The projects of this program have ttwo components including:。1.Molecular identification of tomato genotype resistant to Fusarium oxysporum: tomato fusarium wilt is one of the important fungal disease.The molecular markers linked to fusarium wilt resistant genes I-1 and I-3 have been developed. Specific primer pair I1-A2 was developed to amplify DNA fragment from vary allelic at I-1 locus. A 150 bp DNA fragment was amplified from both resistant and susceptible tomato accessions that corresponding to the presence of I-1 and i-1 alleles.After sequencing of PCR products, G is determined at the nucleotide sequence position no. 80 in resistance tomato accession, where as C is determined in the susceptible tomato accession. The different nucleotide sequence between both of resistance and susceptible tomato accession provide the possibility for development of SNP marker. In addition, the I3-P7-CAPS marker was also developed PCR following restriction enzyme (NsiI) digestion. The 380 bp DNA band was obtained from the tomato accession with resistance genes I-3, however, the susceptible tomato accession with gene i-3 was none band. The results are also confirming the previously study of AVRDC- The World Vegetable Center and other seed companies.。2.Development of molecular marker linked to tomato mosaic virus (ToMV)resistance: ToMV, a member of Tobamovirus, is one of the important tomato viruses, and ToMV-0, ToMV-1, ToMV-2 are three major ToMV strains. The Tm-1, Tm-2 and Tm-22 genes were also identified for ToMV resistance. In the project, we selected the molecular markers Tm1-CAPS and Tm22-SCAR for Tm-1 and Tm-22 based on AVRDC provided tomato lines,and developed a method for specific detection of ToMV-0, 1, and 2. The results showed that a 550 bp DNA band was obtained of PCR product amplified by molecular marker Tm1-CAPS from resistance genes Tm-1 following restriction enzyme (HaeIII) digestion, however 350 bp DNA band was obtained from susceptibility gene tm-1. The Tm22-SCAR could amplify DNA products 255 bp and 214 bp in length from resistance genes Tm-2 and Tm-22, respectively, however, no product was amplified from the susceptible lines. We has also been established molecular marker could be amplified a 2.2 kb of DNA product from ToMV-0, 1, and 2. Those developed molecular markers could help breeder screening and confirmation disease resistant genotypes in early stage to accelerate the breeding and promotion of disease-resistant tomato lines.。
The projects of this program have ttwo components including:。1.Molecular identification of tomato genotype resistant to Fusarium oxysporum: tomato fusarium wilt is one of the important fungal disease.The molecular markers linked to fusarium wilt resistant genes I-1 and I-3 have been developed. Specific primer pair I1-A2 was developed to amplify DNA fragment from vary allelic at I-1 locus. A 150 bp DNA fragment was amplified from both resistant and susceptible tomato accessions that corresponding to the presence of I-1 and i-1 alleles.After sequencing of PCR products, G is determined at the nucleotide sequence position no. 80 in resistance tomato accession, where as C is determined in the susceptible tomato accession. The different nucleotide sequence between both of resistance and susceptible tomato accession provide the possibility for development of SNP marker. In addition, the I3-P7-CAPS marker was also developed PCR following restriction enzyme (NsiI) digestion. The 380 bp DNA band was obtained from the tomato accession with resistance genes I-3, however, the susceptible tomato accession with gene i-3 was none band. The results are also confirming the previously study of AVRDC- The World Vegetable Center and other seed companies.。2.Development of molecular marker linked to tomato mosaic virus (ToMV)resistance: ToMV, a member of Tobamovirus, is one of the important tomato viruses, and ToMV-0, ToMV-1, ToMV-2 are three major ToMV strains. The Tm-1, Tm-2 and Tm-22 genes were also identified for ToMV resistance. In the project, we selected the molecular markers Tm1-CAPS and Tm22-SCAR for Tm-1 and Tm-22 based on AVRDC provided tomato lines,and developed a method for specific detection of ToMV-0, 1, and 2. The results showed that a 550 bp DNA band was obtained of PCR product amplified by molecular marker Tm1-CAPS from resistance genes Tm-1 following restriction enzyme (HaeIII) digestion, however 350 bp DNA band was obtained from susceptibility gene tm-1. The Tm22-SCAR could amplify DNA products 255 bp and 214 bp in length from resistance genes Tm-2 and Tm-22, respectively, however, no product was amplified from the susceptible lines. We has also been established molecular marker could be amplified a 2.2 kb of DNA product from ToMV-0, 1, and 2. Those developed molecular markers could help breeder screening and confirmation disease resistant genotypes in early stage to accelerate the breeding and promotion of disease-resistant tomato lines.。