Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India.
Plant Mol Biol. 2012 Nov;80(4-5):503-17. doi: 10.1007/s11103-012-9964-4. Epub 2012 Sep 8.
A20/AN1 zinc finger domain containing Stress Associated Proteins (SAP) are involved in diverse stress response pathways in plants. In the present study, a novel banana SAP gene, MusaSAP1, was identified from banana EST database and was subsequently characterized by overexpression in transgenic banana plants. Expression profiling in native banana plants showed that MusaSAP1 was up-regulated by drought, salt, cold, heat and oxidative stress as well as by treatment with abscisic acid. Cellular localization assay carried out by making a MusaSAP1::GFP fusion protein indicated that MusaSAP1 is incompletely translocated to nucleus. Copy number analysis performed using real time PCR and Southern blotting indicated that MusaSAP1 occurs in the banana genome in a single copy per 11 chromosome set. Transgenic banana plants constitutively overexpressing MusaSAP1 displayed better stress endurance characteristics as compared to controls in both in vitro and ex vivo assays. Lesser membrane damage as indicated by reduced malondialdehyde levels in transgenic leaves subjected to drought, salt or oxidative stress pointed towards significant role for MusaSAP1 in stress amelioration pathways of banana. Strong up-regulation of a polyphenol oxidase (PPO) coding transcript in MusaSAP1 overexpressing plants together with induction of MusaSAP1 by wounding and methyl jasmonate treatment indicated possible involvement of MusaSAP1 in biotic stress responses where PPOs perform major functions in multiple defense pathways.
A20/AN1 锌指结构域包含应激相关蛋白 (SAP),参与植物中多种应激反应途径。本研究从香蕉 EST 数据库中鉴定出一种新型香蕉 SAP 基因 MusaSAP1,随后通过在转基因香蕉植物中过表达对其进行了表征。在天然香蕉植物中的表达谱分析表明,MusaSAP1 受干旱、盐、冷、热和氧化应激以及脱落酸处理的上调。通过构建 MusaSAP1::GFP 融合蛋白进行的细胞定位分析表明,MusaSAP1 不完全转位到核内。使用实时 PCR 和 Southern 印迹进行的拷贝数分析表明,MusaSAP1 在香蕉基因组中以每条 11 号染色体的单个拷贝存在。与对照相比,持续过表达 MusaSAP1 的转基因香蕉植物在体外和体内试验中表现出更好的应激耐受特性。干旱、盐或氧化胁迫下转基因叶片中丙二醛水平降低表明 MusaSAP1 在香蕉应激缓解途径中发挥重要作用。在过表达 MusaSAP1 的植物中,多酚氧化酶 (PPO) 编码转录物的强烈上调以及创伤和茉莉酸甲酯处理诱导 MusaSAP1 的表达表明,MusaSAP1 可能参与生物胁迫反应,其中 PPOs 在多种防御途径中发挥主要功能。