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超氧化物歧化酶缺失突变体在普通豆(Phaseolus vulgaris L.)中的表现:遗传控制、同工酶的差异表达以及对砷的敏感性。

Superoxide-dismutase deficient mutants in common beans (Phaseolus vulgaris L.): genetic control, differential expressions of isozymes, and sensitivity to arsenic.

机构信息

Department of Botany, RPM College, University of Calcutta, Uttarpara, West Bengal, Hooghly 712 258, India.

出版信息

Biomed Res Int. 2013;2013:782450. doi: 10.1155/2013/782450. Epub 2013 Aug 28.

Abstract

Two common bean (Phaseolus vulgaris L.) mutants, sodPv 1 and sodPv 2, exhibiting foliar superoxide dismutase (SOD) activity of only 25% and 40% of their mother control (MC) cv. VL 63 were isolated in EMS-mutagenized (0.15%, 8 h) M2 progeny. Native-PAGE analysis revealed occurrence of Mn SOD, Fe SOD, Cu/Zn SOD I and Cu/Zn SOD II isozymes in MC, while Fe SOD, and Mn SOD were not formed in sodPv 1 and sodPv 2 leaves, respectively. In-gel activity of individual isozymes differed significantly among the parents. SOD deficiency is inherited as recessive mutations, controlled by two different nonallelic loci. Gene expressions using qRT PCR confirmed higher expressions of Cu/Zn SOD transcripts in both mutants and the absence of Fe SOD in sodPv 1 and Mn SOD in sodPv 2. In 50 μM arsenic, Cu/Zn SODs genes were further upregulated but other isoforms downregulated in the two mutants, maintaining SOD activity in its control level. In an F2 double mutants of sodPv 1 × sodPv 2, no Fe SOD, and Mn SOD expressions were detectable, while both Cu/Zn SODs are down-regulated and arsenic-induced leaf necrosis appeared. In contrast to both mutants, ROS-imaging study revealed overaccumulation of both superoxides and H2O2 in leaves of double mutant.

摘要

两个普通菜豆(Phaseolus vulgaris L.)突变体,sodPv 1 和 sodPv 2,其叶片超氧化物歧化酶(SOD)活性仅为其母本对照(MC)cv. VL 63 的 25%和 40%,是在 EMS 诱变(0.15%,8 h)的 M2 后代中分离得到的。Native-PAGE 分析表明,MC 中存在 Mn SOD、Fe SOD、Cu/Zn SOD I 和 Cu/Zn SOD II 同工酶,而 sodPv 1 和 sodPv 2 叶片中则没有形成 Fe SOD 和 Mn SOD。同工酶的个体酶活在亲本间存在显著差异。SOD 缺乏是由两个不同的非等位基因座控制的隐性突变遗传的。使用 qRT-PCR 的基因表达证实,两种突变体中 Cu/Zn SOD 转录物的表达较高,而 sodPv 1 中没有 Fe SOD,sodPv 2 中没有 Mn SOD。在 50 μM 砷处理下,两种突变体中 Cu/Zn SODs 基因进一步上调,但其他同工酶下调,使 SOD 活性维持在对照水平。在 sodPv 1×sodPv 2 的 F2 双突变体中,检测不到 Fe SOD 和 Mn SOD 的表达,而两种 Cu/Zn SODs 都下调,并且出现砷诱导的叶片坏死。与两种突变体不同的是,ROS 成像研究显示,双突变体叶片中超氧化物和 H2O2 的积累过度。

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