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Expression analysis of the two ferrochelatase genes in Arabidopsis in different tissues and under stress conditions reveals their different roles in haem biosynthesis

机译:拟南芥中两个铁螯合酶基因在不同组织中和在胁迫条件下的表达分析表明,它们在血红素生物合成中的作用不同

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The Arabidopsis thaliana genome has two genes (AtFC-I and AtFC-II), encoding ferrochelatase, the terminal enzyme of haem biosynthesis. The roles of the two enzymes in the synthesis of haem for different haemoproteins was investigated using reporter gene analysis. A 1.41 kb fragment from the 5' upstream region of the AtFC-II gene was fused to the luciferase gene, and then introduced into tobacco plants, followed by luciferase activity measurements. AtFC-II-LUCwas expressed in all aerial parts of the plant, and was highest in flowers, but it was not expressed in roots. It was unaffected by viral infection, and considerably reduced by wounding or oxidative stress. Similarly, a 1.76 kb region of the AtFC-I promoter was fused to the uidA gene encoding beta-glucuronidase. AtFC-I-GUS was expressed in all tissues of the plant, but was higher in roots and flowers than in leaves or stems. It was induced by sucrose, wounding and oxidative stress and, most markedly, by plants undergoing the hypersensitive response to TMV infection. Levels of endogenous ferrochelatase activity were increased in pea chloroplasts isolated from wounded leaves, indicating that the induction in promoter activity is likely to result in increased haem biosynthetic potential. Salicylic acid, but not methyl-jasmonate was able to replace the stress treatment in induction of AtFC-I expression, suggesting that the requirement for haem synthesis is part of the defence response. The implications of the results for the different roles of the two ferrochelatases in haem biosynthesis are discussed.
机译:拟南芥基因组有两个基因(AtFC-I和AtFC-II),编码铁螯合酶,即血红素生物合成的末端酶。使用报告基因分析研究了两种酶在不同血红蛋白血红素合成中的作用。将来自AtFC-II基因5'上游区域的1.41 kb片段与萤光素酶基因融合,然后引入烟草植株中,随后进行萤光素酶活性测量。 AtFC-II-LUC在植物的所有空中部分表达,在花中最高,但在根中不表达。它不受病毒感染的影响,并因受伤或氧化应激而大大减少。类似地,将AtFC-1启动子的1.76kb区域与编码β-葡糖醛酸糖苷酶的uidA基因融合。 AtFC-I-GUS在植物的所有组织中均有表达,但在根和花中高于叶或茎。它是由蔗糖,伤口和氧化应激引起的,最明显的是由对TMV感染产生超敏反应的植物诱导的。从受伤叶片分离的豌豆叶绿体中内源性铁螯合酶活性水平增加,表明启动子活性的诱导可能导致血红素生物合成潜力的增加。水杨酸,但茉莉酸甲酯不能替代应激处理,从而诱导AtFC-1表达,这表明血红素合成的要求是防御反应的一部分。讨论了结果对两种铁螯合酶在血红素生物合成中的不同作用的影响。

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