Primer for detecting vitality of strawberry seedlings and application of the primer
A technology of viability and strawberry, applied in the field of plant biology, can solve the problem of inaccurate judgment of vigor of strawberry seedlings, achieve beneficial evaluation, high repeatability, and reduce workload
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Embodiment 1
[0042] 1. Sampling of experimental materials: 'Toyonoka' strawberry seedlings were collected from Jiangsu Academy of Agricultural Sciences. Put the strawberry seedlings in good growth condition in the greenhouse, and plant 20 plants in pots respectively after 0h, 6h, 12h, 18h, 24h, 30h, 36h, and 48h, and take sample leaves and stolons at the same time, and freeze them in liquid nitrogen , stored in a -80°C ultra-low temperature freezer. After 10 days, the survival rate of each treated seedling was counted.
[0043] 2. DNA extraction from strawberry seedlings: Genomic DNA was extracted by the improved CTAB method, and the specific operation was as follows: fresh leaves (0.1 g) were crushed in liquid nitrogen, and quickly transferred into a 1.5 mL centrifuge tube. Add 600 μL of pre-warmed CTAB (2% CTAB: 100 mM Tris-HCl, pH 8.0; 20 mM EDTA, pH 8.0; 1.4 M NaCl; add 0.4% β-mercaptoethanol before use) extraction buffer. Incubate at 65°C for 30 minutes, and invert several times dur...
Embodiment 2
[0058] Toyonoka ('Toyonoka') strawberry seedlings in good growth condition were placed in a greenhouse without any protective measures, and allowed to wilt naturally, and then respectively after 0h, 6h, 12h, 18h, 24h, 30h, 36h, and 48h, Get 20 plants and plant them in pots, water them, and count the survival rates of strawberry seedlings in each treatment period after planting in suitable environmental conditions for 10 days.
[0059] Then compared with the Ct values obtained by the fluorescent quantitative PCR of GAPDH1 and TIM1 genes in Example 1 corresponding to each time period (0h, 6h, 12h, 18h, 24h, 30h, 36h, 48h). The results are shown in Table 3.
[0060] Table 3 The relationship between the Ct values of GAPDH1 and TIM1 genes and the vigor of strawberry
[0061] GAPDH1 gene Ct value
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