SNP locus combination for detecting resistance of tomato yellow leaf curl virus disease and application thereof
A technology for tomato yellowing and curving leaves and viral diseases, applied in the field of plant biology, can solve the problems of linkage redundancy, insufficient versatility of SNP sites, and invalid selection of target genes, etc., achieve accurate detection, reduce manual inoculation identification and field transplanting The workload and the effect of improving breeding efficiency
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Embodiment 1
[0080] Example 1 Screening of SNP site combinations
[0081] 1. Experimental materials
[0082] The variant group data of 660 representative tomato germplasm resources from different sources worldwide were selected for SNP site screening in this embodiment. The source of the tomato variant group data is mainly based on the previous work of the project team where the inventor belongs. (Lin,T.,Zhu,G.,Zhang,J.et al.Genomic analyzes provide insights into the history of tomato breeding[J].Nat Genet,2014,46:1220~1226; Tieman D,Zhu G,Resende M F R , et al. A chemical genetic roadmap to improved tomato flavor [J]. Science, 2017, 355(6323): 391). Among them, the genotype data of some tomato germplasm resources and the resistance phenotype data of Ty-1 / Ty-3 and Ty-2 genes come from the existing public database (https: / / solgenomics.net / ).
[0083] 2. Screening of SNP site combinations
[0084] Using the target gene Ty-1 / Ty-3 gene (Solyc06g051170, Solyc06g051180, Solyc06g051190, databa...
Embodiment 2
[0088] Example 2 Primer Synthesis and Kit Preparation
[0089] 1. Primer design and screening
[0090] According to the flanking sequences of the Ty1-SNP01 site ~ Ty1-SNP08 site and the Ty2-SNP01 site ~ Ty2-SNP08 site provided in Example 1, for each SNP site, the principle of design and development according to the KASP marker (target product 80-150bp, the primers specifically match the target region on the reference genome and are located in non-SNP dense regions, avoiding complex sequence regions with high A\T or G\C content, etc.), use Primer3.0 software to design two upstream SNP sites A forward primer and a reverse primer were designed downstream.
[0091] Tomato samples to be tested include some germplasm resources of known genotypes and yellow leaf curl virus disease resistance phenotypes and 21 samples randomly selected from breeding materials as resistant, susceptible or heterozygous controls, and finally added 3 ddH 2 O is used as the NTC blank control, a total of ...
Embodiment 3
[0109] Example 3 Verification and Breeding Application of Tomato Yellow Leaf Curl Virus Resistance Genes Ty-1 / Ty-3 and Ty-2 High Efficiency KASP Markers
[0110] This example is based on the SNP site provided in Example 1, using the kit provided in Example 2 (including primer combinations) to carry out tomato anti-yellow leaf curl virus disease gene Ty-1 / Ty-3 and Ty-2 high-efficiency KASP Validation of markers and breeding application work.
[0111] Select 381 representative tomato samples including mainstream commercial varieties, core public germplasm of the National Resource Bank, breeding intermediate materials, new hybrid combinations, etc., involving large fruit tomatoes, cherry tomatoes, cluster tomatoes, fresh tomatoes, processed tomatoes and Various types of cultivated tomatoes such as farm species (local varieties), 154 of which obtained Ty-1 / Ty-3 and Ty-2 genotype data through third-party commercial institutions using closely linked SCAR markers (including 41 yellow...
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