Functional markers related to drought resistance in budding stage of rapeseed and its application
A technology of drought resistance and rapeseed, applied in the fields of rapeseed breeding and molecular biology, can solve the problems of long cycle, heavy workload of drought resistance identification, easy to be affected by the environment, etc., and achieve the effect of improving selection efficiency and speeding up the pace of genetic improvement.
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Embodiment 1
[0053] Example 1 Obtainment of the functional marker BnDM-413 related to the drought resistance of rape bud stage
[0054] 1. Group construction:
[0055] The strong drought-resistant rapeseed variety Qinyou No. 8 (leaf wilting index of 0.06, known and commonly used variety) was used as the female parent, and the drought-sensitive rapeseed variety Huyou 16 (leaf wilting index of 0.94, known and commonly used variety) was used as the male parent. A drought-resistant segregating F 2:3 There are 183 strains in total in the generation family group.
[0056] 2. DNA extraction
[0057] (1) Collection of parents and F 2 From the young leaves of 183 individual plants in the population, the total DNA of the leaves was extracted by the CTAB method. The specific extraction steps are as follows:
[0058] (1) Put 0.5 g of leaves fresh or frozen at -20°C into a 1.5ml centrifuge tube, grind to a homogenate with a glass rod, add 800 μl of CTAB extract (50mmol / L Tris-HCl, pH8.0; 20mmol / L ...
Embodiment 2
[0089] Example 2 Application of Functional Marker BnDM-413 in Identifying Drought Resistance of Rapeseed Budding Stage
[0090] 1. Group construction:
[0091] The strong drought-resistant rapeseed variety Qinyou No. 8 (leaf wilting index of 0.06, known and commonly used variety) was used as the female parent, and the drought-sensitive rapeseed variety Huyou 16 (leaf wilting index of 0.94, known and commonly used variety) was used as the male parent. F 1 generation group, F 1 F 2 generation groups.
[0092] 2. DNA extraction
[0093] from F 2 100 individual plants were randomly selected from the generation population, and their DNA was extracted. The specific operation steps were the same as the DNA extraction method in Example 1.
[0094] 3. PCR amplification
[0095] Using the above-mentioned extracted DNA as a template, the primer BnDM-413-F and primer BnDM-413-R of the functional marker BnDM-413 were used for PCR amplification,
[0096] Primer BnDM-413-F is: 5'-GCC...
Embodiment 3
[0112] Example 3 Application of Functional Marker BnDM-413 in Identifying Drought Resistance of Rapeseed Budding Stage
[0113] With reference to the method of Example 2, the scope of identification was expanded. A total of analyzes detected F 2 183 individual plants of the population, the results showed that F 2 There are 76 individual plants with bands in the population, and 76 F 2:3 The average wilting index of the generation family group was 0.305, which was significantly lower than the group's average value of 0.567 (P<0.001), that is, the drought resistance was significantly improved. The above results fully demonstrate that the marker can indeed be used for prediction, identification and screening of drought resistance.
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