Over-expression of a protein disulfide isomerase gene from Methanothermobacter thermautotrophicus, enhances heat stress tolerance in rice
Over-expression of a protein disulfide isomerase gene from Methanothermobacter thermautotrophicus, enhances heat stress tolerance in rice
复制标题
嗜热甲烷杆菌中蛋白质二硫键异构酶基因的过度表达可增强水稻的热应激耐受性
DOI：
10.1016/j.gene.2018.10.064
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发表时间：
2019
期刊：
影响因子：
3.5
通讯作者：
Peng Xiaojue
中科院分区：
文献类型：
--
作者：
Wang Xin;Chen Jie;Liu Changai;Luo Junling;Yan Xin;Ai Aihua;Cai Yaohui;Xie Hongwei;Ding Xia;Peng Xiaojue
高温胁迫是限制全球农业生产的主要环境胁迫。发现和利用促进作物耐高温的基因是提高作物耐热性的关键。与分子伴侣和蛋白质折叠相关的蛋白质在植物的高温胁迫反应中起着重要的作用。MTH 1745（MtPDI）是一种来源于嗜热甲烷杆菌（Methanothermobacter thermautotrophicus delta H）的具有分子伴侣功能和二硫键异构酶活性的二硫键异构酶样蛋白（disulfide isomerase-like protein，PDI）。通过实时荧光定量PCR和western blot的分子鉴定，证明MtPDI基因在转基因水稻中得到了稳定表达。转基因水稻幼苗的耐热性和存活率显著提高。同时，转MtPDI基因水稻的脯氨酸含量、超氧化物歧化酶（SOD）和过氧化物酶（POD）活性增加，丙二醛（MDA）含量降低。此外，还发现转基因植物中巯基含量增加。这些结果表明，异源表达极端微生物MtPDI可通过积累脯氨酸含量、协同提高抗氧化酶活性和增加巯基的产生来增强转基因水稻的热胁迫耐受性，从而减轻氧化损伤。
High temperature (HT) stress is a major environmental stress that limits agricultural production worldwide. Discovery and application of genes promoting high temperature tolerance is essential to enhance crop tolerance to heat stress. Proteins associated with chaperone and protein folding plays an important role in the high temperature stress response of plants. MTH1745 (MtPDI), a disulfide isomerase-like protein (PDI) with a chaperone function and disulfide isomerase activity from Methanothermobacter thermautotrophicus delta H, was selected for studying the heat stress tolerance using an ectopic expression method in rice. Through molecular identification via quantitative real-time PCR and western blot, we demonstrated that theMtPDIgene was expressed stably in transgenic rice. Heat stress tolerance and survival ratio were significantly improved in seedling transgenic rice. At the same time, proline content, superoxide dismutase (SOD) and peroxidase (POD) activities were increased inMtPDItransgenic rice with a reduced malondialdehyde (MDA) content. Moreover, increased content of thiols group was discovered in transgenic plants. These results indicate that heterologous expression ofMtPDIfrom extremophiles could confer heat stress tolerance of transgenic rice through the accumulation of proline content, the synergistic increase of the antioxidant enzymes activity and elevated production of more thiols group, which finally ameliorated the oxidative damage.