Method for creating wheat seedling drought premature senility physiological phenotype by using cell autophagy inhibitor
An autophagy inhibitor, wheat technology, applied in the field of botany, can solve the problems of long time, cumbersome operation steps, high experimental cost, and achieve the effect of easy operation and observation, and low experimental cost.
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Embodiment 1
[0023] A method for creating the physiological phenotype of drought premature aging of wheat seedlings by using an autophagy inhibitor, the specific operation steps are as follows:
[0024] 1. Select wheat seeds with plump grains, the same size, and harvested in the same year, and place them in a constant temperature light incubator for 10 days; Set the culture temperature at 25°C, 16 hours of sunshine and 8 hours of darkness;
[0025] 2. 20 wheat seedlings with consistent growth are divided into treatment group and control group, each group of 10;
[0026] 3. The treatment group was placed in 20g / mL PEG-8000 solution, and 2mM, 5mM and 8mM 3-methyladenine were added as three treatment groups, while the control group was only placed in 20g / mL PEG-8000 solution. 8000 solution, the incubation time is 72 hours;
[0027] 4. After the experimental treatment, observe the differences in wheat plant phenotype between the treatment group and the control group.
Embodiment 2
[0029] A method for creating the physiological phenotype of drought and premature aging of wheat seedlings by using an autophagy inhibitor, and a method for determining the phenotypic difference, the specific detection method is:
[0030] 1. According to the conventional method, extract the total protein of the wheat leaves of the treatment group and the control group in Example 1 respectively.
[0031] 2. SDS-PAGE gel electrophoresis of the total protein of wheat leaves, the concentration of the used separating gel is 12%, the voltage is 100 volts, and the electrophoresis time is 1.5 hours.
[0032] 3. Perform Western blot detection on the protein in the above gel according to the existing method, use the polyclonal antibody of wheat ATG8 to identify the target protein in the wheat leaf in the gel, and detect the amount of ATG8-PE formed. The weaker the band, it means The more obvious the inhibitory effect is, the more obvious the drought effect is, and the physiological phen...
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