Plant salt tolerant protein and encoding gene and application thereof
A technology that encodes genes and plants, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve problems such as limited economic value, and achieve the effect of improving salt tolerance and high application value
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Embodiment 1
[0029] Embodiment 1, the acquisition of plant salt tolerance protein and its coding gene
[0030] The ESTs obtained from the cDNA library constructed from the leaves of soybean variety Suinong 14 were analyzed, and an EST with Na + / H + Antiporter genes have high homology. The full-length cDNA sequence of the gene was obtained from Wenfeng No. 7 (unified number: ZDD02611; China National Germplasm Bank) treated with salt for 6 h by RT-PCR and 3' RACE. The specific method is as follows:
[0031] Soak the roots of soybean variety Wenfeng 7 seedlings grown for 2 weeks after germination with 200mM NaCl solution for 6 hours, harvest the leaves to extract RNA and reverse transcribe it into the first strand of cDNA, the first strand of cDNA is synthesized according to the ReverseTranscription System Kit (Promega) According to the instructions of the kit, the cDNA was used as a template to carry out PCR amplification with primers N21F (5'-CAGTTGTGTTGAGGGACACA-3') and N21R (5'-GCACTCG...
Embodiment 2
[0041] Example 2, Functional Verification of Plant Salt Tolerance Protein and Its Encoding Gene
[0042] 1. Construction of overexpression vector in Arabidopsis
[0043] The ORF of GmNHX2 was cloned into the pBI121 vector and placed under the control of the 35S promoter. The specific method is:
[0044] Primers were designed according to the above GmNHX2 cDNA sequence, the sequence is as follows:
[0045] BamN2: 5'-GCC GGATCC ATGGCCTCGGAATTAGAAATTTC-3' (underlined is the BamH I enzyme recognition site);
[0046] SacN2: 5′-GCG GAGCTC TCATGATGAATAGTGGTTCTGGC-3' (the Sac I enzyme recognition site is underlined).
[0047] The roots of soybean variety Wenfeng 7 seedlings grown for 2 weeks after germination were soaked in 200mM NaCl solution for 6 hours. RNA was extracted from the leaves and cDNA was obtained by reverse transcription. As a template, PCR amplification was performed using BamN2 and SacN2 as primers.
[0048] A fragment of 1605bp was amplified by PCR, and seq...
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