Plant responding low-phosphor and high-salt stress protein, encoding gene and uses thereof
A technology for coding genes and plants, which is applied in the field of proteins and their coding genes and applications in response to low phosphorus and high salt stress in plants, and can solve problems such as unclear functions
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Embodiment 1
[0034] Example 1. Acquisition of proteins and genes encoding them in response to low-phosphorus and high-salt stresses in plants
[0035] Total RNA was extracted from 10-day-old Arabidopsis (Columbia type) seedlings and reverse-transcribed into cDNA. Using the cDNA as a template, a pair of primers respectively provided with KpnI and SacI restriction sites are used for PCR amplification to obtain the full-length cDNA of drought tolerance related genes. The primer sequences are as follows:
[0036] Primer1: 5'-ggtaccATGGACAGAGGATGGTCTGGTCTC-3'(KpnI);
[0037] Primer2: 5'-gagctcCTATTGATTTTTGTTGTTTCCTTCGCCGTCGT-3' (SacI).
[0038] The high-fidelity Pyrobest DNA polymerase from Takara Company was used for amplification. The amplification system was 50 μL, containing 5.0 μL of 10×Pyrobest PCR buffer, 1.0 μL of 10 mM dNTP mix, 1.0 μL of each primer, 0.5 μL of Pyrobest enzyme, and 3.0 μL of template. (0.1ug), supplemented with sterilized ultrapure water to 50 μL, and the reaction s...
Embodiment 2
[0045] Example 2. Functional verification of proteins and genes encoding them in response to low phosphorus and high salt stress in plants
[0046] 1. Obtainment of AtLPST overexpression plants (35S::ATLPST-9)
[0047] The recombinant vector pMD18-AtLPST containing the AtLPST gene obtained in Example 1 and the pBIB-Super vector (the construction method of the pBIB-Super vector: utilize primers aagcttGTGGGCCTGTGGTCTCAAGAT and tctagaCTAGAGTCGATTTGGT from plasmid pMSP-1 (Ni M, Cui D, Einstein J, Narasimhulu S, Vergara C, Gelvin SB. Strong and tissue specificity of chimaeric promotors derived from the octopine and mannopine systha
[0048] Super::AtLPST was transformed with Agrobacterium to infect Columbia wild-type Arabidopsis plants. In order to obtain AtLPST overexpression T1 transgenic Super::AtLPST Arabidopsis plants. After screening the positive plants of transgenic Super::AtLPST Arabidopsis on MS medium containing 50 mg / L hygromycin and performing segregation ratio statis...
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