Molecular cloning of a 14-3-3 protein gene from Lilium regale Wilson and overexpression of this gene in tobacco increased resistance to pathogenic fungi
Molecular cloning of a 14-3-3 protein gene from Lilium regale Wilson and overexpression of this gene in tobacco increased resistance to pathogenic fungi
复制标题
百合 14-3-3 蛋白基因的分子克隆以及该基因在烟草中的过度表达增加了对病原真菌的抵抗力
DOI：
10.1016/j.scienta.2013.12.034
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发表时间：
2014-03-26
影响因子：
4.3
通讯作者：
Chen, Chaoyin
中科院分区：
文献类型：
--
作者：
Li, Hongli;Liu, Diqiu;Chen, Chaoyin
14-3-3蛋白存在于所有真核生物中，是调节与多种生物学过程相关的多种信号通路的重要调节因子。此外，它们还参与保护植物免受生物和非生物胁迫的分子网络，据报道，许多14-3-3在植物对病原菌的防御反应中发挥着重要作用。为了分离与病原侵染过程中的自我防御有关的14-3-3基因，从百合根部分离到一个新基因LR14-3-3，LR14-3-3是一种野生百合，对植物病原菌、病毒和干旱具有很强的抗性。LR14-3-3在百合正常发育的根、茎、叶中表达水平较低，而在L百合与尖孢镰刀菌不亲和互作以及感病东方百合‘西伯利亚’与尖孢镰刀菌亲和互作中表达显著上调。而在尖孢镰刀菌侵染过程中，L王朝的LR14-3-3转录水平明显高于‘西伯利亚’。此外，LR14-3-3的基因表达受乙烯处理的影响。此外，为了验证LR14-3-3的功能，构建了LR14-3-3的植物表达载体，并将其导入烟草(Nictiana Tabacum L Cv Xanthi)中。定量逆转录-聚合酶链式反应(QRT-PCR)分析表明，LR14-3-3基因在转基因烟草中稳定表达，与对照相比，抗病相关基因N基因、渗透素、MYC和NADPH氧化酶基因在转基因烟草中表达上调。此外，超氧化物歧化酶、谷胱甘肽S转移酶和抗坏血酸过氧化物酶活性在T1转基因系中显著高于WT系。对转LR14-33基因烟草进行了体外平板和体内接种试验，结果表明，转基因烟草对番茄灰霉病、拟南芥和尖孢镰刀菌的侵染有明显的抗性。(C)2014爱思唯尔B.V.保留所有权利。
14-3-3 proteins exist in all eukaryotes and act as important regulators to modulate diverse signaling pathways associated with a wide range of biological processes. Besides, they are involved in molecular networks protecting plant from damages caused by the biotic and abiotic stresses, and it has been reported that many 14-3-3s play an important role in defense response against pathogens. To isolate 14-3-3 genes involved in self-defense during pathogenic infection, a novel gene Lr14-3-3 was isolated from the root of Lilium regale Wilson which is a kind of wild lily species with high resistance to phytopathogens, viruses, and drought. The Lr14-3-3 was expressed at relatively low level in root, stem, and leaf of L. regale under normal development, by contraries, Lr14-3-3 was significantly up-regulated in an incompatible interaction between L regale and Fusarium oxysporum as well as in a compatible interaction between susceptible Lilium Oriental Hybrid 'Siberia' and F. oxysporum. However, the transcription level of Lr14-3-3 in L regale was evidently higher than that in 'Siberia' during F. oxysporum infection. Moreover, the gene expression of Lr14-3-3 was responded to ethylene treatment. Furthermore, in order to verify the function of Lr14-3-3, the constitutive plant expression vector ofLr14-3-3 was constructed and transferred into tobacco (Nicotiana tabacum L cv Xanthi). The following quantitative reverse transcription-PCR (QRTPCR) analysis demonstrated that the Lr14-3-3 gene was steadily expressed in the Ti transgenic tobacco lines, and compared with the WT, the disease resistance-related genes including N gene, osmotin, MYC, and NADPH oxidase gene were up-regulated in the transgenic lines. Moreover, the antioxidant enzymes, superoxide dismutase (SOD), glutathione S-transferase (GST), and ascorbate peroxidase (APX), showed significantly higher activities in the T1 transgenic lines than that in WT. The antifungal activity of Lr14-33 transgenic tobacco was performed in vitro plates and in vivo inoculation, respectively, and the results showed that the transgenic tobacco plants evidently resisted the infection of Botryosphaeria dothidea, Phomopsis sp., and F. oxysporum. (C) 2014 Elsevier B.V. All rights reserved.