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    Crop Genetics & Breeding·Germplasm Resources·Biotechnology

  • CAO Liru, MA Chenchen, PANG Yunyun, YE Feiyu, WANG Zhenhua, LU Xiaomin
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    bZIP transcription factors are widely found in plants and play an important role in regulating plant growth and development and abiotic stress response.In order to explore the functional role of bZIP transcription factor in maize drought stress response,transcriptome sequencing technology was used to analyze the expression changes of transcription factors in maize seedlings treated with drought stress for 5 days and rehydration for 3 days,and a bZIP transcription factor(ZmbZIP26)was screened from transcriptome data in response to drought and rewatering treatment.Co-expression network analysis revealed that ZmbZIP26 was at the core node of network regulation.The gene contained a 558 bp open reading frame encoding 185 amino acids,which was a hydrophilic protein.Phylogenetic tree and conserved sequence analysis showed that ZmbZIP26 protein had high homology with homologous proteins of sorghum and Miscanthus,and also had the same conserved motifs at the same amino acid positions.Cis-element analysis showed that the upstream 2 000 bp region of the ATG site contained drought response elements,hormone response elements and light response elements.qRT-PCR analysis showed that ZmbZIP26 was a constitutively expressed gene,which was highly expressed in young stems,female panicles and roots.ZmbZIP26 positively responded to drought,high temperature,high salt and nitrogen stress and the process of restoring,which might play an important role in the process of plant resistance.Subcellular localization analysis revealed that ZmbZIP26 was a nuclear protein localized in the nucleus.Protein interaction prediction showed that ZmbZIP26 might interact with zinc finger protein,serine protein,Ca-dependent protein and glutathione transfer protein to construct a regulatory network,which cooperatively regulated maize growth and development and stress response process.

  • TANG Bin, GENG Cunjuan, ZENG Qiang, GUO Huanle, LI Han, CAO Zhongyang, DENG Lichao, PENG Ming, ZHOU Hong, CHEN Zhihui
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    Maize grain development stage is very sensitive to high temperature stress,which seriously affects the yield and quality of maize.To study the difference of gene expression in grain and analyze the gene molecular mechanism of different high-temperature tolerant maize inbred lines in response to high-temperature stress,RNA-Seq technology was used to analysis the differentially expressed genes(DEGs)of XN202(heat tolerant)and CT110(heat susceptible)in maize grains 15 days after pollination under normal and high temperature stress.Compared with the control,XN202 and CT110 detected 1 517 and 1 012 DEGs,respectively,with 142 common DEGs,including 7 transcription factors.There were significant differences in grain response to high temperature stress among different heat tolerant inbred lines.DNA replication,nucleosome,minichromosome maintenance complex,alpha DNA polymerase:primase complex,nutrient reservoir activity,alanine,aspartate and glutamate metabolism were involved in response to high temperature stress through gene ontology(GO)function annotation and kyoto encyclopedia of genes and genomes(KEGG)signal pathway enrichment analysis.According to bioinformatics analysis,a total of 374 DEGs were located in the reported QTL interval of heat tolerance related traits in maize,and 42 DEGs were putative heat tolerance candidate genes.In summary,maize grains could form complex cellular protection and defense system under high temperature stress,and DEGs related to high temperature tolerance,such as AP2/ERF,MYB,bHLH,NAC,HSF and HSP,might play an important role in the molecular regulatory network.

  • WANG Kexin, GUO Zhaoyang, YIN Yuhang, CHEN Shengzhong, SONG Xiyun, ZHAO Meiai
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    The glutasidyl thiotransferase gene(GST)can be involved in the process of plants responding to abiotic stresses.In order to understand the ability of ZmGST in response to salt and drought stress in maize,the ZmGST gene was obtained by RT-PCR cloning using the inbred line CA66 of maize as the material,and the gene was bioinformatically analyzed,and the relative expression of the gene in different tissues of maize and simulated drought and salt stress was analyzed by Real-time fluorescence,and the prokaryotic expression vector was constructed and treated with drought and salt.The results showed that the CDS sequence of ZmGST gene was 384 bp in length and encoded 127 amino acids,and the bioinformatics analysis showed that the gene belonged to the Tau family,was a hydrophilic unstable protein,did not have a transmembrane structure,had 12 phosphorylation modification sites,found no signal peptide,was a non-secreted protein,and subcellular localization predicted that ZmGST was located in the nucleus and cytoplasm.The homologous sequence results showed that ZmGST was the closest related to Sorghum bicolor,and had the highest similarity with ZmGSTU14 gene.The promoter analysis results were found to contain TC-rich repeats and other elements,which are involved in defense and stress response.Real-time fluorescence quantitative results showed that the expression of ZmGST gene was the highest in maize roots and was upregulated by salt stress,which reached the highest level after 24 h of treatment.The results of salt and drought stress at the prokaryotic level showed that the recombinant plasmid pET28a-ZmGST could grow normally in medium with different salt concentrations,and the simulated drought was inhibited.It is speculated that ZmGST gene plays an important role in maize response to salt stress.

  • XIANG Guili, WU Rina, YAMAMOTO Naoki, WU Yichao, JIANG Jin, LIAO Mingli, WEI Shuhong, PENG Zhengsong, YANG Zaijun
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    PEPC catalyzes phosphoenolpyruvate(PEP)to generate oxaloacetate(OAA)to participate the tricarboxylic acid(TCA),which plays an important role in development and stress adaptation of plant.However,there have been no reports of PEPC involvement in plant organ development.Exploration and study on biological function of Tappc3A gene in wheat flower development,and provides new clues to explore the molecular mechanism of homologous conversion of stamens into pistillody in wheat.The Tappc3A gene was cloned from CM28TP and HTS-1 by PCR,and the sequence and phylogenetic tree were analyzed by bioinformatics tools,and the expression level of Tappc3A gene in different developmental stages and different reproductive organs of wheat young spikes was analyzed by using qRT-PCR,and whether the protein function encoded by Tappc3A gene was analyzed by prokaryotic expression.The wheat RNA-Seq database was used to analyze the co-expression of Tappc3A gene and other genes that regulate flower organ development.The ORF of Tappc3A gene was 2 901 bp in length,encoding 966 amino acid residues,with a typical phosphoenolpyruvate carboxylase(PEPase)conserved domain,a conserved serine(Ser,S)reversible phosphorylation site(SIDAQLR)at the N-terminal,a plant-type PEPC protein signature sequence(QNTG)at the C-terminal,and the 774th amino acid was the typical alanine of C3 plant PEPC.Cluster analysis also showed that Tappc3A belonged to the C3 type PEPC family.qRT-PCR analysis,in three stages of wheat young spike development,showed that the expression of Tappc3A gene in HTS-1 was higher than that in CM28TP at the dichotomous stage to the florescence differentiation stage and the pharmacophore period,and the expression of Tappc3A gene was significantly higher in pistils(P)and pistillody stamens(PS)than stamens(S).The prokaryotic expression results showed that the protein encoded by Tappc3A gene,which catalyzes the production of OAA from PEP,and its activity was significantly enhanced after IPTG induction.The gene co-expression analysis showed that Tappc3A gene might be involved in the morphogenesis of wheat floral organs.Tappc3A gene might be involved in wheat pistil development,and its overexpression in stamens might be associated with the homologous transformation of stamens into pistillody trait formation.

  • NIE Lizhen, ZHANG Zhicheng, XIE Rui, CHANG Yue, ZHANG Qionglin, HAN Ping'an, YI Jing
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    Anthocyanins are a kind of important flavonoids produced during the secondary metabolism of plants.Anthocyanidin synthase(ANS)catalyzes the conversion of leucoanthocyanidins to anthocyanidins in the anthocyanin synthesis pathway.Previous studies have shown that the anthocyanidin synthase gene plays an important role during the coloring process of plant organs.To investigate whether the gene can promote anthocyanin synthesis in potato tubers,using the red-skinned and red-fleshed colored potato variety Hongmei as the material,anthocyanidin synthasegene was isolated from tubers of color potato by RT-PCR, the characterization analyzed by bioinformatics and function validated by overexpression into Arabidopsis thaliana.The results showed that sequence 1 406 bp cDNA of the gene was amplified from the potato,including 1 368 bp open reading frame(ORF).The ORF encoded a deduced protein of 455 amino acids.The gene was named as StANS1a by bioinformatics analysis.GenBank accession number is ON512347.The amino acid sequence analysis revealed that the deduced protein contained a DIOX_N'domain located at 52—166 resides and a 2OG_FeⅡ_Oxy domain located at 215—312 resides.Arabidopsis thaliana plants was transformed with constructs overexpression vector StANS1a.The transcriptional expression of StANS1a in transgenic lines were tested by RT-PCR.The results showed that in the same population,the transcriptional expression of StANS1a showed an increase in different transgenic lines compared with the wild-type(Wt).The anthocyanin content was further analyzed in transgenic lines.Compared to the Wt,the anthocyanin contents of the seven lines(T1—T7)were increased from 2.17% to 54.61%. These results strongly demonstrated that overexpression of the StANS1a gene could promote anthocyanin synthesis in transgenic plants.The colored potato StANS1a gene plays an important role in the anthocyanin metabolic pathway.

  • GAO Luyao, CAO Jiajian, WANG Chunhua, WU Tao, DU Yalin
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    GDSL is an important gene affecting the development of cuticle.This study cloned cucumber GDSL lipase gene and analyzed its expression pattern,in order to lay a foundation for the study of cuticle development and the glassiness of cucumber.The gene sequence was cloned to clarify its role in cucumbers with different glossiness by using six cultivars with different glossiness as materials.The promoter of CsGDSL lipase gene was cloned to analyze its functional elements.According to the reference sequence in cucumber genome database,we cloned the GDSL lipase gene of cucumber and analyzed by bio-informatics.The expression of the gene in different tissue parts of cucumber was confirmed by qRT-PCR technique.The CDS of CsGDSL length 1 059 bp which encoded 352 amino acids,and the secondary structure was mainly random curl(45.45%)and α-helix(33.24%).This gene was conservative in the process of evolution and was most closely related to CmGDSL.The expression level was the highest in the male flower of cucumber on the flowering day,and the expression level in the ovary at 3 days after flowering was higher than that at 0 days after flowering.The CDS sequence of CsGDSL gene was conservative in six cultivars. CsGDSL gene was responsive to stress,hormone and light.We obtained the CsGDSL lipase gene in this study,and identified its expression in different tissue parts of cucumber,the gene is relatively conserved in different cultivars,suggesting that CsGDSL may affect cucumber glossiness.

  • ZHOU Qianyi, HUANG Sijie, TIAN Jie
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    Neutral/alkaline invertases,as important substances in plant sucrose metabolism,are mainly involved in processes such as plant growth and development,and response to adversity stress.To investigate the response pattern of AsNI to stress,this study cloned two garlic neutral/alkaline invertase genes,using Ledu purple skin garlic as the test material and they were subjected to bioinformatics and expression characterization.The results showed that the open reading frames of AsNI1 and AsNI2 were 522,1 203 bp,encoded 173,400 amino acids.AsNI1 and AsNI2 were both hydrophilic proteins predicted to be localized in the cytoplasm with a Glyco_hydro_100 structural domain.However,the amino acid sequence similarity between the two was only 25.75%,and AsNI2 contained one glycosylation site,while no glycosylation site was detected in AsNI1,and the two were distantly related.The analysis of the protein interaction network showed that AsNI2 and AsNI1 might participate in different biochemical processes.The promoter sequence analysis revealed that the promoter regions of AsNI1 and AsNI2 contained multiple cis-acting elements related to stress response,with the AsNI2 promoter having a significantly larger number of drought and low temperature stress response elements than AsNI1.The prediction of promoter transcription factor binding sites showed that they contained different kinds and numbers of binding sites,indicated that AsNI1 and AsNI2 could perform different gene functions.The qRT-PCR assay revealed that the expression of AsNI was significantly tissue-specific,with the highest expression of AsNI1 and AsNI2 in the roots and bulbs,respectively.Meanwhile,adversity stress was able to induce AsNI expression,and the response of AsNI2 was stronger than that of AsNI1 under both low temperature and drought treatments.Among them,low temperature stress mainly induced the expression of AsNI2 in leaves,and drought stress mainly induced the expression of AsNI2 in roots.The sequence characteristics and expression pattern of AsNI were analyzed to verify the stress resistance function of AsNI.

  • CHEN Yang, WANG Qi, GAO Yansong, YOU Xue, XIONG Yi, JIN Yifeng
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    In order to further study the molecular mechanism of CIPK gene family in response to abiotic stress and explore the role of CIPK32 gene in Poa pratensis L. under abiotic stress. The CIPK32 gene was cloned from Poa pratensis L. midnight Ⅱ by RT-PCR, and its expression in different tissues and under different abiotic stress conditions was analyzed. On-line softwares such as NCBI-CDD and SWISS-MODEL were used to analyze the protein structure, homology alignment and other bioinformatics. The results showed that the sequence ORF of CIPK32 gene of Poa pratensis L. was 1 320 bp, encoding 439 amino acids. It is predicted that the relative molecular weight of CIPK32 protein was 50.28 ku, the isoelectric point was 6.82, and the protein molecular formula was C2249H3574N608O665S16, which belonged to unstable hydrophilic protein. CIPK32 of Poa pratensis L. contained typical STKc_SnRK3 and CIPK_C domains, and had high homology with amino acids encoded by Aegilops tauschii (XP_020198391.1), Lolium rigidum(XP_047091407.1) and Hordeum vulgare(XP_044980943.1). It also contained many binding sites such as Amidation site, N-glycosylation site, N-myristoylation site and so on. The expression level of CIPK32 gene in Poa pratensis L. was tissue-specific, and the order was leaf>stem>root. Under drought stress, the gene was up-regulated first and then decreased, and the highest value was reached when 10% PEG6000 was treated for 16 h, which was 6.2 times that of 0 h; with the increase of NaCl concentration, the expression of the gene was significantly promoted, and 200 mmol/L NaCl was 6.9 times that of 0 mmol/L NaCl. The expression of CIPK32 gene could be promoted by low concentration of NaNO3 and low concentration of KH2PO4(0.1 mmol/L KH2PO4). To sum up, the CIPK32 gene of Poa pratensis L. has tissue specificity, and the expression of CIPK32 gene can be induced by drought, salt, nitrogen and phosphorus stress. CIPK gene family plays a potential role in abiotic stress.

  • LIU Hui, XU Wenjing, YANG Shuo, ZHANG Wei, ZHANG Hongmei, LIU Xiaoqing, ZHU Yuelin, CHEN Huatao
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    In order to study the organic acid synthesis mechanism of vegetable soybean and provide a theoretical basis for the quality improvement of vegetable soybean,a natural population containing 264 accessions of vegetable soybean was used as experimental materials.The contents of tartaric acid,malic acid and citric acid in this population were determined by high performance liquid chromatography(HPLC)in 2020 and 2021,respectively,and genome-wide association study was conducted based on the genotype data of this population,SNP loci and candidate genes significantly associated with organic acid content were identified.The results showed that the average content of tartaric acid in 2020 and 2021 populations were 4.13,4.16 mg/g,respectively;the average content of malic acid was 7.26,8.99 mg/g,respectively;the average content of citric acid was 7.12,10.88 mg/g,respectively.Correlation analysis showed that citric acid content of 264 materials showed a significant positive correlation between 2020 and 2021.Significant positive correlation coefficients were obtained for the pairs of malic/citric acids,tartaric/malic acids,and tartaric/citric acids were 0.790*,0.695*,0.739*,respectively.The results showed that there were significant differences among different varieties in the test population,with rich genetic diversity.Six specific germplasm resources with high organic acid contents were selected to provide excellent materials for the improvement and breeding of organic acid varieties of vegetable soybean.Genome-wide association study based on the mixed linear model the number of SNP loci significantly correlated with the contents of tartaric acid,malic acid and citric acid were 54,189,43,respectively,and based on the gene functional annotation information,3 and 5 candidate genes were identified which were significantly correlated with the content of tartaric acid and malic acid,respectively.

  • Tillage & Cultivation·Physiology & Biochemistry

  • WANG Meiling, JIANG Wenyue, GE Yuyang, ZHU Xinkai, LI Chunyan, ZHU Min, GUO Wenshan, DING Jinfeng
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    To provide references for the waterlogging-tolerance and yield-stability cultivation of wheat,the experiment was conducted to study the effects of different degrees of waterlogging on root dry weight and vigor in different soil layers,shoot growth,grain yield,and its components using Yangmai 25 and Ningmai 13 as experimental materials.The treatments included short-term mild(SL,10 cm water layer under the soil surface for 3 days),short-term severe(SS,2 cm water layer above the soil surface for 3 days),long-term mild(LL,10 cm water layer under the soil surface for 12 days),long-term severe(LS,2 cm water layer above the soil surface for 12 days)waterlogging treatments at stem-elongation stage,and control treatment(CK,maintaining the relative soil water content of 70%—75%).The results showed that Yangmai 25 showed significantly higher grain yield,1000-grain weight,root dry weight,root activity in the 0—40 cm soil layer,and ratio of root to shoot than Ningmai 13.Compared with CK,SL and SS reduced grain yield by 13.44%—22.45%,while LL and LS reduced grain yield significantly by 28.76%—37.26%.There was no significant difference in grain yield between SL and SS,LL and LS.Under the short-term waterlogging,the biomass of root and shoot were slightly reduced,the root in 0—20 cm soil layer maintained high vigor,and the vigor of root in 20—60 cm soil layer could recover.However,long-term waterlogging significantly decreased root dry weight resulting in growth imbalance of root and shoot.In addition,root activity was low and difficult to recover under long-term waterlogging,and the top-three leaves was prone to premature senescence with the top 3rd leaf showing most obvious.The results suggested that lowering water levels as soon as possible after waterlogging could help the surface roots maintain growth and physiological activity,and reduced the risk of premature leaf senescence,satisfying the photosynthetic needs for grain filling.

  • HE Miaoling, WANG Zhilan, DU Xiaofen, HAN Kangni, LIAN Shichao, LI Yuxin, CHENG Kai, LI Yanfang, WANG Jun
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    Ideal plant architecture can significantly improve crop yield,but the relationship between plant architecture traits and yield traits is still unclear in foxtail millet.To provide a theoretical basis and germplasm resource for plant architecture breeding in foxtail millet,10 plant architectures traits(plant height,main stem length,panicle length,panicle neck length,panicle diameter,tiller number,node number,length of flag leaf,width of flag leaf and area of flag leaf)and 3 yield traits(panicle weight,panicle grain weight and 1000-grain weight)were analyzed via 126 F6 recombinant inbred lines(RIL)from a F1 arrived from a cross between Aininghuang and Jingu 21 under three ecological environment consisting of Changzhi,Yuci and Datong.Based on the best linear unbiased estimation(BLUE),variance analysis,correlation analysis,partial correlation analysis,principal component analysis,multiple regression analysis and cluster analysis were conducted.The phenotype variation analysis showed there were an extremely significant difference between plant height and main stem length in three ecological environments,a significantly or extremely significant difference between panicle neck length and node number in two ecological environments,and the significant or extremely significant differences among panicle length,panicle diameter,panicle weight,panicle grain weight and tiller number in single ecological environment.In RIL population, an abundant variation was observed for 13 traits with the approximately normal frequency distribution,and the variation coefficients ranged from 6.86% to 31.71%.Except for the main stem length in Yuci, other traits showed a transgressive separation in three ecological environments.Correlation analysis and partial correlation analysis indicated that panicle weight and panicle grain weight were extremely significant positive correlated with plant height,main stem length,panicle length,panicle neck length and node number,while they were extremely significant negative correlated with tiller number,panicle weight was significantly positive correlated with flag leaf length.Principal component analysis simplified 13 traits into 4 principal components,and the cumulative contribution rate was up to 93.938%.The fitting degree R2 of multiple regression analysis was 0.614, and main stem length,panicle length and tiller number were the main factors affecting panicle weight.The RIL population was divided into 7 groups via cluster analysis,among which Group Ⅴ consisted of 3 materials with the highest yield, medium plant height and good comprehensive traits,which could be used as elite parents for ideal plant architecture breeding.

  • BAI Xiaorong, MIN Weifang, SHI Yafei, SHE Yangmengfei, TIAN Haotian, LUO Chengke
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    To reveal the mechanism of drought resistance of different resistant rice during germination period,Rice drought-sensitive materials(Calrose,Jingning 10,Shanxing 86)and drought resistance materials(Farry,Songjing 3,Ningjing 36)were studied on the effects of simulated drought stress(15% PEG-6000)on the growth index,physiological indexes and corresponding gene expression of different rice seeds.The results showed that under normal conditions,there were no significant differences in the expression levels of growth indicators and stress-related genes between drought-sensitive and drought-resistant cultivars.However,changes in physiological indicators were shown that there were no significant differences in the activities of superoxide dismutase(SOD) and peroxidase(POD),the contents of soluble sugar(SS) and hydrogen peroxide(H2O2) among different genotypes.The contents of malondialdehyde(MDA) and superoxide anion($\mathrm{O}_{2}^{\bar{.}}$) in the drought-sensitive cultivar Shanxing 86 were significantly higher than those in other materials,and the contents of catalase(CAT),proline(Pro) and soluble protein(SP) of drought resistant Ningjing 36 were significantly higher than those of other materials as well.Under drought stress,the relative germination potential(RGP),relative bud length(RSL),germination drought resistance index(GDRI)and vitality index(VI)of germinating seeds increased by 0.03—0.07 percentage,0.32—0.39 percentage,0.12—0.18 percentage and 92.41%—108.39%,respectively;MDA and reactive oxygen species($\mathrm{O}_{2}^{\bar{.}}$,H2O2) contents in germinating seeds of drought-resistant cultivars decreased by 2.54%—61.64%,19.60%—46.30% and 35.61%—62.02% respectively compared with drought-sensitive cultivars.The contents of osmotic regulating substances(Pro,SS,SP) increased by 5.93%—18.29%,1.08%—7.97% and 3.47%—6.03% respectively.The activities of antioxidant enzymes(SOD,POD, CAT) were increased by 17.29%—33.12%,15.24%—76.06% and 14.68%—18.61% respectively.The relative expression levels of OsP5CS,antioxidant enzyme synthesis genes (OsALM1, OsPOX1, OsCATC) were up-regulated by 2.66%—182.31% and 57.14%—513.27%,0.38%—109.06% and 63.39%—184.25% respectively.Comprehensive analysis showed that drought stress inhibited the germination of rice seeds and affected the physiological characteristics of seeds and the expression of corresponding genes during germination.Under drought stress,vigor index(VI),peroxidase(POD)and peroxidase synthesis gene(OsPOX1)are the key indicators affecting rice seed germination,whether it is drought-resistant or drought-sensitive materials.In addition to the above indicators,soluble protein(SP),proline synthesis gene(OsP5CS)and catalase gene(OsCATC)are other key indicators affecting drought-resistant materials.Relative shoot length(RSL),hydrogen peroxide(H2O2)and superoxide dismutase gene(OsALM1)are other key indicators affecting drought-sensitive materials.

  • LI Xing, MENG Fei, HAO Jiaqi, WANG Cong
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    To investigate the mechanism of chitosan promoting nodulation in vegetable soybean under salt stress,the effects of exogenous chitosan on sucrose metabolism and root accumulation in vegetable soybean under NaCl stress were explored.Effects of chitosan on nodulation,the accumulation of sucrose and reducing sugar in leaves and roots,as well as sucrose metabolism-related enzyme in vegetable soybean under NaCl stress were studied by using the vegetable soybean variety Nippon Green and the fast-growing rhizobia HH103 symbiosis as the test material in an artificial climate chamber.The results showed that the number of root nodules and fresh weight of root nodules in vegetable soybean were significantly decreased under the NaCl stress(T2)(P<0.05),with an average decrease of 36.06% and 29.91% in each stress period,respectively;the sucrose content of leaves and roots was significantly increased(P<0.05),and the reducing sugar content was significantly decreased(P<0.05);and the activities of sucrose synthase(SS)and sucrose phosphate synthase(SPS)were increased,while the activities of neutral convertase(NI)and acid convertase(AI)were decreased.The number of root nodules and fresh weight of root nodules significantly increased(P<0.05),after chitosan treatment(T3)in all stress periods,with average increases of 26.87% and 25.63% of T2;the sucrose and reducing sugar contents and sucrose metabolism-related enzyme activities of leaves and roots increased significantly compared with T2(P<0.05),with average increases of 11.32%,21.32% and 10.22%,11.11%,respectively.That chitosan could promote the nodule formation of vegetable soybean by inducing the high level of sucrose metabolism in the leaves and roots under NaCl stress and accelerating the transfer of sucrose to roots,which may be one of the important reasons for chitosan to improve the nodule formation ability of vegetable soybean under NaCl stress.

  • Resources & Environment·Plant Protection

  • LIN Xiaobing, LIU Kailou, HUANG Shangshu, HE Shaolang, XU Xiaolin, ZHOU Qina, ZHONG Yijun
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    To explore the effects of long-term different fertilization on soil microbial biomass carbon,nitrogen and enzyme activity of double cropping maize in Southern China.Based on a 35 a long-term located experiment in dryland red soil in Jinxian,Jiangxi Province,four treatments including no fertilizer(CK),chemical fertilizer(NPK),chemical fertilizer and fresh pig manure(NPKM),and fresh pig manure(OM).The soil nutrients,microbial biomass carbon,nitrogen and enzyme activities were measured in the 0—20 cm,20—40 cm soil layers at the maturity of double cropping maize and the correlations between them were analyzed.Results showed that long-term fertilization(NPK,NPKM and OM)significantly increased soil microbial biomass carbon,nitrogen and enzyme activity.During the spring maize period,the soil microbial biomass carbon and nitrogen content of fertilization treatment compared with CK increased by 67.05%—159.15% and 3.33%—62.37% in the 0—40 cm soil layer.And the activities of catalase,phosphomonoesterase,urease and sucrase increased by 0.22%—79.71%,9.82%—59.51%,8.73%—82.37%,66.67%—538.89%,respectively.During the autumn maize period,the soil microbial biomass carbon and nitrogen content of fertilization treatment compared with CK increased by 36.30%—136.72% and 17.09%—47.29% in the 0—40 cm soil layer.And the activities of catalase,phosphomonoesterase,urease and sucrase increased by 7.41%—74.55%,22.69%—57.39%,18.85%—58.98% and 51.70%—216.67%,respectively.Among them,the NPKM treatment had the best improvement effect.In general,soil microbial biomass carbon,nitrogen and enzyme activity in 0—20 cm soil layer were higher than in 20—40 cm soil layer,and the soil microbial biomass carbon,nitrogen and enzyme activity in autumn maize period were higher than those in spring maize period.NPKM and OM treatments also significantly increased soil pH,organic carbon,total nitrogen,total phosphorus,total potassium,available phosphorus,available nitrogen and available potassium.After long-term application of organic fertilizer,soil phosphorus accumulation was obvious,while NPK treatment accelerated soil acidification.All treatments could significantly increase maize yield(P<0.05). Compared with CK, it increased by 1.04—15.07 times. In conclusion,organic-inorganic application(NPKM)is the best fertilization measure to enhance soil nutrients,microbial biomass,enzyme activity and yield.

  • LI Shujing, WU Jinzhi, HUANG Ming, WANG Chunping, LI Youjun, WANG Hongtao, ZHAO Wenxin, HUANG Xiuli, LI Wenna, LI Shuang
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    In order to study the differences in yield,accumulation and utilization of nitrogen (N),phosphorus (P) and potassium (K)among wheat varieties from different geographical origins and among high and low yield wheat varieties from the same geographical origin in dryland farming, 165 varieties from China(58),the International Maize and Wheat Improvement Center(CIMMYT,42)and others abroad (65)were planted in the farm of Henan University of Science and Technology in Luoyang,Henan Province, to analyze the yield and its components,the characteristics of accumulation,distribution and requirement at maturity of N,P,K.Compared with others abroad,the grain yield,grains per spike,1000-grain weight,N,P and K physiological efficiency of CIMMYT varieties were significantly increased by 21.3%,39.1%,26.4%,33.2%,22.6% and 36.1%,respectively,while those of Chinese varieties were increased by 10.5%,18.1%,24.3%,29.4%, 12.1% and 24.3%,respectively,however,the spike number,the N,P and K accumulation in stem and leaf,the N,P,K and dry matter requirements of 100 kg grains of CIMMYT and Chinese varieties decreased significantly,moreover,the increase and decrease of CIMMYT varieties were higher than those of Chinese varieties.Compared the low-yield varieties from the same geographical origin,the yield,spike number,grain number per spike and biomass of Chinese high-yield varieties were significantly increased by 122.4%,38.7%,39.3% and 96.1%,respectively;the yield,spike number and biomass of CIMMYT high-yield varieties were significantly increased by 97.6%,68.2% and 71.7%;the yield,spike number,grain number per spike,1000-grain weight and biomass of high-yield varieties of others abroad were significantly increased by 157.2%,33.0%,43.6%,35.9% and 76.4%.For Chinese varieties,compared with the low-yield varieties, the shoot and grain N,P,K accumulations of high-yield varieties were significantly increased by 98.8%,101.0%,83.7% and 118.8%,104.7%,131.2%,as well as the N,P,K uptake efficiencies were significantly increased by 93.6%,84.6% and 70.3%,the nutrient requirements for grain N,P,K content formation were significantly increased by 102.8%,109.5% and 75.9%,but the K and dry matter requirements for 100 kg grains formation were significantly decreased by 16.8% and 11.1% respectively.For CIMMYT varieties,compared with the low-yield varieties, the N,P,K uptake efficiencies in high-yield varieties were significantly increased by 68.5%,71.4% and 63.6%,as well as the nutrient requirements for grain N,P,K content formation were significantly increased by 79.2%,81.7% and 76.5%,while the N and P requirements for 100 kg grains were significantly decreased by 10.7% and 10.3%,respectively.For others abroad,compared with the low-yield varieties, the N,P, K physiological efficiencies in high-yield varieties were increased significantly by 34.7%,30.2% and 60.6%, as well as the nutrient requirements for grain N,P,K content formation were increased significantly by 73.0%,110.8% and 52.1%,while the N,P,K and dry matter requirements for 100 kg grains were significantly decreased by 26.7%,23.6%,36.8% and 24.7%,respectively.To sum up,many characteristics of grain formation and N,P,K absorption and utilization in wheat showed significant differences among different geographical origin varieties.CIMMYT varieties had higher grain number per spike,1000-grain weight,harvest index,grain N accumulation,and P,K physiological efficiency,while others abroad had higher spike numbers,N,P,K accumulation in stem and leaf,N,K accumulation in shoot and uptake efficiency,dry matter and N,P,K requirements for 100 kg grains,and the values of most indexes of Chinese varieties were between CIMMYT varieties and others abroad.Compared with low-yield varieties,high-yield varieties from the same geographical origin had the ability to improve yield components,harvest index and nutrient uptake efficiency,while increased the nutrient requirements for grain N,P,K content formation.Therefore,the N,P,K input should be increased appropriately in the production system which aimed to high yield and high nutrient content.

  • HUANG Shangshu, JIANG Xinfeng, ZHU Tong, LIN Xiaobing, HE Shaolang, WANG Binqiang, WU Yan, LEI Liwen, SUN Yongming
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    In order to clarify the influence mechanism of total replacement of chemical fertilizer with organic fertilizer on the nitrogen fertility of tea garden soil.Based on the 5 a experiment of replacing chemical fertilizer with organic fertilizer in tea garden,single application of chemical fertilizer(CF)and full application of organic fertilizer(OF)were selected as treatments,the effects of total replacement of organic fertilizer with chemical fertilizer on the physical and chemical properties,mechanical stability,aggregate composition,and aggregate inorganic nitrogen distribution characteristics of tea garden soil in rainy and dry seasons were studied,and the influencing factors of aggregate inorganic nitrogen distribution were analyzed.The results showed that:Compared with CF treatment,OF treatment increased the pH value,organic matter content and total nitrogen content of surface soil in rainy and dry seasons.However,there was no significant difference in soil pH between the two treatments in the rainy season(P>0.05);in the rainy season,the cation exchange capacity,ammonium nitrogen content and nitrate nitrogen content in the surface soil of the tea garden under the OF treatment were lower or significantly lower(P<0.05)than those under the CF treatment,while in the dry season,the OF treatment was significantly higher than that under the CF treatment(P<0.05),with an increase of 27.02%, 58.97%, and 266.84%, respectively. Different treatments had significant effects on the mass percentage of 0.25—2.00 mm and<0.25 mm aggregates and GWD;compared with CF treatment, OF treatment increased the mass percentage of 0.25—2.00 mm aggregates and GWD, and decreased the mass percentage of<0.25 mm aggregates; seasons had a significant effect on the mass percentage of aggregates<0.25 mm (P<0.05).The contents of ammonium nitrogen and nitrate nitrogen in aggregates of different treatments were as follows:<0.25 mm aggregates were the highest,0.25—2.00 mm aggregates were the second,and>2.00 mm aggregates were the lowest;in rainy season,the content of ammonium nitrogen and nitrate nitrogen in aggregates at all levels of OF treatment was lower or significantly lower(P<0.05) than that of CF treatment,while in dry season,OF treatment was significantly higher than that of CF treatment(P<0.05);compared with the rainy season,the content of ammonium nitrogen and nitrate nitrogen in aggregates at all levels under CF treatment was significantly reduced in dry season(P<0.05),and the content of ammonium nitrogen and nitrate nitrogen in aggregates at all levels under OF treatment was increased or significantly increased in dry season(P<0.05);under different treatments,the proportion of ammonium nitrogen and nitrate nitrogen storage of aggregates showed that 0.25—2.00 mm aggregates were the highest(51.70% and 51.14%),>2.00 mm aggregates were the second(34.59% and 35.51%),and<0.25 mm aggregates were the lowest(13.71% and 13.34%);compared with CF treatment,OF treatment significantly reduced the proportion of<0.25 mm aggregate ammonium nitrogen storage in rainy and dry season and the proportion of <0.25 mm aggregate nitrate nitrogen storage in dry season(P<0.05),and significantly increased the proportion of 0.25—2.00 mm aggregate ammonium nitrogen storage in rainy season and 0.25—2.00 mm aggregate nitrate nitrogen storage in dry season(P<0.05).Correlation analysis showed that the content of ammonium nitrogen in soil aggregates was mainly affected by soil pH,cation exchange capacity and total nitrogen content,while the content of nitrate nitrogen in soil aggregates in tea garden was mainly affected by soil pH,organic matter content and total nitrogen content;further RDA analysis showed that soil pH and cation exchange capacity were the main factors affecting the distribution of inorganic nitrogen in soil mechanically stable aggregates.To sum up,total replacement of chemical fertilizer with organic fertilizer in tea garden is beneficial to improve the content of soil organic matter,coordinate the supply of soil nitrogen in rainy and dry seasons,and promote the formation of large aggregates and the accumulation of soil inorganic nitrogen.

  • XU Lianzhou, WANG Qi, LIU Danyang, ZHONG Rui, MENG Qingfeng, ZHANG Ruyue, LIU Yang, MA Xianfa, LUO Jingmei, XING Huaming, SONG Bo
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    Based on the long-term improvement experiment of organic fertilizer of soda saline-alkali soil in Songnen Plain,the effects of different improvement years(4,11,15,20 a)on soil colloid components,soil organic carbon components and soil organic-mineral compound degree were studied with no improvement as the control(CK).The results showed that with the increase of improvement years,the colloidal content of soil water dispersive group(G0)decreased significantly(P<0.05),while that of soil calcium-binding complex(G1)increased significantly(P<0.05);there were no significant changes in colloid content of soil iron and aluminum oxide binding complex(G2)and(G0+G1+G2)content of different treatments;the organic carbon content in G0 group,G1 group and G2 group showed an increasing trend.Organic carbon content and heavy fraction organic carbon content of all organic fertilizer treatments were significantly increased compared with CK(P<0.05).In all treatments,organic carbon content in 4 a treatment was the highest,and heavy fraction organic carbon content in 11 a treatment was the highest.The total contribution rate of soil carbon sequestration of the treatments complexes with improved years of 11 a or more was 35.51%—54.64%.Compared with CK,organic-mineral compound degree of soda saline-alkali soil treated with organic fertilizer increased to varying degrees,and the increase was obvious in the treatment of 11 a and above.In conclusion,the long-term application of organic fertilizer promoted the transformation of water dispersive colloid of soda saline-alkali soil to water stable complex,significantly increased the contribution rate of the complex to soil carbon fixation,and significantly improved the degree of organic-mineral composition of the soil.

  • WANG Ben, REN Kaiming, MA Shangyu, FAN Yonghui, ZHANG Wenjing, HUANG Zhenglai
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    In order to select a reasonable fertilization pattern suitable for high-yielding and high-quality production of rice stubble weakly glutenous wheat,this experiment was conducted with weakly glutenous wheat Ningmai 13 and Wanximai 0638 as the test varieties,with no fertilization(CK),conventional fertilization of compound fertilizer+urea(T1),slow-release blended fertilizer(T2),loss-control fertilizer(T3),humic acid compound fertilizer(T4),and wheat formula fertilizer(T5),under the same conditions of nutrient supply,the effects on dry matter distribution and transport,filling,yield and quality of weakly glutenous wheat were analyzed.The results showed that slow-release blended fertilizer and wheat formula significantly increased the post-flowering dry matter production,and increased wheat grain filling rate,effective filling days,and grain weight compared with conventional fertilizer application.The grain yield increased by 9.27%—24.30% and 11.64%—22.98% respectively compared with conventional fertilizer.Slow-release blended fertilizer 2 years two varieties than conventional fertilization treatment of nitrogen fertilizer agronomic efficiency increased by an average of 23.14%, 36.88%, wheat formula fertilizer two years two varieties than conventional fertilization increased by an average of 36.31%, 39.35%. Wheat formula fertilizer treatment of two wheat varieties for testing grain quality had reached the national standard of weak gluten wheat. In summary,wheat formula fertilizer can be used as one of the fertilization patterns for high-yielding and high-quality production of rice stubble weak-gluten wheat in the test area,or appropriately reduce the application amount of slow-release blended fertilizer as the fertilization pattern for high-yielding and high-quality production of rice stubble weak-gluten wheat.

  • HE Lu, ZHANG Liru, YANG Ruoqian, YANG Xi, DU Yufan, ZHENG Boyan, LI Ruiqi, WANG Hongguang
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    In order to determine the optimal irrigation period of winter wheat under the condition of limited irrigation in spring and reduce the adverse effects of water restriction on the individual and population quality of wheat,five irrigation treatments,that irrigation at the 2nd,3rd,4th,5th and 6th spring leaf occurring stage(T2—T6)respective,were set.No irrigation treatment(W0)in the whole growth season was set as control.The Gaoyou 2018,a multi-panicle winter wheat variety was used.The effects of irrigation period on leaf area index(LAI)and SPAD of each leaf layer,canopy apparent photosynthetic rate after anthesis, dry matter accumulation and transportation, and yield were studied.The results showed that under the condition of only irrigation once in spring,the decay rate of canopy LAI by T5 and T6 decreased by 58.93% and 14.37% during 0—10 days and 20—30 days after anthesis,respectively,than that by irrigation at the 2nd, 3rd and 4th spring leaf occurring stage.The decay rate of LAI and SPAD of the top 3rd,4th and 5th leaf layers by T5 or T6 were lower significantly during 0—10 days after anthesis,and the decay rate of the flag leaf and top 2nd leaf layers by T5 or T6 were lower during 20—30 days after anthesis.Compared to early irrigation treatment, when irrigation was delayed to the 5th or 6th spring leaf age, the canopy apparent photosynthetic rate was increased by 19.89%,35.86% and 56.08%,respectively during the early,middle and late grain filling stage,the post-anthesis dry matter accumulation amount and the translocation amount from dry matter pre-anthesis to grains were both increased.The final grain yield and harvest index increased by 9.73% and 18.64%,respectively.In conclusion,under the condition of only irrigation once in spring of this study,irrigation at the 5th or 6th leaf age in spring was beneficial to delay leaf decay and increase population photosynthetic capacity and grain yield of wheat.

  • WANG Huijie, HUO Liguang, LI Sheng, LIANG Suming
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    In order to study the effect of different irrigation methods and ridge planting experiments on water use efficiency and yield of potato in dryland,and to identify an optimal ridge planting and irrigation combination mode of potato with high yield and water use efficiency simultaneously,and further provide a theoretical basis for the production efficiency of dryland potato.The two-factor experiment was persistently performed in Lanxian County Shanxi Province during two potato growing seasons(2019—2020).The three ridge planting,including conventional ridge cultivation(M1),concave ridge cultivation technique(M2)and outdoor planting(M3);the two irrigating including hydraulic driving zonal sprinkler(G1)and drip irrigation(G2),deeply exploring the effects of ridge planting and irrigation and their interaction effects on water use efficiency and yield of dry potato.The results showed that irrigation and ridge planting had obvious regulation effects on water use efficiency and yield of dry potato.Compared with M1 and M3,M2 could significantly increase soil use efficiency water storage consumption.Under G1 treatment,the water use efficiency and yield of M1 and M2 treatments increased by 14.08%,13.58%,and 23.28%,21.92%,respectively,compared to M3;under G2 treatment,the water use efficiency and yield of M1 and M2 treatments increased by 11.88%,11.50%,and 22.05%,20.45%,respectively,compared to M3,and the combination of concave ridge cropping + hydraulic driving zonal sprinkler irrigation treatment combination G1M2 had the highest yield.Under the same ridge planting,the water use efficiency and yield of hydraulically driven zonal sprinkler irrigation(G1)were the highest.Based on the regulating effects of ridge planting and irrigation on water use efficiency and grain yield,the G1M2 treatment could help synchronously to improve the grain yield and water use efficiency.This can be an optimal model of ridge planting and irrigation combination treatment of dry potato area of Shanxi Province.

  • PENG Duozi, HUANG Haoyuan, FAN Zhanhuang, DAI Yue, YUAN Miaoxin, ZHANG Zhenqian
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    Heavy metal pollution in soil has seriously affected the development of agriculture and the safety of agricultural products in China.Phytoremediation is a widely concerned soil remediation method,and rape is an ideal remediation plant.In order to screen rape breeding materials suitable for using as remediation plants.The pot experiment was carried out.With 100% nutrient soil environment as a control,three polluted soil environments(Ⅱ(25% contaminated soil,75% nutrient soil), Ⅲ(50% contaminated soil,50% nutrient soil), Ⅳ(75% contaminated soil,25% nutrient soil)with different proportions of contaminated soil were set up.The physiological characteristics and gene expression differences of three different hybrid types of Brassica napus,159-6(Self crossing species),Fengyou 520(Hybrid),and 159-6×Fengyou 520(Triple cross),were studied under different ratios of heavy metal polluted soil environments.The results showed that under different ratios of heavy metal contaminated soil,159-6×Fengyou 520 had the highest fresh and dry quality and was higher than control group I.Under 25% polluted soil environment,159-6 had the highest chlorophyll content,while the rest had 159-6 chlorophyll content×Fengyou 520 had the highest chlorophyll content.Except for Fengyou 520 with the highest soluble protein content in a 50% polluted soil environment,the rest were 159-6×Fengyou 520 had the highest soluble protein content.Under 50% and 75% polluted soil environment,159-6×Fengyou 520 had the highest SOD activity,with MDA content lower than 159-6 and Fengyou 520;four genes related to heavy metal resistance(BnaA08g04000D,BnaA09g24330D,BnNRAMP1,and BnPri-miR167a)were identified in 159-6×The expression levels in the leaves of Fengyou 520 were higher than those of 159-6 and Fengyou 520;Bna0280620 and Bna049040 genes in 159-6 under 75% polluted soil environment×The expression level in the leaves of Fengyou 520 was also higher than that of Fengyou 520 and 159-6.159-6,Fengyou 520 and 159-6×Fengyou 520 could grow and develop normally in polluted soil environments with different ratios;Sanjiao rapeseed 159-6×Fengyou 520 performs better in environments containing 50% and 75% polluted soil.The study was carried out from the genotype and biological characteristics of breeding materials,and the differences between conventional materials and hybrid materials in heavy metal resistance and between different hybrid types were analyzed.It was found that hybrids were better than conventional materials,while three hybrids were better than hybrids.

  • Animal Husbandry·Fisheries·Veterinarian

  • YAN Zunqiang, JI Yanan, WANG Pengfei, ZHANG Bo, SHI Haixia, GUN Shuangbao
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    The purpose of this paper was to clone the CDS region of StAR gene,perform bioinformatics and detect StAR gene expression level among different tissues using RT-qPCR in order to revel its function.The results showed that the CDS region of Hezuo pig StAR gene was 858 bp in length,encoding 285 amino acids.Compared to the reference sequence of pig,the A at the 572nd point mutation to G(leading to lysine mutation to arginine at the 191st point)was missense mutation and insertion of a base C at the 791st point(causing 20 amino acids mutation)was frame shift mutation,respectively.Molecular mass was about 90.95 ku and theoretical isoelectric point was 8.87;extinction coefficient was 33 835 and unstable coefficient was 41.49.Additionally,it contained one N-sugar-based site and secondary structure was mainly composed of α spiral and irregular curling.The secondary structure was similar to the tertiary structure.Lastly,the similarity of StAR nucleic acid sequences among Hezuo pig and Sus scrofa, Ovis aries, Bos taurus was 99.77%, 90.45% and 91.78%,respectively.The phylogenetic tree result showed that Hezuo pig was closed to pig.StAR gene expressed in these tissues,such as spleen,kidney,lung.Among these tissues,testis have higher expression,indicating that it may be related to the development testis in Hezuo pig.

  • YUE Binglin, YANG Youzhualamu, RAN Hongbiao, WANG Hui, CAI Xin, WANG Jiabo, CHAI Zhixin, PENG Wei, SHU Shi, FU Changqi, WANG Guowen, ZHONG Jincheng
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    The aim of this study was to identify bovine circMYH8 and to explore its regulation effect on the proliferation of primary bovine muscle cells.Two sets of primers,namely divergent primer and convergent primer,were designed and synthesized from circMYH8.The cDNA of bovine longissimus dorsi,the cDNA of bovine longissimus dorsi treated with RNase R and the gDNA of bovine longissimus dorsi were amplified by PCR,respectively,and circMYH8 was identified by sequencing and agarose gel electrophoresis;RT-qPCR analysis of skeletal muscle at different developmental stages,nucleo-plasmic separation,and RNase R tolerance tests were performed on circMYH8;the overexpression vector and interference fragment of circMYH8 were constructed and synthesized,which were then transfected into primary bovine muscle cells,respectively,and RT-qPCR,Western Blot,EdU and flow cytometry were performed to investigate the effects of circMYH8 on the proliferative phenotype of primary bovine muscle cells.The results of sequencing and agarose gel electrophoresis showed that circMYH8 was real circRNA,and RT-qPCR analysis revealed that circMYH8 was overexpressed in early skeletal muscle development;the results of nucleo-plasmic separation showed that circMYH8 was expressed in both nucleus and cytoplasm,and RNase R tolerance test verified that circMYH8 was more stable than its linear transcript;overexpression of circMYH8 inhibited the expression of proliferation marker genes and activity of EdU,and inhibition of circMYH8 promoted the expression of proliferating marker genes,with the increased activity of EdU and the proportion of S phase cells.These results indicated that bovine circMYH8 could significantly inhibit the proliferation of primary bovine muscle cells.

  • YANG Ningning, XU Mingguo, ZHANG Jiangwei, YI Jihai, CHEN Chuangfu
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    To explore the non-structural protein NS3(P80)of Bovine viral diarrhea virus (BVDV),and to obtain high immunogenic NS3 recombinant protein,the amino acid sequence of NS3 protein was analyzed by bioinformatics software.The sequence of NS3 gene was amplified by PCR,and the recombinant expression vector pET-22b(+)-NS3 was constructed by seamless cloning technology.The recombinant expression vector was transformed into competent cells of E.coli BL21(DE3) and induced to express NS3 recombinant protein.The reactivity of NS3 recombinant protein was detected by Western Blotting.BABL/c mice were immunized with the obtained high purity NS3 recombinant protein,and the serum was collected.The levels of IgG,IgG1 and IgG2a antibodies in serum were detected by ELISA,and the ability of neutralizing virus was detected by virus neutralization test.The results showed that there was no signal peptide and transmembrane region in the amino acid sequence of NS3 protein,the secondary structure was mainly random coil,and the NS3 amino acid sequence contained dominant antigen epitopes.The recombinant expression vector pET-22b(+)-NS3 was successfully constructed by PCR and seamless cloning techniques,and the recombinant protein of NS3 with high purity was obtained by inducing expression,with the size of about 75 ku,which was consistent with the theoretical size.Western Blotting results showed that the NS3 recombinant protein had high reactivity.ELISA test showed that mice immunized with NS3 recombinant protein could produce high levels of IgG,IgG1 and IgG2a antibodies.The antibody levels of mice immunized with NS3 recombinant protein were extremely significantly higher than those of PBS negative control group(P<0.01),and neutralizing antibody level was also significantly higher than that of PBS negative control group(P<0.01).In conclusion,this study successfully obtained NS3 recombinant protein with high purity and immunogenicity.

  • SU Zonghua, ZHOU Xiaonan, WANG Xiaowei, YANG Chaoyun, DING Yanling, ZHANG Yanfeng, LI Chenglong, ZENG Ling, MING Wenxuan, SHI Yuangang, KANG Xiaolong
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    Residual feed intake is one of the most important indicators used to evaluate the efficiency of feed utilization in livestock,which is regulated by various factors such as digestive metabolism,intestinal immunity,locomotion and hypothalamic feeding center.lncRNAs are a class of non-coding RNA molecules involved in numerous biological processes and have been extensively studied in phenotypic studies such as diseases,but it is still unclear about the characteristics of hypothalamic lncRNA expression and how it is linked to the regulation of feed intake in cattle.The aim of this study was to screen for RFI-related lncRNAs in the hypothalamus and to investigate the previous regulatory relationship between lncRNAs and feed utilization.The experiment selected extremely high and low RFI Angus cattle as the material,collected hypothalamic tissue samples,and carried out RFI-related differentially expressed lncRNA screening and regulatory pathway analysis after total RNA extraction,library construction and high-throughput sequencing.105 differentially expressed lncRNAs were identified,of which 46 lncRNAs were down-regulated and 59 were up-regulated in the low RFI group.These differentially expressed lncRNAs were predicted to target genes mainly enriched in mitochondria-related oxidative phosphorylation and thermogenesis pathways;these differential lncRNA predicted target genes were mainly enriched in mitochondria-related oxidative phosphorylation and thermogenesis pathways;meanwhile the related genes(ATPase,respiratory chain complex)in the above pathways were mainly associated with differentially expressed lncRNAs(LNC_006637,LNC_007569,LNC_012079,LNC_005449).Some genes(NDUFA4,SDHD,UQCRH)in the network constructed by protein interaction network analysis had also been reported to be associated with livestock feed efficiency.The above results suggested that the hypothalamic tissue associated with residual feed intake in cattle might regulate mitochondrial function and energy utilization in the body mainly through thermogenesis and oxidative phosphorylation signaling pathways,but more cellular experiments were needed to verify this.

  • HAN Haoyuan, LI Tao, LI Shikai, SONG Xiaoyu, LI Jun, HA Si, ZHAO Jinyan, WEI Hongfang, QUAN Kai
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    To investigate the effects of GJB6 and PRKAA1 genes on the reproductive performance of Huai goats,122 Huai goats with different lambing number were selected as research objects.The primers were designed based on the reference sequences of GJB6 and PRKAA1 genes from the NCBI database.Single nucleotide polymorphisms (SNPs)were screened by PCR and sequencing methods.The genetic diversity,population genetic parameters and association analyses between different genotypes and lambing number were conducted.As a result,two SNPs (g.50694819 C>T and g.50694816 T>C) were detected in GJB6 gene,and both SNPs were synonymous mutations.Three SNPs(g.33691489 T>C,g.33693395 A>T and g.33693100 T>C)were detected in PRKAA1 gene,and they were all located in non-coding regions.The results of polymorphic information content indicated that g.50694819 C>T locus of GJB6 gene and g.33693100 T>C locus of PRKAA1 gene showed low degree of polymorphism,g.50694816 T>C locus of GJB6 gene,g.33691489 T>C and g.33693395 A>T loci of PRKAA1 gene showed moderate degree of polymorphism.The χ2 test showed that the five loci were all in Hardy-Weinberg equilibrium.The results of the association analysis between different genotypes and lambing number for the two SNPs from GJB6 gene showed that the differences were not significant,indicating that these two SNPs were not suitable for multiple lambing traits selection in Huai goats.The lambing number produced by goats with the TC genotype at g.33691489 T>C of PRKAA1 gene was significantly higher than goats with the CC genotype,and the lambing number produced by individuals with the AT genotype at g.33693395 A>T was significantly higher than goats with the AA,TT genotypes(P<0.05).Linkage disequilibrium analysis revealed a strong linkage equilibrium between the g.33691489 T>C and g.33693395 A>T loci(D'>0.8,r2>0.33).Haplotype correlation analysis showed that the lambing number of goats with haplotype TCA was significantly higher than that with haplotypes CTA,CTT,and TTA(P<0.05).The results showed that Huai goats with the TC genotype at the g.33691489 T>C locus,the AT genotype at the g.33693395 A>T locus and TCA haplotype of PRKAA1 gene had higher lambing number,indicating that these two SNPs could be used as candidate molecular markers for lambing number selection in Huai goats.

  • MENG Quanlu, ZHANG Mingjun, SHI Jinping, FU Lingjuan, LIU Ting, ZHANG Quanwei, CHENG Shuru
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    To explore the association between DLK1 polymorphism and growth performance in sheep. A total of 240 sheep of different breeds (Gansu alpine fine wool sheep, Mongolian sheep, Tibetan sheep, Small tailed han sheep, 50 sheep each and Tan sheep, 40 sheep) were selected as the experimental subjects. The growth performance indexes of different breeds of sheep at different ages were measured. Blood samples were collected and DNA was extracted. DLK1 gene polymorphisms were analyzed by PCR-SSCP combined with DNA sequencing. SPSS 20.0 software was used to analyze the correlation between different genotypes and growth performance of each sheep breed at different ages. The results showed that DLK1 gene was polymorphic in different breeds of sheep. Two alleles (A and B) and three genotypes (AA, AB and BB) were detected, and the dominant allele and genotype were B and AB, respectively. A single nucleotide mutation (G→A) was detected at the 30 412 bp site in intron 5 of DLK1 gene. The G30412A mutation was associated with the weight and chest circumference of sheep at 1 and 3 months old. The phenotype of BB genotype sheep was better than that of AA genotype sheep. There was no correlation between the mutation sites and the body length and body height. The results of this study showed that the G30412A mutation in intron 5 of DLK1 gene existed in different breeds of sheep, and this mutation significantly affected the body weight and chest circumference of sheep at the 1 and 3 months old, which could be used as a candidate gene for the production traits of Gansu sheep breeds.

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