Genetic Map Construction In Unisexual Vitis Amurensis Rupr.

The cold hardy and pathogen resistant Vitis amurensis Rupr.is a valuable plant material for grapevine breeding.In this study the female amurensis cultivar 'Zuoshanyi' and male accession '043' were used as the crossing parents to establish an F1 mapping population.SSR and SRAP molecular markers were used to construct the V amurensis molecular genetic map.The map could provide some theoretical evidence for further mapping of sex locus.A good foundation can.also be laid for cold hardiness and pathogen resistance breeding.The main results of this study were as follows:1.SSR primers producing both maternal and paternal specific bands were obtained.Authentic identification was done for 148 seedlings,resulting 142 true hybrids with a true hybrid rate of 95.5%.The 142 true hybrids were used as the mapping population.2.The SSR-PCR and SRAP-PCR reaction system was established and optimized.For SSR-PCR,.the 16 ?l reaction system contained 1.5ng template DNA,1.0mmol/L Mg2+,0.6mmol/L dNTPs,0.5U TaqDNA polymerase and 0.5mmol/L of each upstream and downstream primer.For SRAP-PCR,the 16 ?l reaction system contained 0.9ng template DNA,0.8mmol/L Mg2+,1.2mmol/L dNTPs,0.6U TaqDNA polymerase and 0.6mmol/L of each upstream and downstream primer.The reliability of the reaction systems were verified and can be used for hybrid authentic identification as well as genetic map construction.3.153 SSR primers,which were heterozygous in at least one parent,were obtained.They showed up polymorphism between the parents and produced separated bands among the progenies.96 separating sites were obtained in paternal parent,and 107 on maternal parent.167 SRAP primers with good polymorphism were also used for the amplification of the mapping population resulting in 258 separating sites.4.The molecular genetic map of 'Zuoshanyi' was constructed,including 19 linkage groups covering a total length of 1551.6 cM.The average length of each linkage group was 81.7cM.And the average distance between two adjacent markers was 5.6cM.The average marker number for each linkage group was 14.6.The minimun distance between two markers was 0.1 cM.The molecular genetic map of '043' was constructed,including 19 linkage groups covering a total length of 1540.9 cM.The average length of each linkage group was 81.1cM.And the average distance between two adjacent markers was 5.3cM.The average marker number for each linkage group was 15.3.The minimum distance between two markers was 0.3cM.