Drought is one of the key factors limiting the yield of japonica rice.Exploring the genetic mechanism of drought tolerance in japonica rice at the germination stage,screening drought-tolerant candidate genes,and identifying their superior haplotypes provide a theoretical basis for deciphering the drought tolerance mechanism of japonica rice at the germination stage and developing related molecular markers.139 core germplasm resources of japonica rice from Northwest China were used as materials.Japonica rice seeds after radicle emergence were subjected to simulated drought stress using 15% PEG-6000.Six drought tolerance-related traits,including relative germination rate(RGR)and relative root length(RRL),were determined.Combined with 255 501 high-quality single nucleotide polymorphisms(SNPs),genome-wide association studies(GWAS)were conducted using the general linear model(GLM),mixed linear model(MLM),and fixed and random model circulating probability unification(FarmCPU)model.Haplotype analysis was further performed based on drought tolerance phenotypic data to identify potential candidate genes.The results showed that the mean values of the six drought tolerance traits followed a normal distribution,with coefficients of variation(CV)ranging from 3.43% to 56.25%.Among these traits,the relative root and shoot dry weight(RRSDW)exhibited the largest CV and showed a significantly positive correlation with relative root length(RRL),relative shoot length(RSL),and relative root and shoot fresh weight(RRSFW).GWAS for the six drought tolerance traits using high-density SNPs detected a total of 1 164 distinct SNPs,among which 379 were regarded as high-confidence SNPs loci and 6 as quantitative trait nucleotides(QTNs).By integrating the 50 kb upstream and downstream regions of these loci,14 significant SNPs loci were screened out,corresponding to 14 reliable quantitative trait loci(QTL)candidate intervals,which were mainly significantly associated with RRSDW.Genes within these 14 intervals were further predicted and analyzed using databases.With missense mutations in exons as the criterion,the gene LOC_Os03g56130,which encodes lichenase-2 precursor,was identified.Combined with drought tolerance phenotypes,haplotype analysis of this gene revealed that 5 missense mutations in exons constituted 4 haplotypes.Among them,Hap.2 showed a significantly higher RRSDW than the other haplotypes,being identified as the superior haplotype.These findings indicate that LOC_Os03g56130 is a potential candidate gene associated with drought tolerance in japonica rice at the germination stage.

To investigate the effects of salicylic acid(SA)spraying on potato physiology and yield under reduced basal fertiliser conditions,with a view to providing technical reference for hormone regulation to improve potato yield.The potato variety Dongnong 310 was used as the test material,and the field experiment was conducted using the randomised block group method.Five treatments were set:conventional basal fertiliser(T1),50% reduction of basal fertiliser(T2),50% reduction of basal fertiliser+spraying of 6.9 mg/L SA(T3),50% reduction of basal fertiliser+spraying of 13.8 mg/L SA(T4)and 50% reduction of basal fertiliser+spraying of 27.6 mg/L SA(T5).Samples were taken at the early tuber formation,late tuber formation,tuber growth stage and starch accumulation stage,and plant height,stem thickness,relative chlorophyll content,photosynthetic parameters,fluorescence kinetic parameters,canopy coverage and yield were determined.The results showed that under weight loss treatment,spraying salicylic acid could promote potato growth and increase yield per unit area.Among the four SA treatments in the same period,T3 and T4 treatments could increase potato plant height,stem thickness,Net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), photochemical quenching coefficient(qP)and starch content;and T4 and T5 treatments could increase potato canopy coverage,Fv/Fm,maximum electron transport rate(ETRmax), and half-saturated light intensity(Ik).Spraying different concentrations of SA had a significant effect on the yield of potato tuber under the weight-loss treatments,and T1,T3,T4,and T5 treatments were significantly higher than T2,increasing 36.74%,27.35%,29.83%,22.96%.The number of potatoes produced per plant was highest in T1 treatment,followed by T4,and the difference between the two was not significant,and T1,T3,and T4 treatments were 33.47%,28.28%,and 30.29% higher than T2.Single plant yield of T1,T3,T4 and T5 treatments increased by 35.19%,27.87%,28.76%,23.39% compared to T2.The commercial potato rate T3 and T4 increased by 3.67, 3.72 percentage points compared with T2,and the differences were significant.In summary,exogenous SA increased the Pn,Tr and Gs of potato leaves under 50% reduction of basal fertilizer treatment,and increased the yield per unit area,and the spraying of 13.8 mg/L SA in the pre-tuber formation stage could effectively alleviate the adverse effects of reduced basal fertilizer on the growth and yield of potato.

To investigate the responses of different types of wheat to various nitrogen-phosphorus-potassium ratio compound fertilizers in the Huaihe River region,in terms of photosynthetic characteristics,dry matter accumulation and distribution,yield and quality,four wheat varieties(Zhongke 166,Yannong 999,Huaimai 43,Shannong 17)were selected as experimental materials.Two types of compound fertilizers were set up:high-nitrogen compound fertilizer(N-P₂O₅-K₂O=26-13-7)and equal-proportion compound fertilizer(N-P₂O₅-K₂O=18-18-18).The effects of different compound fertilizer treatments on the photosynthetic characteristics,dry matter accumulation and distribution,yield and quality of each wheat variety were analyzed.The results showed that compared with the equal-proportion compound fertilizer,the high-nitrogen compound fertilizer significantly increased the LAI,SPAD value,net photosynthetic rate and photochemical efficiency of the four varieties,delayed leaf senescence,and prolonged photosynthetic time;it also increased the dry matter accumulation at the flowering and maturity stages,optimized the dry matter distribution ratio,and ultimately increased the grain yield by 4.5% to 14.9%,with protein and wet gluten contents increasing by 1.36% to 8.17% and 3.55% to 9.36% respectively.The response of different varieties to the high-nitrogen compound fertilizer varied slightly,with Yannong 999 having the best overall performance,the largest increase in SPAD value and LAI,the highest net photosynthetic rate and yield,the highest dry matter distribution ratio at the maturity stage,and significant quality improvement;Zhongke 166 had the largest yield increase,the highest protein content and wet gluten content;Huaimai 43 had the highest increase in grain dry matter accumulation;Shannong 17 had the weakest response to the high-nitrogen compound fertilizer,but its yield was still 9.3% higher than that of the equal-proportion compound fertilizer.In summary,the high-nitrogen compound fertilizer showed significant advantages in the photosynthetic characteristics,dry matter accumulation and distribution,yield and quality of different types of wheat.Among them,Yannong 999 had the best overall performance.

In order to clarify the effects of climate change and organic fertilization on soil nitrogen cycling,a soybean field experiment was conducted in Hefei, Anhui from June to November 2023. Field warming was simulated using infrared radiation warming equipment, and two temperature levels(T0:Ambient temperature;T1:Warming)and different fertilization managements(SF:Synthetic fertilization;OF:Organic fertilization),and determined the effects of warming and organic fertilization on nitrification and denitrification processes and their drivers by investigating soil physicochemical properties,the relative contributions of ammonia-oxidizing bacteria(AOB)and archaea(AOA)to the nitrification potential,denitrification capacity and N₂O/(N₂O+N₂)ratio,as well as soil respiration in combination with enzyme activity measurements.The results showed that warming significantly decreased soil moisture(2.3 precentage points),organic carbon(22.7%),total phosphorus(23.8%),N${\mathrm{H}}_{4}^{+}$-N(17.5%),while significantly increased NO₃^--N(52.8%);organic fertilization significantly increased soil moisture,organic carbon,total phosphorus,NH₄⁺-N content by 1.2 percentage points,26.5%,31.3%,17.0%,respectively.Warming increased soil respiration(26.4%),nitrification potential(31.7%),AOB contribution(16.4 percentage points),denitrification capacity(49.1%),N₂O/(N₂O+N₂)ratio(10.5 percentage points).Organic fertilization significantly decreased the nitrification potential, AOA contribution, denitrification capacity, and N₂O/(N₂O+N₂)ratio by 39.3%, 18.2 percentage points, 24.7% and 16.2 percentage points,respectively.Warming and organic fertilization significantly increased urease (S-UE), nitrite reductase (S-NiR), and hydroxylamine reductase(HR)activities.The correlation analyses showed that NO₃^--N content,S-UE,S-NiR,and HR activities were significantly positively correlated with AOB contribution rate and denitrification capacity.Therefore,warming increased soil N${\mathrm{O}}_{3}^{-}$-N content and the activities of S-UE,S-NiR and HR,which in turn increased the nitrification contribution of AOB,denitrification capacity and N₂O/(N₂O+N₂)ratio,leading to an increase in the loss of N₂O;while organic fertilization could increase the soil nutrients and regulate the role of AOA and AOB in the nitrification process,reduce the contribution of AOA,denitrification capacity and N₂O/(N₂O+N₂)ratio,and thus decrease the N₂O loss,consequently,mitigating the adverse effects of warming.

In order to further analyze the genetic basis regulating the protein content of super sweet corn stalk,this study used the BLINK model to locate the major SNP loci and predict candidate genes controlling the trait,based on the maize 56K gene chip.The results showed that during the harvest period of super sweet corn,the protein content of stalk ranged from 12.63 to 30.29 g/kg,with an average of 20.76 g/kg,showing a normal distribution.Six significant SNP loci regulating the protein content of super sweet corn stalk were identified by GWAS.These loci were distributed on chromosomes 1,4 and 6,and their contribution rates ranged from 10.07% to 18.17%.Among them,Affx-115330268 on chromosome 6 had the highest contribution rate.A total of 184 candidate genes were detected in the 200 kb confidence interval between the upstream and downstream of the significant association sites,of which 34 were annotated genes,accounting for 18.48%.Through GO functional annotation and KEGG pathway enrichment analysis,candidate genes were mainly involved in biological processes such as biological regulation,cellular processes,and catalytic activity,and were enriched in steroid biosynthesis,tryptophan metabolism,photosynthesis-antenna protein and other pathways.Based on the literature search and comparison,it was found that six key genes (GRMZM2G141473, GRMZM2G007854, GRMZM2G111319, GRMZM2G121024, GRMZM2G010353, GRMZM2G049547) that regulate the protein content of super sweet corn stalk,which are mainly involved in protein and enzyme synthesis.

A novel microdroplet digital PCR(ddPCR)assay was developed for rapid and quantitative detection of Pseudomonas amygdali pv.lachrymans(Pal),the causative agent of cucumber bacterial angular leaf spot disease.A specific primer pair Pal-F/R and probe Pal-Pro targeting the glyceraldehyde-3-phosphate dehydrogenase(gap1)gene were designed.The annealing temperature,primer and probe concentrations,and specificity and sensitivity tests of ddPCR were optimized.The results showed that the optimal annealing temperature was 61.2 ℃ and the optimal final concentrations of the primers and the probe were 600,250 nmol/L,respectively.The ddPCR assay demonstrated high sensitivity,with a detection limit of 2.7 copies/μL for genomic DNA.A total of 32 samples of bacterial ooze,greenhouse roof water droplets,and leaf margin guttation fluids collected from asymptomatic leaf lesions in diseased and healthy cucumber greenhouses were detected via ddPCR assay,Pal was detected in 26 samples,which was 3 more than the results obtained by the TaqMan qPCR,indicating that the ddPCR assay exhibits higher sensitivity for early detection compared to qPCR.The ddPCR assay developed in this study is highly specific and sensitive,providing a reliable technical approach for absolute quantitative detection of Pal and is of great significance for the early diagnosis warning of the cucumber bacterial angular spot disease.

It aimed to investigate the effects of different levels of grape pomace in feed on serum antioxidant indicators,blood metabolites,and metabolic pathways in beef cattle,with the goal of providing theoretical support for enhancing antioxidant function and production performance through dietary intervention.Twenty healthy 12-month-old Simmental beef cattle were randomly divided into four groups,each with five replicates.The control group received a basal diet,while experimental groups were fed basal diets supplemented with 10%(Group A),20%(Group B),and 30%(Group C)grape pomace for 40 days.At the end of the trial,serum antioxidant parameters were measured for each animal,and metabolomic analysis was conducted.Adding 20% grape pomace(Group B)significantly increased superoxide dismutase(SOD)activity while markedly decreasing catalase(CAT)activity.Adding 30% grape pomace(Group C)extremely significantly increased SOD levels and significantly elevated total antioxidant capacity(T-AOC)levels.Compared to the control group,metabolomic analysis of Group C(30%)revealed 16 differentially upregulated metabolites.Among these,five metabolites-fumaric acid,phosphatidylcholine, phosphatidylethanolamine, 4-chlorothiourea acid, and compounds containing unsaturated double bonds.Among the 14 downregulated differential metabolites,those involved in antioxidant functions included methyl gallate,L-phenylalanine betaine,and N,N-diethyl-4-hydroxybenzamide.KEGG pathway enrichment analysis identified six pathways associated with antioxidant responses:amino acid metabolism,oxidative phosphorylation,lipid metabolism,ABC transporter pathways,cofactors and vitamins,and energy metabolism.The results indicate that supplementing the diet with 30% grape pomace significantly enhances the oxidative-antioxidant indicators SOD and T-AOC in Simmental beef cattle.This effect is achieved by regulating metabolites such as fumaric acid and methyl gallate,thereby influencing metabolic pathways including amino acid metabolism and ultimately boosting the cattle's antioxidant capacity.

The aim was to clone the IGF-1 gene of Gannan yak,conduct bioinformatics analysis and detect its expression pattern in different tissues,so as to provide a reference for studying its function.Taking the cDNA of testicular tissue of Gannan yaks as the template,the CDS region of the IGF-1 gene was amplified by PCR,and then bioinformatics analysis was carried out.Subsequently,Quantitative Real-time PCR(RT-qPCR)was used to detect its expression patterns in heart,liver,lung,kidney,spleen,muscle,testicular and adipose tissues.The results showed that the CDS region of the IGF-1 gene was Gannan yak was 567 bp in total length and encoded 188 amino acids.The molecular formula of IGF-1 protein was C₉₀₇H₁₄₆₄N₂₇₂O₂₆₅S₁₇,with an isoelectric point of 9.72 and an instability index of 69.91,which belonged to unstable proteins.The IGF-1 protein was a hydrophilic secreted protein containing signal peptides.It was mainly located outside the cell and contained 10 O-glycosylation and 19 phosphorylation sites.The secondary structure was mainly composed of random coil and α-helix.It interacted with proteins such as IGF1R,IGFBP3,INSR and IGFBP1.The results of RT-qPCR showed that the expression level of IGF-1 gene in the liver of Gannan yak was extremely significantly higher than that in other tissues,and the expression levels in the spleen and fat were extremely significantly higher than those in the lungs,testicles and muscles.

To determine the optimum strip configuration for the intercropping of soybean and maize with different border-row advantages,the field experiments were carried out in two distinct ecological regions,Jian'an District of Xuchang City and Shangshui County of Zhoukou City,Henan Province.The experimental materials included two maize varieties with strong border-row advantage(MC121 and Denghai 3206(DH3206)),and two with weak border-row advantage(Haoyu 16(HY16)and MY73),previously screened,and one soybean variety Jindou 99(JD99).Two strip intercropping patterns were established∶6 rows of soybeans alternating with 4 rows of maize(6S4M)and 4 rows of soybeans alternating with 2 rows of maize(4S2M).Sole cropping of soybeans(CKS)and maize(CKM)were set as the controls.This study investigated the variations in maize agronomic traits,dry shoot weight,yield and its components of each row under different strip patterns,and compared the differences of land equivalent ratio(LER).The results showed that the LER was greater than 1 in 12 of the 16 treatments formed by two strip configurations for four maize varieties in the two ecological regions,and the LER of maize varieties HY16 and MY73 was greater than 1 under each strip configuration at two sites.The LER of MC121 and DH3206,with strong border-row advantage,was higher in strip type of 4S2M than that of 6S4M.In contrast,the LER of HY16,with weak border-row advantage,was higher in strip type of 6S4M than that of 4S2M,while it was opposite for MY73.Under 6S4M planting pattern,stem diameter,canopy light transmittance,chlorophyll a and chlorophyll b contents in ear leaf and leaf below the ear of maize variety DH3206 decreased with the increase of border row order;bottom light transmittance,chlorophyll b content in ear leaf and leaf below the ear of maize variety MC121 decreased with the increase of border row order in two experimental sites.The yield of maize varieties MC121 and DH3206 with strong border-row advantage in Jian'an District and Shangshui County decreased by 23.71%,27.00% and by 20.03%,9.05%,respectively.The dry shoot weight per plant of the edge row-2 at maturity decreased by 15.87%,19.93% and by 13.82%,24.56%,respectively,compared with the edge row-1.In contrast,there was no significant change in yield and dry shoot weight per plant at maturity between the edge row-1 and the edge row-2 of maize varieties with weak border-row advantage.In conclusion,the average LER of varieties with weak border-row advantage is higher than those with strong border-row advantage under the same strip configuration,and maize yield difference between intercropping and monoculture is seriously affected by variety type.Therefore,screening and matching suitable maize varieties is essential for improving the yield and benefit of soybean-maize strip intercropping system.

The mechanism of the effect of phosphorus application rate on the growth,yield formation and phosphorus fertilizer utilization rate of high-yield maize in Xinjiang under the condition of drip irrigation and fertilizer integration was clarified,in order to provide a theoretical basis for the high and stable yield of maize in Xinjiang and the green sustainability of fertilizer reduction and efficiency increase.The experiment was carried out in the maize high-yield demonstration base of Qitai Farm in Xinjiang from 2023 to 2024.Under the condition of drip irrigation and fertilizer integration,six phosphorus application levels were set up:no phosphorus fertilizer(P₀),pure phosphorus 60 kg/ha (P₆₀),90 kg/ha(P₉₀),120 kg/ha (P₁₂₀),150 kg/ha (P₁₅₀)and 180 kg/ha(P₁₈₀,CK).The changes of leaf area index(LAI),photosynthetic potential(LAD),dry matter accumulation,harvest index(HI),ear traits,yield and yield components of maize with water and fertilizer integration of dense planting drip irrigation were analyzed,and the effect of phosphorus application rate on yield and phosphorus fertilizer utilization rate of maize with dense planting drip irrigation in Xinjiang was clarified.The results showed that under the condition of drip fertigation and phosphorus application,the maize yield increased first and then tended to be stable with the increase of phosphorus application in the two years,and the yield was the highest(18.79—20.94 t/ha)at P₁₂₀ treatment.The amount of phosphorus application significantly affected the 1000-grain weight and grain number per ear of maize.The amount of phosphorus application mainly affected the length of maize ear bald tip.Compared with P₁₅₀ and P₁₈₀ treatments,the difference of ear bald tip length was not significant.During the critical growth period of corn,LAI showed a trend of first increasing and then stabilizing with increasing phosphorus fertilizer application rates.The LAI of maize under P₁₂₀ treatment at silking stage was 7.56—7.81,which was not significantly different from P₁₅₀ and P₁₈₀ treatments,but was significantly reduced by 21.54% under P₀ compared with P₁₂₀.In addition,the dry matter accumulation before and after anthesis of maize treated with P₁₂₀ was not significantly different from that of P₁₅₀ and P₁₈₀ treatments,which was 37.91% and 93.88% higher than that of P₀,respectively.In summary,drip fertigation and phosphorus application significantly affected LAI,LAD,pre-anthesis and post-anthesis dry matter accumulation and HI,shortened bald tip length,increased grain number per spike and 1000-grain weight,and then increased maize yield and phosphorus utilization efficiency,and realized the synergistic improvement of maize yield and phosphorus fertilizer utilization efficiency.

In order to clarify the function of the protein phosphatase 2C gene GmPP2C72 in salt stress response and its salt-tolerance molecular mechanism,bioinformatics analysis of GmPP2C72 protein was conducted, GmPP2C72 gene was cloned to construct the overexpression vector,and then was transferred into Lotus japonicus using the cotyledon node genetic transformation method mediated by Agrobacterium tumefaciens.Finally,the phenotype,physiological indicators and the expression levels of salt tolerance related genes of transgenic plants were analyzed under salt stress condition.The results showed that GmPP2C72 gene contained an open reading frame of 1 206 bp,encoding 401 amino acids,and the encoded protein belonged to the member of the class A subfamily of PP2C family.GmPP2C72 gene did not exhibit tissue expression specificity,it was expressed in various tissues of soybean,and the expression of this gene was induced by salt stress.The overexpression vector of GmPP2C72 gene was successfully constructed.GmPP2C72 gene was successfully transferred into Lotus japonicus,and three transgenic lines with GmPP2C72 gene were successfully obtained.Compared with wild type,the three transgenic lines maintained better growth state,with significantly higher contents of chlorophyll and proline and lower relative plasma membrane permeability and malondialdehyde content in leaves of Lotus japonicus under salt stress.In addition,the expression levels of four key salt tolerance related genes,LjLEA, LjCOR,LjRD29B,and LjP5CS were significantly higher compared with wild type.Overall,the overexpression of GmPP2C72 gene enhances the salt tolerance of Lotus japonicus by reducing the degree of membrane lipid peroxidation and the permeability of the plasma membrane,increasing the contents of chlorophyll and proline in the leaves,and enhancing the expression levels of salt tolerance related genes.

To explore the accumulation characteristics of soil phosphorus and its distribution pattern in the profile after long-term application of different organic fertilizers,a long-term field experiment was conducted to investigate the distribution and migration patterns of phosphorus in the soil profile under continuous application of different amounts of livestock and poultry manure organic fertilizers and sludge compost.The results showed that after 11 consecutive years of application of high amounts(60 t/ha per year)of pig manure,chicken manure and sludge,the phosphorus they brought in could obviously migrate to the 30—60 cm soil layer.The application of medium and high amounts(45,60 t/ha per year)of pig manure,chicken manure organic fertilizers and sludge compost could make the available phosphorus migrate to the 60—90 cm soil layer.After 11 years of application of different types of organic fertilizers,the phosphorus activation coefficient of the plough layer soil showed a significant upward trend,and the difference between high and low application rates reached a significant level.The order of soil phosphorus activation coefficient under different organic fertilizer treatments was:pig manure organic fertilizer treatment>chicken manure organic fertilizer treatment>sludge compost treatment.The content of available phosphorus in the plough layer soil after the application of chicken manure,pig manure organic fertilizer and sludge compost conformed to the linear plus plateau regression model with the phosphorus input.Under the condition of equal phosphorus input of organic fertilizer,the contribution of pig manure and chicken manure organic fertilizer to the available phosphorus in the plough layer soil was significantly higher than that of sludge compost.When the phosphorus input was greater than 7.14 t/ha,the contribution of pig manure organic fertilizer to the available phosphorus in the plough layer soil was significantly higher than that of chicken manure.The total phosphorus and available phosphorus contents in the soil profile were significantly positively correlated with the organic carbon content.In the 0—90 cm profile,soil organic carbon and phosphorus might co-migrate.

This study aims to explore the effects of reduced nitrogen application combined with organic fertilizer on soil carbon and nitrogen contents in the 0—60 cm layer of fragrant pear orchards,as well as the yield and quality of Korla fragrant pears,and to propose the optimal rate of reduced nitrogen application and matching ratio of organic fertilizer.Using 10—12-year-old Korla fragrant pear trees as test materials,a 3-year continuous experiment was carried out.The experiment set up a conventional nitrogen application treatment(denoted as N,with 300 kg/ha of nitrogen applied),three reduced nitrogen application treatments(denoted as N1,N2,N3,which reduced nitrogen by 10%,20%,and 30% respectively compared with the conventional nitrogen application treatment),and two organic fertilizer treatments(denoted as OF1 and OF2,with 22 500,33 750 kg/ha of sheep manure applied respectively).These treatments were combined to form six integrated treatments:N1F1,N2F1,N3F1,N1F2,N2F2,and N3F2.The soil carbon and nitrogen contents,soil carbon-to-nitrogen ratio(C/N),as well as the yield and quality of Korla fragrant pears were determined.The result showed that N1F1,N1F2,N2F2,and N3F2 treatments significantly increased the contents of total soil carbon,soil organic carbon,soil inorganic carbon,soil microbial biomass carbon(SMBC),soil nitrate nitrogen,and soil ammonium nitrogen in the orchard.They also increased the soil C/N ratio,the contents of soluble sugar and vitamin C in pear fruits,decreased the total acid content in fruits,and had no significant effect on pear yield.The N2F1 treatment significantly promoted the contents of soil nitrate nitrogen and ammonium nitrogen but had no significant effect on pear yield or quality.The N3F1 treatment significantly inhibited the contents of soil carbon fractions and pear yield and reduced the soluble sugar content in fruits.Correlation analysis showed that soil nitrate nitrogen,ammonium nitrogen,organic carbon,inorganic carbon,total nitrogen,total carbon,SMBC,and soil microbial biomass nitrogen(SMBN)were significantly or extremely significantly correlated with pear quality;the soil C/N ratio was significantly or extremely significantly negatively correlated with pear yield and quality.Among these factors,soil organic carbon,total nitrogen,nitrate nitrogen,and ammonium nitrogen had more prominent effects on pear yield and quality.Therefore,in subsequent fertilization management,improving the contents of soil organic carbon,total nitrogen,nitrate nitrogen,and ammonium nitrogen and regulating the soil C/N ratio can better increase the yield and improve the fruit quality of Korla fragrant pears.In the management of 10—12-year-old Korla fragrant pear trees,reducing nitrogen application by 20%—30% on the basis of the conventional nitrogen application rate while applying organic fertilizer at 33 750 kg/ha is beneficial.This measure regulates soil carbon and nitrogen contents,improves pear quality,reduces chemical fertilizer input to alleviate soil pollution,and ensures the health of the soil environment.

Establishing a rapid and sensitive molecular detection technology for Pectobacterium parvum and P.versatile will lay a technical foundation for the rapid detection,early diagnosis,as well as the transmission routes and dynamics of potato stem rot.Through the sequence analysis of the specific locus gene 10748 of P.parvum and the specific locus gene 1602 of P.versatile,the Real-time PCR and LAMP primer design software were carried out by using Primer 3 and NEB website primer design software.After specificity validation,reaction condition optimization,and sensitivity validation,primers were screened and sample detection was performed.Quantitative Real-time PCR primers XX3-F/R and DY1-F/R were designed and screened for P.parvum and P.versatile.The sensitivity for detecting DNA in artificially inoculated plant samples reached 10,1 pg/μL,respectively.The sensitivity for detecting gDNA of pathogenic bacteria reached 0.1 pg/μL.Design LAMP primers for P.parvum and P.versatile,with detection sensitivity similar to Quantitative Real-time PCR detection technology.Quantitative Real-time PCR and LAMP detection techniques had been established for the detection of potato aerial stem rot pathogens,such as P.parvum and P.versatile,which can detect potato stems and tubers infected by P.parvum and P.versatile,as well as drosophila carrying bacteria.

In previous research,cytochrome P450 oxidoreductase(POR)was identified as a critical host factor facilitating aflatoxin B1(AFB1)-induced hepatotoxicity through CRISPR/Cas9 library screening.To further elucidate the mechanistic role of POR in AFB1-mediated hepatocellular damage,we designed two single-guide RNAs(sgRNAs)targeting exon 5 of the human POR gene,based on its published sequence,using CRISPR/Cas9 gene-editing technology.These sgRNAs were cloned into the lentiCRISPRv2 vector,which was subsequently packaged into lentivirus and utilized to infect THLE-2 human hepatocytes.Monoclonal cell lines were isolated by limiting dilution,and knockout efficiency was assessed at both the mRNA and protein levels using off-target sequencing,reverse transcription quantitative PCR(RT-qPCR),and Western Blot analysis.The findings indicated that V2-sgRNA-POR-1 successfully knocked down POR expression,resulting in a significant reduction at both the transcriptional and translational levels in the selected monoclonal cells.Further experiments involved treating THLE-2 wild-type cells with varying concentrations of AFB1(100,200,400,800 μmol/L)for durations of 24,48,and 72 hours.CCK-8 cell viability assays demonstrated that AFB1 toxicity to hepatocytes was significantly dependent on both concentration and exposure time,with an optimal exposure condition identified at 200 μmol/L for 48 hours[IC₅₀(48 h)=(200.55±44.49)μmol/L].Under these conditions,POR knockout cell lines exhibited marked resistance to AFB1-induced cytotoxicity,with significantly higher cell viability than wild-type cells.These findings suggest that POR plays a critical role in facilitating AFB1-mediated hepatocyte toxicity,and that POR knockout substantially enhances cellular resistance to AFB1.

Cinnamic acid 4-hydroxylase (C4H),a key enzyme in the flavonoid synthesis pathway,plays a crucial role in plant stress resistance.To clarify the bioinformatic functions of the C4H gene family in three medicinal liquorice species (Glycyrrhiza uralensis Fisch.,Glycyrrhiza inflata Bat.,and Glycyrrhiza glabra L.),investigate their expression characteristics under abiotic stress,as well as their relationship with the synthesis and accumulation of liquiritin,we used bioinformatics methods to analyze the C4H gene family members,and combined transcriptome data and RT-qPCR validation to understand their expression under abiotic stress and their relationship with liquiritin content.Results showed that a total of 11 C4H genes were identified in the whole genomes of the three medicinal liquorice species and were named GuC4H1—3,GiC4H1—3,and GgC4H1—5,respectively.These genes were distributed on 2 chromosomes in G.uralensis and G.inflata and on 3 chromosomes in G.glabra.Although the gene members differed in physicochemical properties such as the number of amino acids and isoelectric points,all of them were predicted to be localized in the endoplasmic reticulum and contain a C-terminal heme-binding domain (PFGVGRRSCPG).Expression pattern analysis indicated that the 11 C4H genes were expressed in different tissues of the three medicinal liquorice species,and abiotic stress significantly induced the upregulation of C4H gene expression in the underground parts.Among them,the expression of GgC4H5 was the highest under salt stress for 24 h,which was 1.68 times that of the control group (24 h);the expression of GuC4H3 was the highest under drought stress for 2 h,which was 3.03 times that of the control group (2 h).In addition,moderate abiotic stress significantly increased the liquiritin content in the underground parts of the three liquorice species.Under NaCl stress for 2 h,the increase in liquiritin content in G.inflata was the most significant,which was 2.92 times that of the control group (2 h);under PEG stress for 24 h,the increase in liquiritin content in G.glabra was the most significant,which was 2.31 times that of the control group (24 h).GgC4H5 showed a similar expression trend to liquiritin content under salt stress,while GuC4H2,GuC4H3,GiC4H2,and GiC4H3 showed a similar expression trend to liquiritin content under drought stress.Therefore,these genes are likely to be the key genes involved in the response to abiotic stress and liquiritin synthesis in the underground parts of the three medicinal liquorice species.

To investigate the physiological regulatory role of salicylic acid(SA)in alleviating saline-alkali stress in kidney bean seedlings and to provide a basis for stress-resistant cultivation of kidney beans in typical saline-alkali areas of Shanxi Province,kidney beans were used as the test material.A composite saline-alkali stress system with a ratio of NaCl,Na₂SO₄,NaHCO₃,and Na₂CO₃ of 1∶9∶9∶1 was used to simulate saline-alkali stress.Five concentration gradients of 0(CK),20,40,60,and 80 mmol/L were set to screen for the optimal concentration of salt-alkali stress treatment.Subsequently,based on the selected concentration,three SA concentrations(20,40,and 60 μmol/L)were applied simultaneously to study the mitigating effect of SA on kidney bean seedlings under saline-alkali stress.The results indicated that under 40 mmol/L salt-alkali stress,the growth of red kidney beans was significantly inhibited but did not reach lethal or severe stress levels,making it the optimal salt-alkali stress concentration.Stress at this concentration induced oxidative damage in early seedlings,inhibited nutrient absorption and translocation,increased the activities of arginine decarboxylase(ADC)and ornithine decarboxylase(ODC),and led to excessive accumulation of putrescine(Put).Under 40 mmol/L saline-alkali stress,exogenous application of 40 μmol/L SA reduced malondialdehyde(MDA)content in leaves and roots by 37.63% and 39.76%,respectively;hydrogen peroxide(H₂O₂)content by 38.71% and 21.13%,respectively;and superoxide anion(O₂^-)content by 40.00% and 52.17%,respectively.Root activity increased by 56.14%.The Na content in both leaves and roots significantly decreased,while the contents of K,Cu,Ca,and Fe significantly increased.Furthermore,exogenous application of 40 μmol/L SA reduced the activities of ADC and ODC,increased the activity of S-adenosylmethionine decarboxylase(SAMDC),and promoted the conversion of Put to spermidine(Spd)and spermine(Spm),thereby enhancing the plant's antioxidant capacity and ion homeostasis regulation.In conclusion,exogenous salicylic acid can effectively alleviate the physiological damage caused by composite saline-alkali stress in kidney bean seedlings by activating the antioxidant system,optimizing polyamine metabolism,and promoting the absorption and distribution of mineral elements.

In order to investigate the effects of different concentrations of γ-aminobutyric acid(GABA)on the nutritional quality and antioxidant activity of cherry tomatoes,cherry tomatoes were used as experimental materials.Foliar spraying of GABA solutions with different concentrations(0,5,10,20,and 40 mmol/L,denoted as CK,G5,G10,G20,and G40,respectively)was initiated at 15 days after the anthesis of the 3rd inflorescence.It aimed to explore the impacts of exogenous GABA application on the nutritional quality,endogenous GABA content,and antioxidant activity-related indicators of cherry tomatoes.The results showed that exogenous GABA treatment could significantly improve the nutritional quality and antioxidant capacity of cherry tomatoes.With the increase in treatment concentration,the contents of soluble sugar,total phenols,and total flavonoids,as well as the sugar-acid ratio,increased continuously.Nutritional quality indicators such as vitamin C content,soluble sugar content,and sugar-acid ratio all reached their maximum values under the G40 treatment.The contents of total phenols and total flavonoids in the G40 treatment were significantly higher than those in other treatments except G20,increasing by 24.26% and 91.75% compared with CK,respectively.The lycopene content of cherry tomatoes under the G40 treatment was the highest,which was significantly 85.96% higher than that of CK.The endogenous GABA content first increased and then decreased with the elevation of treatment concentration,reaching the maximum value under the G20 treatment,which was significantly 101.85% higher than that of the control.However,there was no significant difference in endogenous GABA content between the G20 and G40 treatments.Both DPPH free radical scavenging activity and FRAP antioxidant capacity were synchronously enhanced,and both reached their maximum values under the G40 treatment,which were significantly higher than those of CK.Correlation analysis indicated that the antioxidant capacity of cherry tomatoes was extremely significantly positively correlated with total phenols,total flavonoids,and lycopene.In conclusion,exogenous GABA(G40 treatment)can serve as a green and efficient strategy to enhance the nutritional quality and antioxidant activity of cherry tomato fruits.

Based on quantitative trait locus(QTL)mapping of grain weight and grain shape-related traits in a backcross inbred line(BIL)population derived from the cross between indica rice variety Changhui 891 and japonica rice variety 02428,this study provided valuable genetic resources for elucidating the genetic basis of grain development and molecular breeding for high yield and quality in rice.A high-density genetic map was used to conduct QTL analysis for thousand-grain weight,grain length,grain width,grain thickness,and length-width ratio under two environments.Transcriptome data of developing grains at 5,10,and 15 days after flowering from both parents were integrated to analyze candidate genes within QTL regions.Frequency distribution indicated that grain weight and shape traits in the BIL population exhibited typical quantitative characteristics,confirming the suitability for QTL mapping.A total of 37 QTLs were detected across both environments,including 18 specific to Hainan(Sanya),12 specific to Jiangxi(Nanchang),and 7 consistently identified in both locations.These QTLs were distributed on chromosomes 1—5,7,8,11,and 12,comprising 7 for thousand grains weight,5 for grain length,7 for grain width,11 for length-width ratio,and 7 for grain thickness.The analysis showed that the LOD values of these QTLs ranged from 2.79 to 43.10,with a contribution rate of 1.36% to 46.12%.By integrating differentially expressed genes from developing grains of both parents at three stages,28 candidate genes were prioritized from the 37 QTL regions.Among these,LOC_Os11g10480 and LOC_Os11g10100 were identified as key candidates for further functional validation.

To investigate the effects of long-term fertilization on yield formation and its physiological regulation mechanisms in double cropping late rice,it utilized a long-term fertilization experiment(initiated in 1981)in red paddy soils of Jinxian.Four typical treatments were selected:no fertilization(CK),single application of nitrogen-phosphorus-potassium(NPK),double dose of NPK(HNPK),and combined organic-inorganic fertilization(NPKM).Comparisons were made on double cropping late rice yield,dry matter accumulation,chlorophyll dynamics,and differential gene expression in leaves at fullheading and filling stages in the 42nd year of long-term fertilization.Results showed that long-term fertilization resulted in yields ranked as NPKM>HNPK>NPK>CK,with HNPK and NPKM treatments significantly surpassing NPK,showing increases of 29.63% and 57.18% respectively.Compared with NPK,both NPKM and HNPK significantly improved yield components:effective panicles,grains per panicle,and grain density increased by 16.98%—46.42%,8.68%—15.26%,3.69%—7.37%,respectively.Regarding dry matter accumulation,NPKM and HNPK significantly enhanced dry matter weight at all growth stages and promoted translocation of stem-leaf dry matter to panicles from filling to maturity.The contents of chlorophyll in NPKM and HNPK treatments were significantly higher than those in CK and NPK treatments at all stages,and NPKM delayed the decay of chlorophyll from filling stage to maturity stage.Correlation analysis revealed significant positive relationships between yield and dry matter accumulation from tillering to heading(△DM1)and filling to maturity(△DM3),while showed extremely negative correlation with chlorophyll reduction from grain filling to maturity(△S3).Transcriptome analysis demonstrated that long-term fertilization significantly affected gene expression in leaves during fullheading and filling stages,with differentially expressed genes primarily enriched in photosynthesis,carbon/nitrogen metabolism,signal transduction,and stress-related pathways.In conclusion,long-term fertilization regulates gene expression and combined organic-inorganic fertilization improved yield components,enhanced early-stage dry matter accumulation,promoted assimilate translocation from vegetative organs to panicles during grain filling,maintain higher chlorophyll levels,and delayed leaf senescence post-grain filling.This comprehensive mechanism achieves yield enhancement through strengthening source(enhancing photosynthetic capacity),expanding sink(increasing dry matter storage),and facilitating flow(promoting assimilate transport).

In order to explore the effect of straw interlayer on soil respiration and the chemical structural stability of organic carbon components during the remediation of saline-alkali soil,two experimental treatments were set up in the alfalfa-planting farmland at the Saline-Alkali Soil Improvement Experimental Demonstration Base in the Agricultural High-Tech Zone of Dongying City,Shandong Province:a straw interlayer treatment (S,with a 5 cm thick straw layer buried at a 35 cm depth) and a control (CK,without an interlayer). Soil respiration characteristics of the different treatments were analyzed,and comprehensive discussions were conducted in conjunction with soil pH,electrical conductivity(EC),organic carbon component content,and chemical structure characteristics of the soil profile.The results showed that:compared with CK,the S treatment significantly increased the soil respiration rate during the alfalfa growth period, with a maximum increase of 79.84%. Furthermore, the S treatment reduced soil EC in the straw interlayer (35—40 cm) and the overlying soil layer (0—35 cm), effectively inhibiting the upward migration of salt. The 40—50 cm soil layer was identified as a critical zone for organic carbon transformation. In this layer, the S treatment significantly increased SOC content (by 16.67%) and highly significantly elevated particulate organic carbon (POC) and dissolved organic carbon (DOC) contents (by 208.07% and 83.41%, respectively) compared with CK. Characterization of the molecular structure of organic carbon in the 40—50 cm layer revealed distinct responses. For DOC, the S treatment reduced the magnitude-weighted averages of unsaturation (by 30.60%) and aromaticity index (by 4.84%) compared with CK, decreasing the proportion of unstable carbon. For POC, the S treatment increased the relative abundances of alkyl C(10.32 percentage points)and O-alkyl C (8.39 percentage points) while reducing carboxyl C (14.24 percentage points), thereby enhancing POC structural stability. However, for bulk SOC, the S treatment decreased the proportions of alkyl C(3.14 percentage points) and aromatic C(3.38 percentage points) while increasing O-alkyl C(5.17 percentage points)and carboxyl C(1.56 percentage points). This shift indicated a decrease in recalcitrant carbon and an increase in labile carbon, resulting in reduced SOC structural stability. Correlation analysis showed that the significant increased in soil respiration was highly significantly and significantly positively correlated with POC and SOC contents in the critical soil layer, respectively. Specifically, the accumulation of O-alkyl C(a labile component) in the chemical structure of SOC reduced the stability of organic carbon,which was the main reason for the increase in soil respiration.In conclusion,incorporating a straw interlayer significantly increased soil respiration in the short term,which was closely related to the reduction in the stability of the organic carbon chemical structure in the key soil layer.Under future "carbon neutrality" strategies,research on the selection of interlayer materials should be considered.

The screening of aromatic rice germplasm resources and the rapid identification and utilization of aromatic genes have practical significance for the genetic breeding of aromatic rice.In order to clarify the aroma types of aromatic rice materials in Heilongjiang Province and improve the breeding efficiency of aromatic rice varieties,180 aromatic rice varieties approved and promoted in Heilongjiang Province were sequenced.Compared with the reference genome Nipponbare,the sequencing results were divided into two types of mutations.The first type was the 8 bp deletion of exon 7 and three base substitutions(Badh2-E7 type),including 177 varieties.The second type was the 7 bp deletion of exon 2(Badh2-E2 type),which contained only three varieties.According to these two mutation types,two specific KASP molecular markers were designed to accurately detect the aroma types of E7 and E2 in rice.Through the identification of 180 aromatic rice varieties,the typing results were consistent with the sequencing results.Two hybrid populations were constructed by using E7 aromatic rice variety Wuyoudao 4 and E2 aromatic rice variety Songkejing 134 with non-aromatic japonica rice varieties Weinong 101 and Dongfu 114,respectively.The F₂ generation of the hybrid population was screened and identified for aroma genes,and the identification results were consistent with 100%,which verified the accuracy of KASP molecular markers.The 620 japonica rice parents created and collected by our laboratory were further identified by KASP markers,and 248 aromatic rice materials were screened.It was found that these parents were all Badh2-E7 type genes.At the same time,the existing 1 220 F₇ generation materials with E7 aromatic rice as parents were genotyped,including 293 aromatic rice materials,906 non-aromatic rice materials and 21 heterozygous rice materials.Five aromatic and non-aromatic rice materials were randomly selected for sequencing verification.The sequencing results verified that the KASP molecular marker typing was correct.This study provided new markers and materials for breeding new fragrant rice varieties in cold regions by using molecular marker assisted selection.

Bacillus thuringiensis(Bt)produces multiple insecticidal proteins and serves as a vital biological control agent.This study aims to identify Bt strains with insecticidal activity against lepidopteran pests,thereby alleviating pest resistance pressure and expanding resource reserves.A total of 109 Bt strains were selected from soil of Hebei Province.The shape of parasporal crystals was observed under an oil microscope,and the insecticidal protein genotypes of the Bt strains were identified by PCR using 40 pairs of primers(including cry,cyt,and vip genotypes).The insecticidal activity bioassay was performed using five lepidopteran pests,including Plutella xylostella and Spodoptera litura.The molecular weight of crystal proteins was detected via SDS-PAGE.The shape of parasporal crystals from 90 strains was spherical,and the crystals from the remaining 19 strains were diamond-shaped.The detection rate of insecticidal protein genotypes reached 93.6%,with 29 distinct cry genotypes,1 cyt genotype,and 3 vip genotypes identified.A total of 91 strains possessed combinations of at least two insecticidal protein genotypes,among which 26 strains contained six or more,predominantly exhibiting cry+vip patterns.We found that 19 Bt strains showed high insecticidal activity against various lepidopteran pests such as P.xylostella. These strains primarily expressed proteins at 130,65 ku,encompassing lepidopteran-killing genotypes such as cry1,cry2,and cry15,consistent with PCR identification results.Here,we identified broad-spectrum insecticidal and highly toxic Bt strains against lepidopteran pests,and revealed its insecticidal genes.Our work provides an important candidate for the development of bioinsecticide.

In order to explore the structure and biological function of Ras related nuclear protein gene(RAN)in yak and resolve how it regulates the proliferation of yak cells and participates in protein synthesis,the CDS region sequence of RAN gene of Sangsang yak was cloned by RT-PCR using Sangsang yak kidney tissue cDNA as template,and bioinformatics analysis was carried out by a variety of software and online tools,the expression of the RAN gene in seven tissues of Sangsang yaks was detected by qPCR technology.The results showed that the CDS region of RAN gene in Sangsang yak was 651 bp in length,encoding 216 amino acids.Through homology comparison,it was found that the genetic relationship between Sangsang yak and wild yak and zebu was the closest,with a similarity of 99.2%,and the farthest from chicken,reaching 86.6%.The prediction results of RAN protein analysis showed that the molecular weight of the protein was 24.423 11 ku,the theoretical isoelectric point was 7.01,the total number of atoms was 3 449,and the molecular composition was C₁₁₀₉H₁₇₂₅N₂₉₅O₃₁₃S₇.RAN protein had no transmembrane structure and no potential sites for N-glycosylation,with 36 phosphorylation sites.The affinity and hydrophobicity were predicted and the instability coefficient was calculated,and it was found that the protein was a stable hydrophilic protein.According to the subcellular localization,the protein was found to be present in the Golgi apparatus,mitochondria,nucleus and cytoplasm in yak cells.Predictions of the RAN protein structure revealed that its higher-order structure consisted mainly of α-helices and did not contain β-turns.Protein interaction network results showed that there was an interaction between the RAN protein and RAN Binding Protein 1(RANBP1),RAN Binding Protein 2(RANBP2),RANGTPase activating protein 1(RANGAP1)and other proteins of Sangsang yak,and there was also an interaction between them.The expression level of the RAN gene in yak testicular tissue was significantly higher than in other tissues,while no expression was detected in muscle tissue.It successfully cloned the CDS region of the RAN gene and completed its bioinformatics analysis,and the expression of this gene in the tissue of Sangsang yak was also studied,and it was found that it played an important role in the development of the reproductive system,cell proliferation,disease prevention and control,and participation in protein synthesis.

The aim is to explore the biological function of the Lon gene in Glaesserella parasuis and its impact on the pathogenicity of the bacterium.A fusion gene fragment containing the homologous arms of the Lon gene and the kanamycin resistance gene selection marker was amplified using overlapping PCR,and a recombinant suicide plasmid pToPo-LR-Kana containing this gene fragment was constructed.The pToPo-LR-Kana was transformed into the parent strain ZJ1208 using natural transformation.The Lon gene-deleted strain ZJ1208-ΔLon was identified by kanamycin resistance screening,PCR,and sequencing.The differences between the wild-type strain ZJ1208 and the gene-deleted strain ZJ1208-ΔLon were compared through determination of growth rate,observation by transmission electron microscopy,stress resistance test,ultraviolet resistance test,serum resistance test,the biofilm formation assay,determination of capsule polysaccharide content and virulence test.The results showed that the Lon gene deletion strain ZJ1208-ΔLon was successfully obtained.The deletion strain had a significantly longer cell size,but its growth rate and number of outer membrane vesicles were similar to those of the wild-type strain ZJ1208.ZJ1208-ΔLon had significantly decreased tolerance to osmotic stress,oxidative stress,and heat stress,as well as significantly decreased resistance to ultraviolet radiation. Compared with the parent strain ZJ1208,the biofilm production capacity of ZJ1208-ΔLon and ZJ1208 was similar,but the capsular polysaccharide content of ZJ1208-ΔLon increased significantly,and the serum resistance and virulence in mice decreased significantly.The above results indicate that the deletion of Lon gene has an impact on multiple biological characteristics of G.parasuis,providing new information for further analysis of the biological function of Lon gene in G.parasuis.

This study aimed to investigate the effects of acidic soil on the growth metabolism and photosynthesis of Hainan green sweet orange (Citrus sinensis L.)and to elucidate its antioxidant regulatory mechanisms under acid stress.A pot experiment was conducted to analyze the physiological metabolic changes,antioxidant responses,and expression patterns of key antioxidant enzyme regulatory genes in green sweet orange under acid stress.The results showed that the plant height growth,leaf area and fresh weight of green sweet orange plants decreased significantly under a strongly acidic soil environment,with the most serious effect of pH 3.5 treatment;the weakly acidic(pH 6.5)soil could promote the growth of plants,which manifested as positive growth in the amount of each growth index.Strong acid treatment significantly impaired photosynthesis and normal growth metabolism of green sweet orange leaves,manifested as negative growth in transpiration rate(Tr)and stomatal conductance(Gs)were significantly reduced.Superoxide dismutase(SOD)and phenylalanine ammonia-lyase(PAL)activities increased by 6.60-fold and 5.90-fold at 24 h post-treatment under pH 3.5.The malondialdehyde(MDA)content in leaves treated with pH 3.5 increased by 6.47-fold compared to the initial time point(0 h),significantly higher than that under pH 6.5.Principal component analysis(PCA)and redundancy analysis(RDA)revealed a significant negative correlation between enzyme activity and physiological-biochemical indices(R²=0.82).Gene expression analysis demonstrated that CsSOD and CsPAL reached peak levels at 24,48 h post-stress respectively and remained upregulated within 72 h,showing 2.33-fold and 2.64-fold increases compared to 0 h,respectively.In summary,strongly acidic soil inhibits the growth and development of green sweet orange by reducing photosynthetic pigment content and metabolic efficiency.SOD and PAL serve as key antioxidant enzymes in response to acid stress,with CsSOD and CsPAL genes dynamically regulating enzymatic activity to mitigate stress.

To identify the suitable sowing date and planting density for the synergistic improvement of yield and quality of strong gluten wheat,the strong gluten wheat variety Kexing 3302 was used as the experimental material.Two sowing dates(October 18(S1)and October 28(S2))and five densities(basic seedlings of 180×10⁴ plants/ha(D180),225×10⁴ plants/ha(D225),270×10⁴ plants/ha(D270),315×10⁴ plants/ha(D315),and 375×10⁴ plants/ha(D375))were set.Field experiments were conducted in 2021—2023 to investigate the effects of sowing date and density on physiological indicators,yield,and quality of wheat plants.The results showed that the relative chlorophyll content(SPAD)of wheat leaves generally decreased with the increase of planting density,and reached its maximum at most growth stages under low-density treatment.With the delay of the sowing date,the relative chlorophyll content of leaves showed an upward trend,with S1 sowing date being lower than S2.As planting density increased,the normalized difference vegetation index(NDVI)generally showed an upward trend,and the NDVI value at S2 sowing date was significantly lower than that at S1 sowing date.The number of spikes and grain yield decreased with the delay of the planting density,but increased with the increase of planting density,with the highest yield under D375 treatment.With the delay of the sowing date,both protein content and wet gluten content generally showed a slight downward trend,but the grain protein content of all treatments was above 13.5%.The total starch content of wheat grains generally decreased with the increase of density at S1 sowing date,but increased with the increase of density at S2 sowing date.In addition,the peak viscosity,trough viscosity,final viscosity,and breakdown value generally increased with the increase of density under the two sowing dates.Parameters such as dough stability time and water absorption rate did not exhibit obvious regular pattern under different sowing dates and planting density,but all met the standards for strong gluten wheat.In summary,at sowing date S1,the planting density of Kexing 3302 at 315×10⁴—375×10⁴ plants/ha can achieve high quality and high yield.

OFP is a class of plant-specific transcription factors that play important roles in the regulation of plant organ morphogenesis and response to abiotic stresses.In order to study the characterization of soybean OFP transcription factor family members and their roles in drought stress and salt stress,bioinformatics methods were applied to identify and analyze soybean OFP family members.The results showed that:a total of 41 GmOFPs,named GmOFP-1—GmOFP-41,were identified in soybean;these genes were unevenly distributed on 19 chromosomes of soybean,encoding 152—414 amino acids;subcellular localization predicted that soybean OFP proteins were mainly localized in nucleus,chloroplasts,and mitochondria;a total of 10 conserved motifs were identified in soybean OFP proteins,conservative Motifs 1 and 2 were present in all OFP members.Phylogenetic analysis classified soybean and Arabidopsis OFP proteins into five subfamilies ClassⅠ—Class V,of which soybean OFP family genes were mainly distributed in ClassⅠ and Class Ⅲ.The collinearity analysis revealed that 75 pairs of genes in the soybean genome had collinearity,four pairs of genes had tandem duplications,and only two genes,GmOFP-2 and GmOFP-39,did not have collinearity,which indicated that gene fragment duplication was the main reason for the increase in the number of soybean OFP family members.The expression patterns of GmOFP gene family members under drought stress and salt stress treatments were analyzed by qRT-PCR,and the results showed that,compared with the control,16 members out of 41 GmOFP genes exhibited significant differences in gene expression levels after drought treatments,with significant up-regulation of the expression of GmOFP-15,GmOFP-17,and GmOFP-32,while the GmOFP-4,GmOFP-5,GmOFP-6,GmOFP-9,GmOFP-12,GmOFP-21,GmOFP-23,GmOFP-25,GmOFP-26,GmOFP-27,GmOFP-38,GmOFP-39,and GmOFP-40 were significantly down-regulated after drought treatment.Eight members of GmOFPs showed significant differences in gene expression levels after salt treatment,among which GmOFP-7,GmOFP-14,GmOFP-31,GmOFP-32,GmOFP-36,and GmOFP-40 were significantly up-regulated,and GmOFP-1 and GmOFP-15 were significantly down-regulated.The above results suggest that the soybean OFP gene family may have important functions in response to drought stress and salt stress.

To explore the disparities in overwintering rates and yields between Brassica napus L.and Brassica rapa L.in the cold and arid regions of the north,as well as to examine the correlation between diverse agronomic traits,yield,and cold resistance-enhancing traits,it utilized 45 B.napus L.and 22 B.rapa L.varieties as subjects.It conducted a statistical analysis of their overwintering rates,average yields and agronomic traits.It also studied seeding density experiments with B.napus L.and B.rapa L.varieties that exhibited similar yield levels,compared root traits between B.napus L.and B.rapa L..The results indicated that among the 67 varieties tested,the average overwintering rate of B.rapa L.(97.59%) was significantly higher than that of B.napus L.(65.87%).The average yield potential of B.napus L.was higher than that of B.rapa L..For B.rapa L.,which was capable of stable overwintering,increasing the seeding density significantly elevated the number of effective plants but did not augment the average yield.Conversely,for B.napus L.,its lower overwintering rate constrained the increase in the number of effective plants,thereby limiting the average yield.B.napus L.with a high overwintering rate exhibited agronomic traits such as higher branching positions and angular density,fewer secondary branches,total branches,and effective siliques per plant after overwintering.Comparison of root traits between B.napus L.and B.rapa L.showed that enlarged root systems,short hypocotyls,and growth points positioned below the ground surface were advantageous traits contributing to the robust cold resistance of B.rapa L..It proposes that breeding density-tolerance B.napus L.with root traits akin to those of B.rapa L.and with growth points situated below the ground surface represents a pivotal breeding direction for enhancing the overwintering rate of B.napus L.,fostering increased rapeseed yield,and ensuring oil supply stability.

The aims were to study the effect of maize straw returning on soil fertility and productivity under wheat-maize rotation system in North China,and to explore the best way and suitable amount of maize straw returning.From 2021 to 2023,a field split-plot experiment was conducted in Quzhou County,Hebei Province to compare the effects of different maize straw returning methods and amounts on wheat growth,yield composition and soil organic matter content.There were two treatments in the main area,which were cutting and crushing.The sub-area was the amount of straw returning to the field,4 levels,which were 0,0.5,1.0,and 1.5 times of the amount of maize straw in the year.The results showed that compared with straw crushing,the yield of wheat increased by 4.3% under straw cutting treatment,the number of spikes increased by 7.6%,and the harvest index increased by 1.4%,with all significant difference in 2023.The N content of wheat straw was significantly increased by 0.04 percentage points,the N and P uptake of wheat aboveground were significantly increased by 10.8% and 14.3%,respectively(2022),but the K content of wheat straw was significantly reduced by 0.13 percentage points(2023).Cutting treatment could significantly promote the growth of wheat before and after winter,and the NDVI value at jointing stage significantly increased by 11.9%(2022).Soil pH increased by 0.22 units(2023).With the increase of maize straw returning amount,wheat yield and NDVI value at jointing stage showed parabolic model of first increased and then decreased,the N,P and K absorption of above ground wheat showed a significant downward trend,and soil organic matter showed a continuous significant increase trend.Based on the growth potential and yield of wheat,the suitable returning ratio of straw cutting treatment was 58%—62%,and the suitable returning ratio of crushing treatment was 29%—42%.Under the conditions of this experiment,maize straw cutting returning has obvious advantages in promoting wheat growth,and increasing wheat yield.It can be popularized and applied in wheat-maize rotation areas that 58%—62% maize straw is returned into the soil.

To clarify the alterations of rice straw decomposition,nutrients release and chemical components in coastal saline paddy soils under different nitrogen (N) application rate,for optimizing the technology of straw returning and realizing the efficient utilization of straw resources in coastal areas.The experimental site was located in Caofeidian District,Tangshan City,Hebei Province.The decomposition characteristics of rice straw and its lignocellulose,as well as the nutrient release characteristics of N,phosphorus (P) and potassium (K) were studied,using a 360-day straw-bag burying method with four different N fertilizer levels,including N0 (0 kg/ha),N1 (225 kg/ha),N2 (300 kg/ha) and N3 (375 kg/ha).Pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) method was used to study the dynamic alterations of principal chemical components of the rice straw residues.The results showed that:the decomposition period of rice straw was divided into three stages,namely,rapid decomposition (0—30 d),slow decomposition (30—210 d) and slow decomposition (210—360 d),and the average decomposition rate of rice straw was 72.5% after 360 days with different N application rates.Increasing N application significantly increased the decomposition rate of rice straw.Compared with the N0 treatment,the N1,N2 and N3 treatments,increased the straw decomposition rate by 6.1, 7.4 and 9.2 percentage points,respectively.The trend of straw carbon (C) release rate was similar to that of straw decomposition rate,while the C release rate was only 43.2% at the end of the experiment.The nutrient release rates of rice straw were as follows:K>P>N.The N,P and K rapid release periods of rice straw was in the 0—30th (38.4%),0—60th (63.7%) and 0—15th (76.7%) days after straw decomposition,respectively.Both N and P of rice straw were enriched during the decomposition period.N application significantly increased the release of N from straw during the decomposition period,P in the early (0—15 d) and late (150—360 d) period,and K in the early period (0—15 d).Compared with the N0 treatment,the N1,N2,and N3 treatments,increased the straw N release by 6.6, 11.1, and 14.7 percentage points,P release by 2.2, 4.0, and 5.6 percentage points,and K release by 1.4, 2.1, and 2.8 percentage points,respectively.The lignocellulose decomposition rates of rice straw were as follows:hemicellulose>cellulose>lignin.Increasing N application significantly promoted the cellulose and hemicellulose decomposition rate of rice straw from day 0—90 and the lignin decomposition rate after day 90.Compared with the N0 treatment,the N1,N2,and N3 treatments,increased the cellulose decomposition rate of rice straw by 5.4, 7.3, and 8.4 percentage points, hemicellulose decomposition rate by 4.9, 6.4, and 7.4 percentage points,and lignin decomposition rate by 2.1, 5.1, and 5.7 percentage points, respectively.2-methoxy-4-vinylphenol,hydroxyacetone,2,3-dihydrobenzofuran,acetosyringone,eugenol,n-hexadecanoic acid,p-methylphenol,2,6-dimethoxyphenol,guaiacol,p-ethylphenol,and stigmasta-3,5-diene were the major (>1% relative) chemical components of straw residues during the decomposition period.Correlation analyses showed that straw decomposition rate,C release rate and cellulose,hemicellulose and lignin decomposition rates,were significantly positively correlated with eugenol,acetosyringone and 2,3-dihydrobenzofuran,while significantly negatively correlated with hydroxyacetone;straw P release rate was significantly positively correlated with hydroxyacetone and significantly negatively correlated with p-ethylphenol,eugenol and acetosyringone;straw K release rate was significantly correlated with p-ethylphenol,eugenol,acetosyringone and 2,3-dihydrobenzofuran,while significantly positively correlated with hydroxyacetone.In conclusion,increasing N application could promote the decomposition rates of rice straw and its lignocellulosic cellulose,and the nutrients release of straw N,P and K in coastal saline paddy field.The recommended optimal N application rate was 300 kg/ha under straw returning 10 500 kg/ha in coastal saline soils.p-ethylphenol,eugenol,acetosyringone,2,3-dihydrobenzofuran,hydroxyacetone,and stigmasta-3,5-diene could indicate the process of straw decomposition in straw residues.Py-GC-MS technique shows a good capability to monitor the chemical components alterations of straw residues,further deepening the understanding of straw decomposition mechanism.

To investigate the effects of climate warming and combined application of organic and inorganic fertilizer on soybean growth and yield,field experiments were conducted using infrared radiation warming systems to simulate temperature elevation.The experimental design included two temperature levels(T0:Ambient temperature;T1:Warming by 2.9 ℃)and two fertilization regimes(SF:Sole inorganic fertilizer;OF:Combined organic-inorganic fertilizer),aiming to examine the impacts of warming and fertilization practices on soybean dry matter accumulation,photosynthetic rate,leaf enzyme activities,and yield.Results demonstrated that compared to ambient temperature,warming shortened the pre-flowering and post-flowering growth periods by 3—4 d,5—6 d,respectively.Warming significantly reduced leaf nitrate reductase(NR)and glutamine synthetase(GS)activities during the pod-filling stage,decreased leaf area index(LAI)by 9.0%—13.7%,and suppressed net photosynthetic rate,collectively leading to 4.1%—19.3% reductions in post-flowering plant height and aboveground dry matter accumulation.In contrast,the combined organic-inorganic fertilization enhanced NR and GS activities,improved post-flowering photosynthetic efficiency,promoted plant growth,and increased the aboveground dry matter accumulation by 4.1%—15.3% compared to sole inorganic fertilization,with a significant interaction observed between NR and alanine aminotransferase activities.Yield analysis revealed that warming caused 9.7%—16.6%,13.3%—19.0% declines in pods per plant and grains per plant,respectively,resulting in 13.7%—21.1% yield reductions.Conversely,organic-inorganic fertilization increased grains per plant by 12.5% and pods per plant by 9.9%—10.5%,achieving 7.6%—10.8% yield improvements.Notably,significant positive correlations were detected among LAI,photosynthetic rate,nitrogen metabolism enzyme activities,and final yield.These findings demonstrate that climate warming inhibits post-flowering photosynthetic efficiency and aboveground growth in soybeans,while integrated organic-inorganic fertilization partially mitigates warming-induced yield losses through enhancing enzyme activities and photosynthetic performance,providing critical insights for adaptive agricultural practices under global warming scenarios.

This study aims to establish a biocontrol bacteria screening system based on root exudates to screen biocontrol strains with strong rhizosphere colonization ability.Using this system,bacterial strains capable of synergistic control of cucumber Fusarium wilt in combination with spent mushroom substrate(SMS)were screened.The chemical composition of cucumber root exudates was modulated by pathogen stress and SMS induction.Chemotaxis,metabolic proliferation,and biofilm formation,which were colonization-related factors,were used as quantitative indicators for the tested strains.Efficient screening of rhizosphere-colonizing bacteria was achieved using capillary assays,96-well plate cultures,and biofilm formation tests.Among 200 bacterial strains isolated from the cucumber rhizosphere,significant differences were observed in chemotaxis,metabolic proliferation,and biofilm formation in response to root exudates.Using this screening system,Bacillus velezensis MTC-5 was screened as an outstanding strain across all screening indicators.Greenhouse experiments demonstrated that MTC-5 strain combined with SMS exhibited enhanced rhizosphere colonization ability and showed significant synergistic effects in disease suppression and plant growth promotion.Field trials revealed that the combined application of MTC-5 strain and SMS achieved a disease control efficacy of 74.5% against cucumber Fusarium wilt,which was 10.7,24.6 percent points higher than the efficacy of SMS or MTC-5 strain alone,respectively.It established a high-throughput biocontrol bacteria screening system based on root exudates,highlighting the critical role of root exudates in bacterial recruitment.The MTC-5 and SMS combination developed through this system demonstrated significant synergistic effects in controlling cucumber Fusarium wilt,providing new insights and technical support for biocontrol bacteria screening and application.

Intramuscular fat deposition,as a key factor affecting meat quality(e.g.tenderness,flavor,and juiciness),is closely related to the regulation of lipogenesis and lipid metabolism by long non-coding RNA(lncRNA).Based on the previous finding that the intramuscular fat content of Tan sheep was significantly higher than that of Dorper sheep and Small-tailed Han sheep,the dorsal longest muscle of these three sheep breeds was used as the target,and differentially expressed mRNAs(DE mRNAs)and lncRNAs(DE lncRNAs)and their related pathways were screened by transcriptome sequencing,with the aim of revealing molecular mechanism of high intramuscular fat deposition in Tan sheep.The results showed:836 DE mRNAs and 832 DE lncRNAs were identified in the Small-tailed Han sheep vs.Tan sheep group;578 DE mRNAs and 700 DE lncRNAs were screened in the Dorper sheep vs.Tan sheep group.The intersection of the two groups fielded a total of 226 DE mRNAs and 308 DE lncRNAs.Target gene prediction for the 308 DE lncRNAs identified 1 475 potential targets,including 687 co_expression(co-expressed)target genes and 788 co_localization(co-localized)target genes.GO and KEGG enrichment analysis of DE mRNAs revealed fat deposition-related pathways such as regulation of actin cytoskeleton and prolactin signaling pathway,with genes like FGF14 and FGF9 were reported to participate in animal fat metabolism.Enrichment analysis of co-expressed and co-localized target genes identified fat metabolism-related pathways,including the cAMP signaling pathway,prolactin signaling pathway,and insulin signaling pathway.Further functional analysis screened key genes associated with fat metabolism(CREBBP,ATP1B3,PPP1R3C, and PIK3R1),which may play crucial regulatory roles in ovine intramuscular fat deposition.The qRT-PCR results of 6 randomly selected DE mRNAs and 6 randomly selected DE lncRNAs were consistent with transcriptome sequencing data.

To investigate the effect of polymorphisms in the glycogen branching enzyme 1(GBE1)gene on meat quality and flavor traits of Bashang long-tailed chickens.Target fragments were amplified using polymerase chain reaction(PCR)and sequenced directly to identify single-nucleotide polymorphisms(SNPs)in the GBE1 gene.Correlation analysis was performed between the SNPs and meat quality traits.In this study,analysis of 60 individuals revealed four SNP loci in the GBE1 gene(g.96619229T>C,g.96619243T>G,g.96619350C>T,and g.96619544C>T),each exhibiting three genotypes with dominant genotypes identified as TT,TT,CC,and CT,respectively.Chi-square tests confirmed that all loci were in Hardy-Weinberg equilibrium,with polymorphism information content(PIC)values of 0.262,0.262,0.332,and 0.373,indicating moderate polymorphism(0.25<PIC<0.50).Association analysis demonstrated significant correlations between GBE1 genotypes and meat quality traits.At g.96619229T>C,the TC genotype had higher leg muscle myristic acid content,and CC genotype showed significantly higher mean values of L^* in leg muscle compared to other genotypes.At g.96619243T>G,the TG genotype was associated with higher leg muscle myristic acid content,and the GG genotype displayed significantly higher mean values of L^* than others.At g.96619350C>T,the CT genotype had increased myristic acid content and abdominal fat rate but reduced mean values of L^* in leg muscle At g.96619544C>T,the CT genotype showed higher levels of various fatty acids in the leg muscle and superior slaughter performance traits,including carcass weight,eviscerated weight,abdominal fat weight,and leg muscle weight,while the TT genotype exhibited higher postmortem 24-hour pH in leg muscle compared to other genotypes.Linkage disequilibrium analysis identified strong correlations among the first three SNP loci,resulting in six major haplotypes,with TTC being the predominant haplotype(0.708).These findings indicate that SNPs in the GBE1 gene are significantly associated with meat quality traits of Bashang long-tailed chickens.The strong linkage disequilibrium among intronic SNPs and the identification of TTC as a favorable haplotype provide valuable insights for breeding programs aiming to enhance meat quality in this chicken population.

To elucidate the molecular mechanisms underlying peanut pod responses to water stress,this study employed pot experiments combined with transcriptomic analysis.Using well-watered conditions as the control, we systematically investigated the effects of periodic drought and waterlogging stress during the flowering-pegging stage on yield,quality,and gene expression in peanut pods. Results demonstrated that both drought and waterlogging stresses significantly reduced peanut pod yield(by 26.43% and 77.69%,respectively)and crude fat content in kernels (by 9.46, 6.71 percentage points,respectively).Transcriptomic analysis further revealed 1 525 and 1 382 differentially expressed genes(DEGs)in the drought-stress and waterlogging-stress groups compared to the control, respectively, with down-regulated expression being predominant in both sets of DEGs. Specifically, drought stress suppressed six key metabolic processes related to lipid and fatty acid metabolism in peanut pods,with 88.38% of associated genes showing downregulated expression,indicating that lipid metabolic disruption may be the primary cause of yield and quality reduction under drought. Waterlogging stress predominantly interfered with pod metabolism and defense functions by downregulating genes associated with catalytic activity,transmembrane transport,redox reactions,and biosynthesis of secondary metabolites.Moreover, KEGG enrichment analysis indicated that metabolic pathways and biosynthesis of secondary metabolites were significantly affected under both water stress conditions. Key gene validation via qRT-PCR corroborated the RNA-seq data, confirming the reliability of the transcriptomic findings. In summary, this research elucidates the molecular basis of peanut pod response to water stress at the transcriptome level, demonstrating that lipid metabolic disruption is the primary factor underlying yield reduction and quality deterioration under drought stress, whereas peanut pods mitigate the adverse effects of waterlogging mainly by modulating the expression of genes associated with redox homeostasis and metabolic pathways.

To elucidate the mechanisms of cold response and explore superior cold-tolerance gene resources in wheat,this study employed transcriptome sequencing to uncover key regulatory networks underlying wheat cold response,and performed functional validation of the candidate gene TaGGCT18-6A.The results demonstrated that 4 ℃ cold treatments induced 10 893 and 18 784 differentially expressed genes(DEGs)in wheat seedlings after 6 h and 24 h cold treatment,respectively.KEGG analysis revealed significant enrichment of DEGs in pathways including MAPK signaling transduction and glutathione metabolism.A γ-glutamyl cyclotransferase gene TaGGCT18-6A was cloned through screening key genes in glutathione metabolism.This gene had a length of 1 772 bp,encoding 218 amino acids with conserved GGCT-like superfamily and ChaC core domain.Promoter cis-acting element analysis identified stress-responsive elements such as low-temperature-responsive(LTR)and dehydration-responsive (DRE)elements; consistently,expression pattern analysis showed sustained upregulation of TaGGCT18-6A under 4 ℃ cold treatments. Functional validation in transgenic rice revealed that overexpression lines OE#1, OE#2 and OE#3 exhibited significantly enhanced survival rate,plant height,and biomass. Overexpression lines OE#1 and OE#2 exhibited significantly enhanced plant height, while overexpression lines OE#1, OE#2 and OE#3 exhibited significantly reduced relative electrolyte leakage under -4 ℃ freezing treatments.After 4 ℃ treatment,overexpression lines accumulated higher levels of osmolytes(proline and soluble sugars),decreased malondialdehyde(MDA)content,and increased activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT).These findings collectively demonstrated that TaGGCT18-6A enhanced plant cold tolerance by regulating glutathione metabolism to improve antioxidant capacity.This study will provide a theoretical foundation and valuable genetic resources for molecular breeding of cold-tolerant wheat.

In order to study the effects of different nitrogen application rates on rice yield,nitrogen uptake and soil fertility,the effects of different nitrogen application levels on rice yield,nitrogen use efficiency and soil tillage fertility in the Yellow River Irrigation Area were studied from 2021 to 2023 using rice variety Fuyuan 4 as the test material.Compared with other treatments,the average yield of 360 kg/ha was 9.4 t/ha,which was 182.94% higher than that of no nitrogen treatment,and 40.72% and 26.34% higher than that of 210,240 kg/ha.From the perspective of yield components,the increase in yield was mainly due to the increase of grain number per spike and the number of effective spikes,and excessive nitrogen application would cause the decrease of 1000-grain weight.The average of the 3-year results showed that the nitrogen use efficiency was up to 26.93% with 240 kg/ha of nitrogen,and the partial productivity of nitrogen fertilizer decreased with the increase of nitrogen application rate.The highest average value of organic matter was 18.60 g/kg after nitrogen application of 210 kg/ha.Compared with the non-nitrogen application treatment,the soil total nitrogen content of each nitrogen application treatment increased by 7.61%—15.67%,and the soil total nitrogen content of the 360 kg/ha nitrogen application treatment decreased with the increase of the experimental year.The total phosphorus content of soil was gradually reduced in the 360 kg/ha nitrogen application treatment,and the total phosphorus content in other nitrogen application treatments was maintained at about 0.85 g/kg.The total potassium content of nitrogen fertilization treatments gradually decreased with the increase of nitrogen application rate,and the total potassium content of the 360 kg/ha nitrogen application treatment decreased by 23.74%,22.16% and 8.85%,respectively,compared with the nitrogen fertilization treatments of 120,210 and 240 kg/ha.Soil available phosphorus increased with the years of the experiment,and the variation range between treatments increased with time.With the increase of nitrogen application rate,soil available phosphorus increased first and then decreased,and the soil available phosphorus of the 240 kg/ha treatment increased significantly in 2023 compared with other treatments.The soil available potassium of each treatment did not change significantly with the test years.By considering crop yield,nitrogen use efficiency and soil fertility,nitrogen application of 240 kg/ha could balance the environment and production requirements.

In order to explore the effect of partial replacement of chemical fertilizer by organic fertilizer on the ecological stoichiometric characteristics of soil carbon,nitrogen and phosphorus in the Loess Plateau of Eastern Qinghai,a field experiment was conducted with continuous cropping of potato(Qingshu 9).This field experiment was established in 2022 with five treatments,i.e.no fertilizer(CK),100% chemical fertilizer(T1),30% organic fertilizer+70% chemical fertilizer(T2),50% organic fertilizer+50% chemical fertilizer(T3),and 70% organic fertilizer+30% chemical fertilizer(T4).Soil contents of organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus,microbial biomass carbon,nitrogen,phosphorus and soil-microbial biomass ecological stoichiometry were determined.The results showed that compared with 2022,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus in each fertilization treatment in 2023 showed an increasing trend,while the CK treatment showed a decreasing trend.Among different soil layers,the content of each index gradually decreased with the increase of soil depth.Among different treatments,according to the content of each index,the order was T3>T4>T2>T1>CK.In the 0—30 cm soil layer,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus under T3 treatment were 6.88%,17.65%,17.88%,84.71%,77.01%,65.67%,80.07%,54.91% higher than those of CK,respectively.The soil C∶N and C∶P in each soil layer were the highest in the CK treatment.Among the four fertilization treatments,soil C∶N,microbial biomass C∶N and C∶P were the lowest in the T3 treatment.In addition to the 0—10 cm soil layer in 2023,the soil C∶P in each soil layer was also the lowest in T3,while the microbial biomass N∶P in the T1 treatment was the lowest among all fertilization treatments.Soil C∶N and C∶P increased with soil depth.Correlation analysis showed that organic carbon,total nitrogen,total phosphorus,available nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus were significantly positively correlated with each other.In summary,the substitution of organic fertilizer for chemical fertilizer can not only change the soil nutrient and soil microbial biomass content,but also change the soil ecological stoichiometry characteristics,and the treatment with 50% organic fertilizer+50% chemical fertilizer has the best effect.

To explore the expression pattern of potato gene StEXLB1a and provide a theoretical basis for the resistance to potato disease,based on the previous research of the Laboratory of Plant Resources and Molecular Biology at Northeast Agricultural University,the study using the Atlantic variety of potato as material, cloned the StEXLB1a gene, and preliminarily analyzed its bioinformatics, expression pattern, and disease resistance. The results showed that the full-length cDNA sequence of StEXLB1a gene was 768 bp,encoding 255 amino acids.The annotation indicated that it belonged to the extend protein family genes.Subcellular localization showed that the protein was localized to the cell wall.Among different tissues of potato,StEXLB1a gene expression was highest in leaves,followed by roots and stems,and lowest in flowers and tubers.The expression level of StEXLB1a gene changed significantly under various stress treatments,such as hormone(indoleacetic acid,gibberellin,abscisic acid),stress(high temperature,low temperature,salt,drought),fungal disease dry rot(Fusarium avenaceum),bacterial disease soft rot(Erwinia carotovora subsp,Ecc)and bacterial wilt(Ralstonia solanacearum,RS).The overexpressed transgenic potato plants were inoculated with the dry rot pathogen Fusarium avenaceum,and the transgenic plants were more seriously infected than the wild-type plants.The enzyme activities of active oxygen scavenging system in overexpressed plants were significantly changed,POD and SOD activities were inhibited,MDA content was higher than that of wild-type plants,and the damage degree of plants was aggravated.The results showed that transgenic potato plants with overexpression of StEXLB1a had lower resistance to dry rot than those with wild type.

Catalase(CAT)plays a critical role in plant resistance against biotic stresses.To investigate the functions of StCAT1 and StCAT3 in potato defense against Alternaria solani infection,this study cloned the cDNA sequences of StCAT1 and StCAT3 via RT-PCR.Sequencing results demonstrated that both genes were 1 479 bp in length,encoding 493 amino acids.Phylogenetic tree construction revealed that StCAT1 shared high homology with SlCAT1 from tomato,while StCAT3 exhibited high homology with both SlCAT3(tomato)and SpCAT3 from wild tomato.Following A.solani infection for 6 days,the expression of StCAT1 and StCAT3 was significantly upregulated by approximately 3.9-fold and 8.7-fold,respectively.A co-silencing vector targeting both StCAT1 and StCAT3 was constructed,and transient co-silencing was achieved using virus-induced gene silencing(VIGS).qRT-PCR screening identified an effective co-silenced potato line(VIGS-3)with suppressed StCAT1 and StCAT3 expression.Upon A.solani inoculation,the silenced line exhibited 42% larger lesion areas compared to the control,indicating that co-silencing StCAT1/StCAT3 compromised potato resistance to early blight.DAB(3,3'-diaminobenzidine)staining further demonstrated that H₂O₂ accumulation in co-silenced leaves was significantly elevated,with staining intensity being 3.8-fold higher than that in the control.These results demonstrate that StCAT1 and StCAT3 regulate H₂O₂ homeostasis to mediate potato resistance against A.solani,providing a theoretical foundation for elucidating the molecular mechanisms of CAT genes in potato-pathogen interactions.

To explore the association between STAT3 single nucleotide polymorphism and lactation traits of sheep,414 lactating sheep were selected as test subjects,the expression of STAT3 in seven tissues including mammary gland was detected by RT-qPCR,the single nucleotide polymorphism of STAT3 was detected by sanger sequencing,STAT3 was typed by Penta-primer amplification refractory mutation system PARMS,and the effect of the nucleotide sequence variation of the gene on milk performance traits was analysed.RT-qPCR results revealed that STAT3 expression was highest in lungs,followed by liver and mammary tissues,and lowest in spleen.A total of five novel SNPs,c.22+537 C/T,c.22+658 C/T,c.318+126 C/G,c.417+360 G/A and c.417+395 T/C,were identified in the sheep STAT3,and three genotypes were detected at each SNP locus.Association analysis revealed that ewes with the genotype CC at the c.318+126 C/G locus had a higher milk fat rate than those with GG,and CC-type at the c.22+537 C/T locus,CC-type at the c.22+658 C/T locus,GG-type at the c.417+360 G/A locus,and TT-type at the c.417+395 T/C locus all had higher milk fat percentage and dry matter content.The haplotype combination H1H7 exhibited significant positive effects on milk fat percentage,dry matter content,and ash content.Additionally,the average daily milk yield of individuals carrying the haplotype combination H1H2 was increased by 0.266 kg compared to those carrying H1H7.In conclusion,the five nucleotide variation sites of STAT3 screened in this study may provide a theoretical basis for molecular selection of milk performance traits in milk sheep.

In order to clarify the regulatory network of oil accumulation in Brassica napus and breed oilseed rapeseed varieties with high oil content.The seed transcriptome data of four rapeseed inbred lines at 25,35,and 45 days after flowering were used to identify candidate genes affecting oil content by transcriptome analysis and correlation analysis.Consequently,a total of 1 530 genes were identified exhibiting differential expression across all three period,comprising 986 up-regulated genes and 544 down-regulated genes.GO enrichment analysis of these differentially expressed genes detected 83 lipid biosynthesis genes,79 lipid degradation genes,21 lipid transporter genes and 80 transcription factors.To further analysis of these differentially expressed transcription factors,genes including BnTT8,BnGL2,and BnNAC082 were identified.Combined with the oil content data of 50 semi-winter rapeseed in three different regions over two years,four SNPs were identified in the exons region of the BnNAC082-A03 significantly associated with oil content using genome-wide association studies.Two haplotypes were detected in the region of this gene and BnNAC082-A03_Hap1 corresponding accessions showed significantly higher oil content than that of Hap2.Additionally,it utilized these transcriptomic data to construct co-expression analysis network,and in the sub-network revealed that BnNAC082-A03 was directly connected with BnTT8-A09 and BnGL2-C06,forming a potential molecular regulatory network affecting seed lipid accumulation.

To address issues such as shallow soil layers,upward movement of the plowpan,and soil salinization in the saline-alkali soils of the West Liaohe Plain,field trials were conducted in Huatugula Town,Horqin Zouyi Middle Banner,Tongliao City,Inner Mongolia Autonomous Region,from 2020 to 2021.Two tillage methods(traditional rotary tillage and powder ridge plowing),two irrigation quotas(2 100,2 700 m³/ha),and mulching and shallow burial measures were set up,resulting in six experimental treatments:2 100 m³/ha irrigation quota+traditional rotary tillage+shallow burial(CK×NM),2 100 m³/ha irrigation quota+traditional rotary tillage+mulching(CK×DM),2 100 m³/ha irrigation quota+powder ridge plowing+shallow burial(FA×NM),2 100 m³/ha irrigation quota+powder ridge plowing+mulching(FA×DM),2 700 m³/ha irrigation quota+powder ridge plowing+shallow burial(FB×NM),2 700 m³/ha irrigation quota+powder ridge plowing+mulching(FB×DM).The effects of powder ridge plowing and mulching treatments on soil properties,structure,saline and alkaline content,and maize yield in the 0—40 cm soil layer under different irrigation quotas were analyzed. The result showed that compared to the CK×NM treatment, in the 0—40 cm soil layer, the soil bulk density decreased by 8.4%—22.9%, the total soil porosity increased by 4.9—14.8 percentage points, and the soil three-phase ratio R value decreased by 34.6%—88.2% under powder ridge plowing + mulching treatment,among them, the bulk density, total porosity, and three-phase ratio R value of the soil treated with FB×DM treatment were significantly reduced by 20.0%,-13.1 percentage points, and 88.2%, respectively;soil moisture content after sowing increased by 5.5—12.1 percentage points in the 20—40 cm soil layer, soil hardness increased by 33.4%—397.5% in the 7.5—17.5 cm soil layer,among them, the soil moisture content, hardness of the FB×DM treatment increased significantly by 12.1 percentage points, 214.3%,respectively;CO₂ flux of the FB×DM treatment increased significantly by 496.4%.Compared to the CK×NM treatment, the powder ridge plowing+mulching treatment reduced the soil pH value, total alkalinity, electrical conductivity, and total salt content in the 0—40 cm soil layer, with reduction rates of 0.7%—10.9%, 2.5%—67.5%, 24.3%—68.7%, and 10.3%—81.0%, respectively. Among them, the soil pH value, total alkalinity, electrical conductivity, and total salt content of the FB×DM-treated soil were significantly reduced by 10.9%, 48.2%, 59.2%, and 80.0%, respectively.Maize germination rate, ear fresh weight, and yield were increased by 13.2—20.1 percentage points,52.5%—68.2%, and 22.4%—45.5%, respectively,among them, the germination rate, ear fresh weight, and yield of the FB×DM treatment were significantly increased by 20.1 percentage points,68.2%, and 45.5%, respectively, compared to the CK×NM treatment. Considering the comprehensive improvement effects and maize yield, it is concluded that the 2 700 m³/ha irrigation quota+powder ridge plowing+mulching(FB×DM) is a more suitable cultivation mode for saline and alkaline land in the West Liaohe Plain.

To investigate the effects of delayed sowing on the growth,development,grain yield and quality of wheat,Yunong 907(YN907)and Yunong 922(YN922),two new high yield and high quality wheat varieties,were selected as experimental materials.Three sowing dates were established:October 22nd(S1,the conventional sowing date),October 31st(S2,delayed sowing by 9 days),and November 9th(S3,delayed sowing by 18 days).The study investigated the effects of delayed sowing on the phenological stages,grain yield and flour quality of wheat,and analyzed the relationship between temperature characteristics and wheat yield,and quality under delayed sowing conditions.The results indicated that compared with the conventional sowing date, the full growth period of YN907 and YN922 was shortened with the delay of sowing date in both years. The accumulated temperature before winter and the average daily temperature before heading decreased with the delay of sowing date, while the average daily temperature and effective accumulated temperature after heading increased. The effective accumulated temperature of the full growth period showed a downward trend. The effective accumulated temperature of the full growth period of YN907 and YN922 in S3 treatment was 243.95,222.10 ℃·d lower than that in S1 treatment in 2022—2023, and 136.30,189.40 ℃·d lower in 2023—2024, respectively. The yield and its components decreased with the delay of sowing, with a yield reduction of 6.45% to 17.26%. Compared with the conventional sowing date, the wet gluten content and protein content increased under the delayed sowing conditions.Under the background of climate warming and the scale-up of the agricultural operation,the sowing date of wheat can be adjusted to adapt to the temperature change.YN907 and YN922 had the highest yield on October 22nd.Delaying the sowing date to October 31st could maintain the higher yield level and increase the grain protein content.When the sowing date continues to be postponed,the grain protein content will be significantly increased but the yield will be significantly decreased.

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.

The effects of different nitrogen reduction and efficiency enhancement measures on ammonia volatilization and crop yield in fluvo-aquic soil of wheat-maize rotation system were studied to provide guidance for rational fertilization and agricultural environmental protection.A long-term nitrogen reduction was carried out since 2016 at the experimental station of Xuchang fluvo-aquic soil area in Henan Province.No nitrogen fertilizer control(CK),conventional nitrogen fertilizer(100N),20% nitrogen reduction(80N),and 20% nitrogen reduction combined with straw returning(80NS),nitrification inhibitor(80NI),and biochar(80NB)were set up.The soil physical and chemical properties,annual ammonia volatilization characteristics and wheat and maize yield of different treatments were studied from 2021 to 2022.In the wheat season,the pH value of 80NS,80NI and 80NB treatments was significantly higher than that of 100N treatment.The organic matter content significantly increased, while the soil bulk density significantly decreased in the 80NS and 80NB treatments, compared with the 80N and CK treatments, respectively. At the base fertilizer stage of wheat,the ammonia volatilization accumulation of 80NS,80NI and 80NB treatments was significantly lower than that of 100N by 28.71%,35.61% and 22.99%,respectively.During the topdressing stage of wheat season,the ammonia volatilization accumulation of 80NS and 80NB treatments was significantly lower than that of 100N by 14.94% and 17.58%,respectively.The ammonia volatilization accumulation of 80NS and 80NI treatments was significantly increased by 22.27% and 27.69% compared with 80N.During the whole growth period of wheat,the accumulation of ammonia volatilization in different nitrogen treatments accounted for 1.31%-2.47% of the nitrogen application rate,showing 100N> 80NB> 80NS> 80NI> 80N.In the maize season,compared with the accumulation of ammonia volatilization under 100N treatment,80N and 80NS treatments significantly decreased by 37.14% and 29.63%,respectively,and 80NI treatment significantly increased by 60.83%.Compared with 80N treatment,the accumulation of ammonia volatilization in 80NI and 80NB treatments increased significantly by 155.79% and 44.05%.The accumulation of ammonia volatilization in maize growth period accounted for 5.81%-14.86% of nitrogen application rate,showing 80NI> 100N> 80NB> 80NS> 80N.The wheat yield results indicated that compared with 100N treatment,80N treatment significantly reduced the yield by 16.67%,while 80NS,80NI and 80NB treatments did not significantly reduce the yield.Maize yield data showed that there was no significant difference between 100N treatment and four nitrogen reduction treatments.In summary,the application of nitrification inhibitors,straw and biochar on the basis of reducing nitrogen by 20% in the experimental fluvo-aquic soil can effectively improve soil fertility and stabilize crop yield.However,nitrification inhibitors and biochar significantly increase the accumulation of ammonia volatilization in maize season,which needs special attention in actual production.

To explore the effects of nitrogen application rate on nitrogen uptake and translocation characteristics,nitrogen use efficiency,and the formation mechanisms of yield and quality in soft wheat under weak light stress during the after anthesis stage,under pot conditions,the soft wheat variety Quanmai 725(QM725)was used as the material,and the ¹⁵N tracer method was used.Two nitrogen rates were set in the experiment,namely N1(N 120 kg/ha),N2(N 180 kg/ha),and two shading treatments were set at the wheat filling stage(7-35 d after anthesis)under each nitrogen application rate,namely CK(no shading),SH(30% shading).The relationship between nitrogen application rate and grain yield and quality of soft wheat under weak light after anthesis was analyzed,and the effects of different nitrogen application rates on nitrogen accumulation,transport,grain yield and quality of soft wheat under weak light after anthesis were studied.The results showed that compared with the control,under the conditions of N1 and N2,the nitrogen accumulation of plants at flowering stage and vegetative organs at maturity stage was significantly reduced by weak light treatment after anthesis,and the proportion of nitrogen from fertiliser was significantly higher than that from soil nitrogen,while the proportion of nitrogen accumulation in grains at maturity stage from soil nitrogen was significantly higher than that from fertiliser nitrogen.Under the same nitrogen application rate,the proportion of basal nitrogen fertiliser was greater than that of topdressing nitrogen fertiliser.Under the same weak light treatment conditions,compared with N1,N2 increased the accumulation of fertiliser nitrogen at flowering stage,the accumulation of total nitrogen and fertiliser nitrogen at maturity stage,and the accumulation of total nitrogen,fertiliser nitrogen and soil nitrogen at maturity stage.Under N1 and N2 treatments,the nitrogen harvest index,nitrogen harvest index,nitrogen production efficiency,grain number per spike,1000-grain weight and grain yield of wheat decreased significantly with the decrease of light intensity after anthesis.The content and accumulation of protein and starch in soft wheat grains increased significantly with the increase of nitrogen fertilizer.However,under the same nitrogen application rate,weak light stress reduced the starch content,protein and starch accumulation in grains.Weak light stress after anthesis significantly affected the nitrogen accumulation of soft wheat plants,reduced the transport efficiency of storage substances from vegetative organs to grains after anthesis,resulting in a decrease in the contribution rate of vegetative organs to grains,which was not conducive to the overall nitrogen transport efficiency of plants.With the increase of nitrogen application rate,the nitrogen harvest index,nitrogen harvest index,nitrogen production efficiency and nitrogen use efficiency of wheat were significantly improved.Under the same nitrogen application rate of N1 and N2,after anthesis weak light stress significantly reduced the accumulation of protein and starch in soft wheat grains,which in turn affected the formation of grain weight,resulting in a decrease in yield.

Fusarium head blight(FHB)is a devastating fungal disease that seriously threatens the safety of wheat production.Marker-assisted selection(MAS)and pyramiding of resistance genes represent efficient strategies for FHB-resistant breeding.To establish a high-throughput screening system for FHB resistance genes and enhance wheat resistance in Sichuan Province,we performed genome-wide genotyping using a 100K SNP array on 14 Sichuan wheat varieties(lines)along with three FHB-resistant genetic materials.Based on the reported genetic linkage intervals of major FHB resistance genes(Fhb2,Fhb4,Fhb5),we identified SNPs co-segregating with Fhb5 or linked to Fhb2,Fhb4,and subsequently developed kompetitive allele-specific PCR(KASP)markers.Results showed that the genetic relationship of 17 wheat varieties(lines)could be clustered into two major groups:two northern wheat-derived resistant materials(NMAS070 and NMAS069)formed an independent cluster distinct from the Sichuan varieties(lines)while the remaining 15 varieties(lines)were clustered together and subdivided into two subgroups.Functional gene profiling revealed FHB-resistant parents carried superior resistance loci,whereas agronomic parents harbored favorable alleles for yield and quality traits.Through SNP screening,we identified 8 critical SNPs within the linkage intervals of Fhb2,Fhb4 and the co-segregation region of Fhb5.These SNPs enabled the successful development of 4,2,and 2 high-specificity KASP marker systems for Fhb2,Fhb4 and Fhb5,respectively.Validation experiments confirmed all KASP markers achieved precise genotyping and were effectively implemented in molecular breeding for FHB-resistance.This study established a high-efficiency KASP marker system for Fhb2,Fhb4 and Fhb5,providing a robust technical platform for improving FHB resistance breeding of wheat varieties in Southwest China.

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.