Mapping Of Sex Locus Based On BSA Analysis In Vitis Amurensis

Vitis amurensis Rupr.is one of the most cold resistant species in grape genus.It has strong resistance to diseases that are susceptible to a variety of Eurasian populations.It is currently a cold-resistant and disease-resistant breeding grape in China and abroad.The main germplasm resources.The wild grape is generally dioecious and hermaphroditic is extremely rare.In this study,we used ‘Zuo shan yi’ and ‘043’ as the test materials.We used SSR molecular marker technology combined with BSA(cluster separation analysis method)to study the sex of Vitis amurensis Rupr.Related molecular markers to enable early identification of grape sex and speed up the grape breeding process.This study has achieved the following main results:1.Two pairs of SSR primers with parental specific bands were screened by SSR-PCR technology,and 117 hybrids were identified for authenticity of the hybrids.There were clear and stable parental specific types in 108 hybrids.With the combination of field morphological observations,the true hybrid rate was determined to be 92.3%.2.Using SSR combined with BSA technology to carry out the gender difference table and screening of the hybrid F1 generations of ‘Zuo Shan Yi’ and ‘043’ hybrids.First,the male and female gene pools(20 plants)were constructed and then scanned with 190 pairs of microsatellite primers.It was found that three pairs of primers(VMC3B10F,Y-2-11 B,VMC2C10-1)could be amplified between the female gene pools.Specific bands,both appearing in the male and female gene pools.Three pairs of primers were individually verified with 40 plants that constructed a gene pool,and the three pairs of primers were able to amplify bands in 40 individuals.These three pairs of primers all originated from the grape No.2 linkage group.Therefore,it is presumed that the gene controlling the S.vitis sex locus is located in the No.2 linkage group.3.Primers were designed on linkage group 2 and 9 specific markers were screened on the parents “Zuo Shan Yi” and “043”.The 9 pairs of primers were microsatellite markers on108 individuals in the F1 generation.Joinmap3.0 performs linkage positioning analysis.There are four markers linked to the S.giganteus sex linkage group.The two markers closest to the sex locus are the markers VMC3B10 and Y-2-11 B,and the distance from the sex locus is 2c M and 7c M,respectively.4.Based on the SSR molecular markers and the re-sequencing BSA analysis results,itwas shown that the sex determination site of the mountain grape is located on the 2nd linkage group.Combined with the results of SSR molecular markers and re-sequencing BSA analysis,the region targeted by the sex locus can be reduced to between 5,082,666 bp and 574,001 bp,and within a range of approximately 650 kb.