| In order to provide reference basis for the protection and utilization of Chinese wild grapevine germplasm resources, phylogenetic relationship and genetic diversity of grapevine germplasm resources were analyzed by SSR molecular markers in this research. Fifteen pairs of nSSR primers were used to analyze the genetic relationship of79grapevine resources collected from grapevine test base of Nanjing Agricultural University. Due to the chloroplast complete sequence of grapevine, seven pairs of cpSSR primers were designed to identify the female parent of a high-quality seedling’2004-2-5’combined with fifteen nSSR primers. Eight pairs of nSSR primers were used to analyze the genetic diversity of9populations (286individuals) of Vitis bryoniaegolia Bge. collected from Nanjing region and3populations (40individuals) of Vitis davidii Foex. collected from Huangshan region in Anhui province.1.15pairs of polymorphic SSR primers were used to analyze the phylogenetic relationship of79grapevine germplasm resources. A total of158alleles were amplified with the percentage of polymorphic loci was100%.13.9%of the alleles belonged to Chinese wild grapevine germplasm resources as special alleles.15pairs of SSR primers could effectively identified67varieties, with a ratio of discriminate84.8%. The genetic similarity index ranged from0.72to1.00.79grapevine varieties were divided into two clusters at the genetic similarity index of0.72through the unweighted pair-group method arithmetic mean (UPGMA), with one group including rootstock varieties’99R’(V. berlandieri Planch.×V. rupestris Scheels.) and ’5BB’(V. berlandieri Planch.×V. riparia Michx.), Chinese wild grapevine germplasm resources V. davidii Foex. and V. bryoniaegolia Bge., and the other one including all the V. vinifera, V. vinifera×V. Labrusca, and three high-quality seedlings. The results indicated a long genetic distance between rootstock grapevine varieties and table grapevine.2.’2004-2-5’, both parents unknown, was a high-quality grapevine seedling selected through the seedling selection method. Due to the complete sequence of grapevine chloroplast,26pairs of cpSSR primers were designed, among which7pairs were selected with good amplification effect and polymorphisms.15pairs of nSSR primers were used to analyze the LOD score between’2004-2-5’and its55candidate parents. The results indicated that the parent combination of’Seto Giants’ and’Christmas Rose’had the maximum possibility. The second parent combination was ’Wink’ and ’Seto Giants’.’Seto Giants’was a male sterility variety, having the maximum possibility to be the female parent. Cpssr analysis showed that ’Wink’ was different from’2004-2-5’, so ’Wink’ could be excluded from the female parent candidates.3. Wild Vitis bryoniaegolia Bge. grapevine germplasm resources were widespread in Nanjing region.8pairs of SSR primers were used to analyze the genetic diversity of nine Vitis bryoniaegolia Bge. populations (286individuals) in Nanjing region. The parameters indicated that99alleles were amplified from the286individuals by8pairs of primers, the percentage of polymorphic loci was100%; the average effective number of alleles (Ne) was3.127; observed heterozygosity (Ho) was0.451; expected heterozygosity (He) was0.581; Nei’s diversity index (H) was0.580; Shannon diversity index (Ⅰ) was1.392and polymorphism information content (PIC) was0.559; fixation index ranged from-0.047to0.750. The results above indicated that the wild Vitis bryoniaegolia Bge. grapevine germplasm resources as investigated were at high genetic diversity level. Among these nine populations, the results were showed as follows:average observed number of alleles (Na) ranged from3.125～9.000, and average effective number of alleles ranged from2.072～3.124; observed heterozygosity (Ho) ranged from0.359to0.486; expected heterozygosity (He) ranged from0.417to0.591; Nei’s diversity index (H) was0.391～0.588; Shannon diversity index (Ⅰ) was0.728～1.328; Fst and Nm were0.025～0.154and1.375～9.793, respectively; genetic identity (Ⅰ) and genetic distance (D) ranged from0.647～0.974to0.027～0.435. Among populations, there were widely gene flow and a low level of genetic differentiation. Geographic distance might be the principle but not the only ingredient affected the genetic identity and genetic distance. Sometimes the objective factors such as habitat and human activity might also be the influencing factors.4. Eight microsatellite (SSR) markers were used to analyze the genetic diversity of40individuals of wild Vitis davidii Foex. collected from Huangshan region, Anhui province. The results showed that36alleles were amplified by the eight SSR loci used, with a range of2-10alleles per locus. The parameters indicating the diversity of40individuals of V. davidii Foex. were showed as follows:the effective number of alleles (Ne) ranged from 1.285to5.378; observed heterozygosity (Ho) ranged from0.250to0.775; expected heterozygosity (He) ranged from0.224to0.824; Nei’s diversity index (H) ranged from0.222to0.814; Shannon diversity index (I) ranged from0.417tol.884and polymorphism information content (PIC) ranged from0.202to0.789. Based on the geographic location,40individuals collected from Huangshan region were divided into3groups (POP1, POP2, POP3). Among these three groups, the genetic identity and genetic distance ranged from0.829to0.912and0.093to0.188, respectively, indicating a low level of genetic differentiation among populations of these three locations. The clustering dendrogram of40individuals from Huangshan and3individuals from Hunan and Fujian was constructed through UPGMA method.43individuals were divided into two groups while the genetic similarity coefficient was0.62.3individuals from Hunan province and Fujian province were clustered as one group. These results indicated that the gene flow of V. davidii Foex. might be limited by the distance and the principle reproductive mode was sexual reproduction. |
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