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| 年度 | 2014 番茄抗萎凋病與菸草嵌紋病毒分子標誌建立與應用 The establishment and application of specific molecular markers for TMV and fusarium wilt resistance in tomato 103農科-9.1.1-種-X1 103AS -9.1.1-SS-X1 生物技術課 Biotechnology Section 孫永偉 Yung-Wei Sun 1. 番茄抗萎凋病基因型之分子鑑定：番茄萎凋病為重要真菌性病害，本試驗建立抗萎凋病基因I-1及I-3之分子標誌。利用I1-At2引子組可同時擴增番茄抗病(I-1)與感病(i-1)基因150 bp之DNA條帶，將抗感病品種進行PCR產物解序後，於序列位置80出現差異點，該位置感病核苷酸序列為C，抗病核苷酸序列為G，應可作為判斷番茄抗感萎凋病基因I-1之SNP標誌。利用I3-P7-CAPS引子組配合限制酶NsiI酶切反應，可擴增抗病基因(I-3) 380 bp之DNA條帶，感病基因(i-3)無DNA條帶出現。此分子標誌檢測結果與亞洲蔬菜中心及國內育種者或種子公司已知抗感性品種吻合。2. 番茄抗菸草嵌紋病毒分子標誌之建立：菸草嵌紋病毒為菸草鑲嵌病毒屬(Tobamovirus)，為番茄三大病毒之一。本研究利用亞洲蔬菜中心提供番茄抗感病品種進行研究，本試驗針對抗病基因(Tm-1及Tm-22)與TMV病毒各篩選專一性分子標誌(Tm1-CAPS、Tm22-SCAR與TMV-012)。利用Tm1-CAPS分子標誌配合HaeIII限制酶反應後，可擴增抗病基因550 bp之DNA條帶、擴增感病基因350 bp之DNA條帶。利用Tm22-SCAR分子標誌，可擴增Tm-2抗病基因255 bp之DNA條帶、擴增Tm-22抗病基因214 bp之DNA條帶，感病品種無DNA條帶出現。TMV-012分子標誌可擴增嵌紋病毒株(TMV-0、TMV-1、TMV-2) 2.2 kb之DNA條帶。上述分子標誌可協助育種者早期篩選抗病植株、確認抗感病基因型及病毒感染情形，提高育種效率。 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 |
| 計畫名稱_中文 | G is determined at the nucleotide sequence position no. 80 in resistance tomato accession |
| 計畫名稱_英文 | 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 |
| 計畫編號_中文 | 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 |
| 計畫編號_英文 | however |
| 主辦單位_中文 | 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 |
| 主辦單位_英文 | a member of Tobamovirus |
| 主辦人_中文 | is one of the important tomato viruses |
| 主辦人_英文 | and ToMV-0 |
| 中文摘要 | ToMV-1 |
| 英文摘要 | ToMV-2 are three major ToMV strains. The Tm-1 |