Ren Jiang-Ping, Yin Jun, Niu Hong-Bin, Wang Xin-Guo, Li Yong-Chun
National Engineering Research Center for Wheat, Henan Agricultural University, Zhengzhou 450002, China.
Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2007 Aug;33(4):325-32.
To clarify the function mechanism of antisense-thioredoxin s (anti-trxs) gene in transgenic wheat, the expression pattern of endogenous trxh gene in transgenic line 01TY70-1-17-5 and non-transgenic cultivar 'Yumai 70' were detected by semi-quantitative RT-PCR using wheat actin gene as the endogenous control. The results of analysis of transgenic and non-transgenic seeds in different maturation periods, different tissues and different germinating processes indicated that the mRNA transcript amounts of trxh gene in transgenic line seed were lowered distinctly, though the trxh gene mRNA transcript level varied greatly in different developing and germination stages. The mRNA transcript amounts of trxh gene in transgenic line seed were significantly lower than the control seeds by 20.1% 15-30 d after anthesis. The lowest mRNA transcript amount of trxh gene appeared at 25 day after fluorescence and the difference was significant at the 0.05 level. The analysis of gene expression in different tissues also indicated that the transcript levels of trxh gene in transgenic seed were significantly lower than control seeds in 25 d and 30 d after anthesis. The lowest amounts of mRNA transcript of trxh gene was from the endosperm 25 d after anthesis followed by embryo and then by whole seed. During seed germination, the mRNA transcript amounts of trxh gene in transgenic seed were lower than control seed after imbibing 24 h, but the difference was not significant. The above result demonstrated that foreign antisense trxs gene directly interferes with the expression of the endogenous gene.
为阐明反义硫氧还蛋白s(anti-trxs)基因在转基因小麦中的作用机制,以小麦肌动蛋白基因作为内参基因,采用半定量RT-PCR技术检测转基因株系01TY70-1-17-5和非转基因品种‘豫麦70’中内源trxh基因的表达模式。对转基因和非转基因种子在不同成熟时期、不同组织及不同萌发过程的分析结果表明,尽管trxh基因的mRNA转录水平在不同发育和萌发阶段差异很大,但转基因株系种子中trxh基因的mRNA转录量明显降低。花后15 - 30天,转基因株系种子中trxh基因的mRNA转录量显著低于对照种子20.1%。trxh基因的mRNA转录量在开花后25天最低,差异在0.05水平显著。不同组织中基因表达分析还表明,花后25天和30天,转基因种子中trxh基因的转录水平显著低于对照种子。trxh基因的mRNA转录量最低值出现在花后25天的胚乳中,其次是胚,然后是整粒种子。在种子萌发过程中,转基因种子吸胀24小时后trxh基因的mRNA转录量低于对照种子,但差异不显著。上述结果表明,外源反义trxs基因直接干扰了内源基因的表达。