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SsCHLAPXs 的过表达赋予转基因拟南芥对高光诱导的氧化应激的保护作用。

Overexpression of SsCHLAPXs confers protection against oxidative stress induced by high light in transgenic Arabidopsis thaliana.

机构信息

Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University, Jinan 250014, Shandong, China.

出版信息

Physiol Plant. 2011 Dec;143(4):355-66. doi: 10.1111/j.1399-3054.2011.01515.x. Epub 2011 Oct 3.

Abstract

To evaluate the physiological importance of chloroplastic ascorbate peroxidase (CHLAPX) in the reactive oxygen species (ROS)-scavenging system of a euhalophyte, we cloned the CHLAPX of Suaeda salsa (SsCHLAPX) encoding stromal APX (sAPX) and thylakoid-bound APX. The stromal APX of S. salsa (Ss.sAPX) cDNA consists of 1726 nucleotides including an 1137-bp open reading frame (ORF) and encodes 378 amino acids. The thylakoid-bound APX of S. salsa (Ss.tAPX) cDNA consists of 1561 nucleotides, including a 1284-bp ORF, and encodes 427 amino acids. The N-terminal 378 amino acids of Ss.sAPX are identical with those of Ss.tAPX, whereas the C-terminal 49 amino acids differ. Arabidopsis thaliana lines overexpressing Ss.sAPX and Ss.tAPX were constructed using Agrobacterium tumefaciens transformation methods. Under high light (1000 µmol m⁻² s⁻¹), malondialdehyde (MDA) content was lower in transgenic plants than in the wild type. Under high light, Fv/Fm and chlorophyll contents of both overexpressing lines and the wild type declined but were significantly higher in the overexpressing lines than in the wild type. The activities of APX (EC 1.11.1.11), catalase (CAT 1.11.1.6) and superoxide dismutase (SOD EC 1.15.1.1) were higher in the overexpressing lines than in the wild type. The transgenic plants showed increased tolerance to oxidative stress caused by high light. These results suggest that SsCHLAPX plays an important role in scavenging ROS in chloroplasts under stress conditions such as high light.

摘要

为了评估真盐生植物活性氧(ROS)清除系统中质体抗坏血酸过氧化物酶(CHLAPX)的生理重要性,我们克隆了盐地碱蓬(Suaeda salsa)的 CHLAPX,该基因编码基质 APX(sAPX)和类囊体结合 APX。盐地碱蓬基质 APX(Ss.sAPX)cDNA 由 1726 个核苷酸组成,包括一个 1137bp 的开放阅读框(ORF),编码 378 个氨基酸。盐地碱蓬类囊体结合 APX(Ss.tAPX)cDNA 由 1561 个核苷酸组成,包括一个 1284bp 的 ORF,编码 427 个氨基酸。Ss.sAPX 的 N 端 378 个氨基酸与 Ss.tAPX 的相同,而 C 端 49 个氨基酸不同。使用根癌农杆菌转化方法构建了过表达 Ss.sAPX 和 Ss.tAPX 的拟南芥株系。在高光(1000 µmol m⁻² s⁻¹)下,转基因植物中的丙二醛(MDA)含量低于野生型。在高光下,过表达株系和野生型的 Fv/Fm 和叶绿素含量均下降,但过表达株系显著高于野生型。过表达株系的 APX(EC 1.11.1.11)、过氧化氢酶(CAT 1.11.1.6)和超氧化物歧化酶(SOD EC 1.15.1.1)活性均高于野生型。转基因植物对高光引起的氧化胁迫表现出更高的耐受性。这些结果表明,SsCHLAPX 在高光等胁迫条件下,在叶绿体中清除 ROS 中发挥重要作用。

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