SSR markers linked with resistance genes Bs_t of physiological races No. 0 and No. 1 of tobacco black shank and applications thereof
A technology of tobacco black shank and physiological race, applied in the field of SSR marker, can solve the problems of chain burden, medium resistance, difficulty in maintaining the quality of flue-cured tobacco, etc., and achieve the effect of low cost and fast cost
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Embodiment 1
[0034] Screening of SSR markers linked to Bs_t genes in tobacco race 0 and race 1 for resistance to black shank by segregating population analysis (BSA)
[0035] 1. Experimental materials
[0036] Flue-cured tobacco Honghua Dajinyuan, which is good in quality but susceptible to tobacco black shank physiological race 0 and 1, was used as the female parent, and the flue-cured tobacco breeding material GH14-62 (its The resistance of race 0 and race 1 to black shank disease is controlled by the Bs_t gene (nucleic acid fragment), and Nicotiana japonica ( N. rustica ) was formed by transferring the genome fragments of races 0 and 1 resistant to black shank into flue-cured tobacco safflower Dajinyuan) as the male parent, crossed and selfed to obtain F1 and F2 populations respectively.
[0037] 2. Identification of resistance to black shank race 0 and 1 in parents and F2 segregation populations
[0038]The test materials were transplanted to the field after they became seedlings, w...
Embodiment 2
[0047] Map Distance of Linkage Markers and Its Validation in Individual Plants of F2 Population
[0048] 1. Data Analysis
[0049] First, the tobacco genomic DNA was extracted according to the method described in Example 1, and the F2 population was cultured and multiplied for the identification of resistance to race 0 and race 1 of blackleg disease and the analysis of SSR markers. Secondly, 300 individual plants in the F2 population were genotyped using markers TM51207 and TM55805 screened by BSA method. Finally, carry out data statistics on the band type of each individual plant. The resistant homozygous band is marked as "B", the resistant heterozygous band is marked as "H", the susceptible band is marked as "A", and the band Unclear or no amplified bands were recorded as "U".
[0050] 2. Calculation of genetic distance of linkage markers
[0051] Using JoinMap 4.0 software combined with the identification data of black shank race 0 and race 1 resistance of individual F2...
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