Bioinformatics and expression patterns of Dicer-like(DCL),Argonaute(AGO)and RNA-dependent RNA polymerase(RDR)gene families in the whole genome of Solanum habrochaites were analyzed,so as to provide references for further study on the functions of DCL,AGO and RDR gene families in the response of S.habrochaites to abiotic and viral infection.Using Arabidopsis thaliana DCL,AGO and RDR genes as reference sequences,the genome of S.habrochaites LA1777 was searched by local perl language and software such as Pfam and SMART,and the members of ShDCL,ShAGO and ShRDR gene families were determined.Bioinformatics analysis of DCL,AGO and RDR family genes in S.habrochaites was carried out by means of ExPASy,GSDS 2.0,MEGA,Tbtools and SWISS-MODEL.According to abiotic stress treatment,Tomato chlorosis virus(ToCV)treatment and Real-time Fluorescence Quantitative PCR technology,the expression patterns of these genes were analyzed.Seven ShDCL,15 ShAGO and 6 ShRDR genes were identified from S.habrochaites,which were distributed on chromosome 5,7 and 6 respectively.The encoded proteins were similar in structure to DCL,AGO and RDR in other plants,and all of them contained conserved domains unique to this family.Phylogenetic analysis showed that these genes were divided into 4 subgroups,and there were high structural and functional similarities between S.habrochaites and S.lycopersicum.ShDCL2a,ShDCL2c,ShDCL3,ShDCL4,ShAGO1b,ShAGO3,ShAGO4b,ShAGO5,ShAGO7,ShAGO10a,ShAGO10b,ShRDR1,ShRDR2,ShRDR3a,ShRDR6a and ShRDR6b were significantly up-regulated after various abiotic stresses and ToCV infection.It is speculated that these genes play important roles in abiotic stress and virus infection.
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The folding mode of Chinese cabbage leaf ball is the main character that determines the appearance shape,taste and stress resistance of commercial organs.In order to explore the internal molecular mechanism of the formation of the folding mode of Chinese cabbage,we cloned the full length sequence of the transcription factor BrPIF5 gene from overlaping and outward-curling Chinese cabbage as experimental materials,and conducted bioinformatics analysis,constructed the plant overexpression vector,and used Agrobacterium to mediate the transformation into tobacco to obtain positive transformation plants.The expression level of BrPIF5 gene in tobacco was detected by qRT-PCR.The results showed that the protein encoded by BrPIF5 gene was a hydrophilic protein with a continuous and complete open reading frame of 634 bp,containing 210 amino acids.The protein was composed of more α-helical structure and random curl,including an AP2/ERF domain.BrPIF5 protein and the other 9 gene family members contained a conserved motif 1,and the position was different from that of other gene family members,which was located in the front of the protein sequence.Phylogenetic tree showed that BrPIF5 gene had close evolutionary relationship with SoPIF15,BhPIF1,BoPIF4,AtPIF4 and BrPIF4 family members.The tobacco strain with overexpression of BrPIF5 was obtained by Agrobacterum-mediated genetic transformation,and the leaves of the tobacco positive transformation strain showed inward curling.qRT-PCR showed that the expression level of BrPIF5 gene in the overlaping Chinese cabbage was higher than that in the outward-curling Chinese cabbage,and the gene expression level in the positive tobacco plants was higher than that in the control.It was further proved that BrPIF5 gene controlled the inward curling of Chinese cabbage leaves,thus promoting the formation of leaf ball folding type.
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Temperature stress is one of the main nonbiological stresses that affect the quality and yield of spinach.Investigating the molecular response mechanism of spinach to temperature stress is crucial for spinach stress tolerance breeding.To provide a theoretical basis for the research of the mechanism of spinach resistant to cold stress and heat stress,this study used the cold-tolerant inbred line D3 and the heat-tolerant inbred line M10 of spinach as experimental materials and analyzed their transcriptomes and metabolomes under cold and heat stress to explore the transcriptional and metabolic mechanisms underlying spinach tolerance.Transcriptomic analysis showed that the pathways in which the DEGs were the most enriched in D3 and M10 were essentially the same under cold stress and heat stress.Metabolomics analysis showed that under cold stress,they were coenriched in the pyrimidine metabolism and lysine degradation in KEGG pathways.Under heat stress,these were mainly enriched in the tryptophan metabolism,toluene degradation,biosynthesis of various other secondary metabolites,and glycine,serine and threonine metabolism in KEGG pathways.The joint transcriptomic and metabolomic analysis indicated that through data analysis and gene annotation.SpADH(sov2g036390),SpSHMT(sov1g001130)and SpALDH-1(sov4g007150)were identified as the candidate genes for cold stress tolerance in spinach.SpALDH-1(sov4g007150),SpALDH-2(sov1g043320)and SpNPC(sov1g040610)were identified as candidate genes for heat stress in spinach.Among them,SpALDH-1(sov4g007150),which may be a regulatory gene for spinach stress tolerance,was significantly expressed under both cold and heat stress.
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In order to investigate the function of watermelon calcium-dependent protein kinase (CDPK) in grafted seedlings and abiotic stress environments, this study used RT-PCR technology to clone the ClCDPK(Cla97C01G019720) gene from watermelon grafted seedlings and performed bioinformatics analysis on it. Further designed specific primers with Kpn Ⅰ and Sal Ⅰ enzyme cleavage sites based on the ClCDPK sequence,conducted amplification and double enzyme cleavage, and connected with pCAMBIA1300 to successfully construct the expression vector pCAMBIA1300-35S-ClCDPK for the target gene.Using RT-qPCR technology, the gene expression levels of ClCDPK were measured in self rooted seedlings (ZG) and grafted seedlings (JJ) after being subjected to salt and drought stress, respectively.The results showed that the ORF of ClCDPK gene was 1 647 bp, encoding 548 amino acids. Its protein contained STKc_CAMK and FRQ1 functional domains, and was a hydrophilic protein. Subcellular localization prediction showed that the protein was located in the nucleus. Evolutionary tree analysis of ClCDPK with CDPK from six other plants revealed that it was closely related to CDPK from Cucurbitaceae melons and pumpkins, with protein sequence homology alignment exceeding 92.64%, indicating high homology.The RT-qPCR expression results showed that the expression level of ClCDPK in grafted seedlings was significantly higher than that in self rooted seedlings. With the duration of stress, the expression levels of ClCDPK in grafted and self rooted seedlings first increased and then decreased, and under the same stress treatment, the expression level of ClCDPK in grafted seedlings was higher than that in self rooted seedlings.This study indicated that ClCDPK responded positively to salt and drought stress, and the ability of grafted seedlings to resist stress was higher than that of self rooted seedlings. It is speculated that ClCDPK is one of the key factors in watermelon's response to grafting, thereby improving the salt and drought resistance of watermelon grafted seedlings.
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Serine hydroxymethyltransferase(SHMT),as an important enzyme involved in basic metabolism,plays an important role in plant cell metabolism,photorespiration and defense activities.To understand the bioinformatics function of the SHMT gene family in watermelon,explore its gene expression characteristics under abiotic stress,and provide a basis for the functional development of watermelon SHMT and the breeding of watermelon stress-resistance genes.Bioinformatics methods were used to identify SHMT family,and RT-qPCR was used to analyze the expression patterns of ClSHMTs in different tissues and abiotic stresses.The results showed that 8 ClSHMTs gene family members were identified in the whole genome of watermelon,which were unevenly distributed on 6 chromosomes and named ClSHMT1—ClSHMT8 in turn.There were some differences in the physical and chemical properties of each gene family member,such as the number of amino acids,molecular weight,isoelectric point.The protein contained 471—585 amino acids,with molecular weight of 51.87—65.00 ku and isoelectric point of 6.57—8.52,all of which were hydrophilic proteins.The subcellular localization prediction was mainly distributed on mitochondria.Gene structure and protein conserved motifs analysis showed that the ClSHMTs structure consisted of 4—15 exons and 3—14 introns,and all ClSHMTs contained conserved SHMT domains.Furtherly,phylogenetic analysis with 6 species such as cucumber and wheat showed that 50 SHMTs were divided into 3 sub-families,Group Ⅰ—Ⅲ.Promoter of ClSHMTs contained cis-acting elements related to light response,plant hormone response and stress response.The expression pattern analysis showed that 6 ClSHMTs were expressed in different tissues of watermelon,and the expression levels of ClSHMT1,ClSHMT4,ClSHMT5,ClSHMT8 in leaves were significantly higher than those in other tissues.Under low temperature,drought and salt stress,the expression abundance of ClSHMTs varies,but the expression was mainly up-regulated.In conclusion,this study systematically analyzed the SHMT gene family in watermelon,and will provide a reference for the further study of the biological functions of ClSHMTs.
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In order to study the role of phototrapping pigment binding protein gene in potato photosynthesis,the phototrapping pigment binding protein gene StCP24 (LOC102586836)was bioinformatics analyzed and cloned into potato variety Dongnong 310 to obtain transgenic potato.The morphological indexes,leaf physiological indexes,fluorescence parameters,and gene relative expression of transgenic and control lines were measured under different light intensities after clonal propagation.The function of StCP24 gene was analyzed.The effect of this gene on chloroplasts was determined by ultrastructural observation.The results of bioinformatics analysis showed that StCP24 protein was a hydrophilic protein,and the promoter of StCP24 gene contained elements such as photoresponse,defense and stress response.Phylogenetic analysis showed that the amino acids encoded by StCP24 gene in potato were closely related to those encoded by StCP24 gene in cultivated tomato.Under different light intensity treatments,StCP24 gene overexpressed potato significantly increased stem diameter,leaf area,leaf weight,root length,chlorophyll a,b content,Fv/Fm,ETR,ETRmax and qP compared with wild-type potato.The overexpression of StCP24 gene can make the lamellar stacking of thylakoid granules more tightly and to a higher degree.In conclusion,the overexpression of StCP24 gene can increase the content of chlorophyll a and b in leaves,improve the electron transfer rate in photo cooperative photoreaction,and promote the growth of transgenic potato plants.
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Tiller-related traits are important characteristics of wheat plant type,which determine plant structure and affect grain yield.In order to understand the inheritance and drought resistance of tiller-related traits in wheat under different water conditions,and to excavate the loci related to tiller-related traits,240 wheat varieties (lines) were selected as the subjects of this study,based on the phenotypic identification of tiller angle,effective tiller number and yield per unit area under normal irrigation (NI) and drought stress (DS) conditions,and the comprehensive evaluation of drought resistance,combined with 90K gene chip,genome-wide association study (GWAS) was to identify genetic loci for tiller-related traits and to screen for superior germplasm.The tiller angle,effective tiller number and yield per unit area showed significant difference,and the coefficient of variation ranged from 0.07 to 0.33.According to D-value,the drought resistance of Zhongyou 206 was the best.A total of 54 stable genetic loci significantly associated with tiller angle and other traits were detected, distributed on all chromosomes except 3D, 4D and 5D. Three identical stable loci were commonly detected under both treatments, located on chromosomes 2B, 4B, and 6B. Additionally, four pleiotropic loci were commonly detected in different traits, located on chromosomes 2B, 2D, and 5B.At the same time,the haplotype analysis of Ra_c491_902 (R2=5.45%—17.91%),which was significantly correlated with tiller angle on chromosome 2B,showed that there were three haplotypes:TA-Hap1,TA-Hap2 and TA-Hap3,the haplotypes (lines) containing TA-Hap1 were mainly derived from Huanghuai winter wheat regain.Five candidate genes related to tiller angle were screened by screening the stable genetic loci detected under different treatments.Gene annotation of the genes selected on Ra_c491_902 showed that the genes encoding cytochrome P450 family protein can be used as important genes such as regulating tillering angle,plant drought resistance and defense,to explore the association between genes and phenotype,and lay the foundation for the genetic improvement of tiller-related traits in wheat.
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To investigate the characterization and function of LbaHY5 gene of Lycium barbarum,explore the role of LbaHY5 in the degree of stem straightening in Lycium barbarum.Using Ningqi No.1 as the test material,a preliminary analysis was conducted on the structure and function of the LbaHY5 gene using bioinformatics,subcellular localization,and qRT-PCR methods,the results indicated that the LbaHY5 gene cloned from Ningqi No.1 had a full-length open reading frame of 483 bp,encoding 160 amino acids,with a molecular weight of 17.52 ku and an isoelectric point of 9.69,classifying as a hydrophilic protein.Conserved domain and multiple sequence alignment analysis showed that LbaHY5 protein contained a type bZIP domain,which belonged to the bZIP gene family.Evolutionary tree analysis showed that the LbaHY5 protein in goji berries was highly homologous to the HY5 protein in Solanaceae plants such as tomatoes and potatoes.In addition,promoter analysis showed that the promoter region of this gene was rich in multiple functional elements,which were respectively related to processes such as light signaling,hormone signaling pathways,and abiotic stress response.The subcellular localization indicated that LbaHY5 was located on the nucleus.The transcript level of LbaHY5 gene was the highest in the flower and the lowest in the stem.During the fruit development period,the expression level of LbaHY5 gradually increased.The results speculated that LbaHY5 gene might play an important role in the growth and development of floral organs and fruits in Lycium barbarum.
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In order to elucidate the sequence characteristics and expression pattern of PatGTL1,a member of Trihelix transcription factor family,and lay a foundation for the further function study of PatGTL1,the total RNA of Pogostemon cablin (patchouli)was extracted and reverse-transcribed to cDNA.The specific primers were designed according to the transcriptome sequence of PatGTL1,and the cDNA was used as a template to clone PatGTL1 gene.The bioinformatics analysis was further performed to reveal the structural,physicochemical properties,and phylogenetic relationships of PatGTL1.Besides,the pAN580-PatGTL1-EGFP vector was built and transformed into Arabidopsis protoplastsa to detect the subcellular localization of PatGTL1.The Real-time Quantitative reverse transcription PCR(qRT-PCR)was also carried out to investigate the expression patterns of PatGTL1 gene in different tissues of patchouli and under treatments of methyl jasmonate(MeJA),salt stress,drought stress,and cold stress.The results showed that the open reading frame of the PatGTL1 gene was 1 497 bp,encoding 498 amino acids.The PatGTL1 protein was an unstable hydrophilic protein that contained 42 serine phosphorylation sites and was predicted to be located in the nucleus without transmembrane domains and signal peptides.Multiple sequences alignment showed that PatGTL1 contained two GT1 conserved domains,and was classified into GT-2 subfamily of Trihelix transcription factor family.According to the phylogenetic tree,PatGTL1 was clustered closely with Sesamum indicum SiGTL1.The qRT-PCR analysis indicated that PatGTL1 expressed in the root,stem,and leaf in patchouli,particularly with the highest expression in the young leaf.The PatGTL1 expression was significantly upregulated by MeJA and showed an increasing trend at 3—24 h after treatment,but downregulated by drought stress.The highest expression of PatGTL1 was showed at 3 h after salt treatment,and at 12 h after cold treatment.These results enrich the Trihelix transcription factor family study of patchouli.
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The seed yield per spike and spike axis dry weight affect yield per plant.In order to reveal the genetic basis,QTLs were mapped with populations F2 and BC1 (F1×P2) derived from the cross combination 9048×16-201 using CIM and ICIM-ADD methods.A total of 7/10 (CIM/ICIM-ADD) QTLs were detected in the F2 population,including 3/6,3/3 and 1/1 QTLs conferring PSSY,PSADW and PBSADW respectively,with a contribution rate of 15.81%/24.09%,2.84%/8.65% and 6.49%/6.56%,respectively.And 1/3 QTLs were identified in the BC1 population,including 0/2 and 1/1 QTLs underlying PSSY and PBSADW,with a contribution rate of 0/10.28% and 11.60%/10.22% respectively.Among all the detected QTLs,qPSSY3.1 and qPBSADW3.1 were stable QTLs,and the latter was also the main-effect QTL,with contribution rates ranging from 3.76%—7.00% and 6.49%—11.60%,respectively.They overlapped in the marker interval RCM920—RCM950,forming a QTL cluster (i.e.,QTL-cluster4).Additionally,a dozen pairs of epistatic QTLs were detected with the epistatic effect of these traits accounting for more than 19% of the total contribution rate,indicating that the epistatic effect was an important genetic component of seed yield per spike and spike axis dry weight in castor.The results not only provided a reference for the understanding of the genetic structure of seed yield per spike and spike axis dry weight but also offered available molecular markers for marker-assisted selection.
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In order to study the problem of strong winter/spring Brassica napus seed germination and flowering period under different winter sowing dates.Two strong winter rapeseeds and two spring rapeseeds provided by Gansu Agricultural University were used as materials.The experiment was carried out in the experimental field of Gansu Agricultural University from October 2022 to August 2023.The winter rapeseeds was carried out on October 11,2022.The winter/spring rapeseeds was sown every 20 days from December 10,2022,and the sowing ended on February 8,2023.The flowering period was recorded,and the germination seeds of winter rapeseed were sampled every 20 days to determine their physiological and biochemical characteristics and analyze the expression characteristics of vernalization genes(FLC,VRN2,FRI,FT).The results showed that the flowering period of winter/spring rape seeds was different by 22—34 days.The difference of flowering time between autumn sowing and spring sowing was 4—7 days.The flowering time of winter rapeseed in autumn sowing(October 11 th)was close to that of spring rapeseed under different winter sowing dates(December 10th,December 30th,January 19th,February 8th),and the flowering overlap time was as long as 15—20 days.With the delay of the sowing date,the relative expression levels of FLC,FRI and FT genes in germinating seeds of winter sowing were down-regulated.The relative expression of VRN2 gene was down-regulated in the early vernalization and up-regulated in the late vernalization.The activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and the contents of soluble protein(SP),gibberellin(GA3)and salicylic acid(SA)in germinating seeds were increased in the early vernalization,but those were decreased in the late vernalization.The contents of malondialdehyde(MDA)and abscisic acid(ABA)were increased in rapeseed germinating with the increase of vernalization time.
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This study explored the effects of intercropping and rotation on the growth,yield and quality of continuous cropping peanut,to provide theoretical basis for achieving high yield in peanut production.From 2022 to 2023,sweet potato-peanut rotation system(PSP)and maize-peanut intercropping and rotation system(PMP)were set up in the experimental farm of Henan University of Science and Technology on the basis of continuous cropping peanut for 2 years and 11 years respectively,with continuous cropping peanut as control(CCP1 and CCP2,respectively).The effects of PSP and PMP on photosynthetic characteristics,root characteristics,dry matter accumulation and distribution and yield of peanut were studied.The results showed that compared with CCP1,the leaf area index(LAI)of rotating peanut in PSP system(SRP)was significantly increased by 35.08%—53.68% and 24.32%—33.52% at pod-setting stage(PSS)and full pod maturity stage(PMS),respectively.The SPAD value at PSS and pod bulking stage(PBS)increased by 11.93%—18.55% and 5.95%—9.63%,respectively.Compared with CCP2,the LAI of rotating peanut in PMP system(MRP)increased by 46.81%—57.96% and 27.00%—61.78% at PSS and PMS,respectively.At PSS and PBS,compared with CCP2,the SPAD value of MRP and intercropping peanut(MIP)increased by 3.32%—3.69%,7.50%—8.64% and 5.47%—18.37%,15.73%—31.11%,respectively.At PSS and PBS,compared with CCP1,the net photosynthetic rate of SRP increased by 23.68%—41.31% and 26.52%—32.55%,and compared with CCP2,MRP increased by 12.77%—17.81% and 16.88%—62.07%,respectively.They both significantly improved the root length and root tip number,and promoted the dry matter accumulation and the distribution to pods during PMS,and the yields increased by 31.42%—47.36% and 54.12%—75.09%,respectively.Compared with CCP2,MIP reduced the LAI,net photosynthetic rate,root length,root tip number,as well as dry matter accumulation and yield of peanut under the influence of maize shading.At the same time,the content of peanut oleic acid and oleic acid-linoleic acid ratio was significantly increased after rotation.Among them,SRP increased by 1.63—1.65 percentage point and 6.59%—10.52%,respectively,compared with CCP1,and MRP increased by 1.95—2.82 percentage point and 9.75%—14.16% compared with CCP2,respectively.In summary,sweet potato-peanut rotation and maize-peanut rotation increased the peanut yield compared with continuous cropping peanut,the reason was that sweet potato-peanut and maize-peanut rotation promoted peanut root growth,delayed the leaf senescence,and increased photosynthetic rate,especially the photosynthetic rate during late growth period,which promoted the dry matter accumulation and distribution to seeds.Besides that,they could improve the quality of peanut to a certain extent.
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In order to explore the improvement effect of calcium fertilizer and ARC microbial agent on red earth dry land with low-calcium,the peanut variety Xianghua 522 was used as the experimental material,and two levels of calcium hydroxide fertilizer(0,750 kg/ha,code Ca0 and Ca50)and three levels of ARC microbial agent(0,30,60 kg/ha,code A0,A2 and A4)were set to form six treatments to carry out pot experiment.It measured soil nutrient and soil enzyme activity in 0—20 cm arable layer soil at peanut seedling stage,flowering stage,pod setting stage and pod filling stage,and pod economic characters and yield were measured at harvest.The results showed that:single application of calcium fertilizer and the combined application of calcium fertilizer and ARC microbial agent could significantly improve the pH of soil at all growth stages,but ARC microbial agent had little effect on it.Compared with CK(Ca0A0),Ca50A2 and Ca50A4 significantly increased the content of hydrolyzable nitrogen in soil in the whole stage and the content of available phosphorus in soil in the first three growth stages;the content of available potassium in soil of Ca50A4 was higher than CK in general,and it was significant at seedling stage and pod setting stage;compared with CK,Ca50A0 significantly increased the content of exchangeable calcium in soil in four stages,with an increase of 23.78%—56.21%;the content of calcium ion in soil with calcium fertilizer application was significantly higher than that without calcium fertilizer application(the flowering stage was not significant),and it was little affected by ARC microbial agent;the content of organic matter in soil remained stable in the whole growth stage,but Ca50A4 and Ca50A2 were significantly higher than CK in each growth stage.Compared with CK,the soil sucrase activity of soil each treatment was significantly increased in four stages,and the increase was the largest in Ca50A4,ranging from 50.79% to 162.56%;the protease activity of soil was significantly increased by Ca50A2 in four stages with an increase of 26.58%—244.63%;the acid phosphatase activity of soil was significantly increased by Ca50A4 and Ca0A2 during the whole growth stage;the catalase activity of soil in all treatments showed a decreasing trend in general.All treatments could increase the yield of peanut pod in different degrees,and the effect of calcium fertilizer application was greater than that of ARC microbial agent,among which Ca50A4 had the best effect,with the pod weight per plant increasing by 12.29%,mainly increased the pod number per plant and the full pod number per plant.To sum up,the combined application of calcium fertilizer and ARC microbial agent has a good interaction effect on improving soil nutrient content,stimulating soil enzyme activity and increasing peanut yield,and the best effect is 750 kg/ha calcium fertilizer+60 kg/ha ARC microbial agent(Ca50A4),which can provide a theoretical basis for green and high yield cultivation of peanut.
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To explore the planting patterns of mechanical harvesting sesame and achieve goals of high yield,stable yield and deep combination of agricultural machinery and agronomy,the split plot design with two factors was carried out to investigate the effects of different planting pattern and density on the photosynthetic characteristics,biomass,yield and mechanical harvesting characteristics of Yuzhi ND837,in order to provide theoretical foundation and technical guidance for the full mechanization production of sesame.The planting pattern included wide-and narrow-row spacing planting(Z1,wide-row:60 cm,narrow-row:20 cm),banding planting of 4 rows(Z2,line spacing:30 cm,banding spacing:60 cm),banding planting of 8 rows(Z3,line spacing:30 cm,banding spacing:60 cm)and equidistant row planting(Z4,line spacing:30 cm)as primary area.The planting density included 180(M1),225(M2),270(M3)and 315 thousand plants/ha(M4)as vice-area.The results showed that planting pattern and planting density both had significant effects on the photosynthetic characteristics,substance accumulation,yield and mechanical harvesting characteristics of sesame.The net photosynthetic rate(Pn),SPAD value and biomass per plant descended with the order of M1>M2>M3>M4,while the biomass of population increased at first and then descended with the planting density increasing under the same planting pattern.Under different planting patterns,the Pn,SPAD value,biomass per plant and biomass of population all showed Z1>Z2>Z3>Z4.Under the comprehensive effects of planting pattern and density,the Pn and SPAD value of Z1M1 were the highest,and the biomass per plant was also the highest in Z1M1(59.76 g/plant),but the biomass of population and yield were the highest in Z1M3(13 032.97,1 719.87 kg/ha,respectively).The uniformity of plants tended to be the same,the diameter of stem and the ability of capsule formation per plant decreased,but the lodging percentage at maturity also gradually decreased with the increase of planting density(M1>M2>M3>M4).The lodging percentage showed Z1>Z2>Z3>Z4 among different planting patterns.Under the comprehensive influence of planting pattern and density,the lodging percentage of Z3M1(17.51%)was the highest and Z4M4(7.97%)was the lowest.Under the conditions of the experiment,Z1M3 has the biggest yield,and the agronomic traits and mechanical harvesting characteristics at maturity are also better.
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This study systematically investigates the effects of silicon-modified biochar (MSC) on the chemical properties of acidic soil,organic carbon and silicon fractions,and the growth of tomato plants.Silicon-modified biochar was prepared,with a focus on investigating its impacts on carbon and silicon chemical fractions,and the availability in acidic soils;tomato growth and soil microbial activity were also evaluated.The results showed that silicon-modified biochar significantly increased soil pH,cation exchange capacity,electrical conductivity,available phosphorus and potassium.MSC also raised the levels of water-soluble sodium and iron in the soil and enhanced the activities of hydrogen peroxidase and sucrase enzymes,thereby improving soil quality.Both biochar modification and unmodified biochar significantly increased the content of different carbon fractions in the soil.Compared with unmodified biochar,silicon-modified biochar significantly increased soil microbial biomass carbon(21.9%) and water-soluble organic carbon (898.3%).Furthermore,silicon-modified biochar significantly increased the contents of soil available silicon,water-soluble silicon,free silicon,active silicon,iron-manganese-bound silicon and amorphous silicon by 362.6%,158.9%,18.1%,34.9%,193.8%,and 74.1%,respectively.Meanwhile,the application of biochar promoted the growth of tomato plants and the absorption of silicon nutrients,with modified biochar showing more pronounced effects.The accumulation of plant dry matter,silicon content,and absorption rate increased by 82.0%,98.9%,and 261.5%,respectively.In summary,silicon-modified biochar significantly affected the carbon and silicon chemical forms and transformation in the soil,increased soil effectiveness and enzyme activity,thereby promoting nutrient absorption and growth of crops,demonstrating its good potential application in agricultural production.
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To explore the effects of soil phosphorus levels and exogenous microbial agents on soil functioning(soil physico-chemical properties,soil enzyme activities,and arbuscular mycorrhizal fungi(AMF)community composition and structure)of the rhizosphere soil of maize,a two-factor interaction experiment was conducted to study the effects of two levels of low phosphorus and normal phosphorus and four treatments of inoculation with AMF,phosphate-solubilizing bacteria(PSB),AMF-PSB and no inoculation of exogenous agent(CK)on rhizosphere soil indexes in different growth stages of maize.The results showed that the soil total phosphorus content was the highest at jointing stage and silking stage under the condition of normal phosphorus level+AMF treatment,while the soil available phosphorus content was the highest at jointing stage and mature stage under the condition of normal phosphorus level+AMF treatment.At jointing stage and silking stage,alkaline phosphatase activity was the highest under low phosphorus+AMF treatment,and the activity of alkaline phosphatase under low phosphorus+AMF treatment and low phosphorus+PSB treatment was higher than that under normal phosphorus level.Under low phosphorus level,total phosphorus content and available phosphorus content were higher than CK treatment,and the increase rate was higher than that under normal phosphorus level.At low phosphorus level,soil pH value was lower than CK treatment under the condition of inoculated foreign bacteria,and also lower than CK treatment under normal phosphorus level at jointing stage.Under the low phosphorus level,the CK treatment had the highest observed number of AMF community OTUs and α-diversity indexes,while under the normal phosphorus levels,the observed number of OTUs and α-diversity index in the CK treatment group were lower than those in the AMF and PSB treatments.Non-metric multidimensional scaling analysis indicated that the composition and structure of soil AMF communities were convergently regulated by the types of inoculated exogenous microorganisms.In summary,inoculating AMF and PSB agents in low-phosphorus alkaline maize fields in northern China can improve soil physicochemical properties and increase soil alkaline phosphatase activity,but it also increases competition and exclusion among soil AMFs.
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To study the photosynthetic characteristics,fluorescence parameters and yield response to phosphorus in maize,and to clarify the optimal phosphorus application rate for maize under drip irrigation and water fertilization technology.Providing solid theoretical basis and technical support for high-yield and high-efficiency cultivation of maize in Ningxia region.The experiment was carried out at Pingjipu Farm,Yinchuan,Ningxia,from 2019 to 2020,with six phosphorus treatments in the order of 0(P0),60(P1),120(P2),180(P3),240(P4),and 300 kg/ha(P5).Analysis of the changing patterns of photosynthetic and fluorescence parameters of spring maize leaves and their correlation with yield under different phosphorus fertilizer treatments.In two years,during the big bell mouth stage,leaf area index (LAI) was increased by 4.21% to 12.78% and 4.68% to 15.60% for P3 compared to other treatments,respectively.Phosphorus fertilizer at 180 kg/ha was most effective in promoting leaf area index and photosynthetic potential(LAD) of maize.LAD was significantly increased by 14.42% under P3 treatment compared to no phosphorus fertilizer treatment during the full two year period.The photosynthetic characteristics of maize responded differently to the intensity of phosphorus application,and as the intensity of phosphorus application increased,the net photosynthetic rate (Pn) all reached the maximum value after the stamen pumping stage,and at the R1 stage of the 2 years,the Pn was significantly increased by 10.68% under the P3 treatment as compared to the no-phosphorus-fertilizer treatment.The coefficient of determination (R2) was 0.926 5,0.889 9,and 0.832 0,respectively.Phosphorus application increased the maize photosystem Ⅱ composite performance index (PI),which had its maximum peak at the R1 stage in 2 years,and PI increased by 1.12% to 8.50% and 8.47% to 15.40% under the P3 treatment compared with the other treatments,respectively.The maximum yield was obtained at 180 kg/ha of phosphorus application,which was 17.27% higher as compared to no phosphorus treatment.Based on the analysis of the yield fitting equation,it was shown that the maximum corn yield of 13 823.84 kg/ha was reached at 179.34 kg/ha of phosphorus applied.Pearson's correlation analysis showed that appropriate leaf area index significantly affected maize yield in the late stage,and the photosynthetic parameters all had highly significant effects on maize yield completion;principal component analysis showed that the P3 treatment had the highest composite score for the optimization effect on maize yield.Reasonable transportation of phosphorus fertilizer can effectively ensure higher SPAD value,PSⅡ reaction center activity,improve the capture and utilization of light energy in spring maize,and promote photosynthesis,so as to improve the yield and economic benefits of maize.
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To compare the effects of different nitrogen enhanced fertilizer on apple growth and to identify suitable nitrogen enhanced fertilizer and application methods for apple cultivation,this study examined the impact of four different types of nitrogen enhanced fertilizer on apple growth,yield,quality,and soil nitrogen supply capacity.The experiment consisted of seven treatments:no nitrogen fertilizer(CK),regular application of ordinary urea by farmers(U),coated urea mixed with ordinary urea 3∶7(CU1),basal application of coated urea and follow-up application of ordinary urea(CU2),loss-control urea(KSU),stabilizing urea(WDU),and humic acid stabilizing urea(FZU).A field experiment was conducted in Qixia City,Shandong Province,to analyze the effects of different fertilization treatments on the growth,yield,and quality of 6-year-old Golden Crown apple trees,as well as nitrogen utilization and soil nitrogen supply capacity.The results demonstrated that the application of diverse synergistic nitrogen fertilizers could markedly enhance spring and autumn growth,elevate leaf SPAD,and enhance apple yield and quality.Among these,the yield of the FZU treatment exhibited a significant increase of 8.89% in comparison to that of the control.The Vc content of the fruits of the CU1 treatment was found to be significantly increased by 66.73% in comparison to the U treatment.Furthermore,the CU1 treatment demonstrated the most pronounced impact on the improvement of growth in both spring and autumn.Nitrogen enhanced fertilizer had been demonstrated to significantly enhance nitrogen accumulation in new shoots and fruit nitrogen accumulation,nitrogen fertilizer agronomic efficiency,nitrogen fertilizer bias productivity and nitrogen utilization in the middle and late stages of apple fertility.Of these,the FZU treatment had been observed to exert the most pronounced effect.The nitrogen accumulation of new shoots in FZU treatment was significantly increased by 37.25%,15.91% and 37.85% compared with that in U treatment at bud differentiation,fruiting and ripening stages,respectively.The CU2 treatment was found to have the most beneficial effect on nitrogen utilization.Nitrogen enhanced fertilizer treatments demonstrated the capacity to significantly enhance the nitrate and ammonium content of soil.Among these treatments,the FZU treatment exhibited the most pronounced effect.The FZU treatment was observed to enhance soil urease activity during the flowering and bud differentiation stages.Additionally,the WDU and FZU treatments were found to significantly reduce the nitrification of soil ammonium and nitrogen loss.The -N of the FZU treatment was concentrated at a depth of 20—60 cm soil,which was consistent with the distribution of apple roots in the soil and reduced the risk of -N leaching.A comprehensive analysis demonstrated that the FZU treatment exhibited clear advantages in promoting apple growth,increasing apple yield and quality,and enhancing soil nutrient supply capacity.Consequently,the FZU treatment was identified as the optimal treatment.
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Improper application of soybean fertilizer can lead to late ripening,lodging,worsening of diseases and pests,and a decrease in fertilizer and yield benefits,to improve fertilizer efficiency and fully tap into the potential of soybean yield and quality,therefore,this article investigates the effects of seven fertilization modes F1(basic application of 3 000 kg/ha organic fertilizer+300 kg/ha chemical fertilizer),F2(basic application of 3 000 kg/ha organic fertilizer+150 kg/ha chemical fertilizer+topdressing 150 kg/ha chemical fertilizer),F3(basic application of 3 000 kg/ha organic fertilizer+225 kg/ha chemical fertilizer+topdressing 75 kg/ha chemical fertilizer),F4(basic application of 1 500 kg/ha organic fertilizer+75 kg/ha chemical fertilizer+topdressing 225 kg/ha chemical fertilizer),F5(basic application of 300 kg/ha chemical fertilizer),F6(basic application of 150 kg/ha chemical fertilizer+topdressing 150 kg/ha chemical fertilizer)and F7(basic application of 225 kg/ha chemical fertilizer+topdressing 75 kg/ha chemical fertilizer)on soybean growth,development,photosynthesis,yield,and quality through field experiments using a single factor experimental design.The results showed that increasing the application of organic fertilizer significantly enhanced the dry weight of soybean roots when compared to applying chemical fertilizer alone,applying chemical fertilizer alone with a base to top dressing ratio of 3∶1 was the most beneficial for increasing root dry weight.The combination of organic fertilizer and chemical fertilizer resulted in higher fresh leaf weight,dry leaf weight,fresh stem weight,dry stem weight per plant,as well as higher fresh weight and dry weight per plant when compared to single application of chemical fertilizer,and there were no significant differences in the above indicators among the treatments treated with single application of chemical fertilizer.The combination of organic fertilizer and chemical fertilizer increased the number of leaves per plant and leaf area index of soybean at pod bearing stage and filling stage,under the condition of no organic fertilizer application,the base to top dressing ratio of 3∶1 of chemical fertilizer was beneficial for improving the number of leaves and leaf area index of soybean.Compared to single application of chemical fertilizer,combined application of organic fertilizer and chemical fertilizer increased the photosynthetic rate at flowering and filling stages by 16.69%—21.66% and 14.99%—30.66%,respectively,unreasonable base to top dressing ratio of chemical fertilizer or reducing organic fertilizer dosage could affect the chlorophyll content and photosynthetic rate of soybean in the middle and later stages of growth and development.Increasing organic fertilizer application amount could increase soybean yield and protein content,while reduce fat content,applying chemical fertilizer alone with a base to top dressing ratio of 3∶1 was the most conducive to increasing yield.Therefore,it can be concluded that the combination of organic fertilizer and chemical fertilizer is significantly better than that of single application of chemical fertilizer in improving soybean growth and development,photosynthesis,yield,and quality,in addition,single application of chemical fertilizer performs better with a base to top dressing ratio of 3∶1.
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This study investigates the influence of high and low nodule density on the composition and functionality of the soybean rhizosphere microbiome.The study used the wild-type soybean Williams 82 and a high-nodulation mutant(HiN)derived from this line to assess the impact of nodule variation on associated microbial populations.A controlled pot experiment was conducted,comparing the growth of both genotypes at the flowering stage,followed by the collection and high-throughput sequencing of 16S rDNA from the rhizosphere microbes.The high-nodulation mutant exhibited significant increases in plant height,aboveground biomass,and nodule count,with respective enhancements of 41.22%,37.46%,and 119.23% over the wild type.Comparative analyses revealed distinct differences in microbial community structure and composition between HiN and WT,with an elevated relative abundance of Bacteroidetes in the HiN rhizosphere.At the genus level,several key genera,including Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium,Bradyrhizobium,Chryseobacterium,Microbacterium,and Nocardioides,were more abundant in HiN.LEfSe analysis highlighted the enrichment of Actinobacteria and Bacteroidetes in HiN,contrasting with the predominance of Firmicutes in WT.GUS-labeled Bradyrhizobium further substantiated the enhanced capacity of HiN to enrich Bradyrhizobium.Collectively,these findings elucidate the regulatory role of nodule number on the symbiotic interactions within the soybean rhizosphere microbiome.
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In order to clarify the role of the Zn(Ⅱ)2Cys6 transcription factor gene VDAG_ 04814 in the growth, development and pathogenicity of the Verticillium dahliae. It constructed a VDAG_04814 gene knockout mutant using homologous recombination mediated by polyethylene glycol. Wild-type and mutant strains were inoculated separately onto PDA media supplemented with hydrogen peroxide, sodium chloride, potassium chloride, sorbitol, sodium dodecyl sulfate, Congo red, as well as onto media overlaid with sterile cellophane, to analyze their levels of resistance to oxidative stress, salt stress, osmotic stress, stress on cell wall and plasma membrane integrity, and strain penetration ability. Their pathogenicity was assayed, and the fungal biomass in potato plants was detected. After hygromycin selection and PCR validation, the correct knockout transformants were able to amplify DNA bands of 1 500 bp upstream and downstream, respectively, as well as the full-length 4 500 bp knockout fragment sequence. The results demonstrated that the growth rate and melanin formation ability of the ΔVDAG_04814 mutants were significantly reduced. On media subjected to oxidative stress and salt stress with the addition of hydrogen peroxide, sodium chloride, and potassium chloride, ΔVDAG_04814 showed a higher inhibition rate compared to the wild-type. On osmotic stress media with sorbitol, the growth inhibition rate of ΔVDAG_04814 was significantly lower than the wild type. No growth inhibition was observed for ΔVDAG_04814 on media subjected to cell wall and membrane integrity stress with the addition of sodium dodecyl sulfate and Congo red. On media overlaid with sterile cellophane, no colonies grew for ΔVDAG_04814, whereas the wild-type strain produced normal colonies. Pathogenicity tests indicated that the wilting index of ΔVDAG_04814 was significantly reduced compared to the wild-type, with wilting index ranging from 47.22 to 55.56. It has been demonstrated that VDAG_04814 can regulate the growth, development, stress resistance, penetration ability and pathogenicity of V. dahliae towards potato. This study provides a new target for the control of potato Verticillium wilt disease.
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Investigating the predominant genotypes of Bovine viral diarrhea virus(BVDV)infecting cattle in Tongliao,Inner Mongolia,to provide reference for the BVDV epidemiology and prevention and control.In the preliminary phase of the experiment,fecal samples from diarrheic calves were collected at a cattle farm in Tongliao,Inner Mongolia.These samples were tested using PCR to detect BVDV positivity.Positive fecal samples were then inoculated into madin-darby bovine kidney cells(MDBK)for isolation.The isolated strains were identified using RT-PCR and indirect immunofluorescence staining.Subsequently,the full-length genome of the isolates was sequenced,followed by genetic evolution analysis and genotype determination based on sequences of the 5'UTR,Npro,and E2 genes.The results indicated that this experiment successfully isolated a strain of BVDV,designated as NM-21.Inoculation of NM-21 into MDBK did not induce cytopathic effects,indicating it was a non-cytopathic strain(NCP).Both RT-PCR and indirect immunofluorescence staining confirmed its positivity,with a virus titer of 10-3 TCID50/mL.Based on the full-length genomic sequence,and homology and genetic evolution analysis of the 5'UTR,Npro,and E2 gene sequences,the isolate NM-21 showed the highest nucleotide homology with the BVDV-1c subtype strain NM2103(GenBank accession number ON337882.1)from Inner Mongolia,China.
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To study the structure and function of phosphotyrosine interaction domain 1 (PID1) gene in yak,and to explore its expression in various tissues.The CDS region of Sangsang yak PID1 gene was cloned using Sangsang yak adipose tissue cDNA as template,and the sequence was analyzed bioinformatically.Meanwhile,the relative expression level of PID1 gene was detected in seven tissues of yak namely heart,liver,spleen,lung,kidney,longissimus dorsi muscle and adipose tissue by Real-time Fluorescence Quantitative PCR(RT-qPCR).The results showed that the PID1 gene in yaks had a coding region length of 654 bp,which encoded 217 amino acids.Homology comparison showed that yak and wild yak were closely related,and the similarity reached 100%.The molecular weight of yak PID1 protein was about 24.84 ku and the theoretical isoelectric point was 6.30.According to the calculation results of instability coefficient,the instability of the protein was high (47.96),and it belonged to an unstable protein.The protein had one N-glycosylation site and 23 phosphorylation sites,with no signal peptide or transmembrane structure.The results of RT-qPCR showed that the expression of PID1 gene could be detected in all tissues,with the highest expression in the lung.The yak PID1 gene was cloned and its protein structure was analyzed.The expression of PID1 gene in yak tissue was also studied.Further study on the role of PID1 gene in yak fat deposition provided preliminary data.
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The highly contagious gastrointestinal infectious disease caused by Porcine epidemic diarrhea virus(PEDV)has led to significant economic losses in China's pig industry.It aimed to establish a basis for PEDV antibody detection methods and functional research of the N protein through the screening of specific nanobodies(Nbs)using phage display technology.Peripheral blood lymphocytes were isolated from a camel immunized with the N protein,and total RNA was extracted and reverse transcribed into cDNA.The variable domain of the heavy chain of heavy chain antibodies(VHH)was amplified by PCR,subcloned into the pCANTAB5E-ccdb vector,and electroporated into ER2738 competent cells to construct the VHH phage antibody display library.Subsequently,the library was subjected to four rounds of panning against the PEDV N protein,and positive phage clones were cloned into the pET-30a vector.The binding affinity and specificity of the Nbs were determined by indirect ELISA and Western Blot.The results showed that after the fifth immunization,the antibody titer reached 1∶25 600.The constructed phage display library had a capacity of 4.72×108 and an abundance of 4.3×1010 cfu/mL,with a 93.75% positive rate.After four rounds of screening,16 Nb clones with different amino acid sequences were obtained,and Nb45 was validated to possess excellent specificity and binding ability to the PEDV N protein.This study successfully screened and obtained specific N protein-targeting Nbs,providing biological materials for the establishment of PEDV detection methods and foundational research.
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The aim of this study is to analyze the structure and biological function of ORF2 protein of Avian hepatitis E virus (aHEV)CaHEV-GDSZ01 strain,and the recombinant ORF2 protein was expressed in E.coli prokaryotic expression system and polyclonal antibodies against ORF2 protein were prepared to verify the immunogenicity and reactivity of the recombinant protein.The physicochemical properties,structure and function of ORF2 proteins of CaHEV-GDSZ01 strain were analyzed by bioinformatics software.The prokaryotic expression plasmids of pET-32a-ORF2-His and pET-32a-ORF2 were constructed by digesting and ligating pET-32a(+)vector with ORF2 gene of CaHEV GDSZ01 strain,and then transformed into DE3 competent cells for induction and expression,and the expression form was analyzed.The recombinant ORF2 protein was purified by gel cutting,its reactivity was verified by Western Blot test,and then rabbits were immunized to obtain rabbit anti-ORF2 protein polyclonal antibody.The polyclonal antibody was identified and analyzed by Western Blot,and the antibody titer was detected by indirect ELISA.The results of bioinformatics analysis showed that the ORF2 protein of CaHEV-GDSZ01 strain was hydrophilic and unstable,and the probability of the existence of signal peptide was as high as 93.59%.The secondary structure of the protein was mainly irregular curl and had the structural conditions for binding antibodies.And the overall prediction score of protective antigen of ORF2 protein showed that it had good immunogenicity.ORF2 gene prokaryotic expression plasmids was successfully constructed,and the target protein was successfully expressed.SDS-PAGE showed that the protein was expressed as insoluble protein,and the protein was proved to have good reactivity by Western Blot.The titer of the prepared polyclonal antibody was 1∶102 400,and it could specifically recognize immunogen ORF2 protein,non-immunogen ORF2-His protein and truncated expressed sORF2 protein.The physicochemical properties and some biological functions of ORF2 protein was compared and analyzed,the related proteins were expressed successfully,and the rabbit polyclonal antibody against ORF2 protein was prepared.
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Bimonthly, Started in 1962
CN 13-1101/S
ISSN 1000-7091
CODEN: HHUOA6
Responsible Institution: Hebei Academy of Agriculture and Forestry Sciences
Sponsored by: the Academy of Agricultural Sciences and Agricultural Association of Hebei, Beijing, Tianjin, Shanxi, Henan and Inner Mongolia.
Editor-in-chief: Qiang Zhang
Edited and Published by: Editorial Department of Acta Agriculturae Boreali-Sinica
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