Being effective only to the traits controlled by large-effect QTL,conventional marker-assisted selection(MAS)can hardly improve the quantitative traits that are controlled by many small-effect QTL.Later on, the genomic selection (GS) technology proposed in 2001 solves the problem of improving complex traits controlled by the minor polygenic effects due to its high prediction accuracy through estimating the breeding value of individual with high-density molecular markers. At present,it has been successfully applied to the genetic improvement of quantitative traits in animals and plants such as dairy cattle,pigs,sheep,maize,and wheat in the US,Canada,Australia,Germany,France and so on.As a breaking-through breeding technology and a hot spot in research and application now. In this review,we summarized the factors affecting predictive accuracy of GS and the research progress of GS in breeding maize,wheat,rice and rapeseed at home and abroad,and finally discussed the existing problems in its application. This review will provide some reference for GS of the crops in the future.

Wheat yellow dwarf disease,caused by Barley yellow dwarf viruses(BYDVs),is an important viral disease in wheat production.BYDV GAV has become the main pathogen causing wheat yellow dwarf disease.Till now the studies on the function of BYDV GAV encoded proteins P1,P2 and CP are lacking.We focued on the function of P1,P2 and CP,which could lay the foundation for the pathogenic mechanism of BYDV GAV.Phylogenetic analysis of BYDV GAV encoded P1,P2 and CP was conducted using Mega 7.0.We constructed the YFP expression vector of P1,P2 and CP,and then transformed them into GV3101 and infiltrated the leaves of Nicotiana benthamiana.The subcellular localization of P1,P2 and CP was observed using confocal laser scanning microscopy(CLSM).We constructed the biomolecular fluorescence complementation assay(BiFC)vectors of five coding proteins,and then transformed them into GV3101 and infiltrated the leaves of Nicotiana benthamiana.CLSM was used to observe the interaction of P1,P2 and CP and other viral proteins in vivo.Furthermore,we constructed the Potato virus X(PVX) expression vectors of P1,P2 and CP,transformed them into GV3101 and infiltrated the leaves of Nicotiana benthamiana.At 5 days post inoculation(dpi)the symptom formation of PVX infection was observed.The systemic leaves were collected for detection of viral accumulation to determine the effects of the pathogenicity of P1,P2 and CP.Results showed that BYDV GAV was most closely related to BYDV PAV at the nucleotide level.Subcellular localization of P1,P2 and CP was cytoplasm and nuclear.P1 interacted with itself in vivo using BiFC.In the pathogenicity assay,the systemic leaves of PVXCP infection showed chlorosis at 5 dpi,and PVX accumulation was detected,while PVX,PVXP1 and PVXP2 infection showed no symptoms in systemic leaves and PVX accumulation was undetectable,which was detected at 10 dpi,indicating that CP promoted the formation of PVX symptoms.In brief,P1 possibly involves in viral infection via self-interaction in vivo,and CP can promote viral infection.

To investigate the function of ATP-binding cassette transporter subfamily G11(ABCG11)in xerophytic economic shrub Lycium barbarum ssp. Bianguo,using the Lycium barbarum ssp. Bianguo as the material. The cDNA sequence of Lycium barbarum ssp. Bianguo LbABCG11 gene was obtained by RT-PCR and RACE method, the sequence patterns were analyzed by bioinformatics software. Quantitative Real-time PCR(qRT-PCR)was used to analyze the expression patterns of LbABCG11 in different organs of Lycium barbarum ssp. bianguo under salt treatment,osmotic stress and abscisic acid(ABA)treatments.The results showed that LbABCG11 gene contained 2 971 bp in full length and a 2 130 bp open reading frame,encoding 710 amino acids.The protein contained six transmembrane regions, the calculated molecular weight, isoelectric point, adipose index, instability index and average hydropathicity was 79.39 ku,8.42, 89.63, 35.52, 0.070, the encoded protein chemical formula was C₃₅₉₀H₅₅₇₇N₉₃₅O₁₀₂₇S_35. LbABCG11 is a stable alkaline hydrophobic protein.The protein contained a nucleotide-binding domain(NBD)and a transmembrane domain(TMD),which were arranged in the form of NBD-TMD.LbABCG11 was a white-brown complex (WBC)half-size molecular transporter.The secondary structure of the protein was mainly composed of α-helix and random coil, accounting for 45.21% and 33.66%, respectively, which was consistent with the predicted results of the tertiary structure. Phylogenetic analysis showed that LbABCG11 had closer genetic relationship with ABCG11 from Solanum lycopersicum and Capsicum barbarum, but has low homology with ABCG11 from Arabidopsis thaliana and Chenopodium quinoa. LbABCG11 gene was expressed in some organs of Lycium barbarum ssp. bianguo,including leaves,stems and roots.The LbABCG11 expression level was induced by NaCl,osmotic stress and ABA,manifesting that LbABCG11 was involved in the stress response of xerophytes.

In order to have a clearer understanding of the research progress on cold resistance of winter rapeseed in cold and arid regions of China,and to provide theoretical support for breeding and production.The research progress of cold-resistance and cold-resistance breeding of winter rapeseed were summarized respectively from six aspects such as selection index and method,genetic law,breeding method,screening effect,germplasm classification and molecular mechanism research,and four aspects such as breeding method,breeding varieties,cold resistance classification and adaptability screening of varieties.Based on the analysis of the existing problems,six measures were put forward,such as creating germplasm resources by multiple ways,screening out core germplasm by comprehensive evaluation,carrying out research on cold-resistance mechanism and construction of innovation system of molecular design breeding,constructing breeding system and carrying out cold-resistance breeding by multiple ways,improving the adaptability of varieties by comprehensive screening,promoting varieties yield level and improving benefits by carrying out matching cultivation techniques,in order to promote the development of rapeseed production in cold and arid regions of our country.

Grapes are often subjected to various abiotic stress during their growth and development,such as drought,salt,heat or chilling,among them,drought stress is one of the most important abiotic stresses inhibiting vegetative and reproductive growth of grape.Although grapes are more resistant to drought stress,about half of our country's main grape cultivation area belongs to arid and semi-arid climates,drought is one of the main factors restricting the development of our country's grape industry.To ensure the healthy development of our country's grape industry,the research on the effects of drought stress on grape,the formulation of rational irrigation system and the breeding of drought-resistant varieties have become the research hotspots in recent years.Due to the extensive effects of drought stress on grape,in older to cope with drought stress,grapes have evolved many regulatory mechanisms to balance the effects of drought stress in grape growth.Firstly,the water demand degree of grape in different growth period was analyzed according to the law of grape water consumption,and the effects of drought stress on main physical and chemical indexes were exhibited,including photosynthesis,osmotic regulation and reactive oxygen regulation.And then,the quality and yield of grape fruit were analyzed under drought stress,the effects of drought stress on grape fruit quality were reviewed.We give the prospect for how to use drought stress strategy to improve grape quality and how to further study the molecular mechanism of grape response to drought stress at last.

In order to further definite the genetic basis of black rot resistance in cabbage,and breed high-quality disease resistant cultivars,F₁ progenies were obtained by crossing the inbred line 4674(high resistance to race 1)as male parent,with the inbred line 4673(high susceptibility to race 1)as female parent.The F₂ population containing 152 individuals were obtained by F₁ self-crossing. The F₂ population was inoculated by spraying at seedling stage. After 12-14 days, the phenotype of F₂ population was identified according to the identification method of cabbage seedling stage.A total of 175 markers with good polymorphism and clear bands were selected from 404 pairs.Subsequently, the 175 molecular markers were used for genotyping of F₂ population and constructing a genetic linkage map. Finally, the QTL for black rot resistance of cabbage was mapped with phenotypic data and genetic map.The results showed that there were 154 molecular markers linked to nine chromosomes,including 110 pairs of InDel markers and 44 pairs of SSR markers and covering a length of 714.29 cM,and the average distance between markers was 4.64 cM.Seven QTLs were located,of which three major QTLs were qBR-7-2,qBR-7-3 and qBR-4-3,mapped on the genomic markers(CG842110-CG842482 and M29-M39)of chromosome 7,and on the genomic markers between CD838151 and BOE417 of chromosome 4,respectively.The explainable phenotypic variation of QTLs were 16.0%,9.2% and 10.0%,and their LOD values were 5.75,3.20 and 3.47,respectively.

In order to investigate the reasons for the differences in plant height between SSSL-B50 and HJX74,an F₂ population was constructed by backcrossing SSSL-B50 with HJX74 for genetic analysis and gene mapping of the plant.The genetic analysis showed that F₁ plants exhibited tall plant height.And the separation ratio within F₂ was 3:1(χ²=0.18<3.84)between the tall plant individuals and the semi-dwarf ones,which indicated that the tall plant height of SSSL-B50 was a dominant trait.Based on the linkage analysis between the marker genotypes and phenotypes of plant height in the F₂ population,the gene controlling tall plant height was mapped between markers S18 and X161 within the 38.38—39.07 Mb interval of the substituted segment on chromosome 1.Moreover,it was found that the known gene of "Green revolution",SD1,fell to the mapping interval.Through amplification and sequence alignment of SD1 gene in Oryza glumaepatula,SSSL-B50,and HJX74,it showed that a 280 bp deletion in CDS in HJX74 compared with Oryza glumaepatula and SSSL-B50.The qRT-PCR results showed that the expression level of SD1 was significantly higher in the second,third,and fourth sections of SSSL-B50 stem than in HJX74.The comparison of the SD1 gene sequence of Oryza glumaepatula with the previous reported results showed that the base substitutions occurred at two sites (E100G, Q339R) in the encoded amino acid.The results revealed that the tall plant height of SSSL-B50 was controlled by SD1.Furthermore,a new allelic type of SD1, SD1^Glu,was identified in Oryza glumaepatula.

It explored the ratio of organic fertilizer replacing nitrogen fertilizer in the piedmont plain of Hebei Province,in order to provide a basis for reducing the amount and increasing the efficiency of nitrogen in wheat in this area.Field experiments were carried out in Boyuan farm in Yongnian,Hebei Province for two consecutive years,and five organic and inorganic fertilizer combination treatments were set up.The results showed that organic fertilizer instead of 20% and 40% chemical fertilizer could significantly improve the number of grains per spike and yield.Compared with the high nitrogen and saving nitrogen treatment of single chemical fertilizer application,the yield increased by more than 4.0%,and the number of grains per spike increased by 3.6—5.6.Most of the grain quality indexes for organic fertilizer instead of 20% and 40% chemical fertilizer treatment,and saving nitrogen treatment were better,and the stabilization time increased by 2.2—2.7 min,the tensile area increased by 10.5—17.5 cm²,and the maximum tensile resistance increased by 28.0—75.5 EU.Various nitrogen efficiency indicators of treatment for organic fertilizer instead of 20% were higher.The nitrogen fertilizer efficiency,nitrogen utilization efficiency,and nitrogen harvest index increased 109.3%,9.3% and 11.3% respectively compared with high nitrogen treatment,and 6.9%,8.5% and 8.3% respectively compared with the saving nitrogen treatment.When organic fertilizer replaced chemical fertilizer in different proportions,nitrate nitrogen in 0—20 cm soil appeared "surface accumulation",and the content of nitrate nitrogen increased,which was more than 38.5% higher than that of the saving nitrogen treatment.The nitrate nitrogen in 20—40 cm soil was significantly higher for the saving nitrogen treatment and the high nitrogen application treatment.Organic fertilizer instead of 20% nitrogen fertilizer treatment had the best yield and grain quality,significantly improve the nitrate nitrogen content in 0—40 cm soil,improve the nitrogen absorption and utilization of wheat,and finally obtain higher environmental benefits.

Grain specific promoter with high transcription activity can regulate the specific and high-level expression of target genes in plant grains.In order to explore specific promoters of maize grain,the dominant expression gene GRMZM2G006585 was screened from the published data of maize expression profile microarray.The DNA sequence about 2 000 bp upstream of its coding region was cloned and named PZm2G006585.By using the online websites New PLACE and PlantCARE to analyze its promoter elements,we found that it contained multiple grain specific related elements such as E-box and P-box.It was preliminarily considered that the upstream sequence of the cloned coding region was a grain specific promoter from maize.In order to verify its function,the expression vector of GUS gene was constructed and transformed into plants.GUS histochemical staining results of transgenic rice showed that the expression pattern of exogenous genes driven by the promoter was grain specific and embryo dominant.The results of GUS activity detection in T₃ seeds of transgenic Arabidopsis thaliana single copy line showed that the GUS activity driven by PZm2G006585 was 909.52 nmol/(min·mg).The discovery and functional verification of the grain-specific promoter PZm2G006585 can provide candidate promoter resource for specific expression of target genes in maize,rice and other monocotyledons.

To investigate the potential functions of DUF760s in rice growth and development,the genome-wide identification,classification,promoter sequence and expression profile analysis of the DUF760 gene family were performed.This research identified 6 and 8 members of DUF760 gene family in rice and Arabidopsis respectively by bioinformatics analysis.Phylogenetic tree analysis divided these members into two subfamilies,there were also some characteristic differences between the two subfamilies in protein conserved motif and gene structure.Multiple cis-acting elements responding to stresses and phytohormones existed in the promoter regions of rice DUF760 family genes.ABRE(abscisic acid response)element was present in the promoter sequence of all DUF760 family members,the promoter region of OsDUF760-1 possessed 9 abscisic acid(ABA)related response elements.The transcriptional expression level of OsDUF760-1 in rice was significantly down-regulated after ABA treatment,while OsDUF760-3 was significantly up-regulated.The expression change patterns of these two genes in rice after drought stress treatment were consistent with that after ABA treatment,which indicated that these two genes may participate in rice drought stress response through ABA signaling pathway,and play different roles.In addition to strong responses to ABA and drought stress treatments,members of rice DUF760 family also showed relatively strong expression changes in response to JA(Jasmonic acid),low temperature and M.oryzae treatments.

Gymnocypris eckloni is an important cold-water indigenous fish from the upper reaches of the Yellow River.Because of its slow growth and development and late sexual maturity,its population has been declining for many years.MSTN(Muscle Growth Inhibitor) is a factor that negatively regulates muscle growth.This study cloned the MSTN-1 gene of G.eckloni and examined its expression characteristics to provide basic information on the molecular regulation mechanism of growth retardation in G.eckloni.The full-length cDNA sequence of MSTN-1 gene was obtained by PCR,5'-RACE and 3'-RACE.qPCR was used to detect the expression characteristics of MSTN-1 gene in different tissues of G.eckloni and in muscle tissues of different Cyprinidae fishes.The full length of the cDNA sequence of MSTN-1 gene was 2 190 bp,with an open reading frame of 1 128 bp,a 5'UTR of 96 bp,and a 3'UTR of 966 bp,encoding a total of 375 amino acids.MSTN-1 was an unstable hydrophilic secretory protein containing a signal peptide and no transmembrane structure,which was mainly distributed in the nucleus,mitochondria and cytoplasm.The secondary structure of MSTN-1 protein was dominated by random curls,and exists in two TGF-β structural domains:namely,the TGF-β prepeptide region (37—268 aa) and the TGF-β functional region (281—375 aa).MSTN-1 protein contained one the proteolytic site RIRR and nine conserved cysteinee in the TGF-β functional region.Amino acid sequence homology analysis showed that the amino acid sequence of MSTN-1 from G.eckloni had high similarity with other Cyprinidae fishes MSTN-1s and low similarity with mammalian and avian MSTN-1s.Phylogenetic analyses showed that the MSTN-1 of the G.eckloni clustered in the same evolutionary branch with other Cyprinidae fishes.The results of Real-time Fluorescence Quantitative PCR showed that MSTN-1 was expressed in brain,muscle,gill,heart,liver and kidney tissues of the G.eckloni,but the expression was higher in brain and muscle tissues.The gene was expressed in the muscle tissues of different Cyprinidae fishes,with the highest expression in the muscle tissues of the G.przewalskii,followed by the higher expression in the muscle tissues of the G.eckloni and the Cyprinuscaurpio.The full-length cDNA sequence of the MSTN-1 gene of G.eckloni was obtained by cloning,and bioinformatics analysis and qPCR assay were performed.The characteristic differences in the expression of MSTN-1 in the muscle tissues of these indigenous fishes can help to further understand the molecular mechanism of growth retardation in highland fishes.

WRKY transcription factor is a family of transcription factors unique to plants.Studies have demonstrated that WRKY transcription factor played an important role in plant growth and development and in plant response to biological and abiotic stress.In order to reveal the function of tomato WRKY gene,two inbred lines of tomato with high resistance to bacterial wilt Hm 2-2(R)and high susceptibility to bacterial wilt BY 1-2(S)were used as experimental materials based on the preliminary transcriptome data,and a WRKY transcription factor SlWRKY75 gene(Solyc05g015850.3)was cloned.The structure,expression pattern and function of the gene and its encoded protein were analyzed by bioinformatics analysis,multiple alignment of amino acid sequences,phylogenetic tree construction,Real-time Quantitative PCR(qRT-PCR)and virus induced gene silencing(VIGS).The results showed that the full length of the cDNA of this gene was 653 bp,its maximum open reading frame was 519 bp,encoding 172 amino acids,the relative molecular weight of the protein was 19.878 51 ku,the theoretical isoelectric point was 9.32.The protein belonged to the hydrophilic non-secreted protein,and there was no transmembrane structure.Meanwhile,the protein had a highly conserved WRKY domain and a CX₄CX₂₃HXH zinc finger motif,which belonged to the Class Ⅱ family.Phylogenetic tree analysis showed that SlWRKY75 was closely related to Solanum pennellii SpWRKY75 and clustered into a group with other Solanaceae,while it was far related to Hevea brasiliensis HbWRKY75 and Gossypium hirsutum GhWRKY75 and was in different branches in the phylogenetic tree.The results of qRT-PCR analysis showed that the expression of SlWRKY75 gene were tissue-specific and could be induced by Ralstonia solanacearum,salicylic acid and jasmonic acid.VIGS result showed that silencing SlWRKY75 reduced plant resistance to bacterial wilt,indicating that SlWRKY75 positively regulated tomato resistance to bacterial wilt.These results suggested that SlWRKY75 gene played an important role in regulating tomato resistance to bacterial wilt.

In order to study the effects of OsBAK1P,a precursor of Brassinosteroid insensitivity 1-associated receptor kinase 1,on the agronomic characteristics of rice.A target sequence fragment with a CDS fragment size of 651 bp was amplified from the rice panicle cDNA using the Zhonghua 11 japonica rice variety as the material according to the specific primers designed by the gene;the PTCK303-OsBAK1P overexpression vector and the PTCK303-OsBAK1P RNA interference vector were successfully constructed by restriction enzyme digestion and ligation methods;the Agrobacterium tumefaciens EHA105 strain was transformed into the correct expression vector plasmid,and the positive Agrobacterium tumefaciens clones were screened out by using CDS amplification primers;the callus of japonica rice cultivar Zhonghua 11 was infected by Agrobacterium tumefaciens genetic transformation to obtain transgenic plants;finally,compared to Zhonghua 11,two overexpressed and RNA-interfered T₁ transgenic plants with similar phenotypes were selected to measure and analyze the agronomic characteristics such as plant height,panicle length,and leaf angle,changes in root length and coleoptile length at the early stage of germination,and response to brassinolide(BL).The results showed that the plant height of OE-OsBAK1P transgenic rice was dwarfed,the panicle length was shorter,and the leaf angle decreased,meanwhile,the root length increased and the seedling length shortened after seed germination,and the leaves were not sensitive to BL.However,the plant height,panicle length and leaf angle of RNAi-OsBAK1P transgenic rice increased,meanwhile,root length shortened and seedling length increased after seed germination,and the leaves were sensitive to BL.To sum up,these results provide theoretical support for changing rice plant structure to increase grain yield and may provide a reference for further studies on other functions of OsBAK1 and its precursor OsBAK1P.

In order to provide a theoretical basis for rational application of organic fertilizers and ensure the healthy development of soil ecosystems in corn fields,the effects of soil fungal communities under different levels of organic fertilizer application for the composition,diversity,and functional characteristic were studied.In the meantime,a field experiment was used,with a total of 4 treatments were set up,and the high-throughput sequencing and FUNGuild method were used to analyze the soil fungal diversity,community structure and functional groups in different fertilization rates.The results showed that the increased application of organic fertilizer could increase the content of soil organic matter,available phosphorus,available potassium,urease,catalase,etc.,and it was positively correlated with the application amount of organic fertilizer.Applied organic fertilizers could increase the diversity of soil fungal communities and reduce their richness.The dominant fungal communities in the soil under different fertilization rates were Ascomycota,Mortierellomycota,Basidiomycota,and Olpidiomycota,the relative abundance of Ascomycota and Basidiomycota was significantly higher in the treatment with a medium amount of cattle manure than in the control without organic fertilizer;from the perspective of fungal genera,the dominant genera include Fusarium,Humicola,Olpidium,and Microidium.The application of organic fertilizer increased the relative abundance of symbiotic and saprophytic nutrients,and with the increase of organic fertilizer application,the abundance of pathological nutrients showed a decreasing trend;the number of woody saprophytic fungi in the treatment with increased organic fertilizer application was significantly higher than that in the treatment without organic fertilizer application,while the number of plant and animal pathogens was lower than the control.Therefore,it is believed that increasing a certain amount of organic fertilizer application could optimize the soil microbial environment and promote the increase of corn plant yield.

Auxin response factor(ARF)is a class of transcription factors with B3 domain,which is a direct molecule regulating auxin response and controlling gene expression.A gene,ZmARF10,which encoded ARF protein and actively participated in drought-rehydration stress response,was previously screened in maize by analyzed transcriptome data.To further research the molecular mechanism of ZmARF10 regulating drought resistance of maize,and also provide a new idea for molecular breeding of drought resistance,the gene was firstly analyzed by bioinformatics software.Secondly,Quantitative Real-time polymerase chain reaction(qRT-PCR)was used to detect the expression pattern of ZmARF10 in different tissues,under high temperature,drought,high salt,ABA and restoring treatments,and in different inbred lines.Finally,the function of ZmARF10 was analyzed using CRISPR/Cas9 technology.The results showed that ZmARF10 was located on chromosome 3 of maize,with a total length of 2 127 bp,and encoded 708 amino acids with a typical B3 domain.The upstream 2 kb region of ATG of this gene contained response elements related to methyl jasmonate,auxin,abscisic acid and low temperature.The phylogenetic tree showed that the protein encoded by the ZmARF10 gene was closely related to sorghum.qRT-PCR results showed that ZmARF10 was a constitutive expression gene,and the expression level of ZmARF10 was the highest in mature corn roots of maize.Under high temperature,drought,high salt and ABA treatments,the expression of this gene was significantly up-regulated,and the up-regulation ratio was up to 8.2 times after drought stress.After drought stress,the expression level of ZmARF10 gene was significantly higher in the drought-resistant inbred line Zheng 36 than that in the drought-sensitive inbred line B73.Investigation of Arabidopsis wild type and ARF10-deficient mutants showed that,compared with wild type,the mutant plants showed leaf wilting and even dry death,roots curled,root branch number decreased,and lateral root growth and development were hindered under drought conditions.Determination of physiological and biochemical indexes showed that the relative water content,chlorophyll content and net photosynthetic rate of the deficient mutants were significantly lower than those of the wild-type plants after drought stress,indicating that the drought resistance of Arabidopsis decreased after ARF10 gene was knocked out.

The bioinformatic analysis of SAUR genes aims to unveil their structure,function,and evolutionary relationships.Simultaneously,through subcellular localization,the goal was to understand their position within the cell,thereby inferring their biological functions.Such analyses contribute to a deeper understanding of the gene's mechanistic role and potential applications within an organism.This held particular theoretical and practical significance for regulating the germination of windproof seeds,reducing seed dormancy duration,and facilitating green cultivation and breeding of wind-resistant plants.In order to explore the biological characteristics and functions of SAUR genes,SdSAUR32 and SdSAUR23 were cloned from Saposhnikovia divaricata,and bioinformatics analysis was conducted to analyze the expression and subcellular localization of these two genes in different dormancy stages of Saposhnikovia divaricata seeds.The findings indicated that the SdSAUR32 protein had a molecular weight of approximately 14.48 ku(pI 6.45),while the SdSAUR23 protein had a molecular weight of around 10.29 ku(pI 8.00),confirming that both proteins were hydrophilic.The conserved domain analysis of SdSAURs gene encoded protein showed that SdSAUR32 and SdSAUR23 had the same conserved domain,which belonged to auxin induced superfamily members.The phylogenetic analysis showed that both SdSAUR32 and SdSAUR23 encoded protein was the closest relative to Daucus carota. The subcellular localization results indicated that both SdSAUR32 and SdSAUR23 were found within the nucleus.The SdSAUR32 and SdSAUR23 genes were found to be expressed during different stages of Saposhnikovia divaricata seed development.It was observed that the expression levels of these two genes were significantly higher during the germinal stage compared to the release dormancy stage and dormancy stage.This suggested that SdSAUR32 and SdSAUR23 may have a crucial role in the germination,growth,and development of Saposhnikovia divaricata seeds.This study offers not only insights for investigating the role of the SAUR gene family but also establishes atheoretical framework for examining the function of other genes in the parsnip family at the molecular level.

In order to clarify the effects of high temperature stress during grain filling stage on leaf senescence and yield,a field experiment from 2019 to 2020 was conducted with employing heat-sensitive variety(XY335)and heat-resistant variety(ZD958). Simple plastic greenhouse was constructed for heat treatment(HT)during 12-25 days after silking,the natural temperature was set as control(CK). The two years results showed that from 12 to 25 days after silking,high temperature stress significantly reduced the grain weight and yield of summer maize. For ZD958,the grain yield of HT was reduced by 18.8% and 19.3% than CK and thousand kernels weight(TKW)was decreased by 23.4% and 9.1%,respectively.For XY335,the grain yield and TKW of HT were reduced by 29.4%,26.3% and 20.1%,14.2%,respectively.High temperature during grain-filling stage accelerated leaf senescence. The two years results showed that leaf senescence rate(LSR)of ZD958 and XY335 under HT were increased by 76.4%,140.6% and 135.1%,139.6%,respectively.Leaf area duration(LAD)after silking with HT was reduced by 17.7%-36.5% compared with CK. Ear leaf photosynthesis under HT was lowered by 20.0%-42.9% than CK.Correlation analysis results indicated that accelerated LSR under HT had negative correlation with grain yield but without statistical significance(P=0.064 7).LAD after silking,photosynthesis,and SPAD significantly reduced under HT,which showed significant positive correlation with grain yield(P< 0.05). Taken together,heats tress during grain filling significantly accelerated leaf senescence,thus lowering kernel weight and resulting in yield loss. Under high temperature stress,heat-resistant maize variety had higher regulation ability than high temperature-sensitive maize variety,which could significantly mitigate yield loss caused by high temperature stress.

In order to study the biological function of NbEHD1 in Nicotiana benthamiana,its gene structure,conserved domains,phosphorylation sites,subcellular localization and evolutionary relationship were predicted by bioinformatics methods.The encoding sequence of NbEHD1 was 1 638 bp,and its genomic sequence contained 16 exons and 15 introns.NbEHD1 protein was predicted located in the cytoplasm,which had 42 phosphorylation sites and had no signal peptide and transmembrane region.NbEHD1 belonged to the P-loop_NTPase superfamily and had a conserved domain specific to the EHD family.Phylogenetic relationship showed that NbEHD1 was closely related to EHD sequences of Solanum tuberosum and Solanum lycopersicum.Blast analysis against N.benthamiana sub-data base in SNG was performed to obtain the predicted full length of NbEHD1,then its full nucleic acid sequence was amplified by gene-specific primers.After obtaining NbEHD1 CRISPR/Cas9 gene editing construct,the vector was successfully transformed into the leaves of N.benthamiana by Agrobacterium-mediated genetic transformation.Eventually 16 T₀-generation plantlets were successfully identified,which provided materials for further research to determine the biological function of NbEHD1.

JAZ protein plays a key role in plant growth and development and stress signaling pathways.In order to explore the regulatory mechanism of JAZ protein in wheat late spring cold,TaJAZ6 gene was cloned from young spike of wheat,and its molecular characteristics,expression characteristics and subcellular localization were analyzed.The results showed that the full-length CDS sequence of the gene was 549 bp,encoding 178 amino acids.The predicted molecular weight of the encoded protein was 18.376 ku,the theoretical isoelectric point was 9.37,and the instability coefficient was 62.44,so it was an unstable protein.The protein encoded by this gene had a TIFY domain and a CCT_2 domain.Phylogenetic tree analysis showed that the protein had the closest relationship with TIFY 11b proteins of Triticum dicoccoides and Triticum urartu.In addition to the basic response elements such as CAAT-box and TATA-box,the promoter region of TaJAZ6 gene also contained hormone response elements,light response elements,low temperature response elements,defense and stress response elements.Real-time Quantitative PCR analysis showed that TaJAZ6 gene was expressed in roots,stems,leaves and young panicles,with the highest expression in roots.The expression of TaJAZ6 gene was also induced by low temperature and methyl jasmonate (MeJA).Under low temperature stress,the expression trends of TaJAZ6 in roots,stems and leaves of Aikang 58 (tolerant to late spring cold ) and Zhengmai 366 ( sensitive to late spring cold ) were the same trend,which significantly increased.After spraying 300,350 μmol/L MeJA,the expression of TaJAZ6 in plants treated with low temperature decreased significantly in both wheat varieties.The expression level of TaJAZ6 in the young panicles after low temperature stress showed an opposite trend.The expression level of TaJAZ6 in the young panicles of Aikang 58 decreased significantly,and increased significantly in the young panicles of Zhengmai 366.It was speculated that the gene might negatively regulate the defense response of wheat to late spring cold stress.By spraying MeJA,the relative expression of TaJAZ6 gene in young spikes of two wheat varieties under low temperature stress was significantly reduced,and the grain number of wheat was increased.Subcellular localization assay showed that TaJAZ6 protein was localized in the nucleus.The results above indicate that TaJAZ6 may play an important role in the response of wheat to late spring cold stress.

Nitrogen is an essential element for plant growth and development.It is the main limiting factor of the primary productivity of terrestrial ecosystem and has important significance for plant growth and development.Mycorrhizal fungi form a symbiotic relationship with plants,promote plants to absorb nitrogen from the soil,reduce the restriction of nitrogen on plant growth,and enhance the nitrogen exchange between soil,mycorrhizal fungi,and plants,playing an important role in the nitrogen cycle.It reviewed the utilization of different forms of nitrogen by mycorrhizal fungi,their effects on plant nitrogen metabolism,and their ecological significance on soil nitrogen cycle.Based on the current research status of mycorrhizal fungi' impact on plant nitrogen utilization,it is suggested to combine genomics,transcriptome,proteomics and environmental genomics technologies,focus on studying the nitrogen transport mechanism of mycorrhizal fungi plant symbiosis,analyze the main path of nitrogen exchange and gene interaction network between mycorrhizal fungi, plant, and soil,so as to promote plant nitrogen utilization rate,reduce nitrogen fertilizer application,and promote sustainable agricultural development.

In order to clarify the similarities, differences and biological functions of Nodule inception(NIN)genes, which expressed core transcription factors NIN involved in all the biological processes from rhizobia infection to root nodule formation and nitrogen fixation in soybean, and integrates nodulation and autoregulation of nodulation signals to dynamically control the number of nodules, methods of bioinformatics were used to analyse the phylogeny, protein sequence, gene sequence and promoter elements of the four GmNINs, and Real-time PCR were carried out to verify the tissue expression pattern.The results showed that the four GmNINs proteins with similar protein sequences belonged to the specific NIN protein family in Legumes, located in the nucleus, had multiple phosphorylation sites and similar core tertiary structure with significant difference in corner. Acorrdingly, the above results indicate that the four GmNINs proteins may be redundant in core functions, but have differences in specific expression patterns.Analysis of the upstream-3 kb promoter sequence of GmNINs showed that the promoter sequence of GmNINs contained many elements such as ABRE, DRE1 and other elements related to stress and hormone response, except NBS and CYC elements.Transcriptome analysis and Real-time PCR results showed that GmNINs were highly expressed in nodules and inhibited in high nitrogen condition; they responded to salt and drought stress of varying degrees, among which GmNIN2a and GmNIN2b were more sensitive and significant in responding to abiotic stress, and GmNIN2a and GmNIN2b may play an important role in the process of plant response to abiotic stress. In short, the results revealed that GmNINs not only regulate the nodule number of soybean and nodulation, but also participate in the process of root response to abiotic stress. The discovery of this regulatory mechanism provides essential clues for breeding new varieties with high quality and high yield.

In order to explore the gene expression regulation mechanism in the formation of reactive oxygen species (ROS) induced by heat stress,heat stress treatments were set at seedling,heading and filling stage to investigate the dynamic change of ROS accumulation in rice,respectively.And the Quantitative Real-time PCR (qRT-PCR) was used to analyze the expression pattern of nine respiratory burst oxidase homologue (Rboh) encoding genes (OsRboh1-OsRboh9) in rice under different growth stages.Results showed that the ROS accumulation in rice leaves and grains significantly increased with the extension of heat stress.The ROS content increased slowly after 7 days of heat stress at seedling stage, while increased continuously after heat stress during the heading stage and early filling stage (1-10 d) in rice grains. Expression levels of the nine OsRboh family genes continuously increased with the extension of heat stress at seedling and heading stages,and OsRboh7 and OsRboh5 showed higher expression levels under heat stress.The expression levels of OsRboh1,OsRboh5 and OsRboh9 were continuously increased,while other genes showed a change tendency of increasing initially and then decreasing under heat stress at filling stage.The expression levels of OsRboh7 and OsRboh5 were all reached to a high level at seedling,heading and filling stage under heat stress.Furthermore,higher expression levels of OsRboh7 and OsRboh5 were showed in various tissues and organs of rice such as seedling leaves,flag leaf,floret,lemma,palea,stamen,pistil and grain.The higher induced range of gene expression levels in OsRboh7 and OsRboh5 by heat stress was shown in seedling leaves,floret,stamen,pistil and grain.Taken together,OsRboh7 and OsRboh5 were mostly remarkably responsive to heat stress at different growth stages among the OsRboh family genes in rice,which indicated that OsRboh7 and OsRboh5 played a vital role in the regulation of ROS formation pathway in rice under heat stress condition.

In order to determine the suitable sowing date of green manure for multiple cropping rape in Hetao irrigation area,field experiments were carried out in Hangjinhou Banner,Bayannur City,Inner Mongolia Autonomous Region in 2019 and Dalad Banner,Ordos City in 2022.There were five sowing dates in Hangjinhou Banner experimental site from July 26th to August 15th,2019,and one sowing date was set every five days,which were sown on July 26th,July 31st,August 5th,August 10th and August 15th,respectively.Similarly,from July 22nd to August 11th,2022,the experimental site in Dalad Banner was set up with five sowing dates,and one sowing date was set every five days to study the effects of different sowing dates on the green manure biomass,nutrient content and nutrient accumulation of rape.The results showed that although the biomass level and climate conditions of the two experimental sites were different,the overall trend showed a decline with the delay of sowing date.Compared with the first sowing stage,the rape biomass in the fifth sowing stage decreased by 90.3% and 75.4% respectively,and the average active accumulated temperature,effective accumulated temperature and sunshine hours in the two-point growth period decreased by 469.9 ℃,409.9 ℃ and 179.1 h,respectively.At the same time,when sowing in the first,second and third stages,compared with the nutrient input (N 27.3 kg/ha,P₂O₅ 34.5 kg/ha),the nutrient input of rape was higher than the input,and the amount of carbon and potassium returned to the field was at least C1 800 kg/ha and K₂O 200 kg/ha.Therefore,considering the biomass of rape green manure and the amount of nutrients returned to the field,rape could be sown early after wheat harvest in Hetao irrigation area,making full use of light and temperature resources to promote rape growth and nutrient accumulation.In order to achieve more than 5 t/ha of rape returning to the field and obtain a higher amount of nutrients returning to the field,at least 1 300 ℃ of active accumulated temperature,1 100 ℃ of effective accumulated temperature and 640 h of effective sunshine hours should be accepted during the rape growth period.

As one of the main products of photosynthesis,sucrose has a variety of physiological functions in plants.As a group of glycosyltransferase enzymes,sucrose synthase plays an important role in the process of sucrose metabolism.In plants,sucrose synthase reversibly catalyzes the decomposition of sucrose and nucleoside diphosphate(NDP)to produce nucleoside diphosphate glucose(NDPG)and fructose.This reaction is the most important rate-limiting step in sugar metabolism.Sucrose synthase exists in the form of a gene family in plants.Phylogeny shows that there are three subgroups of sucrose synthase,and different subgroups have different expression patterns.Recent studies have shown that sucrose synthase has several major physiological functions in plants,such as participating in the synthesis of macromolecular sugars,response to adversity,participating in reproductive growth and regulating fruit quality.This article reviews the research progress of sucrose synthase in recent years,and looks forward to future research directions in order to provide a reference for future research.

In order to explore the interaction between ScPRT1 and Sporisorium scitamineum genes in sugarcane,this study used Saccharum spontaneum and Saccharum as research objects.The ScPRT1(GenBank Accession Number:MT747433)gene was cloned by RT-PCR and was analyzed by bioinformatics,quantitative Real-time PCR and subcellular localization.Bioinformatics analysis showed that the full-length cDNA of the ScPRT1 gene was 1 621 bp,including a complete open reading frame with 1 260 bp in length and encodes 419 amino acids.The ScPRT1 protein with two RING domains and one ZZ domain,was 46.56 ku in molecular weight and was an acidic and unstable hydrophilic protein with nuclear localization signal and no signal peptide.The higher structural elements of protein were mainly random coil.The results of qRT-PCR analysis showed that ScPRT1 was higher expressed in stem pith than in bud,epidermis and leaf.And ScPRT1 was significantly up-regulated under abscisic acid(ABA)stress and in the smut-resistant variety but significantly down-regulated in the smut-susceptible variety under Sporisorium scitamineum stress.The result of subcellular localization showed that ScPRT1 was localized in the nucleus.In the co-expression network between ScPRT1 and Sporisorium scitamineum genes,three genes that encoded effectors with aromatic amino acid residues at the N-terminal were found co-expressing with ScPRT1,and their proteins may interact directly.It is speculated that ScPRT1 plays an important role in sugarcane hormone signal transduction and response to smut infection.

In order to clarify the sequence characteristics of the MYB62 transcription factor,expression changes after stress,and further explore the biological function of the MYB62 transcription factor,kohlrabis were used as plant materials.MYB62 transcription factors were obtained by homologous cloning method and bioinformatics was analyzed.Spatiotemporal expression analysis of MYB62 and its expression after stress were performed by Real-time PCR.Results of gene cloning showed that gDNA length of BocMYB62 gene was 1 353 bp,the length of CDS was 837 bp,which contained four exons and three introns and encoded 278 amino acids.The sequence structure analysis showed that BocMYB62 was a hydrophilic protein with two SANT-MYB domains,belonging to the R2R3-MYB type in the MYB transcription factor family.The spatial structure prediction revealed a typical α-helix structure.Phylogenetic analysis indicated that BocMYB62 was closely related to MYB62 in Brassica napus.The spatiotemporal expression results showed that BocMYB62 was consistently higher in green kohlrabi than that in purple with clear tissue specificity.The expression of BocMYB62 was significantly increased during drought stress with the highest expression under 12 h.The BocMYB62 expression was significantly lower than control and the lowest at 4 ℃ cold stress.These results inferred that BocMYB62 may be involved in the regulation of anthocyanin biosynthesis and the regulation of stress.It provides a theoretical basis for the subsequent functional identification of MYB62 transcription factors.

To clarify the effects of drought stress and cultivars on the net photosynthetic rate(Pn),the activities of photosynthetic enzymes in flag leaf in the afternoon(FLA)during grain filling stage and grain yield of winter wheat,a pond culture experiment with four water levels and two winter wheat cultivars was conducted under the condition of rainproof pond cultivation in 2019—2021.The four water treatments included severe drought(W1),moderate drought(W2),mild drought(W3),and suitable water supply(W4).The Pn,and the activities of Rubisco,RCA,PEPC,ATPase and PPDK in flag leaf from 14:00 to 16:00 during the early(EGFS)and medium(MGFS)grain filling stage,and the grain yield at maturity of strong drought resistance cultivar Jinmai 47(JM47)and weak drought resistance cultivar Yanzhan 4110(YZ4110)were investigated.Drought stress decreased the FLA Pn and activities of most photosynthetic enzymes,and the grain yield of wheat.The decrease of these indexes increased with drought stress degree,but the effects were different among varieties and years.Compared with W4,the FLA Pn of JM47 under W1,W2 and W3 decreased by 33.6%—40.6%,12.0%—30.5% and 5.0%—13.5%,as well as YZ4110 decreased by 44.0%—52.0%,22.5%—38.1% and 11.5%—20.5%,respectively.Compared with W4,the FLA Rubisco activity decreased during EGFS but increased during MGFS for JM47,while it decreased by 13.3%—25.6%,7.1%—14.0% and 11.2%—11.6% for YZ4110,respectively,under W1,W2 and W3.Compared with W4,the FLA RCA activity significantly decreased under most drought treatments during EGFS,while increased under W2 and W3 for JM47 and decreased under W1 and W2 for YZ4110 during MGFS.Compared with W4,the FLA ATPase activity of JM47decreased under W1 but increased under W3,while that of YZ4110 decreased under W1,W2 and W3 by 19.3%—48.7%,7.2%—24.2% and 0.1%—8.9%,respectively.The FLA PEPC activity under different treatments varied with growing seasons and varieties.Compared with W4,the FLA PPDK activities of JM47 and YZ4110 under W1 were decreased by 12.4%—18.8% and 16.7%—18.2%.Compared with YZ4110,in most conditions,the FLA Pn and photosynthetic enzyme activities of JM47 had no significant difference under suitable water supply(W4),but increased under drought treatments(W1,W2 and W3).The results of correlation analysis showed that yield,FLA Pn were significantly positively correlated to FLA ATPase activity during EGFS and MGFS,as well as FLA PEPC activity during EGFS.Thus,increasing the FLA ATPase and PEPC activities during the grain filling period is conducive to the increase of FLA Pn and grain yield of wheat.

To analyze the same protein of whey and milk fat globule membrane (MFGM)of jersey yak crossbreeds milk and yak milk by proteomics method. The milk yield of jersey yak crossbreeds was higher than that of yak. If the function of jersey yak crossbreeds milk is like that of yak milk,it will improve the economic level of local herdsmen. Whey and milk fat were separated by centrifugation,and proteins were extracted respectively. The same protein was obtained by qualitative and quantitative analysis of protein by tandem mass tag (TMT)technique. GO functional annotation,KEGG metabolic pathway,protein interaction and other bioinformatics analysis were performed on the same protein. Result analysis indicated that 651 whey proteins and 990 MFGM proteins were identified in the peak lactation period of jersey yak crossbreeds milk and yak milk by label quantitative proteomics,and 298 whey proteins and 283 MFGM proteins were screened. GO annotation analysis showed that the same proteins of jersey yak crossbreeds milk and yak milk whey were mainly involved in the negative regulation of endopeptidase activity,immune response and immunoglobulin production. The same proteins of MFGM were mainly involved in the negative regulation of endopeptidase activity and complement activation. The cell components of the same proteins of whey and MFGM in the classical pathway were mainly extracellular space and extracellular region. The molecular functions involved were mainly guanosine 5'-triphosphate (GTP)enzyme activity and GTP binding. KEGG enrichment analysis showed that the main enrichment pathways of the same proteins in jersey yak crossbreeds milk and yak milk whey and MFGM included complement and coagulation cascades,phage and protein processing in endoplasmic reticulum. During the peak lactation period,the proteins with higher abundance in the milk of jersey yak crossbreeds and yak showed excellent performance in immunity and promoting nutrient absorption. However,the milk yield of jersey yak crossbreeds was higher than that of yak,which would increase the economic income of local herdsmen and provide more valuable plateau dairy products for people.

To explore the influence of straw returning years to the field after deep turning back on soil bacterial community in continuous cropping corn fields in the West Liaohe Plain,and provide theoretical guidance for continuous straw returning to improve soil fertility.This study takes corn straw leaving the field as a control (CK),based on the high-throughput sequencing technology of bacterial 16S rDNA V3-V4 region,combined with bioinformatics,we analyzed the characteristics of organic matter,nitrogen nutrients content,soil bacterial community structure and diversity changes in 0-20 cm and 20-40 cm soil layers of corn fields with continuous straw returning for 2(SR2),5(SR5) and 10 a (SR10).The results showed that SR2 had no significant difference with CK in soil organic matter and nitrogen nutrients content,while SR5 and SR10 significantly increased soil organic matter and nitrogen nutrient content;under different straw returning years,soil bacterial diversity was different and each had its own unique OTU,among which SR10 OTU had the largest number;alpha diversity index showed that the relative abundance and diversity of soil bacterial community were not significantly different between SR2 and CK,SR5 and SR10 increased significantly,and SR10 and CK had the largest difference;a total of 51 bacterial groups were obtained at the phyla level,including Actinobacteriota,Proteobacteria,Acidobacteriota,Crenarchaeota and Chloroflexi,whose relative abundance was more than 5%.The relative abundance of Proteobacteria,Acidobacteriota and Chloroflexi increased with the increase of returning years.RDA clustering analysis showed that the soil bacterial community structure of 0-20 cm soil layer SR2 was similar to CK,SR5 was more similar to SR10,20-40 cm soil layer,different straw returning years,treatments were different from CK,SR2 and SR5 were similar,SR10 had significant differences in community structure compared to SR5 and SR2.The contents of soil {\mathrm{NO}}_3^{-}-N, {\mathrm{NH}}_4^{+}-N,OM,and TN all had effects on bacterial flora,and they were soil {\mathrm{NO}}_3^{-}-N> {\mathrm{NH}}_4^{+}-N>OM>TN.The contents of soil organic matter and nitrogen nutrient had no significant difference between continuous straw returning for 2 a and CK,while continuous straw returning for 5 and 10 a were significantly increased.In summary,the microbial diversity and richness of bacteria showed minimal changes in the soil layers of 0-20 cm and 20-40 cm after continuous straws returning for 2 a.After continuous straws returning for 5 and 10 a,both soil layers significantly increased;the bacterial community structure continuous straw returning for 2 a was similar to CK in the 0-20 cm soil layer,while the bacterial community structure continuous straw returning for 2,5,10 a were changed in the 20-40 cm soil layer.

The plant NADPH oxidase Rbohs(Respiratory burst oxidase homologs) is the main source of reactive oxygen species (ROS),which participate in various physiological processes such as plant growth,development,stress resistance and plant-microorganism interaction.In order to explore the function and mechanism of Rbohs in symbiotic nitrogen fixation,GmRbohL,a member of soybean Rbohs gene family,was cloned in this study.The gene expression pattern,protein subcellular localization and gene function were studied by molecular biology,cell biology and genetics,respectively.The results revealed that: GmRbohL gene was induced by rhizobia and expressed specifically in soybean roots and nodules.Subcellular localization analysis indicated that the gene-encoded protein GmRbohL was a membrane protein.The plant gene silencing (RNAi) vector of GmRbohL was constructed,and the transgenic hairy roots were obtained by the transformation of soybean hairy root mediated by Agrobacterium rhizogenes K599.Gene silencing of GmRbohL resulted in a significant reduction in the number of nodules of transgenic hairy roots,and the production of ROS was also inhibited. Gene silencing of GmRbohL reduced the infection of rhizobia at the stage of root nodule organogenesis,and the expression level of nodulation marker genes also decreased with the decrease of GmRbohL expression.The root nodule tissue sections showed that gene silencing of GmRbohL significantly reduced the number of symbionts in the infected area of root nodules,and the nitrogenase activity of root nodules also decreased accordingly.The above data indicated that gene silencing of GmRbohL significantly inhibits the symbiotic nodulation process of soybean by reducing the production level of ROS.It is speculated that GmRbohL may play an important positive regulatory role in organogenesis of soybean nodules and regulation of nitrogen fixation function.

Genomic-wide identification of systematic work on the SKS gene family in cotton has not been reported.Identifying SKS (Skewed5 similar) gene family members and analyzing evolutionary relationship in Gossypium hirsutum,and analyzing the mechanism of GhSKS13 gene regulating cotton disease resistance are expected to provide a new direction for the cultivation of resistant plants.Here,the members of upland cotton SKS family were identified by bioinformatics at the genome-wide based on published genomic data of upland cotton genetic standard line TM-1,and G.hirsutum variety CRI-14 was used as the material.And chromosome distribution,evolutionary relationships,gene structure,and collinearity of SKS family members were predicted.Then,the expression pattern of GhSKS13 was analyzed by Real-time Quantitative PCR and virus-induced gene silencing was employed for preliminary investigation of the function of GhSKS13 in cotton resistance to Verticillium dahliae.A total of 48 upland cotton SKS genes were identified,unevenly distributed on 19 chromosomes,clustered into 5 subgroups with highly conserved gene sequences.Collinearity analysis revealed that the upland cotton SKS gene family was subject to purifying selection.Expression pattern analysis showed that GhSKS13 was predominantly expressed in upland cotton root tissues and significantly up-regulated by V.dahliae infection. GhSKS13-silenced reduced plants resistance to V.dahliae and suppressed the expression of the pathogen-related genes including GhPR1, GhPR2,GhPR3 and GhPR5 compared to control plants. GhSKS13-silenced plants invaded by V.dahliae showed significantly lower hydrogen peroxide (H₂O₂) deposition compared to the control,suggesting that GhSKS13 promotes the formation of reactive oxygen species (ROS).In conclusion,this study clarified the phylogenetic relationships,chromosomal distribution characteristics and gene structure characteristics of upland cotton SKS family members,and elucidated the involvement of GhSKS13 in upland cotton plant resistance to V.dahliae.

In order to reveal the molecular mechanism of MiPDCD6 protein suppressing tomato PTI immunity,the MiPDCD6 overexpression seedlings of tomato variety Xinjinfeng 1 were used as experimental materials,and the tissue culture seedlings of tomato variety Xinjinfeng 1 were used as control.Transcriptome sequencing was performed on tomato MiPDCD6 overexpressing seedlings and control seedlings,respectively.With tomato cultivars Heinz 1706,comparing the genome as a reference genome,FPKM method was used to calculate quantity of gene expression,set parameters(|log₂ FC|>1 and P<0.05)in screening the differentially expressed genes(DEGs).Gene ontology(GO)database was used to analyze the GO functional enrichment of DEGs,count the number of DEGs in each GO term,calculate the significance of gene enrichment,and find out the functional term with significant enrichment.KEGG database was used for Pathway enrichment analysis of DEGs,and hypergeometric distribution test was used to calculate the significance of enrichment of DEGs in each Pathway.The enrichment degree of KEGG was measured by FDR and gene number.Combined with gene differential expression analysis and functional enrichment analysis,the effect of MiPDCD6 protein on tomato PTI immune-related pathway genes was studied.The results showed that there were 2 366 DEGs in MiPDCD6 overexpressed tomato plants compared with wild-type tomato,including 1 354 up-regulated genes and 1 012 down-regulated genes.In these DEGs,a large number of differentially expressed genes were concentrated in KEGG pathways such as plant hormone signal transduction(sly4075),plant-pathogen interaction(sly04626),plant MAPK signal pathway(sly04016)and procycloid biosynthesis(sly00940)through GO and KEGG annotation.SA biosynthesis pathways included ICS and PAL.In the MiPDCD6 overexpressed tomato plants,PAL1 and PAL-like genes in SA synthesis pathways and TGA9,TGA10-like and PR1a2 genes in SA signal transduction pathways were significantly down-regulated,suggesting that MiPDCD6 may inhibit SA synthesis thus inhibiting plant PTI immunity.

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.

In order to promote the breeding of high quality grain shape Japonica rice varieties,a natural population consisting of 295 Japonica rice varieties collected from around the world was used as test material for phenotypic analysis of five grain shape-related traits,including grain length,grain width,grain thickness,aspect ratio and thousand grains weight,between 2018 and 2019,combined with 788,396 SNPs obtained by resequencing,and genome-wide association analysis using the MLM model of TASSEL 5.0.Haplotype analysis was performed on all the genes in the QTL intervals that were jointly detected during the two years and controlled multiple shape-related traits, and candidate genes for high quality rice grains were mined by combining the results of previous studies and functional annotation of the genes. The results showed that there was extensive phenotypic variation for the five grain shape-related traits,all of which were approximately normally distributed,and most of the grain shape-related traits were significantly or highly significantly correlated with each other. And a total of 221 QTLs significantly associated with rice grain shape-related traits were detected under the threshold condition of P≤5.46×10^-6,which were distributed on all 12 chromosomes of rice,and phenotypic contribution was 8.62%-20.73%, of which, seven QTLs were jointly detected in two years, and combined with the haplotype analysis and functional annotation of the genes, it was hypothesized that LOC_Os12g 44290 was a new candidate gene for rice grain shape. In summary,this study used genome-wide association analysis for QTL localization and candidate gene analysis for grain type-related traits in 295 Japonica rice varieties,providing a theoretical basis for breeding Japonica rice varieties with high quality grain types.

In order to reveal the influence mechanism of sowing date adjustment on crop yield formation, a field experiment was conducted to adjust the sowing date of the same variety of summer maize from 2019 to 2021 at Hebei Gucheng Agricultural Meteorology National Observation and Research Station in Northern north China, which was set up in four sowing date, as 10 d early sowing, 10 d late sowing, 20 d late sowing and control. By monitoring the growth period changes, plant dry matter accumulation, leaf photosynthetic characteristics and grain filling characteristics, and sampling at maturity to determine the yield components, the results showed that:the sowing date of summer maize was appropriate to be advanced,the seedling stage,ear stage and the whole growth period were extended,especially,the duration of effective filling was extended with the advance of sowing date.The effective grain filling duration days were extended by 4.7 days for every 10 days ahead of the sowing date.The average filling rate of winter wheat was 4.04% higher on the sowing date was advanced by 10 days, compared with the sowing date was deferred by 10 or 20 days and the control.Grain filling accumulation increased,and 100-grain weight increased by 5.459 g.The main yield components of summer maize,such as grain number per ear,grain weight per ear and 100-grain weight showed significant differences during sowing,and increased with the sowing date.The theoretical yield increased by 1 395.4 kg/ha with 10 days ahead of the sowing date during the experimental period from June 8 to July 8.The average net photosynthetic rate(Pn)of leaves in the critical development period of summer maize increased by 0.764 μmol/(m²·s)with every 10 days ahead of sowing date,and the average Pn of sowing date 10 days earlier was increased by 7.31% compared with the control and late sowing 10 and 20 days.With the increase of photosynthetic rate,dry matter production,accumulation and transfer to grain were increased,and the grain weight per ear and 100-grain weight increased by an average of 24.01% and 18.00%,respectively,compared to the control and late sowing 10 and 20 days.Advancing sowing date resulted in low plant height,strong stems,lodging resistance,large individual green leaf area,high leaf area index(LAI),and high leaf photosynthesis ability,above-ground dry matter allocation rate at maturity stage every 10 days in advance of sowing date:grain weight improved by 2.26%,the source—pool distribution of dry matter between plant vegetative organ and ear changed,ear weight per ear,100-grain weight and grain yield increased.The research showed that the double cropping area of winter wheat—summer maize in north China could effectively improve the yield per unit area by making full use of the heat resources increased by climate warming,reasonably allocating stubble,sowing summer maize early at the appropriate time,and extending the development period and grain filling time.

12-oxo-phytodienoic(OPR) acid reductase is a flavin mononucleotide-dependent oxidoreductase,a key enzyme for the synthesis of jasmonic acid,which is important for plant growth, development and defence regulation.In order to study the disease resistance effect of OPR gene in maize, bioinformatics methods were used to identify OPR family members in 31 different maize inbred lines and the expression pattern after infestation by S. turcica was analyzed.The results showed that eight OPR genes were identified in maize line B73 and they were unevenly distributed on seven chromosomes.In addition,these expressed proteins of OPR genes were rich in acidic amino acids.Further analysis revealed that the maize OPR family contained only one structural domain,Oxidored_FMN,and all OPR members contained 10 protein conserved motifs identified.Phylogenetic relationship analysis of OPR family members from maize,wheat,rice and Arabidopsis using MEGA software revealed significant differences in the evolutionary relationships of OPR gene families in these plants.The OPR proteins of maize and those of rice were the closest in evolution.Homology analysis using OrthoFinder software revealed that the maize OPR gene family was highly conserved,with all OPR genes being core genes,but with slightly different functions.According to the previous RNA-seq data of Hebei key Laboratory of Plant Physiology and Molecular Pathology,we analyzed the expression pattern of OPR genes of maize B73 in response to S.turcica infection and verified the gene expression pattern of OPR genes through qRT-PCR.It was found that there were three different expression patterns in the process of S.turcica infection.This study systematically identified the maize pan-genomic OPR family genes and their expression patterns in response to S.turcica infection.

Photoperiod is one of important environmental factors affecting plant bolting and flowering which are regulated by plant phytochrome proteins.The signal network mediated by PHYB has an important inhibitory effect on plant bolting and flowering.Previous studies revealed that there were large segment insertion/deletion differences between the PHYB gene promoter of the Chinese cabbage late-bolting line 06-247 and the easy-bolting line He102.In order to further investigate the impact of promoter mutation on PHYB and the key genes of its downstream pathways,this study was conducted.Based on bioinformatics methods,the redundancy characteristics of phytochrome genes in genome Chinese cabbage were firstly analyzed.It was found that the Chinese cabbage genome contained six phytochrome genes,of which PHYA had two copies,and PHYB,PHYC,PHYD,and PHYE all had only one copy.Then amino acid sequence alignment was used for screening of the specific sequence of PHYB.Antigenic determination clusters were designed based on the specific sequence and the antibodies against PHYB was prepared.The fluorescence quantitative RT-PCR technology and Western Blot technology were used to study the relative content of PHYB in 06-247 and He102.At the same time,the dynamic changes of key regulatory genes such as CCA1,FLC,CO and FT which in the downstream of PHYB pathway were also compared.The results showed that the promoter mutation caused significant differences both in level of mRNA in 06-247 and then significantly increased the protein level of PHYB.At the same time,the downstream regulatory genes such as CCA1,FLC,CO and FT were highly expressed in 06-247,which had an important impact on bolting resistance of 06-247.

In order to study the response mechanism of pepper CaWRKY30 transcription factor and Tomato spotted wilt orthotospovirus,it was experimental materials with pepper Xiangyan 11.The CaWRKY30 coding sequence was obtained by RNA extraction,RT-PCR,split gel and cloning.Biological information analysis results showed that CaWRKY30 full length was 1 122 bp,encoding 373 amino acids,the gene encoded protein contains 1 WRKY conservative domain and 1 C₂H₂ domain,belonged to a typical Ⅱ(e)subfamily member.System evolution analysis showed that the relative relationship with the potato StWRKY22 amino acid sequence was recently.It was found that CaWRKY30 was positioned in the nucleus and cell membranes in its cigarette seedlings,and leads to cell membranes.The results of Real-time fluorescence quantitative PCR analysis showed that the viral accumulation of Tomato spotted wilt orthotospovirus mechanical friction-vaccination was found that the viral accumulation was gradually increased from 1 to 14 days after inoculation,and virus accumulation reached its maximum in 14 days,after inoculation 14 days,viral accumulation gradually declined.At the same time,CaWRKY30 was induced by Tomato spotted wilt orthotospovirus,when the inoculation 1—14 days,the CaWRKY30 expression was raised,and the peak was reached in 14 days,the expression in 14 days gradually decreased.In summary,it obtained the CaWRKY30 transcription factor gene sequence,which was located in the nucleus and cell membrane,and preliminarily explained the expression trend of CaWRKY30 transcription factors under the stress of Tomato spotted wilt orthotospovirus.

In order to study the changes of genes and metabolites pre- and post-flowering in Aurea helianthus,transcriptome and metabolome techniques were used to detect flower buds and flowers of Aurea helianthus. Through transcriptomic analysis,206 636 Unigenes were identified and 42 618 differentially expressed Unigenes were identified,including 63 differentially expressed transcription factor families and 24 transcription regulatory factor families. GO analysis showed that the differentially expressed genes were mainly enriched in biological processes such as retrograde vesicle-mediated transport. KOG functional annotation showed that the function of differentially expressed genes was mostly general function prediction only,followed by signal transduction mechanism,posttranslational modification,protein turnover,chaperones,and carbohydrate transport and metabolism. KEGG enrichment analysis showed that the differentially expressed genes were mainly enriched in metabolic pathways such as metabolism,plant hormone and signal transduction,starch and sucrose metabolism. A total of 135 metabolites were detected by metabolomics analysis,including lipid,flavonoids,amino acids and derivatives. KEGG enrichment analysis showed that differences metabolites mainly enriched in glycerophospholipid metabolism,glycosylphosphatidylinositol(GPI)-anchor biosynthesis and biosynthesis of secondary metabolites-unclassified,notably,valine,leucine and isoleucine degradation appeared simultaneously in the genes and metabolites KEGG enrichment Top 20 metabolic pathways.

The purpose of this study was to preliminarily analyze the promoter activity and transcription regulatory elements of porcine CRYBB2. Using bioinformatics analysis,PCR amplification,gene cloning,cell transfection,double luciferase activity analysis and other methods,we obtained the sequence characteristics of CRYBB2 promoter region,constructed double luciferase reporter gene vectors of CRYBB2 promoter region with different fragment lengths,analyzed its luciferase activity,and then determined the core promoter region and key regulatory region of CRYBB2.Finally,the transcription factors and their binding sites in key regulatory regions were predicted. The results showed that the candidate promoter region of CRYBB2 might contain four core promoters and one CpG island.-52—-3 bp might be the core promoter region of CRYBB2 gene,and-505—-19 bp might be the key regulatory region of CRYBB2 gene promoter,which played a positive regulatory role,while-2 060—-505 bp didn't have any regulatory elements that affected the activity of CRYBB2 gene promoter.The key regulatory region of CRYBB2 promoter contained multiple potential transcription factor binding sites,such as TBP,NFIA,FOXP1,NKX2-8,KLF4,Tcf3,Crx,SNAI3,Rfx1 and CREB1. This study laid a foundation for further study on the expression mechanism of porcine CRYBB2.