[Effects of antisense-thioredoxin s gene on expression of endogenous thioredoxin h gene in transgenic wheat seed].

植物生理与分子生物学学报 Pub Date : 2007-08-01
Jiang-Ping Ren, Jun Yin, Hong-Bin Niu, Xin-Guo Wang, Yong-Chun Li
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Abstract

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.

[反义硫氧还蛋白s基因对转基因小麦种子内源硫氧还蛋白h基因表达的影响]。
为明确反义硫氧还蛋白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为内源对照,采用半定量RT-PCR技术检测了内源trxh基因在转基因小麦品系01TY70-1-17-5和非转基因品种‘育麦70’中的表达谱。对转基因和非转基因种子在不同成熟期、不同组织和不同萌发过程的分析结果表明,转基因系种子中trxh基因mRNA转录量明显降低,但trxh基因mRNA转录量在不同发育和萌发阶段差异较大。转基因系种子trxh基因mRNA转录量在开花后15 ~ 30 d显著低于对照种子20.1%。trxh基因mRNA转录量在荧光后25 d最低,差异在0.05水平上显著。基因在不同组织中的表达分析也表明,在开花后25 d和30 d,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mRNA转录量最低的是花后25 d的胚乳,其次是胚,最后是全种子。在种子萌发过程中,转基因种子吸收24 h后trxh基因mRNA转录量低于对照种子,但差异不显著。以上结果表明外源反义trxs基因直接干扰内源基因的表达。
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