Genetic Diversity And Hybid Purity Identification Of Lycopersicon Esculentum Var. Cerasiforme Germplasms By RAPD Analysis

In this paper,the technique of RAPD molecular markers was firstly used to study genetic diversity and molecular identification of 32 native and abroad Lycopersicon Esculentum var.Cerasiforme Alef cultivars from Fujian Agricultural Academy of Science and to analyse their genetic relationship and germplasm resources in order to offer theoretical foundation for identification, selection, and hybridization breeding.An optimized PCR reaction system for RAPD analysis was established: PCR was performed in a 25ul reaction mixture with 25ng DNA, 0.15mmol.L^-1 of each dNTP, 0.4μmol.L^-1 random primer,1U Taqpolymerase, 1.5mmol.L^-1 MgCl₂ and 2.5μL10XPCR buffer The temperature profile used for PCR was 95℃ for 5min,followed by 45 cycles of 94℃ for lmin,38℃ for lmin,72℃ for 2min,and was terminated with a 10 min DNA extension step at 72 ℃.The detected materials included thirty-two cultivars. Twenty-seven primers selected from two handred primers amplified 214 RAPD fragments and 139 bands that occupied 64.95% of the whole amplified showed polymorphisms. Each primer could amplify 2 to 12 polymorphic bands, with an average of 7.93 bands. The length of most amplified fragments ranged from 270 to 2000 bp. Cluster analysis showed that the 32 cultivars could be classified into 3 groups at the level of D 0.33.which evidently showed some variation of some important traits of Lycopersicon Esculentum var.Cerasiforme Alef.The primers, which were suitable for the purity identification of Lycopersicon Esculentum var.Cerasiforme Alef hybrid seeds, were screened and the fingerprints of RAPD were established among the parents and their hybrid seeds. 18 primers were screened from 27 the ones that could be beneficial to the purity idenetification of Lycopersicon Esculentum var.Cerasiforme Alef hybrid seeds. The results showed that there were 4 primers of partial female type,5 primers of partial male type,4 primers of complementary type, 2 primers of peculiar,3 type primers of shortage type. It was found there were 2 fasle seeds among 21 Lycopersic Esculentum var.Cerasiforme Alef hybrid...