Molecular Markers Development And Gene Cloning Of Heading Time And Plant Height Genes From Foxtail Millet

Foxtail millet is high photosynthetic efficiency,drought-tolerance and rich nutrition crop,but the molecular breeding research still lags far behind other staple crops such as rice and wheat.The exploring of millet molecular markers and positional cloning of time to flowering and plant height genes has very important significancefor the permanent development of millet industry.In this study,we use highthroughput sequencing technology to obtain whole genome sequencing date of Jingu21,GBS,B168 and the F2 population derived from a cross between a dwarf and early heading(GBS)and a high and late heading(B168)millet parents,such as extreme early heading pool,late heading pool,dwarf plant pool and high plant pool.The results are as follows.(1)169037 Indel and 1167555 SNP molecular marker were indentified based on Jingu 21 resequencing data,14578 In Del sites were suitable for agarose gel detection,69 In Del and 1 SNP locus were selected and verified.These results showed that the In Del and SNP markers obtained by secondgeneration sequencing technology were true and reliable,and they can be used for identification of hybrid species and germplasm resources,such as green bristlegrass and foxtail millet.2G5509176,a specific In Del marker of Jingu 21 was developed for the identification of Jingu 21 and its derivatives.(2)Based on the analysis of genome resequencing data to GBS,B168,extreme early heading pool and late heading pool.The heading date gene was located in the 3.42 Mb interval of chromosome 9,which was annotated with 19 site of frameshift mutation among parents.We further fine map heading time genes by the mapping cloning technology.The heading time gene was located in the 511.5 kb interval of chromosome 9.(3)The plant height gene was located in the 4.27 Mp region of chromosome 5.There were 40 nonsynonymous mutations among parents in this region.The candidate gene Si SD1 related to plant height was identified according to the bioinformatics analysis of GBS,B168,extreme high plant pool and drawf plant mix pool.In addition,the mutant gene from GBS was named sisd1.The G base deletion located in the third exon at 2266 in sisd1,resulting in a frameshift mutation and premature termination,which ultimatelyresults in the deletion of 83 amino acids.Further,Si SD1 and sisd1 genes were cloned,two vector of p BASi SD1(WT)and p BA-sisd1(GBS)were successfully constructed.These two vector were transferred into grobacterium GV3101,which were transfected into wild-type Arabidopsis col and dwarf mutant Arabidopsis cs62 and SALK-016701 C by inflorescence infection method for verificate function of gene,and T0 seed was obtained.The research can provide theoretical foundation for future molecular marker-assisted breeding and genome-modified breeding.