cad2突变体中NTR基因被RNAi下调会影响拟南芥的发育

T. Bashandy, J. Reichheld
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引用次数: 0

摘要

NADPH- thioredoxin系统(NTS)和NADPH谷胱甘肽系统(NGS)是两个主要的硫醇还原系统,在维持细胞氧化还原稳态和一些植物发育过程中起关键作用。通过TRX还原酶(ntra ntrb)和谷胱甘肽生物合成(cad2)突变,研究了这两种硫醇还原系统之间的串扰。cad2突变体揭示了一个与花分生组织发育相关的新表型。不幸的是,这个突变体是不受精的,因此它不能维持在纯合子阶段。在这项研究中,我们使用RNAi技术获得了与该突变体接近的相似表型,但具有可育性。RNAi策略是通过在cad2突变体的遗传背景下引入含有两个NTRA基因首尾拷贝的RNAi构建物来下调两个NTR基因的表达。所获得的转化植株与ntrb cad2突变体相比表现出较弱的表型。值得注意的是,没有植物表现出ntra ntrb cad2突变体特有的针状表型。然而,有些植株看起来肥沃,但表现出顶端优势减少。另一些则更受影响,开出不育的花朵。我们的数据表明,RNAi策略是一种有效的策略,可以产生具有下调NTS和NGS还原系统的可育植物,并研究这两个硫醇系统之间的串扰。
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DOWN-REGULATION OF NTR GENES BY RNAi IN THE cad2 MU-TANT IMPAIRS PLANT DEVELOPMENT OF Arabidopsis thaliana
The NADPH-Thioredoxin System (NTS) and NADPH Glutathione System (NGS) are the two major thiol reduction systems that play a key role in the maintenance of cellular redox homeostasis and several plant developmental processes. Crosstalk between these two thiol reduction systems has been studied by associating TRX reductase (ntra ntrb) and glutathione biosynthesis (cad2) mutations. Triple ntra ntrb cad2 mutant revealed a new phenotype related to flower meristem development. Unfortunately, this mutant is unfertile and therefore it cannot be maintained at a homozygous stage. In this study, we used the RNAi technique to obtain close similar phenotype to this mutant, but that are fertile. RNAi strategy is performed by down-regulating the expression of both NTR genes by introducing RNAi construct harbouring two head-to-tail copies of the NTRA gene in the genetic background of the cad2 mutant. The transformed plants obtained exhibit attenuated phenotypes compared to the ntra ntrb cad2 mutant. Remarkably, no plants exhibit the characteristic pin-like phenotype of the ntra ntrb cad2 mutant was obtained. However, some plants looks fertile but show a decrease of the apical dominance. Others are more affected and show unfertile flowers. Our data show that the RNAi strategy is an efficient strategy to generate fertile plants with down-regulated NTS and NGS reduction systems and to investigate the crosstalk between these two thiol systems.
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