[Tobacco gamma-tubulin gene silencing mediated by recombinant potato virus X vector].

植物生理与分子生物学学报 Pub Date : 2007-10-01
Qi Wang, Shuang Zhao, Lu Wang, Xiao-Dan Wang, Kun Yang, Guo-Lei Wen, Mu Zhuang, Xiao-Wu Wang, Yan-Hong Li
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Abstract

Gamma-tubulin gene is a very important house-keeping gene and plays key roles in cell division, microtubule nucleation and cell cycle regulation in both plants and animals. In this paper, we studied the gamma-tubulin gene silencing in Nicotiana tabacum var. Samsun NN using the recombinant PVX vector with D fragment of gamma-tubulin cDNA. Special phenotype appeared from recombinant PVX-vector-infected plants, the different morphological leaves grew alternatively in leaf layers (Figs.3-5), and all the buds fell off gradually (Fig.4), resulting in failure of all the microspores to develop to the tetrad stage (Fig.8). The silencing initiated from the apical tissue and kept on until it was mature (Figs.3, 4). There was the increasing of target gene mRNA obviously during gene silencing, even to exceed the positive control (Fig.6). The PVX vector was also suppressed strongly and could fluctuate to some degree (Fig.9), which was probably correlated with target gene mRNA expression.

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[重组马铃薯病毒 X 载体介导的烟草γ-微管蛋白基因沉默]。
γ-微管蛋白基因是一种非常重要的看家基因,在动植物的细胞分裂、微管成核和细胞周期调控中起着关键作用。本文利用带有γ-微管蛋白cDNA D片段的重组PVX载体,研究了烟草变种Samsun NN中γ-微管蛋白基因沉默的情况。重组 PVX 载体感染的植株出现了特殊的表型,不同形态的叶片在叶层中交替生长(图 3-5),所有的芽逐渐脱落(图 4),导致所有的小孢子都不能发育到四分体阶段(图 8)。沉默从顶端组织开始,一直持续到成熟(图 3、图 4)。在基因沉默过程中,目的基因 mRNA 明显增加,甚至超过了阳性对照(图 6)。PVX载体也受到强烈抑制,并有一定程度的波动(图9),这可能与目的基因mRNA的表达有关。
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