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毛竹(Phyllostachys edulis)镁原卟啉 IX 单甲酯环化酶基因 PeMPEC 的分子特征与初步功能分析

Molecular characterization and primary functional analysis of PeMPEC, a magnesium-protoporphyrin IX monomethyl ester cyclase gene of bamboo (Phyllostachys edulis).

机构信息

Key Laboratory on the Science and Technology of Bamboo and Rattan, International Center for Bamboo and Rattan, Beijing, 100102, People's Republic of China.

Institute of Gene Science for Bamboo and Rattan Resources, Beijing, 100102, People's Republic of China.

出版信息

Plant Cell Rep. 2015 Nov;34(11):2001-11. doi: 10.1007/s00299-015-1846-1. Epub 2015 Jul 28.

Abstract

A homologous gene of MPEC from Phyllostachys edulis was isolated and characterized. Its overexpression in Arabidopsis thaliana increased chlorophyll concentration and photosynthesis efficiency, indicating it is involved in chlorophyll biosynthesis. Magnesium-protoporphyrin IX monomethyl ester cyclase (MPEC) is an essential enzyme in the biosynthesis of chlorophyll, which plays an important role in photosynthesis. However, limited information is available on the roles of MPEC gene in bamboo. A homologous gene, PeMPEC was identified from Phyllostachys edulis, which comprised 1474 bp and contained an open reading frame encoding 415 amino acids. PeMPEC was transcribed abundantly in leaf blade where photosynthesis predominantly occurs, which agreed with the protein accumulation pattern confirmed by Western blotting. The PeMPEC transcription was promoted by continuous darkness for 24 h, and was suppressed by increasing light intensity (100-1500 µmo1 m(-2) s(-1)) and high temperature (42 °C). However, transcription was induced within 0.5 h and thereafter declined with prolonged treatment (up to 12 h) under low temperature (4 °C). Although PeMPEC expressed weakly in etiolated leaves, transcript levels increased gradually with subsequent light treatment (200 µmol m(-2) s(-1)). Overexpression of PeMPEC in Arabidopsis thaliana resulted in increased chlorophyll concentration and photosynthesis efficiency in sense transgenic plants compared with a reduction in antisense transgenic plants. These changes were consistent with the transcript levels of PeMPEC. These results indicated that PeMPEC might be involved in chlorophyll biosynthesis and play important roles in maintaining the stability of photosystems, and provide a basis for the study of chlorophyll biosynthesis and dissection of photosynthesis in bamboo.

摘要

从毛竹中分离并鉴定了一个与 MPEC 同源的基因。该基因在拟南芥中的过表达增加了叶绿素浓度和光合作用效率,表明其参与了叶绿素的生物合成。镁原卟啉 IX 单甲基酯环化酶(MPEC)是叶绿素生物合成中的一种必需酶,在光合作用中起着重要作用。然而,关于 MPEC 基因在竹子中的作用的信息有限。从毛竹中鉴定出一个同源基因 PeMPEC,它由 1474bp 组成,包含一个开放阅读框,编码 415 个氨基酸。PeMPEC 在叶片中大量转录,而光合作用主要发生在叶片中,这与 Western blot 验证的蛋白积累模式一致。连续黑暗 24 小时可促进 PeMPEC 的转录,而增加光照强度(100-1500µmol m(-2) s(-1))和高温(42°C)则抑制其转录。然而,在低温(4°C)下,转录在 0.5 小时内被诱导,随后随着处理时间的延长(长达 12 小时)而下降。虽然 PeMPEC 在黄化叶片中表达较弱,但随着随后的光照处理(200µmol m(-2) s(-1)),其转录水平逐渐增加。在拟南芥中过表达 PeMPEC 导致 sense 转基因植物的叶绿素浓度和光合作用效率增加,而 antisense 转基因植物的叶绿素浓度和光合作用效率降低。这些变化与 PeMPEC 的转录水平一致。这些结果表明 PeMPEC 可能参与叶绿素的生物合成,并在维持光合作用系统的稳定性方面发挥重要作用,为研究竹子中的叶绿素生物合成和光合作用的解析提供了基础。

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