Development Of SNPs For Leaf Mould Resistance Gene Cf-5 In Tomato And Screen Of Germplasm Resources

Leaf mould (Fulvia fulva) is one of the serious diseases on tomato under protective cultivation and also a wide-spread disease of tomato in the world. It can cause heavy losses of tomato production and friuts'quality. If choosing and identification resistance breeding by people, it needs much time and nicety is not high. So, developed technology of molecular markers for leaf mould, assisted breeding breed out tomatoes which contain genetic resistance to leaf mould, resolve the problem of leaf mould disease in fundamental is the trend of resistance leaf mould breeding.In this study, the Cf-5 gene were isolated from four resistance of leaf mould tomatoes and four feeling sick tomatoes. Then, genome-specific primer set were designed and two SNPs were detected by sequencing. Finally, a stable SNP molecular marker was designed. We aimed to develop SNPmarkers about Cf-5 gene related the resistance to leaf mould in tomato. Established and developed a feasible way and technology for tomato SNP markers. The results as follows:1,Material of tomato resistance to leaf mould provided by our research group. Applying resistant parent 06885, susceptible parent 06919, build F2 segregating population 258 single plants, by Identified the resistance character between parents and separate groups of F2, the results indicated that the genetic pattern consistent with Bangladesh Dell's dominant single-gene inheritance.2,Designed three nested primers for Cf-5 gene, and compared Cf-5 sequence of all eight materials with the Cf-5 gene in the GenBank. Finally, we detected two SNPs: T-C; G-T.3,Firstly, for different types of nucleotide mutation, the method of introducing proper mismatched bases at the second or the third sites of the 3'-end of the primers may facilitate getting ideal allele-specific primers; Secondly, introducing mismatched bases at the second and the third sites of the 3'-end of the primer can repressed PCR completely ; Thirdly, the mismatched base pairs C/T is stronger than G/T, and the A/G is stronger than T/G.4,Used the SNP markers to identify 42 laboratory tomato material in laboratory, obtained 2 of the germplsam resource which related to the leaf mould. Combined with artificial inoculation research, consistent, further confirmed the value of this marker. This research provided a Theoretical basis for tomato molecular marker assisted selection breeding.