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采用cDNA-SRAP方法筛选出的ATP合酶CF1α亚基基因(CTL-spn)的表达与红花的多刺性相关。

Expression of ATP synthase CF1 alpha subunit gene (CTL-spn) as screened by the cDNA-SRAP approach is correlated with spininess in Carthamus tinctorius L.

作者信息

Guo Dan-dan, Guo Qing-hua, Gao Yue, Guo Mei-li

出版信息

Yao Xue Xue Bao. 2015 Aug;50(8):1052-9.

Abstract

The safflower floret is a traditional Chinese medicine used to promote blood circulation and remove obstruction in the channels. The spines on its bracts are considered a handicap when manual harvest is involved. In this study, cDNA-SRAP was used to systematically investigate which genes are associated with the spines. Sixty pairs of possible primer combinations were used on two cDNA pools representing spininess and spinelessness. Six transcript-derived fragments were identified, of which two with low recombination were sequenced successfully and named as GPY-1 and GPY-2. By using the RACE method, the full-length cDNA of GPY-2 is cloned and named as CTL-spn. The full-length cDNA of CTL-spn was 1 679 bp long with a 1 524 bp ORF encoding a 508 aminoacid protein. The deduced amino acid sequence of the CTL-spn gene shared a high homology (97%) with other known ATP synthase CF1 alpha subunits. Semiquantitative RT-PCR analysis revealed that the mRNA of GPY-1 and GPY-2 accumulated in only spiny lines. Considering the important role of ATP synthase CF1 alpha subunit in plants, it may directly take part in the formation process of spininess and enhancing resistance reaction of spiny safflower. Also, our results provide the important insights for breeding spineless cultivars of safflower.

摘要

红花小花是一种传统中药,用于促进血液循环和疏通经络。当涉及人工采收时,其苞片上的刺被认为是一个障碍。在本研究中,利用cDNA-SRAP技术系统地研究了哪些基因与刺有关。在代表有刺和无刺的两个cDNA文库上使用了60对可能的引物组合。鉴定出6个转录衍生片段,其中2个重组率低的片段被成功测序,命名为GPY-1和GPY-2。通过RACE方法,克隆了GPY-2的全长cDNA,命名为CTL-spn。CTL-spn的全长cDNA长1679bp,开放阅读框1524bp,编码一个508个氨基酸的蛋白质。CTL-spn基因推导的氨基酸序列与其他已知的ATP合酶CF1α亚基具有高度同源性(97%)。半定量RT-PCR分析表明,GPY-1和GPY-2的mRNA仅在有刺品系中积累。考虑到ATP合酶CF1α亚基在植物中的重要作用,它可能直接参与有刺性状的形成过程和增强有刺红花的抗性反应。此外,我们的结果为红花无刺品种的选育提供了重要的见解。

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