Genetic Analysis Of Resistance Gene And Spotted Leaf Gene In Wheat N0883 And Cloning And Expression Analysis Of TaGeBPL Gene

Stripe rust and powdery mildew occur frequently in our country,which have a serious impact on wheat production.It is the most effective measure to explore new resistance genes of wheat and breed resistant varieties to solve this problem.Studies have found that the spotted leaf mutants in plants can produce necrosis or death spots similar to the HR response phenotype of resistance genes.Spotted leaf genes can activate immune system of plants and improve the resistance of plants to pathogens.The regulation of stripe rust resistance genes,powdery mildew resistance genes,and spotted leaf genes in wheat all belong to programmed cell death.To study resistance genes（stripe rust resistance gene,powdery mildew resistance gene）and spotted leaf genes in wheat,N0883（both resistance to stripe rust and powdery mildew;spotted）,Feng 1718（susceptibility to stripe rust;no spot）,ST34（susceptibility to stripe rust;no spot）,Yuanfeng 175（susceptibility to powdery mildew;no spot）,N0658（susceptibility to powdery mildew;no spot）as the parent,the offspring populations constructed by the five parents was used to perform genetic analysis on these three genes to test the alleles of resistance genes and spotted leaf genes.Meanwhile,the gene locations were analyzed by molecular markers and SNP gene chips.Based on the previous transcriptome data of RILs（N13039H/N）,phenotyping spotted Vs.normal leaves,the TaGeBPL gene was cloned from wheat using RT-PCR technology,and its expression analysis,subcellular localization,and transcription factor self-activation detection were performed.The main results of this study were as follows:1.Two F₂ populations（N0883/Feng1718,N0883/ST34）were constructed from the parents N0883,Feng 1718 and ST34,and four different phenotypes were isolated from the two F₂ populations:resistance to stripe rust and no spot,resistance to stripe rust and spots,susceptibility to stripe rust and no spots,susceptibility to stripe rust and spots.There was a segregation of traits in the F₂ population,which indicated that the stripe rust resistance gene and the spotted gene were located on different chromosomes.2.The F₂ population of the N0883/Feng 1718 hybrid descendants and the BC₁F₁population of the N0883/Feng 1718//Feng 1718 backcross descendants were subjected to genetic analysis with chi-square test analysis for resistance to stripe rust.The results showed that the resistance of N0883 was determined by a single dominant gene.55K SNP gene chips and SSR molecular marker screening results showed that the resistance to stripe rust gene was located on the second homologous group of wheat.The genetic analysis of powdery mildew resistance was performed on the F₂ population of the N0883/Yuanfeng175 hybrid with chi-square test,and the results showed that the resistance to powdery mildew was a single recessive gene in N0883.660K SNP gene chips analysis showed that there were 2286 different SNPs between powdery mildew resistant and susceptible mixed pools,mainly distributed on 1B,2A,5A,5B,6A chromosomes.There were 1240differential SNPs on chromosome 2A and 238 differential SNPs on chromosome 6A,accounting for 51.6%and 9.9%of the total number of differential SNPs.It was preliminarily speculated that the powdery mildew resistance gene might be located on chromosome 2A or 6A.3.Based on the previous transcriptome data of RILs（N13039H/N）,phenotyping spotted Vs.normal leaves,in this study,a GeBP（GLABROUS1 enhancer-binding protein）gene was cloned from wheat“Xinong N13039H”.The CDS coding region of this gene was1527 bp,encoding 508 amino acids,which contained a DUF573 domain and 3 nuclear localization signals.Inter Pro comparison showed that the encoding protein belonged to the GeBP family and then the gene was named TaGeBPL.The results of q RT-PCR indicated that TaGeBPL gene might be involved in the related pathway to pathogen response under the infection condition of powdery mildew bacteria E09.The subcellular localization results showed that the TaGeBPL was localized in the nucleus.The yeast transcription activation experiments indicated that the TaGeBPL transcription factor had no transcriptional self-activation activity.