Application of Arabidopsis idd14 gene in improving plant drought stress tolerance
A technology of IDD14 and drought stress, applied in the field of Arabidopsis genes that improve plant drought stress tolerance, can solve problems such as blindness in molecular breeding for drought resistance, and achieve the effect of improving drought stress tolerance
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Embodiment 1
[0034] Embodiment 1: screening overexpression IDD14 gene plant
[0035] The mutant idd14-1D used in the present invention is a mutant with abnormal leaf development screened from an Arabidopsis T-DNA mutant library containing an activation tag (Cui et al., PLOS Genetics, 2013, 9: e1003759), with obvious Leaf roll down phenotype. The previous genetic analysis showed that idd14-1D is a dominant mutant with a single T-DNA insertion, and the phenotype of mutant idd14-1D is caused by the overexpression of IDD14.
Embodiment 2
[0036] Example 2: IDD transcription factor subfamily positively regulates drought stress tolerance response in Arabidopsis
[0037] Tolerance responses to drought stress of idd14-1D mutant plants overexpressing IDD14 gene and control wild-type plants were analyzed using drought treatment experiments, determination of water loss rate of detached leaves, and comparative analysis of leaf stomatal density.
[0038] (1) The idd14-1D mutant plants overexpressing the IDD14 gene have strong drought stress tolerance
[0039] A certain amount of seeds of idd14-1D mutant plants and wild-type Arabidopsis plants were weighed, and after disinfection, the seeds were sown on MS medium for 2-3 days at a low temperature of 4°C, and cultured under continuous light for 7 days, and transplanted. Mix vermiculite: nutrient soil at a ratio of 1:1, weigh the same weight of nutrient soil (about 70-85 grams) in a small pot with a flat bottom, water the bottom to make the soil completely wet, and plant 9...
Embodiment 3
[0050] Embodiment 3: Isolation and cloning are used to construct the DNA fragment of IDD14 gene plant expression vector
[0051] Total RNA was extracted from Arabidopsis leaves using TRIzol reagent (purchased from Invitrogen). The specific steps are as follows: pre-cool the centrifuge, take 50-100 mg of sample and add liquid nitrogen to grind thoroughly, add 1 mL of TRIzol solution, and let stand at room temperature for 5 minutes; add 200 μL of chloroform, shake vigorously for 15 seconds, and let stand at room temperature for 3 minutes; 4°C, 12,000 g Centrifuge for 15 minutes; draw the supernatant into a new 1.5mL centrifuge tube (RNase-free, ribonuclease-free) (purchased from AXYGEN), add 1 times the volume of isopropanol, mix well, and let stand at room temperature for 10 minutes; Centrifuge at 12,000 g at 4°C for 10 min; discard the supernatant, add 1 mL of 70% ethanol solution, and pop the pellet; centrifuge at 7,500 g at 4°C for 5 min; discard the supernatant, centrifuge ...
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