摘要
红花的花是传统的活血化瘀中药,但因其花序周围苞叶有刺非常不利于人工采摘。本研究采用cDNA-SRAP技术分析了与红花苞叶刺性状紧密连锁的基因。在分别建立红花F2代c DNA无刺池和有刺池的基础上,利用60对引物筛选与苞叶刺连锁的基因片段,获得6条与红花刺性状连锁的基因片段,其中基因GPY-1和GPY-2重组率低,与苞叶刺紧密连锁并成功测序。采用RACE技术,克隆了GPY-2 c DNA全长,命名为CTL-spn,生物信息学分析显示:CTL-spn全长1 679 bp,具有1 524 bp的开放阅读框,编码508个氨基酸。同源比对发现:CTL-spn表达蛋白与其他物种的ATP合成酶CF1-α亚基具有很高的同源性(97%)。半定量RT-PCR分析结果显示:GPY-1和GPY-2仅在有刺F2代单株中表达。因此,红花ATP合成酶CF1-α亚基基因(CTL-spn)可能直接参与了红花刺性状的形成从而提高了有刺红花的抗逆性,这一发现为红花无刺品种的选育提供了重要思路。
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 eDNA 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 eDNA 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 CF 1 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.
出处
《药学学报》
CAS
CSCD
北大核心
2015年第8期1052-1059,共8页
Acta Pharmaceutica Sinica
基金
Project supported by the National Natural Science Foundation of China(81173484,81473300)
Shanghai Natural Science Foundation(13ZR1448200)
