Cytoplasmic Genome Analysis Of YS-type Wheat Male Sterile Lines And Identification Of Candidate Genes For Male Sterility

Wheat(Triticum aestivum L.)is an important food crop,and its yield and quality can be effectively improved by using heterosis.Male sterile lines are very important in the utilization of heterosis,but the mechanism of male sterility in wheat is still not explained systematically.This study mainly explored the cytoplasmic sterility genes of K-type wheat male sterile line K519 A and temperature-sensitive male sterile line YS3038,using K519 A and its isotype maintainer line 519 B and temperature-sensitive male sterile line YS3038 as materials,using the second generation High-throughput sequencing technology to sequence,assemble and splice the cytoplasmic genomes of the three materials of wheat,compare and analyze the genome sequence to obtain candidate differential genes,and use the barley stripe mosaic virusinduced gene silencing(BSMV-VIGS)method to perform gene silencing Verification of fertility function,further screening of candidate genes related to male sterility.The main results obtained are as follows:1.Using the second-generation high-throughput sequencing technology,the complete chloroplast genome data of K519 A,519B and YS3038 were obtained,and the assembly and splicing of the data were completed.The chloroplast genome of wheat male sterile line K519 A is 136,996 bp in length,of which the large single copy region(LSC)fragment is 811,36 bp long,the small single copy region(SSC)fragment is 12,776 bp long,and the inverted repeat region(IR)is 21,542 long bp,the genome G/C content is 38.27%.The chloroplast genome of519 B is 136,235 bp in length,LSC is 80,335 bp(LSC),SSC is 12,796 bp and IR is 21,552 bp,and the genome G/C content is 38.28%.The total length of YS3038 chloroplast genome is136,919 bp,LSC is 81,059 bp(LSC),SSC is 12,776 bp,IR is 21,542 bp,and the genome G/C content is 38.28%.2.A total of 104 gene base mutation sites were found after the comparison of K519 A and519B.The mutations occurred in 42 chloroplast genes,of which 16 genes were nonsynonymous mutant genes;among the chloroplast coding genes between K519 A and YS3038 There are no base mutations.3.Using the second-generation high-throughput sequencing technology,the complete mitochondrial genome data of K519 A,519B and YS3038 were obtained and the assembly and splicing of the data were completed.The total length of the K519 A mitochondrial genome is420,543 bp,and the G/C content of the genome is 44.14%;the total length of the 519 B mitochondrial genome is 433,560 bp and the G/C content is 44.14%;the total length of the YS3038 mitochondrial genome is 452,567 bp,and the G/C content is 44.35%.After comparing the mitochondrial genomes of K519 A and 519 B,a total of 14 differential genes and 83 differential open reading frames were found.After comparing the mitochondrial genomes of YS3038 and K519 A,it was found that there were 10 differential genes and 122 differential open reading frames in YS3038 and K519 A.4.After performing qPCR on the candidate genes,it was found that the expression levels of most genes were significantly different between the binucleate stages and the trinucleate stages,which was consistent with the period of wheat fertility transition.After BSMV-VIGS verified the fertility function of the candidate genes,it was found that the chloroplast atp B and ndh H genes may be related to the fertility of K519A;the mitochondrial cox1 gene caused the decrease of YS3038 fertility under fertile conditions.It is speculated that the expression of cox1 gene may be related to YS3038 temperature-sensitive characteristics.