A kind of plant stress resistance-related protein osc3hc4 and its coding gene and application
A coding gene and plant technology, applied in the field of plant stress resistance-related protein OsC3HC4 and its coding gene and application, can solve problems affecting food production and restrictions, achieve broad application space and market prospects, strong salt tolerance and drought resistance The effect of increasing the yield of crops
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Embodiment 1
[0049] Example 1 Acquisition of Rice Stress Resistance-Related Proteins and Their Encoding Genes
[0050] 1. Experimental materials
[0051] Referring to Jan et al. (2013) [Asad Jan, Kyonoshin Maruyama, Daisuke Todaka, Satoshi Kidokoro, Mitsuru Abo, Etsuro Yoshimura, Kazuo Shinozaki, Kazuo Nakashima and Kazuko Yamaguchi-Shinozaki. OsTZF1, a CCCH-Tandem Zinc Finger Protein, Confers Delayed Stressed Sess in Rice by Regulating Stress-Related Genes. Plant Physiology, 2013, 161:1202-1216], the plant leaf material of the rice variety 'Huaidao No. 5' was removed, quick-frozen in liquid nitrogen, and stored at -80°C.
[0052] 2. Extraction and purification of total RNA from leaves
[0053] Take about 2.0 g of the expanded leaves of Huaidao No. 5 seedlings, grind them into powder in liquid nitrogen, add them to a 10 mL centrifuge tube, and use the Applygen Plant RNA Extraction Kit (Applygen Technologies Inc, Beijing) to extract total RNA from sweet potato tubers. Including: Plant RNA...
Embodiment 2
[0061] Example 2 OsC3HC4 Stress expression analysis of genes
[0062] 1. Adversity stress treatment
[0063] The plump seeds of Huaidao No. 5 were surface-sterilized with 1% sodium hypochlorite for 20 min, washed 6 times with distilled water, then soaked in distilled water for 24-36 h, placed on wet gauze, and germinated at 32 °C for about 2 days. Seeds that germinated uniformly were sown on foamed plastic orifice plates with gauze for liquid culture. After 4 weeks of normal growth, the following stress treatments were started.
[0064] Mannitol treatment: rice seedlings were transferred from the nutrient solution to a solution containing 200 mM mannitol, and the roots and leaves of rice were harvested at 0, 3, 6, 12, 24, and 48 h, respectively;
[0065] PEG treatment: the treatment method is the same as that of mannitol, the concentration of PEG6000 solution is 20%, and the roots and leaves of rice are taken at 0, 3, 6, 12, 24, and 48 hours respectively;
[0066] NaCl tre...
Embodiment 3
[0073] Example 3 OsC3HC4 Construction of gene overexpression vector
[0074] The correct DNA fragment containing the nucleotide shown in the sequence table SEQ ID NO 1 identified by sequencing in Example 1 is used Bam H I and Sac I carried out double enzyme digestion, recovered DNA fragments with 1% agarose gel, and passed T 4 DNA ligase will recover the OsC3HC4 The gene fragment was connected with the pYPx245 plasmid containing double 35S promoters, and the rice containing OsC3HC4 Gene recombinant plasmid AH128. The expression vector also contains gusA Reporter gene and intronic kanamycin resistance marker gene, vectors such as figure 2 shown.
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