Increasing Nitric Oxide Content in Arabidopsis thaliana by Expressing Rat Neuronal Nitric Oxide Synthase Resulted in Enhanced Stress Tolerance
Increasing Nitric Oxide Content in Arabidopsis thaliana by Expressing Rat Neuronal Nitric Oxide Synthase Resulted in Enhanced Stress Tolerance
复制标题
通过表达大鼠神经元一氧化氮合酶来增加拟南芥中的一氧化氮含量，从而增强应激耐受性
DOI：
10.1093/pcp/pcr181
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发表时间：
2012-02-01
影响因子：
4.9
通讯作者：
Lu, Ying-Tang
中科院分区：
文献类型：
--
作者：
Shi, Hai-Tao;Li, Rong-Jun;Lu, Ying-Tang
一氧化氮(NO)在植物的许多生理和发育过程中发挥着重要作用，包括生物和非生物胁迫，这些都会对农业生产产生不利影响。然而，由于缺乏关于一氧化氮合酶(NOS)的研究结果，在研究NO在体内的生理作用从而将其用于基因工程方面遇到了许多困难。在此，为了探索调控内源NO水平的可能性，我们在拟南芥中表达了大鼠神经元一氧化氮合酶(NNOS)。利用荧光探针3-氨基-4-氨甲基-2‘，7’-二乙酸二荧光素(DAF-FM DA)和血红蛋白测定，35S：：nNOS植株表现出较高的NOS活性和NO积累。与野生型相比，35S：：nNOS植株表现出更好的耐盐性和抗旱性，这进一步从生理参数的变化中得到证实，包括减少失水率，减小气孔开度，改变脯氨酸和丙二醛含量。实时定量聚合酶链式反应分析表明，在转基因株系中，几个逆境调节基因的表达上调。此外，转基因品系还表现出对紫丁香假单胞菌的抗病能力。番茄(PST)DC3000通过激活防御相关基因的表达。此外，我们还发现，35S：：nNOS系通过调控CO、FLC和LFY基因的表达而延迟开花。综上所述，这些结果表明，通过大鼠nNOS的表达来挖掘植物NO在各种过程中的作用是一种有用的策略。这种方法也可能对环境适应性更强的作物的基因工程有用。
Nitric oxide (NO) plays essential roles in many physiological and developmental processes in plants, including biotic and abiotic stresses, which have adverse effects on agricultural production. However, due to the lack of findings regarding nitric oxide synthase (NOS), many difficulties arise in investigating the physiological roles of NO in vivo and thus its utilization for genetic engineering. Here, to explore the possibility of manipulating the endogenous NO level, rat neuronal NOS (nNOS) was expressed in Arabidopsis thaliana. The 35S::nNOS plants showed higher NOS activity and accumulation of NO using the fluorescent probe 3-amino, 4-aminomethyl-2', 7'-difluorescein, diacetate (DAF-FM DA) assay and the hemoglobin assay. Compared with the wild type, the 35S::nNOS plants displayed improved salt and drought tolerance, which was further confirmed by changes in physiological parameters including reduced water loss rate, reduced stomatal aperture, and altered proline and malondialdehyde content. Quantitative real-time PCR analyses revealed that the expression of several stress-regulated genes was up-regulated in the transgenic lines. Furthermore, the transgenic lines also showed enhanced disease resistance against Pseudomonas syringae pv. tomato (Pst) DC3000 by activating the expression of defense-related genes. In addition, we found that the 35S::nNOS lines flowered late by regulating the expression of CO, FLC and LFY genes. Together, these results demonstrated that it is a useful strategy to exploit the roles of plant NO in various processes by the expression of rat nNOS. The approach may also be useful for genetic engineering of crops with increased environmental adaptations.