAD value,PSⅡ reaction center activity,improve the capture and utilization of light energy in spring maize,and promote photosynthesis,so as to improve the yield and economic benefits of maize.

In order to explore the improvement effect of calcium fertilizer and ARC microbial agent on red earth dry land with low-calcium,the peanut variety Xianghua 522 was used as the experimental material,and two levels of calcium hydroxide fertilizer(0,750 kg/ha,code Ca₀ and Ca₅₀)and three levels of ARC microbial agent(0,30,60 kg/ha,code A₀,A₂ and A₄)were set to form six treatments to carry out pot experiment.It measured soil nutrient and soil enzyme activity in 0—20 cm arable layer soil at peanut seedling stage,flowering stage,pod setting stage and pod filling stage,and pod economic characters and yield were measured at harvest.The results showed that:single application of calcium fertilizer and the combined application of calcium fertilizer and ARC microbial agent could significantly improve the pH of soil at all growth stages,but ARC microbial agent had little effect on it.Compared with CK(Ca₀A₀),Ca₅₀A₂ and Ca₅₀A₄ significantly increased the content of hydrolyzable nitrogen in soil in the whole stage and the content of available phosphorus in soil in the first three growth stages;the content of available potassium in soil of Ca₅₀A₄ was higher than CK in general,and it was significant at seedling stage and pod setting stage;compared with CK,Ca₅₀A₀ significantly increased the content of exchangeable calcium in soil in four stages,with an increase of 23.78%—56.21%;the content of calcium ion in soil with calcium fertilizer application was significantly higher than that without calcium fertilizer application(the flowering stage was not significant),and it was little affected by ARC microbial agent;the content of organic matter in soil remained stable in the whole growth stage,but Ca₅₀A₄ and Ca₅₀A₂ were significantly higher than CK in each growth stage.Compared with CK,the soil sucrase activity of soil each treatment was significantly increased in four stages,and the increase was the largest in Ca₅₀A₄,ranging from 50.79% to 162.56%;the protease activity of soil was significantly increased by Ca₅₀A₂ in four stages with an increase of 26.58%—244.63%;the acid phosphatase activity of soil was significantly increased by Ca₅₀A₄ and Ca₀A₂ during the whole growth stage;the catalase activity of soil in all treatments showed a decreasing trend in general.All treatments could increase the yield of peanut pod in different degrees,and the effect of calcium fertilizer application was greater than that of ARC microbial agent,among which Ca₅₀A₄ had the best effect,with the pod weight per plant increasing by 12.29%,mainly increased the pod number per plant and the full pod number per plant.To sum up,the combined application of calcium fertilizer and ARC microbial agent has a good interaction effect on improving soil nutrient content,stimulating soil enzyme activity and increasing peanut yield,and the best effect is 750 kg/ha calcium fertilizer+60 kg/ha ARC microbial agent(Ca₅₀A₄),which can provide a theoretical basis for green and high yield cultivation of peanut.

Vernalization is a unique and necessary physiological stage in the growth and development of winter wheat,which determines the ecological adaptability and yield of winter wheat.This paper briefly summarizes the production and application value of vernalization of winter wheat.By summarizing and analyzing the structure and function of the key genes TaVRN1,TaVRN2 and TaVRN3 in wheat vernalization and their regulatory networks,it is believed that the study of epigenetic regulation of TaVRN1 is the core work of wheat vernalization mechanism analysis.Combined with the existing research on the expression regulation of TaVRN1,the causes of the difference in vernalization demand of wheat,the mechanism of long-term low temperature perception in winter,the research progress of overwintering memory and overwintering memory reset after vernalization,and the effect of vernalization on the regulation of cold tolerance,spikelet development,tillering and other agronomic traits of wheat were mainly introduced.In addition,this study summarizes the unsolved scientific problems in vernalization,and discusses the future research direction of wheat vernalization,which provides reference suggestions for the research of high yield and rapid breeding of wheat.

The identification and expression pattern analysis of CmPIPs gene family in melon could provide theoretical basis and support for further exploring the function of CmPIPs gene family and the genetic improvement of melon.TBtools,MEME,MEGA X and Plant-CARE tools were used to analyze the bioinformatics of CmPIPs,and the expression level of CmPIP2;7 in the pericarp of melon at different stages after pollination,and the expression level of each member of CmPIPs in different tissues and different concentrations of plant hormone treated young leaves were visualized in the software GraphPad Prism 10.The results showed that CmPIP2;7 and CsPIP2;8 had nearest kinship;the 12 members of the CmPIPs family were mainly distributed on chromosomes 1,3,4,5,9,10 and 11;except that CmPIP2;8 had 3 CDS regions,the other members had 4 CDS regions.The promoter regions of each member of CmPIPs had multiple cis-acting elements,hormone responsive elements,such as auxin,gibberellin,and abscisic acid.The expression level of CmPIP2;7 was significantly up-regulated during rapid development and maturity of melon fruit.Members of various families of CmPIPs were expressed in different tissues of melon.After treatment with 40.0 μmol/L auxin,the expression level of CmPIP2;4 was significantly up-regulated,while the expression level of CmPIP1;1,CmPIP 2;1,CmPIP2;2 and CmPIP2;3 were extremely significantly down-regulated,and when the concentrations of abscisic acid were 0.4,4.0 and 40.0 μmol/L,the expression level of CmPIP1;1,CmPIP2;1,CmPIP2;3,CmPIP2;9 were significantly down-regulated.After treatment with 44.640 μmol/L methyl jasmonate,the expression level of CmPIP2;1 and CmPIP2;5 were significantly down-regulated,while the expression level of CmPIP2;2,CmPIP2;3,CmPIP2;7 and CmPIP2;9 were significantly up-regulated.When the concentration of ethylene glycol was 4.0 mmol/L,the expression level of each member of CmPIPs was significantly up-regulated.The gene structure,sequence characteristics,evolutionary relationship and collinearity of CmPIPs gene family members were clarified,and their expression pattern were analyzed.

DNA damage significantly hinders impact plant growth and development.NBS1 plays an important role in the cellular response to DNA damage,to analyze the functional difference between NBS1 and its alternative splicing isoform,NBS1-3.Specific primers were designed to clone NBS1 and NBS1-3 from the complementary DNA of Arabidopsis thaliana,and the sequences of the NBS1,NBS1-3 and the three-dimensional structure of the proteins were analyzed.Transgenic Arabidopsis thaliana lines overexpressing of NBS1 and NBS1-3 were generated respectively,and nbs1 homozygous mutant plants were identified,detecting the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3.To further elucidate the functional distinction between NBS1 and NBS1-3,wild type,mutant and overexpressed plants were treated with 0.6 mmol/L Methyl Methanesulfonate(MMS)and observed damage area.The results of quantitative showed that the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3 were higher than wild type.PI staining of root tips showed that after 0.6 mmol/L MMS treatment,the mutant showed the highest relative damage area,while the NBS1-3 overexpressed lines exhibited the least damage,followed by NBS1 overexpressed lines and wild type.These results suggest a potentially more important role of NBS1-3 in DNA damage repair compared to NBS1.

Cloning the wheat grain superoxide dismutase(SOD)gene and developing competitive allele-specific PCR(KASP)markers related to SOD activity are of great significance for breeding wheat varieties with high SOD activity.According to the gene ID,specific primers were designed to clone the gDNA sequence of TaSOD-B1 gene.The single nucleotide polymorphism(SNP)loci were obtained by comparing the wheat genome genetic variation and Ensembl Plants database,and the KASP markers closely related to the SOD activity of wheat were developed.The practicability of the markers was verified by the correlation analysis between SOD activity and genotypes of 287 winter wheat varieties(lines).The TaSOD-B1 gene fragment of Chinese spring variety was amplified by six pairs of specific primers,and the TaSOD-B1 gene on chromosome 5B was obtained by splicing.The full length of the gene was 6 491 bp,including an open reading frame(ORF)of 1 650 bp,which encoded a total of 549 amino acids.The predicted molecular weight was 60.80 ku,the gene was composed of 12 exons and 11 introns.The intron conformed to the typical GT-AG structure.The KASP marker was developed based on the 44th base of the first exon of the TaSOD-B1 gene,and was verified by sequencing.The results showed that the genotype of the allelic variation type TaSOD-B1a was AA,which was associated with high SOD activity,and was labeled with the fluorescent gene FAM(shown as blue).The genotype of TaSOD-B1b was GG,which was associated with low SOD activity and was marked with the fluorescent gene HEX(shown as red).The detection of 287 winter wheat varieties(lines)at home and abroad showed that the SOD activity of different genotypes was significantly different.Based on the TaSOD-B1 sequence,a set of KASP markers related to SOD activity was successfully developed and could be used for genetic improvement of SOD activity.

In order to explore a good fertilization model to improve the fertility of yellow-cinnamon soil in Southern Henan,the characteristics of stable and high yield of wheat-maize rotation system and its relationship with soil nutrients under different fertilization treatments on yellow-cinnamon soil were studied.Based on the long-term experiment since 2012,four treatments were set up:no fertilizer(CK),chemical fertilizer(NPK),chemical fertilizer combined with manure(NPKM)and chemical fertilizer combined with straw(NPKS).Plants and soil samples were collected at maturity stage to determine crop yield and soil organic carbon,alkaline nitrogen,available phosphorus and available potassium.The results showed that compared with CK treatment,the crop yield of each fertilization treatment was significantly increased,the yield of wheat was increased by 53.70%—64.50%,and the yield of maize was increased by 44.54%—58.31%.The yield of NPKM treatment was the highest(wheat 8 162.61 kg/ha,maize 8 836.33 kg/ha),and there was no significant difference between NPKS and NPKM treatments.The yield sustainability index(SYI)of NPKM treatment was the highest,and the SYI values of wheat season and maize season were 0.84 and 0.82,respectively.The crop yield and its SYI value both showed NPKM>NPKS>NPK>CK,indicating that chemical fertilizer combined with organic materials could significantly increase crop yield and its sustainability.At the same time,different fertilization treatments could improve soil nutrients to varying degrees,among which NPKM treatment had the most significant improvement effect.The relationship between crop yield and soil nutrients was analyzed.The relationship between crop yield and soil nutrients was analyzed.There was significantly positive correlation between crop yield and soil organic carbon(SOC),available nitrogen and available phosphorus contents,among which the correlation between crop yield and SOC was the most significant.With the increase of soil organic carbon content,the SYI value of crops increased first and then stabilized,and the inflection point was 15.15 g/kg.To sum up,the application of chemical fertilizer combined with manure can significantly increase crop yield and soil nutrients,and maintain high crop yield sustainability.It is a recommended fertilization model to achieve sustainable production of soil-crop system in yellow-cinnamon soil ecological zone.

PHR1 is a crucial factor in balancing plant disease resistance and low phosphorus stress resistance.To investigate the nature and function of the StPHR1 gene in potato and to explore the role of StPHR1 in the process of potato resistance to Alternaria solani infection,the CDS sequence of the StPHR1 gene was cloned by PCR technology using potatoes as the research material,and the structural,physicochemical properties,and phylogenetic relationships of StPHR1 were analyzed and predicted using bioinformatics software,then,the expression level of StPHR1 during the infection of potatoes by A.solani and under different hormone treatments was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the CDS of the StPHR1 gene was 1 353 bp,encoding 450 amino acids.The protein had a molecular formula of C₂₁₄₇H₃₃₉₉N₅₉₅O₇₁₁S₁₈,a molecular weight of 49.51 ku,and a theoretical isoelectric point of 5.07,encoding a hydrophilic,unstable protein with no signal peptide and no transmembrane structure.Its secondary structure consisted mainly of random coil and α-helix.Phylogenetic tree analysis revealed that the StPHR1 protein was most closely related to Arabidopsis thaliana; conservative domain analysis revealed that the StPHR1 protein,like other PHR1s,possesses both MYB-CC and MYB conserved structural domains at its C-terminus.Relative expression analysis found that StPHR1 was significantly induced by A.solani and salicylic acid,and it was hypothesized that StPHR1 played an important role in A.solani infection of potato and in the response to salicylic acid; and the subcellular localization showed that the StPHR1 protein was localized in the nucleus.It is hypothesized that StPHR1 may regulate potato resistance to A.solani through its MYB transcription factor activity and response to salicylic acid.

In order to provide production advice of high-quality upper tobacco leaves,field experiment was carried out to elucidate the effects of nitrogen application on the growth,development,carbon and nitrogen metabolism of upper leaves of different tobacco varieties.Three treatments,37.5(LN),75.0(MN)and 112.5 kg/ha nitrogen application rate(HN)were conducted with NC89 and Yunyan 87 as the test varieties.The effects of different treatments on the agronomic traits,photosynthetic characteristics,leaf tissue structure,key enzymes activity of carbon and nitrogen metabolism,and chemical composition of the upper leaves of were studied.The results showed that the leaf length,leaf width,leaf area,and leaf dry mass of the upper leaves of both varieties were significantly improved with the increased of nitrogen application rates.At 115 days after transplanting(DAT),the leaf area of NC89 and Yunyan 87 of HN were significantly increased by 63.10% and 68.43% compared to LN,respectively.Increasing nitrogen rate improved the leaf chlorophyll content of NC89,the leaf chlorophyll content of HN was 6.67%—37.50% higher than that of LN,respectively.The leaf net photosynthetic rate was notably improved by increasing nitrogen rate of Yunyan 87,especially at 70 DAT and 80 DAT.The leaf palisade tissue,leaf spongy tissue,and leaf thickness were markedly promoted with the increased nitrogen application rate on 85—115 DAT.The palisade tissue and spongy tissue thickness of LN and MN were almost stable from 95 to 115 DAT,but HN improved by 9.82%—14.08% and 10.72%—13.72% respectively.The leaf carbon content and leaf carbon and nitrogen ratio of both varieties were significantly decreased by reduced nitrogen rate,while leaf nitrogen content significantly increased.The activities of invertase,sucrose phosphate synthase,sucrose synthase,and glutamate synthase of both varieties were increased with the increased nitrogen application rate.HN decreased the amylase activity of Yunyan 87,but increased that of NC89.At 115 DAT,the amylase activity of HN of Yunyan 87 was 27.53% lower than MN,while HN increased amylase activity by 33.86% and 21.74% compared to LN and MN of NC89,respectively.The activity of glutamine synthetase of Yunyan 87 significantly increased with the increased nitrogen application rate,while there was no significant difference among different treatments of NC89.The content of flue-cured tobacco leaf reducing sugar and total sugar were reduced,but the content of nicotine and total nitrogen were improved with increased nitrogen application rate.Under the same nitrogen application rate,the content of flue-cured tobacco leaf nicotine,total nitrogen,and potassium of Yunyan 87 were higher than NC89,while the content of reducing sugar,content of total sugar(except LN),sugar and nicotine ratio,and nitrogen and nicotine ratio were lower than NC89.The effect of nitrogen application rate on upper leaves of tobacco varieties is difference.Increasing nitrogen application rate could promote the growth,development and carbon metabolism,reduce sugar and nicotine ratio and nitrogen and nicotine ratio,and improve the coordination of chemical components of NC89.However,increasing nitrogen application rate could lead to excessive nitrogen metabolism,delay the transformation from nitrogen metabolism to carbon accumulation metabolism,and cause late maturity of Yunyan 87.

To explore the effects of soil phosphorus levels and exogenous microbial agents on soil functioning(soil physico-chemical properties,soil enzyme activities,and arbuscular mycorrhizal fungi(AMF)community composition and structure)of the rhizosphere soil of maize,a two-factor interaction experiment was conducted to study the effects of two levels of low phosphorus and normal phosphorus and four treatments of inoculation with AMF,phosphate-solubilizing bacteria(PSB),AMF-PSB and no inoculation of exogenous agent(CK)on rhizosphere soil indexes in different growth stages of maize.The results showed that the soil total phosphorus content was the highest at jointing stage and silking stage under the condition of normal phosphorus level+AMF treatment,while the soil available phosphorus content was the highest at jointing stage and mature stage under the condition of normal phosphorus level+AMF treatment.At jointing stage and silking stage,alkaline phosphatase activity was the highest under low phosphorus+AMF treatment,and the activity of alkaline phosphatase under low phosphorus+AMF treatment and low phosphorus+PSB treatment was higher than that under normal phosphorus level.Under low phosphorus level,total phosphorus content and available phosphorus content were higher than CK treatment,and the increase rate was higher than that under normal phosphorus level.At low phosphorus level,soil pH value was lower than CK treatment under the condition of inoculated foreign bacteria,and also lower than CK treatment under normal phosphorus level at jointing stage.Under the low phosphorus level,the CK treatment had the highest observed number of AMF community OTUs and α-diversity indexes,while under the normal phosphorus levels,the observed number of OTUs and α-diversity index in the CK treatment group were lower than those in the AMF and PSB treatments.Non-metric multidimensional scaling analysis indicated that the composition and structure of soil AMF communities were convergently regulated by the types of inoculated exogenous microorganisms.In summary,inoculating AMF and PSB agents in low-phosphorus alkaline maize fields in northern China can improve soil physicochemical properties and increase soil alkaline phosphatase activity,but it also increases competition and exclusion among soil AMFs.

4-coumaric acid:coenzyme A ligase(4CL),as the main biosynthetic enzyme of flavonoids and lignin,is closely related to the formation of plant flavonoids.In order to explore the relationship between the 4CL gene family of Glycyrrhiza uralensis and the synthesis and accumulation of isoglycyrrhizin,this study determined the content of isoglycyrrhizin in G. uralensis after drought stress,and the expression characteristics and bioinformatics analysis of the 4CL gene family of G. uralensis Fisch.to understand the relationship between the accumulation characteristics of isoglycyrrhizin and the 4CL gene of G.uralensis.The results showed that isoliquiritigenin was mainly accumulated in the roots of G.uralensis,and drought stress could significantly increase the content of isoliquiritigenin in the roots of G.uralensis.The content of isoliquiritigenin after 2 h of polyethylene glycol(PEG)stress was 3.91 times that of the control group(0 h).Drought stress could induce the up regulation of Gu4CL gene in the underground part.The expression levels of Gu4CL2, Gu4CL4 and Gu4CL5 were higher after PEG stress induction,while Gu4CL2 was most significantly up-regulated after PEG stress.The expression of Gu4CL2 was similar to that of isoliquiritigenin.Bioinformatics analysis of Gu4CL gene showed that 11 genes had two conserved polypeptide motifs Box Ⅰ(SSGTTGLPKGV)and Box Ⅱ(GEICIRG),and 11 genes were distributed on 11 Scaffold fragments.Promoter cis-acting element analysis showed that Gu4CL2, Gu4CL4 and Gu4CL5 genes contained more abscisic acid and jasmonic acid response elements than other Gu4CL genes.Therefore,drought stress may induce the synthesis of jasmonic acid and abscisic acid,regulate the expression of Gu4CL2, Gu4CL4 and Gu4CL5 genes in licorice roots,and increase the accumulation of isoliquiritigenin.This study provides a research basis for optimizing the cultivation techniques of G.uralensis,improving the adaptability and quality of cultivated land in saline-alkali land,exploring the role of Gu4CL gene family in G.uralensis,and exploring the synthesis mechanism of isoliquiritigenin.

IQM(IQ-motif containing protein),a plant-specific calmodulin-binding protein,plays crucial roles in plant growth,development,and responses to various stresses.In order to study the characteristics and potential functions of the maize IQM gene family,bioinformatics approaches were used to identify IQM genes in the maize whole genome,and protein properties,phylogenetic relationship,gene structure,chromosome location,gene replication,cis-acting element,tissue-specific expression and expression patterns under various stresses were investigated.A total of 11 ZmIQMs genes were identified in the whole genome of maize,named ZmIQM1 to ZmIQM11 based on their chromosomal locations.ZmIQMs genes could be classified into three subfamilies,with genes within different subfamilies exhibiting similar structures.Segment duplication was found to play a major role in the amplification and evolution of the ZmIQMs gene family.Cis-acting element analysis showed that the promoter region of ZmIQMs gene contained multiple hormone and stress response elements.The expression pattern of ZmIQMs genes was investigated,and it was found that ZmIQMs genes had different expression patterns in different tissues,and the expression levels of several ZmIQMs genes were changed under different abiotic and biotic stresses.qRT-PCR results showed that under drought stress,the expression of ZmIQM3,ZmIQM4 and ZmIQM10 was up-regulated, and ZmIQM3,ZmIQM4,ZmIQM5,ZmIQM10 and ZmIQM11 responded to Cochliobolus heterostrophus infection.The results showed that ZmIQMs genes played an important role in stress response.

In order to accelerate the breeding of pink-tomato varieties,a rapid breeding strategy was used to generate pink-fruited tomato material by CRISPR/Cas9-mediated gene editing of SlMYB12.Two adjacent target sites within the first exon of SlMYB12 were selected to construct the CRISPR/Cas9 binary vector, and then the vector was introduced into red-fruited inbred line R18-10C through Agrobacterium-mediated transformation. The homozygous mutants without exogenous Cas9 were screened using specific primers and their agronomic traits and fruit nutritional quality were analyzed. Sequencing results showed that three homozygous mutants of different mutation types were obtained and all of which were frame shift mutations caused by base deletion. Compared with wild-type red fruit tomatoes, SlMYB12-edited plants grew and developed normally,and there were no significant difference in plant height,single-fruit weight,total yield per plant,fruit total soluble solid content and lycopene level,but the mature fruit showed pink and the flavonoid naringenin chalcone(NarCh)content of tomato peels was significantly reduced in MYB12-edited plants.In summary,the procedure for the generation of pink-fruited tomato plants through CRISPR/Cas9-mediated targeted mutagenesis of SlMYB12 was set up and new pink-fruited tomato germplasms with stable inheritance were obtained.

Currently,traditional breeding methods cannot fully meet the demands of the cotton market and production. Therefore, molecular biotechnology can be employed to accelerate the development of new cotton varieties. Transcription factors have emerged as significant tools in the study of cotton gene function, genetics, and breeding during this period. Among them, the MYB transcription factor family is one of the largest transcription factor families present in many plants, and it performs multiple roles in plant growth and development. Hence, the MYB transcription factor holds high research value. While the function of MYB transcription factors has been extensively studied in model plants, research in non-model plants, particularly cotton, remains limited and is mostly concentrated in upland cotton. To further understand MYB transcription factors, this paper reviews the research progress on MYB transcription factors in various plants and cotton. It covers their classification basis, structural characteristics, evolution, roles in response to biotic and abiotic stress, cotton fiber development, and secondary metabolism. Additionally, it provides statistics on the known functions of cotton MYB transcription factors. This review aims to deepen our understanding of cotton MYB transcription factors and serve as an important reference for future studies on the functions and mechanisms of MYB transcription factors in different cotton species.

To explore the planting patterns of mechanical harvesting sesame and achieve goals of high yield,stable yield and deep combination of agricultural machinery and agronomy,the split plot design with two factors was carried out to investigate the effects of different planting pattern and density on the photosynthetic characteristics,biomass,yield and mechanical harvesting characteristics of Yuzhi ND837,in order to provide theoretical foundation and technical guidance for the full mechanization production of sesame.The planting pattern included wide-and narrow-row spacing planting(Z1,wide-row:60 cm,narrow-row:20 cm),banding planting of 4 rows(Z2,line spacing:30 cm,banding spacing:60 cm),banding planting of 8 rows(Z3,line spacing:30 cm,banding spacing:60 cm)and equidistant row planting(Z4,line spacing:30 cm)as primary area.The planting density included 180(M1),225(M2),270(M3)and 315 thousand plants/ha(M4)as vice-area.The results showed that planting pattern and planting density both had significant effects on the photosynthetic characteristics,substance accumulation,yield and mechanical harvesting characteristics of sesame.The net photosynthetic rate(Pn),SPAD value and biomass per plant descended with the order of M1>M2>M3>M4,while the biomass of population increased at first and then descended with the planting density increasing under the same planting pattern.Under different planting patterns,the Pn,SPAD value,biomass per plant and biomass of population all showed Z1>Z2>Z3>Z4.Under the comprehensive effects of planting pattern and density,the Pn and SPAD value of Z1M1 were the highest,and the biomass per plant was also the highest in Z1M1(59.76 g/plant),but the biomass of population and yield were the highest in Z1M3(13 032.97,1 719.87 kg/ha,respectively).The uniformity of plants tended to be the same,the diameter of stem and the ability of capsule formation per plant decreased,but the lodging percentage at maturity also gradually decreased with the increase of planting density(M1>M2>M3>M4).The lodging percentage showed Z1>Z2>Z3>Z4 among different planting patterns.Under the comprehensive influence of planting pattern and density,the lodging percentage of Z3M1(17.51%)was the highest and Z4M4(7.97%)was the lowest.Under the conditions of the experiment,Z1M3 has the biggest yield,and the agronomic traits and mechanical harvesting characteristics at maturity are also better.

In order to study the synergistic regulation characteristics of plant growth regulators on canopy-root traits of spring maize and to further reveal the lodging resistant mechanism of the plants,in 2021—2022,with the maize varieties Dika 159(DK159)and Xianyu 335(XY335)as experimental materials,at two planting densities of 75 000,90 000 plants/ha,plant growth regulator treatment (PGR) and water control (CK) were set up to analyze and compare the canopy structure,stem basel internode characters,root morphological characteristics and physicochemical properties of root bleeding sap under different treatments.The results showed that PGR had regulatory effects on both the maize canopy and roots.After PGR treatment,plant height,ear height and center of gravity height decreased,the mean leaf tilt angle above ear increased,light transmittance of ear layer increased by 23.59% on average,light transmittance of basel layer increased by 18.60% on average,and stem quality of basal internode was significantly improved.At the same time,the total root number,root length and root dry weight of PGR treatment increased,the root width at 10 cm below the surface increased,the root bleeding sap flow and nutrient flow increased,and the root morphological characteristics and transport capacity were obviously optimized.The flow of CTK and IAA in the root bleeding sap increased,while the flow of GA decreased.PGR effectively reduced the occurrence of stem lodging and root lodging through the synergistic regulation of canopy and root system,the field lodging rate of maize decreased from 13.43% to 6.47%,and the average yield of maize increased by 16.10%,thus achieving stable and high yield.

To understand the member characteristics of the WRKY transcription factor family in hydrangea and their roles in response to leaf spot disease,this study utilized the bioinformatics method to characterize WRKY family members in Hydrangea macrophylla Endless Summer,and systematically analyzed the protein physicochemical features,gene structure,systematic evolution,collinearity and expression patterns under the infection with Corynespora cassiicola of WRKY family members.The results showed that there were 84 non-redundant HmWRKY members in H.macrophylla genome.All of the HmWRKYs were hydrophilic proteins and unevenly distributed on 18 chromosomes of H.macrophylla,encoding 112—1 046 amino acids.In addition,HmWRKY members were divided into 3 subgroups(Group Ⅰ—Group Ⅲ),which contained a conserved DNA binding domain composed of WRKYGQK and C2H2.The sequence length of HmWRKY varies greatly from 512 bp to 40 338 bp,and 8 collinear gene pairs with Ka/Ks ratios less than 1 were detected,indicating that the HmWRKY family experienced purification selection in evolution.18 HmWRKY members showed significantly differential expression after infection with C.cassiicola,of which 9 up-regulated and 9 down-regulated.The results indicated that these HmWRKY genes might play important roles in response to leaf spot disease of hydrangea.

Chalcone synthase(CHS)is an important structural gene that regulates the early biosynthesis of flavonoid pathways in plants,and plays a role in plant growth and development and stress response.Previously,the key genes StCHS4 and StCHS5 for anthocyanin biosynthesis were identified in the potato CHS family by expression analysis.To further explore the function of potato StCHS4 and StCHS5 in the biosynthesis of flavonoids and anthocyanins,the characterization of StCHS4 and StCHS5 proteins was analyzed by online website.The 35S∷StCHS4-GFP and 35S∷StCHS5-GFP recombinant vectors were constructed by homologous recombination method based on the pRI101 binary vector,and then were transformed into Agrobacterium GV3101 strain.The subcellular localization of StCHS4 and StCHS5 proteins was determined by transient transformation of Nicotiana benthamiana.N.tabacum was used as the experimental material for transient overexpression and stable genetic transformation to analyze the content of total flavonoids and anthocyanins after overexpression of StCHS4 and StCHS5 genes.The results showed that the secondary structures of StCHS4 and StCHS5 proteins were mainly α-helix and random coil.StCHS4 was an unstable hydrophilic protein,and StCHS5 was a stable hydrophilic protein.The sequence alignment revealed that StCHS4 and StCHS5 had a close relationship with the CHSs of Capsicum annuum and Solanum lycopersicum,respectively.The results of subcellular localization revealed that StCHS4 and StCHS5 proteins were localized in the cytoplasm and cell membrane.In transient overexpression of tobacco,StCHS4 and StCHS5 genes significantly enhanced anthocyanin accumulation at 3—5 days after injection.Three transgenic N.tabacum lines of StCHS4 and StCHS5 gene were obtained,respectively.Compared with the wild type,the expression of StCHS4 and StCHS5 in transgenic plants was significantly higher,and the contents of total flavonoids and total anthocyanins were higher than those in the wild type.The total flavonoid content in StCHS4-OE3 and StCHS5-OE1 transgenic plants was significantly increased.The anthocyanin content in StCHS5-OE1 and StCHS5-OE2 plants increased by 89%,131%,respectively.The above results demonstrated that StCHS4 and StCHS5 were the key CHS genes in the flavonoid pathway of Solanum tuberosum,and the overexpression of StCHS4 and StCHS5 contributed to the biosynthesis of anthocyanins and flavonoids.

Cucumber powdery mildew is one of the main diseases that adversely impacts cucumber production,posing a significant challenge to its sustainable cultivation.Identifying genes related to cucumber resistance to powdery mildew can help understand the genetic principles and molecular mechanisms of cucumber resistance to powdery mildew,and provide diverse gene resources for disease resistant breeding.This study constructed F₁ and F₂ populations of QK×QG using cucumber resistant inbred line QK and susceptible inbred line QG as parents.Using the extreme trait mixed pool resequencing (BSA-seq) method,the genomic regions harboring the resistance genes of cucumber powdery mildew were preliminarily located.By integrating transcriptome data with gene annotation information,the association interval of the disease phenotype was narrowed,sequence variations were identified,and key genes were screened.The results showed that the resistance of powdery mildew may be controlled by recessive genes,and the population of F₂ showed a continuous normal distribution from resistant to susceptible.The BSA-seq analysis,combining the SNP-Index method and QTG(quantitative trait genomics)-seq method analysis highlighted the 19—21 Mb region of chromosome 5,where there were 77 annotated genes with SNP differences between samples,including 33 non-synonymous mutations.The transcriptome sequencing (RNA-seq) results showed that there were 309 upregulated genes and 697 downregulated genes in the susceptible material.The expression levels of 13 genes within the candidate segment of chromosome 5 showed significant differences after infection.Through a comprehensive analysis of differentially expressed genes and BSA,the candidate genes in this segment were narrowed to 3,and only SNP mutations were detected in the LOC101207011 gene.The candidate gene LOC101207011 was characterized by a mutation resulting in an amino acid change from Valine at position 656 to Leucine.This gene emerges as the primary focus of our investigation due to its potential role in conferring resistance to powdery mildew.

To investigate the transcriptome differences between resistant and susceptible varieties of cauliflower after inoculation with Xanthomonas campestris pv.campestris (Xcc),and to identify genes associated with cauliflower resistance to black rot disease,the susceptible variety Y1-2 and the resistant variety EC-247 of cauliflower were selected as the research subjects.Total RNA was extracted from cauliflower leaves at 0,1,3,and 5 days post-inoculation with Xcc,respectively.High-throughput parametric transcriptome sequencing was then conducted utilizing the Illumina RNA-Seq platform,followed by Real-time Quantitative PCR for validation of selected differentially expressed genes(DEGs).DEGs associated with disease resistance were screened and analyzed.The findings revealed that 6 355 genes exhibited significant differential expression between resistant and susceptible cultivars across the four time points.KEGG enrichment analysis focused on plant disease resistance pathways,identifying 47 genes involved in plant-pathogen interactions and 61 genes related to plant hormone signaling.Cluster analysis of these gene expression levels disclosed specific genes,including one CDPK,four CMLs,one PTK,one CaM,one RLK,and one SGT1 in the plant-pathogen interaction pathway,and three auxin-responsive protein genes,a TIFY gene,an indole-3-acetic acid-amido synthetase gene,two brassinazole-resistant protein genes,and a Shaggy-associated protein kinase zeta gene in the plant hormone signaling pathway.Notably,the expression of these genes was significantly higher in resistant varieties compared to susceptible ones,indicating their active response to pathogen infection at various time points.The results indicated that these differential genes might be related to disease resistance in cauliflower,which provided important genetic resources and scientific basis for molecular breeding of disease resistance in cauliflower.

To explore the function of key genes in photosynthesis, a functional knockout mutant (zmC₄nadp-me) of ZmC₄NADP-ME, the gene encoding the rate-limiting enzyme of the dark reaction of photosynthesis in maize, was obtained. Evolutionary tree analysis showed that ZmC₄NADP-ME and its homologous genes exist in multiple copies in most plants, with diverse expression patterns. Phenotypic analysis revealed that the entire zmC₄nadp-me plant was yellow-green, and its seedling-stage leaves dried up and died rapidly under light. Chlorophyll fluorescence analysis indicated that Y(Ⅱ) and electron transport rate ETR(Ⅱ) of photosystem Ⅱ (PSⅡ) in zmC₄nadp-me decreased significantly, with little change in Y(NPQ), while the Y(NO) increased notably. Measurement of the absorption capacity (P700) of photosystem Ⅰ (PSⅠ) found that both the electron transport rate (ETR(Ⅰ)) and the actual photoelectron efficiency (Y(Ⅰ)) of zmC₄nadp-me dropped substantially, and the gap widened with increasing light intensity. Under specific light intensities, Y(ND) and Y(NA) of zmC₄nadp-me were greater than those of the wild type (WT). In conclusion, ZmC₄NADP-ME is essential for plant growth and development. Disruption of this gene severely stresses PSⅡ, and the plant can't alleviate this stress by increasing Y(NPQ). Meanwhile, at low light intensities, the inhibition of PSⅠ may originate from the electron donor side of PSⅠ, and as the light intensity increases, the inhibition from the electron acceptor side of PSⅠ becomes a key factor.

Elongation of very long chain fatty acids protein 3(ELOVL3),mainly involved in the synthesis of very long chain fatty acids(VLCFA),plays an important role in fatty acid metabolism.The objectives of this study were to clone the protein-coding sequence of ELOVL3 gene from Jiulong yak,to construct the eukaryotic expression vector of ELOVL3 gene,and to predict and analyze its biological functions.The CDS region of the ELOVL3 gene was amplified by PCR and inserted into the pcDNA3.1 vector by homologous recombination to construct a recombinant plasmid.The recombinant plasmid was validated by restriction digest and PCR,and the biological functions of its protein coding sequence were analyzed by online prediction software in combination with sequencing results.In addition,qPCR and Western Blot were performed for the expression of ELOVL3 gene at mRNA level and protein level.The results showed that the protein-coding sequence of Jiulong yak ELOVL3 gene was 813 bp in length,encoding a total of 270 amino acids.Enzymatic cleavage,PCR and sequencing identification confirmed the successful construction of the pcDNA3.1-ELOVL3 eukaryotic expression vector.Bioinformatics analysis showed that the protein encoded by the ELOVL3 gene was a hydrophobic protein,with 6 transmembrane structural domains and 27 phosphorylation sites.The phylogenetic tree showed that the closest relation to the Jiulong yak was the common cow,and the furthest was the original chicken.In addition,the ELOVL3 gene was most highly expressed in the lung tissue of the Jiulong yak.

Plant sodium-hydrogen antiporter(NHX,Na⁺/H⁺ antiporter)plays a crucial role in plant sodium and potassium ion balance and cellular pH regulation.In order to investigate the relationship between salt tolerance and ScNHXs,it was conducted to identify and analyze the ScNHXs by bioinformatics process,and to examine the expression pattern of ScNHXs under salt stress by RT-qPCR,which can provide the reference information for the investigation of the potential functions of ScNHXs as well as the mining of salt tolerance genes in rye.A total of 10 rye NHX gene family members(ScNHX1—ScNHX10)were identified,and the phylogenetic tree analysis showed that they could be divided into two subfamilies,Vac and Endo,containing four and six genes,respectively.Physicochemical property analysis of the encoded proteins showed that most of the molecular weight ranged from 27.92 to 59.72 ku,the number of amino acids from 253 to 546 aa,and the isoelectric point between 5.17 and 8.81,with most of proteins being classified as acidic proteins.Signal peptide prediction indicated the absence of signal peptides in the members,and transmembrane structure analysis revealed that all members possessed transmembrane structures.The subcellular localization prediction indicated that ScNHXs were located in the plasma membrane and vesicles.Spatial structure prediction showed that their secondary structures mainly consisted of α-helices and irregular convolutions.Gene structure and motif analyses revealed that the number of exons of the ScNHXs varied from 13 to 24,and all of them possessed a conserved Na⁺/H⁺ exchange structural domain.In addition,cis-acting element analysis revealed that numerous elements related to hormone response and abiotic stresses were found in the promoter region of ScNHXs.Analysis of rye transcriptome data revealed significant differences in the expression patterns of ScNHXs in different tissues of rye.RT-qPCR analysis showed that ScNHXs responded differently to different concentrations of NaCl stress,and were able to persistently respond to salt stress over a long period of time.In summary,ScNHXs may be involved in the biological regulation during salt stress in rye.

To investigate the effect of applying different gradients of nitrogen reduction on leaf production capacity of Korla fragrant pear,no fertilizer treatment(CK),no nitrogen fertilizer treatment(N0),conventional fertilizer treatment(N),and three nitrogen fertilizer reduction gradients(N1,N2 and N3,which reduced nitrogen use by 10%,20%and 30%,respectively,compared with conventional fertilizer)were set up in a total of six treatments.Leaf nutrient content,net photosynthetic rate,chlorophyll fluorescence,chlorophyll content,leaf area index and yield were compared under different fertilizer application methods based on multi-year fertilizer trials.Nitrogen fertilization significantly increased leaf and branch nutrient content,leaf chlorophyll content,leaf area index,net photosynthetic rate,chlorophyll fluorescence and yield,and soluble solids and VC content in fruit.Nitrogen reduction of 10% had no significant effect on leaf and branch nutrient content,chlorophyll fluorescence,net photosynthetic rate,chlorophyll content,leaf area index and fruit soluble solids,VC,stone cells and total acid content compared to complete N application,and N reduction of 10% to 20% had no significant effect on the orchard yield and was able to maintain it at the normal range level.Net photosynthetic rate,chlorophyll fluorescence parameters,and yield were significantly and positively correlated with N,P,K,Fe,Mn,Cu,and Zn contents in leaves and branches.According to the experimental results and analysis,the recommended range of nitrogen fertilizer reduction for Korla fragrant pear of 10—12 years old is 10%—20%(240—270 kg/ha)on the basis of fully applied nitrogen(300 kg/ha)as the optimal amount of nitrogen fertilizer reduction in fragrant pear orchards.

In order to investigate the effects of conservation tillage techniques on soil nutrient and enzyme activity indicators,as well as ecological stoichiometry characteristics in black soil areas,this study used a 3-year positioning experiment method,with tillage (A-A) as the control,to investigate the changes in total nutrients soil organic carbon(SOC),total nitrogen (TN),total phosphorus(TP) and enzyme activities of β-D-glucosidase(βG),leucine aminopeptidase(LAP),N-1,4-acetylglucosidase(NAG),acid phosphatase(ACP)and their ecological stoichiometry characteristics in black soil under rotary tillage (B-B),conventional no tillage (C-C),and no tillage and straw mulching with original crop (0-0) treatments.The results showed that no tillage with straw mulching significantly increased soil SOC,TN,and TP content compared to tillage;except for C/N,the C/P and N/P ratios of straw mulching in no tillage with straw mulching were higher than those in tillage.The soil SOC and TN content of rotary tillage and conventional no tillage were relatively reduced compared to tillage,while the C/N,C/N,and N:P values all significantly increased in the second year.Compared with tillage,the activities of four enzymes were significantly increased under the no tillage and straw mulching treatment.In the third year,rotary tillage significantly increased ACP and βG by 22.05% and 50.00%,respectively,compared with tillage.Rotary tillage,conventional no tillage,and no tillage with straw mulching all significantly increased soil enzyme C/P.The vector angles of soil enzyme activity under the tested cultivation methods were all less than 45 degrees,indicating that soil microorganisms in the experimental area may be limited by N;the length of enzyme vector increased significantly with age,indicating an increased degree of restriction by C.The results of principal component analysis,grey relational analysis,and correlation analysis showed that no tillage with straw mulching had the most significant impact on soil nutrient content and enzyme activity.In summary,the no tillage and straw mulching with original crop technology measures have a good improvement effect on activating soil nutrients and enzyme activity,as well as maintaining soil ecological stability.

The naked seeds of zucchinii possess significant natural advantages in processing.To investigate the genetic mechanisms underlying the hull-less trait in zucchini,we utilized the hulled 17pu10 (P₁) and the hull-less 17pu08 (P₂) as parental strains,and constructed populations of F₁ (P₁×P₂),F₂,and BC₁.The phenotypes of the zucchini seeds in the progeny population were assessed.The results showed that the number ratio of hulled seeds and hull-less seeds in the progeny population was in line with 3∶1 separation ratio.This indicated that the hull-less trait was regulated by a single gene and that the hull-less allele was recessive.Genetic mapping within this interval revealed that the gene for hull-less in zucchini was located between the markers InDel3157329 and InDel3724121,with genetic distances of 1.4,2.6 cM,respectively,while the physical distance was 0.6 Mb.The annotation and function analysis of 24 genes in the interval showed that 4 genes were directly or indirectly involved in the biosynthesis of cell wall,cellulose and lignin.Further analysis of the expression differences of 4 genes showed that only Cp4.1LG12g04350 and Cp4.1LG12g04370 had significant differences in expression levels during seed development.It was inferred that Cp4.1LG12g04350 or Cp4.1LG12g04370 was the candidate gene controlling the hull-less trait.In addition,InDel markers linked to hull-less genes were developed,which could be used as markers to identify zucchini hull-less traits,so as to accelerate the breeding of high-quality zucchini seed hull-less varieties.

In order to investigate the effect of exogenous 24-epibrassinolide(EBR)on the physiological characteristics of Lycium barbarum seedlings under salt stress,Ningxia L. barbarum Ningqi 7 was used as the experimental material.Five treatments were set up,namely 0 mmol/L NaCl+distilled water spray(CK),150 mmol/L NaCl+distilled water spray(N0 treatment),150 mmol/L NaCl+0.005 mg/L EBR(T1 treatment),150 mmol/L NaCl+0.050 mg/L EBR(T2 treatment),and 150 mmol/L NaCl+0.500 mg/L EBR(T3 treatment).The seedling plant height,basal diameter,aboveground and underground biomass,chlorophyll content,photosynthetic parameters,antioxidant enzyme activity and the content of osmoregulatory were measured on the 7th day,14th day,and 21st day respectively.The results showed that compared with N0 treatment,T1,T2,T3 treatments significantly increased the plant height,basal diameter,aboveground and underground biomass,photosynthetic pigments in leaves,net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), superoxide dismutase (SOD), peroxidase (POD), catalase(CAT)activities,as well as the content of osmoregulatory in L.barbarum seedlings,intercellular CO₂ concentration (Ci) and malondialdehyde (MDA) content significantly decreased,T2 treatment had the best effect.On the 21st day of salt stress,compared to the N0 treatment,the plant height,basal diameter,aboveground and underground biomass of L.barbarum seedlings increased by 23.63%,15.45%,17.70%,and 47.06%,the content of Chlorophyll a,Chlorophyll b,and Chlorophyll a+b increased by 10.68%,12.31%,and 6.57%,respectively;Pn,Tr,and Gs increased by 55.53%,27.83%,and 9.76%,respectively;the Ci value decreased by 14.42%;the activities of SOD,POD,and CAT increased by 13.23%,20.10%,and 9.31%,respectively;the MDA content decreased by 35.28%;the proline content,soluble sugar content,and soluble protein content increased by 45.17%,86.54%,and 57.00%,respectively.In summary,an appropriate concentration of exogenous 24-epibrassinolide can promote the growth of L.barbarum seedlings under salt stress,improve the photosynthetic capacity and antioxidant enzyme activity of L.barbarum seedlings,increase the content of osmoregulatory substances,alleviate the damage of salt stress to L.barbarum seedlings,and enhance the salt tolerance of L.barbarum seedlings.Among them,exogenous 0.050 mg/L EBR has the best effect.

To analyze the variability of the novel Goose astrovirus(GAstV)in Lu'an,Anhui Province and to express the VP27-VP34 fusion protein,the gout samples were collected from a farm in Lu'an.After confirming positivity via RT-PCR,the virus was isolated by passage culture in goose embryos.Then,the isolated strain was subjected to an animal regression test,whole genome amplification sequencing and genetic evolution analysis.Subsequently,the VP27-VP34 fusion protein of the isolated strain was induced and expressed,and purified recombinant protein was used to immunize 6-week-old female BALB/c mice to produce polyclonal antibodies.Serum antibody titers were assessed using agar diffusion methods,and the specificity of the polyclonal antibodies was detected by indirect immunofluorescence(IFA).The specificity of the antibody was detected by indirect immunofluorescence(IFA),and the titer of the prepared antibody was detected by the agar diffusion method.The results showed that one strain of GAstV,named AH-2021 strain,was isolated from clinical samples.The animal regression test showed obvious urate deposition on the surface of the heart and liver of goslings,and the kidney was white and swollen.Genetic evolution results revealed that AH-2021 belonged to GAstV-1,showing 98.0%—99.0% identity with other GAstV-1 strains in GenBank.The recombinant expression vector pCold-TF-VP27-VP34 was induced by IPTG to obtain the target protein,and SDS-PAGE showed that the molecular weight of the recombinant protein was about 110 ku,which mainly existed in the form of supernatant.IFA results showed that the polyclonal antibody was able to specifically recognize the GAstV,and the agar diffusion results showed that the titer of polyclonal antibody was up to 1:16.In conclusion,a strain of novel GAstV AH-2021 was isolated from gouty goslings,and animal regression tests showed that the novel Goose astrovirus was the pathogen causing gout in goslings,and a polyclonal antibody to the VP27-VP34 fusion protein was prepared.

To investigate the characterization and function of LbaHY5 gene of Lycium barbarum,explore the role of LbaHY5 in the degree of stem straightening in Lycium barbarum.Using Ningqi No.1 as the test material,a preliminary analysis was conducted on the structure and function of the LbaHY5 gene using bioinformatics,subcellular localization,and qRT-PCR methods,the results indicated that the LbaHY5 gene cloned from Ningqi No.1 had a full-length open reading frame of 483 bp,encoding 160 amino acids,with a molecular weight of 17.52 ku and an isoelectric point of 9.69,classifying as a hydrophilic protein.Conserved domain and multiple sequence alignment analysis showed that LbaHY5 protein contained a type bZIP domain,which belonged to the bZIP gene family.Evolutionary tree analysis showed that the LbaHY5 protein in goji berries was highly homologous to the HY5 protein in Solanaceae plants such as tomatoes and potatoes.In addition,promoter analysis showed that the promoter region of this gene was rich in multiple functional elements,which were respectively related to processes such as light signaling,hormone signaling pathways,and abiotic stress response.The subcellular localization indicated that LbaHY5 was located on the nucleus.The transcript level of LbaHY5 gene was the highest in the flower and the lowest in the stem.During the fruit development period,the expression level of LbaHY5 gradually increased.The results speculated that LbaHY5 gene might play an important role in the growth and development of floral organs and fruits in Lycium barbarum.

To reveal the sequence characteristics,subcellular localization,and expression pattern of PatASIL2,belonging to the Trihelix transcription factor family,the PatASIL2 gene was first cloned using the cDNA of Pogostemon cablin (patchouli)as a template,and the bioinformatics analysis was further performed.The PatASIL2-EGFP expressed vector was constructed and transformed into Arabidopsis protoplasm to investigate the subcellular localization of PatASIL2.The real-time quantitative reverse transcription PCR(qRT-PCR)was also performed to detect the expression profiles of PatASIL2 gene in different tissues of patchouli and under treatments of methyl jasmonate(MeJA),salt stress,drought stress,and cold stress.The results indicated that PatASIL2 gene contained an open reading frame of 1 035 bp and encoded 344 amino acids.The PatASIL2 protein was an unstable hydrophilic protein without transmembrane domains and signal peptides and had 41 serine phosphorylation sites and one Myb_DNA-bind_4 conserved domain.The phylogenetic analysis showed that PatASIL2 was classified into the SIP1 subfamily of Trihelix transcription factor family,and clustered closely with Sesamum indicum SiASIL2.The subcellular localization result indicated that PatASIL2 was a nuclear-localized protein.The qRT-PCR results indicated that PatASIL2 expressed in the young leaf,mature leaf,old leaf,stem,and root in patchouli,particularly with the highest expression in the old leaf.The PatASIL2 expression was significantly upregulated by MeJA at 12—24 h and salt stress at 3—24 h.The expression of PatASIL2 was significantly upregulated at 24 h after drought treatment,and at 12 h after cold treatment.

As one of the subfamily members of Ca²⁺-ATPase,ACA exerts a vital role in sustaining the intracellular Ca²⁺ concentration equilibrium and modulating plant growth and development in response to abiotic stress.To acquire an in-depth comprehension of the function of the lettuce ACA gene family,bioinformatics methodologies were utilized to identify and dissect the members of the lettuce ACA gene family.The outcomes manifested that 17 ACA genes were identified in lettuce,denominated as LsACA1 to LsACA17;the LsACA genes were unevenly distributed across 8 chromosomes;the subcellular localization prediction results disclosed that all LsACA proteins were localized in the plasma membrane;the number of introns exhibited significant variation(0—32)among the members of the LsACA gene family,a total of 15 conserved domains of LsACA proteins were identified,with amino acids ranging from 21 to 50;the proportion of the secondary structure was alpha helix>random coil>extended strand>beta turn;based on phylogenetic analysis,LsACA proteins were categorized into 5 subfamilies,namely Group Ⅰ to Group Ⅴ;according to collinearity analysis,it was found that there were fragment duplications in 6 pairs of genes,the Ka/Ks of their collinear gene pairs were all less than 1,signifying that purifying selection was the preponderant force in evolution.The expression patterns of LsACA gene family members under different calcium ion concentrations were analyzed by qRT-PCR.The results showed that:compared with the control,the expression levels of 12 LsACA genes in the calcium-sensitive variety Baoshilü under low calcium treatment were extremely significantly downregulated,while the expression levels of 9 LsACA genes in the calcium-insensitive variety Yeluo were extremely significantly upregulated and the expression level of 1 LsACA gene was significantly upregulated.The ACA gene family members of lettuce were identified and analyzed,and the characteristics of the LsACA gene family members were revealed.

This study aims to provide theoretical guidance for the effective use of sister line materials,accelerate the promotion of the wheat Annong 1687 variety,and offer a reference for the genetic improvement of new wheat varieties.Wheat Annong 1687 and its sister lines and parents was used as materials.We combined phenotypic identification of field agronomic traits with genotypic identification of wheat materials using 55K microarrays.The data was analysed using SPSS and mapped it with the RIdeogram package.Significant differences in spike length were found between parents and some sister line combinations.Specifically,parents Annong 1106 and Xinong 822,and sister lines Annong 1687 and Xinong 822,showed very significant differences.The parental lines Annong 1106 and Xinong 822,as well as the sister lines Q6,8 and 105,133,exhibited a total of 909 differential SNP sites across 19 chromosomes.Notably,chromosome 5B contained 500 of these sites,which were concentrated in intervals of 63—87 Mb and 400—410 Mb.Cloning and sequencing of candidate genes within the intervals revealed that two genes encoding growth hormone-inducible proteins,TraesCS5B01G225000 and TraesCS5B01G058700,had missense mutations resulting in amino acid changes.Mutations in TraesCS5B01G225000 resulted in a codon change to the termination codon TGA and termination of amino acid synthesis.The bi-parental and sister line combinations had five and two missense mutations,respectively.It is hypothesized that these two genes may have an effect on the length of wheat spikes.

To explore the impact of organic fertilizer on the physiological growth of flax and the rhizosphere bacterial communities,and to investigate green high-yield cultivation techniques for flax in dryland,a field experiment was conducted using Baxuan No.3 as the material.The study examined the effects of four different fertilization treatments(T0:no application; T1:low quantity of cow manure; T2:medium cow manure; T3:high quantity of cow manure)on the physiological growth changes,nitrogen utilization,dry matter accumulation,and the diversity,community composition,co-occurrence networks,and metabolic pathways of the rhizosphere bacteria of flax,as well as discussing the environmental factors driving the differences in bacterial communities.The results showed that the T3 treatment resulted in higher flax production. Compared to the control, this fertilization condition also had the highest indicators for plant height, capsule fruit number per plant, thousand-grain weight, and nitrogen use efficiency, which form the physiological basis for stable yield following the application of organic fertilizer.The application of organic fertilizer significantly affected the diversity and richness of the rhizosphere soil bacteria of flax,and there were significant differences in the structure of the rhizosphere bacterial community.The population structure of the rhizosphere bacteria of flax was influenced by organic matter,total nitrogen,and available phosphorus.The dominant flora in the rhizosphere of flax was the same across different treatments,but the relative abundance of each dominant flora varied significantly.The rhizosphere of flax was dominated by the phyla Proteobacteria,Actinobacteria,Acidobacteria,Chloroflexi,and Bacteroidetes.The relative abundance of Proteobacteria and Actinobacteria increased with the increase of organic fertilizer treatments,while that of Acidobacteria decreased with the increase of organic fertilizer treatments.WGCNA analysis identified 15 co-expression modules,with the Red and Pink modules showing a significant positive correlation with organic matter content.The application of organic fertilizer increased the complexity of the bacterial network,and seven key OTUs were identified through combined WGCNA analysis.In conclusion,the application of organic fertilizer promoted the growth of flax and altered the structure and network complexity of the bacterial community in the rhizosphere soil of flax.

In order to reveal the expression patterns of PsbS protein in Nicotiana tabacum,the full-length NtPsbS gene sequence was cloned from cDNA of the tobacco cultivar K326,and the multiple protein sequences of NtPsbS gene and PsbS genes of seven crops including rice,tomato and soybean were compared by DNAMAN software.The phylogenetic analysis was carried out by MEGA 11 software,and the phylogenetic tree was established by neighbor-joining method.Tissue expression of NtPsbS gene in tobacco at different growth stages was detected by qRT-PCR.The plant expression vector pS1300-PsbS-GFP was constructed to find subcellular localization of the NtPsbS mature protein.Finally,the mRNA levels of NtPsbS under abiotic stresses were analyzed in K326.The results showed that the total length of tobacco NtPsbS gene was 825 bp,encoding 274 amino acids.The homology of tobacco NtPsbS protein and tomato SlPsbS protein was the highest up to 91%.The NtPsbS gene exhibited the highest mRNA levels in the leaves among the tissues,including leaves,roots,stems,seeds and other parts of K326.The mature NtPsbS protein was located in chloroplasts.For the abiotic stress,it was found that the expression level of NtPsbS was significantly increased under salt stress,cold stress and Abscisic acid(ABA)treatments.In summary,the expression level of NtPsbS was the highest in tobacco leaves at different growth stages,and it was higher when under the treatments of salt stress,cold stress and ABA,indicating that this gene may be involved in the salt and cold stress resistance and ABA metabolic pathway in tobacco,which could provide basis and reference for the functional analysis of the NtPsbS gene in future.

This study aimed to compare and analyze the milk composition,serum biochemical indices and expression levels of Leptin and LF genes during lactation between Bamei pigs and Yorkshire pigs.The six purebred Bamei pigs and Yorkshire pigs with healthy body conditions were selected respectively,which also had close parity,mating and delivery periods(within three days).A total of 50 mL milk,2 mL serum and 2 mL whole blood samples were collected from Bamei pigs and Yorkshire pigs on the 1st,7th,14th,21st and 28th day after delivery,respectively.Then,the milk,serum and whole blood samples were used to detect the milk compositions,serum indexes of alkaline phosphatase(ALP),albumin(ALB),globulin(GLB),triglyceride(TG),urea nitrogen(BUN),alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and the expression levels of Leptin and LF genes.The results showed that on the 1st day of lactation in Baimei pigs,in addition to milk fat(Fat),the content of casein(CS),total protein(TP),total solid matter(TS),non-fat milk solids(SNF),lactose(L),free fatty acid(FFA)and acidity in the colostrum of Bamei pigs were all extremely significantly higher than those in the colostrum of Yorkshire pigs,while,the content of CS,TP,Fat,TS,SNF and L of Bamei pigs on the 21st day after delivery were extremely significantly lower than those of Yorkshire pigs.The serum biochemical indexes of two pig breeds showed different trends during the whole lactation.The content of ALT,AST,GLB,BUN and TG of Bamei pigs were significantly higher than those of Yorkshire pigs on the 1st day after delivery,while the content of ALP and ALB were all significantly lower than those of Yorkshire pigs.There were no different levels of these two genes on the 1st day,however,from the 7th day after delivery,the expression levels of LF in the Bamei pigs were significantly different than those in the Yorkshire pigs,while Leptin gene was extremely significantly expressed in Yorkshire pigs than in Bamei pigs.In general,the milk nutrients of Bamei and Yorkshire pigs showed a decreasing trend as the extension of the lactation periods,and the expression levels of Leptin and LF genes were likely related to the milk yield and quality between the two breeds,which may need to be further verified.

The aim of this study was to explore the effects and regulatory roles of HSP60 on the testes and ovaries,uterus and fallopian tubes before and after ovulation in Bactrian camels at different developmental stages.The CDS region sequence of the HSP60 was cloned from the Bactrian camel testis and its bioinformatics analysis was analyzed using ProParam and MEGA 7.0 software,and its expression was explored by polymerase chain reaction(PCR),hematoxylin and eosin(H&E)staining,immunohistochemistry(IHC),Real-time Quantitative PCR(qPCR)and Western Blot.The cloning results showed that the coding sequence(CDS)region of the HSP60 was 1 722 bp in length and encoded 537 amino acids,and the HSP60 sequence of the Bactrian camel was similar to that of dromedary camel and alpaca.The qPCR results showed that HSP60 was expressed in the testes of Bactrian camels at different developmental stages,and the expression levels in the testes after sexual maturity(3,5 and 7 years old)were significantly higher than that at 3 months of age.The mRNA expression of HSP60 was found in the ovaries,uterus and fallopian tubes before and after ovulation,and the mRNA levels in the ovaries and uterus were significantly higher after ovulation than before ovulation,but there was no significant difference in the fallopian tube before and after ovulation.Western Blot results showed that the expression trends of HSP60 in the testes at different developmental stages were similar to that of mRNA,and the protein expression level increased with age.The protein levels of HSP60 in the ovaries,uterus and fallopian tubes were significantly higher after ovulation than before ovulation.Immunohistochemical staining revealed that the HSP60 protein was primarily located in Sertoli cells,Leydig cells,part of spermatogenic cells,granulosa cells of the ovaries,glandular cells of the endometrium and muscle cells.The results showed that HSP60 was involved in testicular development and spermatogenesis in Bactrian camels,as well as in the regulatory processes of ovulation induction and meiosis.

In order to clarify the role of the Zn(Ⅱ)2Cys6 transcription factor gene VDAG_ 04814 in the growth, development and pathogenicity of the Verticillium dahliae. It constructed a VDAG_04814 gene knockout mutant using homologous recombination mediated by polyethylene glycol. Wild-type and mutant strains were inoculated separately onto PDA media supplemented with hydrogen peroxide, sodium chloride, potassium chloride, sorbitol, sodium dodecyl sulfate, Congo red, as well as onto media overlaid with sterile cellophane, to analyze their levels of resistance to oxidative stress, salt stress, osmotic stress, stress on cell wall and plasma membrane integrity, and strain penetration ability. Their pathogenicity was assayed, and the fungal biomass in potato plants was detected. After hygromycin selection and PCR validation, the correct knockout transformants were able to amplify DNA bands of 1 500 bp upstream and downstream, respectively, as well as the full-length 4 500 bp knockout fragment sequence. The results demonstrated that the growth rate and melanin formation ability of the ΔVDAG_04814 mutants were significantly reduced. On media subjected to oxidative stress and salt stress with the addition of hydrogen peroxide, sodium chloride, and potassium chloride, ΔVDAG_04814 showed a higher inhibition rate compared to the wild-type. On osmotic stress media with sorbitol, the growth inhibition rate of ΔVDAG_04814 was significantly lower than the wild type. No growth inhibition was observed for ΔVDAG_04814 on media subjected to cell wall and membrane integrity stress with the addition of sodium dodecyl sulfate and Congo red. On media overlaid with sterile cellophane, no colonies grew for ΔVDAG_04814, whereas the wild-type strain produced normal colonies. Pathogenicity tests indicated that the wilting index of ΔVDAG_04814 was significantly reduced compared to the wild-type, with wilting index ranging from 47.22 to 55.56. It has been demonstrated that VDAG_04814 can regulate the growth, development, stress resistance, penetration ability and pathogenicity of V. dahliae towards potato. This study provides a new target for the control of potato Verticillium wilt disease.

In order to explore the production performance and soil improvement effect of sweet sorghum in saline-alkali land,Zhongketian 438 sweet sorghum was planted in Southern Xinjiang,and its yield was measured,in addition,0-10 cm,10-20 cm,20-30 cm soil layers were selected on saline-alkali land before planting and after harvesting of feed sweet sorghum varieties,and the effects of soil physical and chemical properties,soil ions and soil enzyme activities were measured and compared.The results showed that the total salinity,pH value and electrical conductivity of the soil decreased after the planting of sweet sorghum.Compared with before planting,the contents of Na⁺,K⁺,HCO₃^-,Cl^-and SO₄^2- and total potassium,total phosphorus,available phosphorus and available nitrogen decreased significantly,while the contents of Ca²⁺,Mg²⁺,total porosity,organic matter,total nitrogen and available potassium increased.The content of Mg²⁺ in soil cation was highest,and the content of SO₄^2- in anion was the higher than that of Cl^-.In soil enzyme activity,the activities of catalase,sucrase and alkaline phosphatase were not significantly different,but the urease activity was significantly different.Compared with before planting,the change of 10-20 cm soil layer in different soil layers was more obvious.The results of correlation analysis showed that there was a significant positive correlation between pH value and total salinity;there was a significant negative correlation between bulk density and pH,total salinity;and a significant positive correlation between sucrase and alkaline phosphatase.The above results showed that the saline-alkali soil in this area is a soil type dominated by Na₂SO₄,and the cultivation of feed sweet sorghum in the saline-alkali soil in this area could not only harvest high-yield forage grass,but also reduce some ions in the soil to improve soil physical and chemical properties.

This study evaluates the efficacy of bio-organic fertilizer in managing Verticillium wilt in sunflowers,examining its effects on plant growth and disease resistance,and exploring its potential as a sustainable disease control strategy in sustainable agriculture.Pot experiment was conducted to investigate the effects of bio-organic fertilizer at different concentrations and under various treatment conditions on the sunflower Verticillium wilt pathogen and its impact on sunflower growth and disease resistance.The results indicated that the treatment significantly reduced the disease index of Verticillium wilt by 14.57% compared to the control group,with a relative control efficacy of 28.54% under indoor conditions.The application of bio-organic fertilizer significantly promoted sunflower growth and development.Specifically,when 1 g of bio-organic fertilizer was mixed with every 50 g of field soil,the sunflower emergence rate increased by 8.67 percentage point,and key physiological parameters,including seedling height,stem diameter,and fresh weight,were significantly enhanced.Further investigation revealed that bio-organic fertilizer fermentation filtrates at various concentrations effectively inhibited the growth of Verticillium dahliae colonies and spore germination,with inhibitory effects diminishing as dilution increased.Additionally,volatile compounds from the fertilizer significantly suppressed the formation of microsclerotia.In terms of disease resistance mechanisms,the bio-organic fertilizer fermentation filtrates enhanced plant resistance by inducing induced resistance.Physiological analysis showed that the filtrates triggered reactive oxygen species (H₂O₂)bursts,elevated the activities of superoxide dismutase (SOD) and peroxidase (POD),reduced malondialdehyde (MDA) accumulation,and increased the activity of phenylalanine ammonia-lyase (PAL),thereby significantly enhancing disease resistance in sunflowers.This study highlights the control potential of bio-organic fertilizer for managing sunflower Verticillium wilt and promoting plant health,providing theoretical and experimental support for optimizing fertilizer formulations and developing effective disease control strategies to enhance agricultural production.

In order to investigate the immune protective effect of apical membrane antigen 1(AMA1)of Eimeria tenella on chickens infected with E.tenella, the AMA1 gene amplified was connected to expression vector pET-32a(+).The recombinant plasmid was transformed into E.coli BL21(DE3)to express recombinant protein.108 chickens were randomly divided into 6 groups with 3 replicates in each group and 6 chicks in each replicate,including non-immune and non-attacking group(negative control group),non-immune attack group(positive control group),Freund's adjuvant group and 12.5,25.0,50.0 μg rEtAMA1 immunized groups.All of the chickens were injected with different concentration of recombinant protein at the age of 14,21 days,and infected orally with 5×10⁴ sporulated oocysts of E.tenella at the age of 28 days and killed at the age of 35 days.The immune protection effect of rEtAMA1 was evaluated by body weight gain,lesion score,oocyst production,immune organ index and levels of cytokines and antibodies.There was no significant difference in average weight gain among the groups.Compared with the positive group,the scores of cecal lesions in 25.0 μg and 50.0 μg rEtAMA1 groups were significantly reduced by 53%,41%,the OPG in 12.5,25.0,50.0 μg rEtAMA1 groups were significantly reduced by 16%,27%,54% respectively,the immune organ index of bursa of Fabricius in 50 μg rEtAMA1 groups significantly increased by 27%,the immune organ index of thymus in 25.0 μg and 50.0 μg rEtAMA1 groups significantly increased by 25%,22% respectively.There was no significant difference in the immune organ index of spleen.Seven days after the second immunization,the levels of IL-2 and IFN-γ in 12.5 μg and 25.0 μg rEtAMA1 groups were significantly higher than the negative control group.The level of serum total IgG concentration in the 50.0 μg rEtAMA1 group significantly increased by 47% than that in the negative control group,and the concentration after the second immunization was obviously higher than that after the first immunization.The results showed that the rEtAMA1 could alleviate cecal lesions,decrease the oocyst production,promote the development of immune organs and increase the levels of cytokines and antibodies in serum,which had a certain immune protective effect on chickens infected with E.tenella.

In order to investigate the effects of different mulching measures on soil moisture and heat status and yield of winter wheat in northwest dry area,winter wheat Kangzhuang 974 was used as test material.From September 2022 to July 2023,three different coverage treatments including wheat straw strip covering three rows(M3),four rows(M4)and five rows(M5)and plastic film covering(PM)treatment were set up in the experimental base of Gansu Agricultural University,The open field(CK)was used as the control.The results showed that:compared with CK,the soil water storage of 0—200 cm during the whole growth period of winter wheat was significantly increased by mulch,the average increase of straw strip mulch was 13.22%,the increase range was M3>M4>M5,and the PM increased by 19.65%.The soil moisture increasing effect of mulch increased gradually with the progress of growth stage,and the maximum increase was 37.53—87.76 mm at maturity stage.It decreased with the deepening of soil layer,and the increase of 0—20 cm was the largest,ranging from 5.10—9.48 mm.Mulching significantly reduced the total water consumption and total water consumption intensity during the whole growth period,and the influence of mulching on water consumption and total water consumption intensity during the later growth period was most obvious.Compared with CK,wheat straw strip mulching significantly reduced the soil temperature of 0—25 cm during the whole growth period by 1.60—2.70 ℃,and M3 treatment had the largest decrease.The maximum decrease occurred at the grouting stage(3.67 ℃),and the maximum decrease between soil layers(3.01 ℃)occurred at 5 cm.Compared with CK,PM significantly increased the soil temperature from 0 to 25 cm during the whole growth period by 1.50 ℃,and the overwintering period and 5 cm had the largest increase,which were 2.20,1.79 ℃,respectively.The temperature of straw mulch increased at 7:00 at wintering stage,jointing stage and ripening stage,and the temperature increased and decreased at other times.PM temperature increased at all time except at 14:00 in grouting stage and ripening stage.Compared with CK, the yield and water use efficiency of M5 and PM were increased by 8.67%, 26.49% and 0.96, 2.94 kg/(ha·mm), panicle number was the most significant factor(CV=17.67%).Yield was significantly positively correlated with spike number(r=0.754^**),WUE(r=0.891^**)and soil temperature(r=0.723^**),and significantly positively correlated with grain number per spike(r=0.522^*).Banded mulching of wheat stalk can achieve both ecological and economic benefits,and M5 is more conducive to the formation of yield.

In order to study the role of phototrapping pigment binding protein gene in potato photosynthesis,the phototrapping pigment binding protein gene StCP24 (LOC102586836)was bioinformatics analyzed and cloned into potato variety Dongnong 310 to obtain transgenic potato.The morphological indexes,leaf physiological indexes,fluorescence parameters,and gene relative expression of transgenic and control lines were measured under different light intensities after clonal propagation.The function of StCP24 gene was analyzed.The effect of this gene on chloroplasts was determined by ultrastructural observation.The results of bioinformatics analysis showed that StCP24 protein was a hydrophilic protein,and the promoter of StCP24 gene contained elements such as photoresponse,defense and stress response.Phylogenetic analysis showed that the amino acids encoded by StCP24 gene in potato were closely related to those encoded by StCP24 gene in cultivated tomato.Under different light intensity treatments,StCP24 gene overexpressed potato significantly increased stem diameter,leaf area,leaf weight,root length,chlorophyll a,b content,Fv/Fm,ETR,ETRmax and qP compared with wild-type potato.The overexpression of StCP24 gene can make the lamellar stacking of thylakoid granules more tightly and to a higher degree.In conclusion,the overexpression of StCP24 gene can increase the content of chlorophyll a and b in leaves,improve the electron transfer rate in photo cooperative photoreaction,and promote the growth of transgenic potato plants.

In order to explore the rapid and scientific improvement model of soil fertility in newly cultivated land,it took the maize wheat rotation system of newly cultivated land as the research object,and conducted different fertilization treatment experiments (including no fertilization control (CK),organic fertilizer 15 t/ha(OF15),chemical fertilizer (MIN),organic fertilizer 7.5 t/ha+fertilizer (OF7.5+MIN),organic fertilizer 15 t/ha+fertilizer (OF15+MIN),and organic fertilizer 30 t/ha+fertilizer (OF30+MIN)).Based on the basic data of soil physical and chemical properties such as total nitrogen,alkaline nitrogen,available phosphorus,available potassium,and organic matter,the integrated fertility index (IFI) of soil nutrients was used to quantitatively and comprehensively evaluate the soil fertility of different fertilizers.The results showed that:on the basis of applying chemical fertilizers to newly cultivated land, organic fertilizers were added to improve soil available nutrients, increasing soil total nitrogen by 108.6%-149.0%, soil alkali hydrolyzed nitrogen by 253.6%-311.0%, soil available phosphorus by 78.8%-171.6%, soil available potassium by 35.6%-80.5%, soil organic matter content by 5.8%-41.9%, soil capacity by 1.0%-8.3%, soil pH value by 3.4%-7.3%, soil fertility comprehensive index by 21.6%-27.7%, and annual yield of corn and wheat by 33.2%-127.1%. The available nutrients, organic matter, soil fertility level, and annual yield of corn and wheat in newly cultivated land all showed an increasing trend with the increase of organic fertilizer application, among which 30 t/ha of organic fertilizer had the best effect. Compared with the application of organic fertilizer alone, the combination of organic fertilizer and chemical fertilizer effectively increased soil organic matter and soil comprehensive fertility index by 12.3% and 22.0%, respectively. In summary, under the conditions of this experiment, the combination of organic fertilizer and chemical fertilizer resulted in higher soil comprehensive fertility and annual crop yield compared to single application of chemical fertilizer or single application of organic fertilizer. The treatment with 30 t/ha of organic fertilizer and 2+chemical fertilizer significantly improved the soil fertility and crop yield of newly reclaimed farmland.

Cd-sensitive wheat CM28 was used as the research object to investigate the effects of spraying exogenous glycine on its physiological and biochemical processes under different concentrations of cadmium stress.The plant height and root length,above-ground transport coefficient and enrichment coefficient of cadmium,chlorophyll content and malondialdehyde(MDA)content,and the enzyme activities of superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)of CM28 under different treatments were determined.The expression levels of PEPC genes in roots and leaves under different treatments were analyzed by qRT-PCR.The results showed that under 30 μmol/L Cd stress,the spraying of exogenous glycine led to a decrease in the above-ground dry and fresh weights,a decrease in the total-chlorophyll content,and an increase in the MDA content of CM28,indicating that plants were affected by more reactive oxygen species.Under 50 μmol/L cadmium stress,glycine could effectively alleviate the negative effects of cadmium on CM28,resulting in an increase in the total-chlorophyll content,an increase in the above-ground dry and fresh weights,and a decrease in MDA content,suggesting that the plant was affected by reactive oxygen species to a greater extent.The antioxidant enzyme activity of CM28 increased with the increase of cadmium concentration,and the spraying of exogenous glycine could enhance the activity of antioxidant enzymes and alleviate the effect of cadmium on the whole.The expression of PEPC genes in roots increased overall with the rise of cadmium concentration,indicating that PEPC played an important role in the process of plant root resistance to stress,and the spraying of exogenous glycine can also effectively alleviate the cadmium stress on CM28 and reduce the expression of PEPC genes on the whole.In summary,cadmium stress negatively affects the physiological and biochemical processes of CM28,and the spraying exogenous glycine can alleviate the effects.

The ubiquitination pathway is one of the key signaling pathways in response to drought stress.In order to clarify the biological function of E3 ubiquitin ligase TaSINA101 gene in response to drought stress,the TaSINA61,TaSINA101 and TaSINA105 genes were cloned from JM47,an excellent drought-resistant wheat cultivar,and their sequence characteristics were analyzed by bioinformatics methods,and the expression levels of the three genes in wheat roots and leaves were detected by qRT-PCR under PEG-6000,NaCl,low temperature and ABA treatments.The heterologous expression of TaSINA101 in transgenic rice was used to analyze the biological function of TaSINA101 in response to drought stress.The results showed that the TaSINA61,TaSINA101 and TaSINA105 genes contained one intron and two exons,and the encoded proteins were composed of 282 amino acids.The qRT-PCR expression analysis showed that the expression of these three genes was induced by various abiotic stresses such as drought stress in roots and leaves.Phenotypic analysis of TaSINA101 transgenic rice under drought stress showed that the leaf fresh weight and dry weight, maximum root length, average root diameter and leaf relative water content of transgenic rice lines OE-1, OE-2 and OE-3 were significantly lower than those of wild type,while the relative conductivity of leaves of transgenic rice lines OE-1 and OE-2 was significantly higher than that of wild type.Therefore,TaSINA101 negatively regulates drought stress tolerance in rice.This study provides a basis for in-depth analysis of the biological function of TaSINA101 gene in wheat.

In order to elucidate the sequence characteristics and expression pattern of PatGTL1,a member of Trihelix transcription factor family,and lay a foundation for the further function study of PatGTL1,the total RNA of Pogostemon cablin (patchouli)was extracted and reverse-transcribed to cDNA.The specific primers were designed according to the transcriptome sequence of PatGTL1,and the cDNA was used as a template to clone PatGTL1 gene.The bioinformatics analysis was further performed to reveal the structural,physicochemical properties,and phylogenetic relationships of PatGTL1.Besides,the pAN580-PatGTL1-EGFP vector was built and transformed into Arabidopsis protoplastsa to detect the subcellular localization of PatGTL1.The Real-time Quantitative reverse transcription PCR(qRT-PCR)was also carried out to investigate the expression patterns of PatGTL1 gene in different tissues of patchouli and under treatments of methyl jasmonate(MeJA),salt stress,drought stress,and cold stress.The results showed that the open reading frame of the PatGTL1 gene was 1 497 bp,encoding 498 amino acids.The PatGTL1 protein was an unstable hydrophilic protein that contained 42 serine phosphorylation sites and was predicted to be located in the nucleus without transmembrane domains and signal peptides.Multiple sequences alignment showed that PatGTL1 contained two GT1 conserved domains,and was classified into GT-2 subfamily of Trihelix transcription factor family.According to the phylogenetic tree,PatGTL1 was clustered closely with Sesamum indicum SiGTL1.The qRT-PCR analysis indicated that PatGTL1 expressed in the root,stem,and leaf in patchouli,particularly with the highest expression in the young leaf.The PatGTL1 expression was significantly upregulated by MeJA and showed an increasing trend at 3—24 h after treatment,but downregulated by drought stress.The highest expression of PatGTL1 was showed at 3 h after salt treatment,and at 12 h after cold treatment.These results enrich the Trihelix transcription factor family study of patchouli.

In order to investigate the impact of different environments on the symbiotic nodulation ability of soybean-rhizobia and to screen suitable culture conditions for symbiotic nodulation,this study established 10 treatments based on 4 environmental factors(culture temperatures:22,26,30 ℃;culture substrates:vermiculite,vermiculiteand nutrient soil;inoculation periods:0 day post-inoculation,inoculation at 10 days;final bacterial concentration:OD₆₀₀=0.5,OD₆₀₀=0.9,OD₆₀₀=1.3),resulting in 36 different culture conditions.Five nodulation-related phenotypes of soybean-rhizobia were measured under these conditions,including the number of nodules per plant,nodule dry weight per plant,nodule size,SPAD of leaf growth at 10 and 24 days post-inoculation(DPI).Principal component analysis,fuzzy mathematics membership function method,and cluster analysis were employed to comprehensively evaluate and classify the symbiotic nodulation ability of soybean-rhizobia under different conditions.A comprehensive evaluation system for the nodulation ability of soybean-rhizobia was established,and the optimal culture conditions were selected based on the comprehensive evaluation index D value.The results showed that culture temperature was the primary environmental factor influencing the symbiotic nodulation of soybean-rhizobia.The maximum values of the number of nodules per plant,nodule dry weight per plant,nodule size,SPAD of leaf growth at 10 and 24 days post-inocubion corresponded to culture temperatures of 30,26,26,30 and 26 ℃,respectively,under,different culture conditions.Under the culture conditions set,the optimal culture temperature for the symbiotic nodulation of soybean-rhizobia was 26 ℃.Multivariate linear regression analysis and grey relational analysis based on the comprehensive evaluation index D value indicated that nodule dry weight and nodule size had a strong correlation with the symbiotic nodulation ability of soybean-rhizobia,serving as important reference indicators for nodulation ability identification.The results of this study established a comprehensive evaluation system for the symbiotic nodulation ability of soybean-rhizobia,providing a theoretical basis for selecting phenotypes and culture conditions for nodulation ability identification.

Calcium-dependent protein kinases(CPK)are a class of protein kinases that respond to calcium signals and participate in regulating the polar growth of pollen tubes,which is crucial for plant reproduction and is precisely controlled by calcium signals and the cytoskeleton.To reveal the function of CPK in the polar growth of pollen tubes in Arabidopsis,bioinformatics methods were used to analyze the expression patterns and evolutionary relationships of CPK family members,and constitutively active(CA)and dominant-negative(DN)CPK17 mutants were constructed.Based on gene gun technology,we observed the phenotypic changes of CA-CPK17 and DN-CPK17 overexpressing pollen tubes,compared the effects of high and low calcium stress on the growth of CPK17 and CA-CPK17 overexpressing pollen tubes,and analyzed the co-expression of CPK17 and its homologous protein CPK34.Additionally,the protein interaction network of CPK17 was predicted,and some of the interacting proteins were validated.The results showed that there were seven pollen-specifically expressed CPKs in Arabidopsis thaliana.Among them,CPK17 and CPK34 had a high similarity (93%),and were highly expressed in mature pollen,and low or no expression in other tissues.Overexpression of CPK17 had no significant effect on pollen tube growth,while CA-CPK17 caused depolarized growth and tip swelling of pollen tubes,and DN-CPK17 had no obvious effect.Overexpression of CPK34 also caused similar defects in pollen tube growth,and co-overexpression of CPK17 and CPK34 aggravated the defects.Calcium stress experiments showed that overexpressing CPK17 pollen tubes were insensitive to low calcium conditions,but sensitive to high calcium conditions.While pollen tubes overexpressing CA-CPK17(lacking the regulatory domain)were insensitive to both high and low calcium conditions.The predicted interacting proteins of CPK17 were mainly ROP negative regulators,but yeast two-hybrid experiments failed to confirm the interaction.These results indicated that the C-terminal regulatory domain of CPK17 plays an important role in the response to calcium signals.CPK17 and CPK34 have synergistic effects on pollen tube growth,possibly by regulating calcium influx and affecting intracellular calcium gradient.

Improper application of soybean fertilizer can lead to late ripening,lodging,worsening of diseases and pests,and a decrease in fertilizer and yield benefits,to improve fertilizer efficiency and fully tap into the potential of soybean yield and quality,therefore,this article investigates the effects of seven fertilization modes F1(basic application of 3 000 kg/ha organic fertilizer+300 kg/ha chemical fertilizer),F2(basic application of 3 000 kg/ha organic fertilizer+150 kg/ha chemical fertilizer+topdressing 150 kg/ha chemical fertilizer),F3(basic application of 3 000 kg/ha organic fertilizer+225 kg/ha chemical fertilizer+topdressing 75 kg/ha chemical fertilizer),F4(basic application of 1 500 kg/ha organic fertilizer+75 kg/ha chemical fertilizer+topdressing 225 kg/ha chemical fertilizer),F5(basic application of 300 kg/ha chemical fertilizer),F6(basic application of 150 kg/ha chemical fertilizer+topdressing 150 kg/ha chemical fertilizer)and F7(basic application of 225 kg/ha chemical fertilizer+topdressing 75 kg/ha chemical fertilizer)on soybean growth,development,photosynthesis,yield,and quality through field experiments using a single factor experimental design.The results showed that increasing the application of organic fertilizer significantly enhanced the dry weight of soybean roots when compared to applying chemical fertilizer alone,applying chemical fertilizer alone with a base to top dressing ratio of 3∶1 was the most beneficial for increasing root dry weight.The combination of organic fertilizer and chemical fertilizer resulted in higher fresh leaf weight,dry leaf weight,fresh stem weight,dry stem weight per plant,as well as higher fresh weight and dry weight per plant when compared to single application of chemical fertilizer,and there were no significant differences in the above indicators among the treatments treated with single application of chemical fertilizer.The combination of organic fertilizer and chemical fertilizer increased the number of leaves per plant and leaf area index of soybean at pod bearing stage and filling stage,under the condition of no organic fertilizer application,the base to top dressing ratio of 3∶1 of chemical fertilizer was beneficial for improving the number of leaves and leaf area index of soybean.Compared to single application of chemical fertilizer,combined application of organic fertilizer and chemical fertilizer increased the photosynthetic rate at flowering and filling stages by 16.69%—21.66% and 14.99%—30.66%,respectively,unreasonable base to top dressing ratio of chemical fertilizer or reducing organic fertilizer dosage could affect the chlorophyll content and photosynthetic rate of soybean in the middle and later stages of growth and development.Increasing organic fertilizer application amount could increase soybean yield and protein content,while reduce fat content,applying chemical fertilizer alone with a base to top dressing ratio of 3∶1 was the most conducive to increasing yield.Therefore,it can be concluded that the combination of organic fertilizer and chemical fertilizer is significantly better than that of single application of chemical fertilizer in improving soybean growth and development,photosynthesis,yield,and quality,in addition,single application of chemical fertilizer performs better with a base to top dressing ratio of 3∶1.

To investigate the impact of saline water irrigation on greenhouse gas emissions,including CO₂,N₂O,CH₄,and the soil microbial community in winter wheat fields,three types of saline water with different salinity levels(1,3,and 5 g/L,denoted as W1,W3,and W5)were employed.Field experiments were conducted at the Nanpi Eco-Agriculture Experimental Station of the Chinese Academy of Sciences from March to June 2023.The results indicated that CO₂ emissions had similar trends in the different saline water irrigation treatments during the wheat growing stages,which were high in the early stage,low in the middle stage,and high in the late stage.N₂O emissions exhibited a trend of high values in the early period and lower values in the later stages.While,CH₄ showed fluctuations between positive and negative emissions.Comparative analysis revealed that the average CO₂ and N₂O emission rates in W3 treatment were significantly lower than in W1,with reductions of 39.4% and 68.9%,respectively.The average CO₂ and N₂O emission rates in W5 treatment decreased by 21.9% and 40.0%,although the difference was not statistically significant.Saline water irrigation with different concentrations minimally affected soil microbial α-diversity but significantly altered community structure.Cluster analysis demonstrated a significant difference in microbial composition between W1 and W5,with W3 positioned between the two treatments.Correlation analysis showed that there was a significant positive correlation between CO₂ and N₂O emission rates and soil TN,while there was a significant positive correlation between N₂O emission rate and soil TN,TOC,DOC,MBC,respectively.Soil N₂O and CH₄ emission rates correlated positively with the abundance of S0134 terrestrial and Sphingomonas and Subgroup 25,respectively.Redundancy analysis(RDA)identified pH,NH₄⁺,EC,and DOC as key physicochemical factors influencing the abundance of Sphingomonas and Subgroup 25.In conclusion,irrigation with 3 g/L mildly saline water can reduce soil respiration rates and decrease agricultural carbon emissions without significantly increasing soil salinity,providing theoretical support for the development and utilization of mildly saline water resources in the North China Plain.

In order to explore the role of NBS-LRR gene RPM1 in the resistance to Verticillium wilt in Solanum,it took the wild Solanum sisymbriifolium Lam.as material,and cloned the homologous sequence of RPM1 gene on the basis of its transcriptional sequencing.The physicochemical properties and molecular structure of the sequence encoded protein were analyzed,the evolutionary relationship tree was constructed,and subcellular localization was performed in tobacco.At the same time,the relative expression level of RPM1 gene in different parts of Solanum sisymbriifolium Lam.was detected,as well as the relative expression level at for time points(0,24,48 and 72 h)after inoculation with Verticillium dahliae(a pathogen of Verticillium wilt).The results showed that the total length of RPM1 gene(SsRPM1)was 2 772 bp,encoding 924 amino acids.SsRPM1 protein,with a total molecular weight of 105.99 ku,was an alkaline hydrophilic protein without transmembrane structure.SsRPM1 protein was mainly composed of α helix and random coil,including LRR,NBC and CC domains.Solanum dulcamara RPM1 protein had the closest relationship with it.Subcellular localization in tobacco found that the protein was located on the cell membrane.SsRPM1 gene was expressed in different organs(root,stem and leaf),among which the stem had the highest relative expression,followed by leaf and root.After inoculation with V.dahliae,in general,SsRPM1 gene expression in both control group and inoculation group showed a trend of first increasing and then decreasing.The relative expression level of SsRPM1 gene was the highest at 24 h after inoculation.Compared with the control group,the relative expression level of SsRPM1 gene in inoculation treatment was lower.It is suggested that SsRPM1 is a negative regulatory gene in response to Verticillium wilt stress.

Cabbage fusarium wilt is a serious soil-borne fungal disease caused by Fusarium oxysporum f.sp.conglutinans (FOC),which affects the yield and quality of cabbage.In order to clarify the biological function of the transcription factor SNT2 in this pathogen,the SNT2 gene knockout mutant ΔSNT2 in F.oxysporum was successfully obtained by homologous recombination and protoplast transformation,and its phenotype and pathogenicity were analyzed.The results showed that SNT2 in F.oxysporum encoded 1 529 amino acids,had a SANT domain that binds to DNA,and the protein belonged to a hydrophilic protein.Compared with the wild type strain,the mycelial growth rate of the ΔSNT2 mutant decreased and the septation increased significantly,and the conidial production decreased significantly.Based on the results of exogenous stress,ΔSNT2 was insensitive to osmotic stress of 1 mol/L sorbitol,but its tolerance to oxygen stress,salt stress and cell wall stress of 0.1% H₂O₂,2 mol/L NaCl and 0.05% Congo red was reduced.At the same time,the pathogenicity test showed that the disease index of ΔSNT2 mutant was significantly lower than that of wild type,and the deletion of SNT2 resulted in a significant decrease in the pathogenicity of F.oxysporum.In conclusion,the transcription factor SNT2 plays an important role in maintaining the integrity of the cell wall during the interaction between the pathogen and the host,and participates in the regulation of the growth and development of F.oxysporum and the expression of pathogenicity.

The establishment of a coloration method for Fusarium verticillioides can clarify its infection pathway in maize plants and further determine its pathogenic mechanism.This study took Fusarium verticillioides as the research object to explore the staining effects of FDA,trypan blue,toluidine blue,acid fuchsin,basic fuchsin,FM4-64,and red fluorescence vector on fungal spores of F.verticillioides and the root system of maize seedlings inoculated with F.verticillioides.A coloration method for F.verticillioides was established.The results showed that 5% basic fuchsin and 1% toluidine blue had good staining effects on F.verticillioides spores,with clear color and clear structure.After staining with 5.00 g/L FDA,0.4% trypan blue,and 0.50 g/L acid fuchsin,the F.verticillioides spores could be stained with a very light color,making it difficult to observe the spore morphology.Staining the root system of maize seedlings with stains showed that 0.4% trypan blue and 1% toluidine blue stained the root system of maize with blue and brown colors,allowing clear observation of the brown area as the site of root rot caused by F.verticillioides infection in maize.However,staining with 0.50 g/L acid fuchsin and 5% basic fuchsin did not allow clear observation of the site of root rot.Observation of the red fluorescence coloration of F.verticillioides spores showed that FM4-64 had a deeper color and unclear structure,limiting observation time.However,after being transferred to a red fluorescence vector,the fluorescence color was obviousr and the structure was clear,without any time limit for observation.This method can be used in future studies on the pathogenic mechanism of F.verticillioides causing kernel rot in maize and wheat.After comprehensive comparison,the F.verticillioides transferred with a red fluorescence vecror had more advantages than other staining methods in terms of research on pathogenic mechanisms.

The highly contagious gastrointestinal infectious disease caused by Porcine epidemic diarrhea virus(PEDV)has led to significant economic losses in China's pig industry.It aimed to establish a basis for PEDV antibody detection methods and functional research of the N protein through the screening of specific nanobodies(Nbs)using phage display technology.Peripheral blood lymphocytes were isolated from a camel immunized with the N protein,and total RNA was extracted and reverse transcribed into cDNA.The variable domain of the heavy chain of heavy chain antibodies(VHH)was amplified by PCR,subcloned into the pCANTAB5E-ccdb vector,and electroporated into ER2738 competent cells to construct the VHH phage antibody display library.Subsequently,the library was subjected to four rounds of panning against the PEDV N protein,and positive phage clones were cloned into the pET-30a vector.The binding affinity and specificity of the Nbs were determined by indirect ELISA and Western Blot.The results showed that after the fifth immunization,the antibody titer reached 1∶25 600.The constructed phage display library had a capacity of 4.72×10⁸ and an abundance of 4.3×10¹⁰ cfu/mL,with a 93.75% positive rate.After four rounds of screening,16 Nb clones with different amino acid sequences were obtained,and Nb45 was validated to possess excellent specificity and binding ability to the PEDV N protein.This study successfully screened and obtained specific N protein-targeting Nbs,providing biological materials for the establishment of PEDV detection methods and foundational research.

In order to obtain Nipah virus nucleocapsid protein(N protein)with similar function and activity to natural proteins,the NiV N protein open reading frame gene was amplified by PCR,cloned into pFastBac^TMHTB vector,transformed into DH10Bac receptive cells,and transfected into sf9 insect cells.The protein was identified by Western Blotting and bioinformatics analysis was used to study the characteristics of N protein.The results showed that the recombinant plasmid pFastBac^TMHTB-NiV-N was obtained through PCR and double enzyme digestion identification.Recombinant protein Bacmid-NiV-N with immune activity was identified by SDS-PAGE and Western Blotting.According to molecular bioinformatics software analysis and prediction,the recombinant protein contained 532 amino acids with a molecular size of 58.34 ku.It was an unstable and hydrophilic protein without a transmembrane region.The secondary structure of the protein was mainly irregular curls,with 5 T cell epitopes and 18 B cell epitopes.The above research provides a material basis for further understanding the function of NiV N protein and establishing corresponding antigen detection methods.