【作者】 李强；

【导师】 张辉； 斯钦巴特尔；

【作者基本信息】 内蒙古农业大学 ， 作物遗传育种， 2008， 硕士

【摘要】 本文利用mRNA差异显示技术,以显性核不育亚麻兄妹交分离出的遗传背景极其相似的可育株和不育株为材料,对可育、不育花蕾中基因差异性表达进行研究,并对差异片段进行反向Northern杂交验证、克隆、测序以及序列分析。主要研究结果如下:(1)建立了适合核不育亚麻的DDRT-PCR体系。(2)共获得差异片段52个。其中36个为不育差异片段,其余的16个为可育差异片段。(3)对这52个差异片段进行反向Northern杂交验证,最终获得了5个不育花蕾阳性片段:G+E07+100bp、G+E07+330bp、G+E07+830bp、G+S273+500bp、A+S267+300bp;2个可育花蕾阳性片段:G+S274-250bp、G+S274-450bp。(4)这些阳性片段经克隆、测序和BLAST序列分析,结果显示:G+E07+100bp与植物GTP结合蛋白高度同源,在细胞信号传导过程中有调节作用,推测可能与核不育亚麻育性有密切关系;G+E07+330bp为亚麻花蕾的一段cDNA,可能与核不育亚麻花粉发育有关;G+E07+830bp与β-木糖苷酶同源性很高,它可能影响了亚麻花药、花粉中碳水化合物的代谢,从而导致了花粉的败育;G+S273+500bp与NAD+ADP核糖转移酶有很高的同源性;A+S267+300bp与分泌蛋白高度同源,推测它可能参与了核不育亚麻信号传导途径、形态发生、细胞凋亡等过程,最终影响了雄性不育的发生;G+S274-250bp和G+S274-450bp与已知植物的碱基序列和氨基酸序列同源性很低,推测它们可能是与显性核不育亚麻育性有关的新基因。更多还原

【Abstract】 The mRNA differential expression was compared by mRNA differential display PCR (DDRT-PCR) between male sterile and fertile flower buds of flax. The positive differential fragments were confirmed by Reverse Northern Dot Blotting, and then sequence analysis of these fragments was carried out with the main results as follows:(1) The optimum conditions of DDRT-PCR system of flax were established.(2) Among the 52 fragments that had been obtained, 36 fragments were from male sterile flower buds; the other 16 fragments were from male fertile flower buds.(3) 9 fragments were confirmed by Reverse Northern Dot Blotting. 5 fragments were specific to male sterile flower buds: G+E07+100bp, G+E07+330bp, G+E07+830bp, G+S273 +500bp, A+S267+300bp; 2 other fragments were specific to male fertile flower buds: G+S274-250bp, G+S274-450bp.(4) The positive fragments were sequenced and analyzed with BLAST. The G+E07+100bp which is homologous to GTP-binding protein of plant suggested that it may play an important role in signal transduction. The G+E07+330bp may be closely related to fertility. The G+E07+830bp, which is most similar to the sequence of Beta-D-xylosidase, may influence carbon hydrate metabolism in flax anther and pollen, resulting in pollen abortion. The G+S273+500bp is homologous to NAD+ADP ribosyltransferase. The A+S267+300bp, homologous to secreted protein, may play roles in signal transduction, Morphogenesis and Apoptosis in flax and finally lead to male sterility. The G+S274-250bp and G+S274-450bp had no significant similarity with any known genes in plants, so they may be newly found genes related to fertility in flax.更多还原