Application of rice glutaredoxin gene osgrxc2 in breeding
A gene and rice technology, applied in the application field of rice redoxin gene OsGrxC2 in breeding, can solve the problems of high seed cost, impact on application, and many promotion links, so as to improve rice yield and quality, increase rice thousand-grain weight, The effect of broad application prospects
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Embodiment 1
[0051] Example 1 OsGrxC2 Construction of Gene Overexpression Vector and Obtainment of Transgenic Rice Plants
[0052] 1. Rice cultivation
[0053] Put the rice seeds in a 50ml triangular flask, sterilize them with 75% alcohol for 90 seconds, then sterilize them with 2.5% sodium hypochlorite for 45 minutes, wash them with sterile water for 5 times, dry them on sterile filter paper, and finally sow them in the sterilization MS solid medium (1.5% sucrose, 0.8% agar, pH 5.8), 16h / 8h (28 ℃) day and night, incandescent light for cultivation.
[0054] 2. Rice RNA extraction: the rice RNA was extracted using the conventional Trizol method in the art.
[0055] 3. Reverse transcription reaction (cDNA synthesis)
[0056] The reverse transcription reaction was performed with the reverse transcription kit of Takara Company, and the reaction system and reaction conditions refer to the kit manual.
[0057] 4. PCR amplification of the target gene
[0058] according to OsGrxC2 For the ...
Embodiment 2
[0080] Example 2 OsGrxC2 Functional identification of genes
[0081] 1, OsGrxC2 Tissue Localization Observation of Genes
[0082] Research OsGrxC2 For the tissue localization of the gene, total RNA was extracted from the corresponding tissues or organs at the 4-leaf stage, tillering stage, and heading stage of rice respectively, and the Osactin RT-qPCR detection as an internal reference, for OsGrxC2 tissue-specific analysis. The result is as figure 2As shown, the expression of this gene can be detected in roots, stems, leaves at tillering stage, flag leaves at heading stage, tillers, inflorescences and seeds, and its expression level is the highest in seeds, especially in wax-ripening stage seeds. About 45 times in the. This result shows that, OsGrxC2 The expression of the gene has no tissue specificity, it is a constitutively expressed gene, and the expression level is higher in the mature stage of the seed. The primers used are as follows:
[0083] OsGrxC2 The 5...
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