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目的:通过同源克隆获得了花生闽花6号的RGA片段,为其抗性的研究及抗性育种提供了参考资料。方法:试验分为两组:其一通过利用抗性基因的NBS保守区所设计的简并引物对花生品种闽花六号进行了RGA片段扩增,其二结合已登录的花生RGA片段序列经过多元比对后设计简并引物进行RGA片段的扩增及序列分析;分析比较两组克隆方法的效果。结果:测序分析表明:前者20条随机测序序列中没有一条与已知RGA片段序列相似;后者20条随机测序序列中有18条为RGA片段序列,其登录号为GenBankEU639668-EU639685。结论:前一种方法克隆扩增RGA基因片段的效率很低,而后一种方法克隆扩增效果更好,这为闽花6号花生的遗传改良提供了理论基础。
OBJECTIVE: The RGA fragment of peanut Minhua 6 was obtained by homologous cloning, which provided references for its resistance and resistant breeding. Methods: The experiment was divided into two groups: one was RGA fragment amplified from peanut cultivar Minhua No.6 by degenerate primer designed by using NBS conserved region of resistance gene, After multiple alignment, degenerate primers were designed for amplification and sequence analysis of RGA fragments. The effects of two cloning methods were analyzed and compared. Results: Sequencing analysis showed that none of the 20 randomly sequenced sequences of the former were similar to the known RGA fragment sequences; 18 of the 20 randomized sequence fragments were RGA fragment sequences with the accession number of GenBankEU639668-EU639685. CONCLUSION: The former method is inefficient in cloning and amplifying RGA gene fragments, while the latter method is more effective in cloning and amplifying, which provides a theoretical basis for the genetic improvement of the peanut variety Minhua 6.