Applications of double-SNP marker in detection of Tomato yellow leaf curl disease-resistant gene
A technology of tomato yellowing curved leaves and disease resistance genes, applied in the field of plant molecular marker preparation, can solve the problems of SNP gene marker reporting without Ty-1 gene, achieve high accuracy and stability, and improve efficiency
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Embodiment 1
[0041] Example 1 Obtaining the cDNA of tomato Ty--1 and ty--1 and the gDNA sequence of ty--1 The establishment of double SNP markers
[0042] (1) Refer to the publication number WO2012125025A1 (http: / / www.google.com.hk / patents / WO2012125025A1?cl=en) to find the cDNA sequence of Ty-1 and its susceptible allele ty-1 (see figure 1 ), the reference (Verlaan MG et al.2013) obtained the gDNA sequence of the susceptible gene ty-1 and its position on the whole genome (chromosome 6 30850904-30879542, composed of Solyc06g051170, Solyc06g051180 and Solyc06g051190), and used for the development of SNP markers.
[0043] (2) Through MultAlin online sequence comparison software (http: / / multalin.toulouse.inra.fr / multalin / multalin.html), the anti-susceptible cDNA sequences were compared, and 54 SNP sites were found ( figure 1 shaded bases). Among them, the bases at positions 313 and 314 of Ty-1 and ty-1 are respectively GT and CG (named NL2) see figure 1 a Bordered bases, the bases at 1159, ...
Embodiment 2
[0058] Example 2 The genotyping and field inoculation verification of the test material with NL2-3 double SNP markers
[0059] (1) NL2-3 test for genotyping of test materials
[0060] The tomato materials used for testing genotyping (not for the purpose of genetic breeding) came from commonly used tomato materials collected by the tomato research group of Huazhong Agricultural University. The results of SNP molecular marker identification are shown in Figure 6 (Where Marker is DS5000, lane 1. is the known disease-resistant homozygous material LA1969, 2. is the known susceptible material Moneymaker (according to the article published by Verlaan M G et al. in 2013), 3. is the known heterozygous disease-resistant material commodity Species F1 generation Huafan No. 11, 4. Swimming lanes 3-19 are the test materials). The screening results showed that lanes 4, 9, and 15 were homozygous disease-resistant genotype Ty-1 / Ty-1, and lanes 2, 5, 6, 8, 10, 12, 13, 14, 16, 17, 18, and 19 w...
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