| 文心兰RFNR的克隆、亚细胞定位及其与LFNR不同的胁迫响应机制研究 |
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| 引用本文： | 李 蓉,吴晓佩,王雪晶,陈裕坤,郭容芳,林玉玲,赖钟雄,徐 涵.文心兰RFNR的克隆、亚细胞定位及其与LFNR不同的胁迫响应机制研究[J].园艺学报,2018,45(11):2164-2176. |
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| 作者姓名： | 李 蓉 吴晓佩 王雪晶 陈裕坤 郭容芳 林玉玲 赖钟雄 徐 涵 |
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| 作者单位： | （1福建农林大学园艺植物生物工程研究所，福州 350002；2法国图卢兹综合科学研究所（IRIT-ARI），法国 31300） |
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| 摘 要： | 以‘小樱桃’文心兰为材料克隆了铁氧还蛋白氧化还原酶（FNR）两种类型基因RFNR（Root-type Ferredoxin-NADP+ oxidoreductase）和LFNR（Leaf-type Ferredoxin-NADP+ oxidoreductase）。生物信息学分析表明，RFNR与LFNR都属于FNR-like超家族，具有黄素腺嘌呤二核苷酸（flavin adenine dinucleotide，FAD）和烟酰胺腺嘌呤二核苷酸（nicotinamide adenine dinucleotide，NAD）功能结合域，在进化过程中比较保守；两种类型的FNR在氨基酸组成、蛋白结构和构型方面存在明显差异，RFNR比LFNR有更强的保守性。FNR蛋白亚细胞定位结果表明文心兰FNR蛋白都定位于叶绿体。实时荧光定量PCR分析表明，LFNR和RFNR具有器官表达特异性：LFNR更多地在叶中表达，RFNR更多地在根中表达；在软腐病胁迫下LFNR下调响应，RFNR上调响应；两种类型的FNR在盐胁迫以及高温胁迫处理时上调响应，但RFNR响应较快且强度更大；LFNR和RFNR在水杨酸（salicylic acid，SA）处理时轻微波动，茉莉酸甲酯（methyl jasmonate，MeJA）处理时呈现单峰反应。文心兰两种类型的FNR基因在不同类型的胁迫中显示相似的和有区别的表达特性，表明其具有不同的胁迫响应分子机理。
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| 关 键 词： | 文心兰 铁氧还蛋白氧化还原酶 胁迫 亚细胞定位 基因表达 |
Cloning and Subcellular Localization of RFNR and the Mechanisms of Stress Induced Response of RFNR and LFNR in Oncidium |
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| LI Rong,WU Xiaopei,WANG Xuejing,CHEN Yukun,GUO Rongfang,LIN Yuling,LAI Zhongxiong,XUHAN Xu,.Cloning and Subcellular Localization of RFNR and the Mechanisms of Stress Induced Response of RFNR and LFNR in Oncidium[J].Acta Horticulturae Sinica,2018,45(11):2164-2176. |
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| Authors: | LI Rong WU Xiaopei WANG Xuejing CHEN Yukun GUO Rongfang LIN Yuling LAI Zhongxiong XUHAN Xu |
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| Institution: | （1Institute of Horticulture of Biotechnology，Fujian Agriculture and Forestry University，Fuzhou 350002，China；2Institut de la Recherche Interdisciplinaire de Toulouse，Toulouse，31300，France） |
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| Abstract: | Two types of FNR genes（Root-type Ferredoxin-NADP+ oxidoreductase，RFNR and Leaf-type Ferredoxin-NADP+ oxidoreductase，LFNR）were cloned from‘Little Cherry’Oncidium. The bioinformatics analysis showed that they both belonged to the FNR-like super-family with flavin adenine dinucleotide（FAD）and nicotinamide adenine dinucleotide（NAD）functional domains and they were conservative in the evolution process，but exhibited significant differences in the composition of amino acids，protein structure and configurations. The RFNR was more conservative than LFNR. The results of FNR protein localization showed their encoded proteins were found mainly in the chloroplasts. The real-time fluorescence quantitative PCR expression analysis showed that LFNR and RFNR had organ-specific expression，i.e. LFNR was mainly in the leaf and RFNR in the root. The soft rot induced the down-regulation of LFNR gene and the up-regulation of RFNR gene. Both LFNR and RFNR showed up-regulation in response to the treatments of high salt concentration and high temperature and RFNR showed more quickly and higher levels of response，as well as both responded slightly to salicylic acid（SA）treatment. When treated with methyl jasmonate（MeJA），both LFNR and RFNR exhibited single-peak expression. There were similar and different expression characteristics under various stress conditions in two types of FNR in Oncidium，which implied different molecular mechanisms in responding to different stresses. |
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| Keywords: | Oncidium FNR（Ferredoxin-NADP+ oxidoreductase） stress subcellular localization gene expression |
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