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Effects of ERA1 Gene Down Regulation Mediated via siRNA on Arabidopsis Drought Tolerance

siRNA介导的ERA1表达下调对拟南芥抗旱性的影响

作 者 ：史金凤1，刘志学1，祝 建1，REINHARD Nehls2，王 泓1,2*

期 刊 ：西北植物学报 2012年 11期 页码：2157~2163

关键词：ERA1；RNAi技术；拟南芥；气孔开度；抗旱性；

Keywords:ERA1, RNAi technology, Arabidopsis thaliana, stomatal aperture, drought tolerance,

摘 要 ：ERA1是控制植物气孔开闭的一个重要基因，根据其保守域构建RNA干扰（RNAi）载体并转化拟南芥，考察转基因植株的生长、气孔导度、离体叶片失水率以及ERA1和相关基因表达，探讨siRNA介导的ERA1表达下调对拟南芥抗旱性的影响。结果表明：转基因拟南芥株系中ERA1的表达受到明显抑制，其离体叶片失水率低于野生型，但并未出现ERA1缺失突变体的负面生长表型；转基因株系对ABA处理比野生型更敏感，其ABA处理株的根长显著变短，气孔孔径更小；转基因株ABI1、ABI2、ATHB6的表达量降低，而RAB18、RD29B、ADH1的表达量升高，siRNA介导的ERA1表达下调可能会激活RAB18、RD29B等逆境响应元件。研究发现，采用RNAi技术可以有效下调ERA1表达，在没有过多负面生长表型的前提下提高拟南芥的抗旱性，且ERA1表达下调可能通过ABA途径正面影响拟南芥的抗旱性。

Abstract:ERA1 is an important gene in control of plant stomata opening.The conserved region of the gene has been chosen to construct RNAi vector to transform Arabidopsis.Growth,stomata aperture,detached leaf water loss and expression of ERA1 and some relative genes in transgenic plants were tested to evaluate how ERA1 down regulated by siRNA and affected Arabidopsis drought tolerance.The transgenic Arabidopsis showed clear repression of ERA1 gene expression.The transgenic plants are more sensitive to ABA comparing to wild type plant:the root is significant shorter and stomata aperture is smaller when treated with ABA.Moreover,the water loss rate of siRNA transgenic plant is lower than that of wild type control.However,siRNA transgenic plant has no visible unwanted growth phenotypes like observed in knock-out mutants.qRT-PCR analysis showed that repression of ERA1 by siRNA may activate some key stress responding gene like RAB18 and RD29B.This study found that use of siRNA to suppress the ERA1 gene can enhance Arabidopsis drought tolerance without any negative growth phenotype and may positively effect Arabidopsis drought tolerance through ABA pathway.

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