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首页> 外文期刊>Planta: An International Journal of Plant Biology >Overexpression of glycerol-3-phosphate acyltransferase gene improves chilling tolerance in tomato
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Overexpression of glycerol-3-phosphate acyltransferase gene improves chilling tolerance in tomato

机译:甘油-3-磷酸酰基转移酶基因的过表达提高番茄的耐寒性

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A tomato (Lycopersicon esculentum Mill.) glycerol-3-phosphate acyltransferase gene (LeGPAT) was isolated. The deduced amino acid sequence revealed that LeGPAT contained four acyltransferase domains, showing high identities with GPAT in other plant species. A GFP fusion protein of LeGPAT was targeted to chloroplast in cowpea mesophyll protoplast. RNA gel blot showed that the mRNA accumulation of LeGPAT in the wild type (WT) was induced by chilling temperature. Higher expression levels were observed when tomato leaves were exposed to 4 degrees C for 4 h. RNA gel and western blot analysis confirmed that the sense gene LeGPAT was transferred into the tomato genome and overexpressed under the control of 35S-CaMV. Although tomato is classified as a chilling-sensitive plant, LeGPAT exhibited selectivity to 18:1 over 16:0. Overexpression of LeGPAT increased total activity of LeGPAT and cis-unsaturated fatty acids in PG in thylakoid membrane. Chilling treatment induced less ion leakage from the transgenic plants than from the WT. The photosynthetic rate and the maximal photochemical efficiency of PS II (Fv/Fm) in transgenic plants decreased more slowly during chilling stress and recovered faster than in WT under optimal conditions. The oxidizable P700 in both WT and transgenic plants decreased obviously at chilling temperature under low irradiance, but the oxidizable P700 recovered faster in transgenic plants than in the WT. These results indicate that overexpression of LeGPAT increased the levels of PG cis-unsaturated fatty acids in thylakoid membrane, which was beneficial for the recovery of chilling-induced PS I photoinhibition in tomato.
机译:分离出番茄(Lycopersicon esculentum Mill。)的甘油3-磷酸酰基转移酶基因(LeGPAT)。推导的氨基酸序列表明,LeGPAT包含四个酰基转移酶结构域,与其他植物中的GPAT具有高度同一性。 LeGPAT的GFP融合蛋白靶向cow豆叶肉原生质体中的叶绿体。 RNA凝胶印迹显示,低温可诱导野生型LeGPAT的mRNA积累。当番茄叶片暴露于4摄氏度4小时时,观察到更高的表达水平。 RNA凝胶和蛋白质印迹分析证实有义基因LeGPAT已转移到番茄基因组中,并在35S-CaMV的控制下过表达。尽管番茄被归类为对寒冷​​敏感的植物,但LeGPAT对18:1的选择性高于16:0。 LeGPAT的过表达增加类囊体膜中PG的LeGPAT和顺式不饱和脂肪酸的总活性。与WT相比,低温处理引起的转基因植物离子泄漏更少。在最佳条件下,低温胁迫下转基因植物中PS II(Fv / Fm)的光合作用速率和最大光化学效率降低得更慢,恢复速度更快。在低温和低辐照度下,野生型和转基因植物中可氧化的P700均明显降低,但转基因植物中可氧化的P700的恢复比野生型中的更快。这些结果表明LeGPAT的过表达增加类囊体膜中PG顺式不饱和脂肪酸的水平,这有利于恢复番茄中冷诱导的PS I光抑制。

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