椰心叶甲CYP4基因的克隆及绿僵菌侵染后的诱导表达研究

›› 2007, Vol. 50 ›› Issue (1): 14-19.

椰心叶甲CYP4基因的克隆及绿僵菌侵染后的诱导表达研究

王九辉，黄俊生^*，方晓东

（中国热带农业科学院环境与植物保护研究所，热带作物生物技术国家重点实验室，海口 571101）

出版日期:2007-01-20 发布日期:2007-02-20
通讯作者: 黄俊生

Identification and induction by Metarhizium anisopliae infection of family 4 cytochrome P450 in Brontispa longissima

WANG Jiu-Hui,HUANG Jun-Sheng, FANG Xiao-Dong

(State Key Biotechnology Laboratory for Tropical Crops, Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China)

Online:2007-01-20 Published:2007-02-20
Contact: HUANG Jun-Sheng

摘要/Abstract

摘要：

根据细胞色素P450家族4（CYP4）的氨基酸保守序列设计1对简并引物，从椰心叶甲Brontispa longissima成虫总RNA中扩增得到5个cDNA片段（GenBank登录号: DQ238840－DQ238844）。以3′-RACE法获得片段BLWH4的3′端序列，推导的氨基酸序列表明其结构中含有CYP家族的特征性保守序列：螺旋K区的ETLR和血红素结合区的F××G×××C×G。以18S 为对照的RT-PCR分析表明，BLWH4在成虫的mRNA表达量远大于幼虫。绿僵菌Metarhizium anisopliae菌株MA-3和MA-4侵染椰心叶甲成虫及5龄幼虫后，BLWH4的mRNA表达增强，提示BLWH4可能具有增强椰心叶甲抵抗绿僵菌侵染的作用。

关键词: 椰心叶甲, CYP4, RT-PCR, mRNA表达, 绿僵菌

Abstract: In order to obtain cDNA clones coding for CYP4 proteins in Brontispa longissima, degenerate primers were designed using the conserved sequences of known CYP4s and used to amplify partial cDNA fragments from adults B. longissima. Five genes were identified (GenBank accession numbers: DQ238840 －DQ238844), and the 3′-end of BLWH4 was cloned by 3′-RACE. The deduced amino acid sequence of BLWH4 has CYP conserved domains of helix K and heme-binding motif. The RT-PCR analysis indicated that the BLWH4 mRNA expression in adults was more than in larvae, and increased when adults and 5th instar larvae were infected with Metarhizium anisopliae MA-3 and MA-4. The results suggested that BLWH4 should play a role in protection against pathogenic fungus.

Key words: Brontispa longissima, CYP4, RT-PCR, mRNA expression, Metarhizium anisopliae

王九辉，黄俊生^*，方晓东. 椰心叶甲CYP4基因的克隆及绿僵菌侵染后的诱导表达研究[J]. , 2007, 50(1): 14-19.

WANG Jiu-Hui,HUANG Jun-Sheng, FANG Xiao-Dong. Identification and induction by Metarhizium anisopliae infection of family 4 cytochrome P450 in Brontispa longissima[J]. , 2007, 50(1): 14-19.

[1]	唐芬芬, 郑亚强, 白兴荣, 陈斌. 短时热激后家蚕对核型多角体病毒的抗性及相关抗菌肽和热激蛋白基因表达水平的变化[J]. , 2017, 60(5): 505-512.
[2]	张志翔, 张美萍, 王桂荣, 刘杨. 绿盲蝽气味受体基因AlucOR40的克隆及功能研究[J]. , 2016, 59(6): 592-601.
[3]	程小娟, 蔡立君, 郑丽双, 覃江梅, 黄宇萍, 尤民生. 小菜蛾气味结合蛋白OBP2基因的克隆、表达谱及其结合特性分析[J]. , 2016, 59(4): 365-376.
[4]	杨丽红, 豆威, 蒋红波, 牛金志, 丁天波, 王进军. 柑橘全爪螨热激蛋白基因PcHsp90的克隆及表达模式分析[J]. , 2016, 59(4): 411-420.
[5]	张中印, 杨珊珊, 孟娇, 赵慧婷, 姜玉锁. 中华蜜蜂嗅觉受体AcerOrco的表达及定位分析[J]. , 2016, 59(2): 185-191.
[6]	唐文超, 肖媛媛, 杨成飞, 郭东东, 朱勇. 氟化物导致的家蚕生殖损伤及氧化应激反应[J]. , 2016, 59(12): 1308-1316.
[7]	刘一鹏, 刘杨, 杨婷, 桂富荣, 王桂荣. 小菜蛾普通气味受体基因 PxylOR9 的鉴定及功能研究[J]. , 2015, 58(5): 507-515.
[8]	李慧, 费东亮, 胡影, 马鸣潇. 中华蜜蜂幼虫原代细胞培养及中华蜜蜂囊状幼虫病毒在培养的细胞中的复制[J]. , 2015, 58(12): 1362-1367.
[9]	陈静, 张道伟. 德国小蠊两个海藻糖合成酶基因的克隆、组织分布及温度诱导表达分析[J]. , 2015, 58(10): 1046-1053.
[10]	刘宁灿，张进，王桂荣，刘杨，严善春. 棉铃虫普通气味受体基因HarmOR9和HarmOR29的克隆和组织表达分析[J]. , 2014, 57(5): 522-529.
[11]	韩岚岚，朱明贺，董天宇，赵奎军，曲忠诚，吕洋，韩晓旭. 热应激和吡虫啉对大豆蚜hsp70和hsc70基因mRNA表达的影响[J]. , 2014, 57(4): 387-394.
[12]	童应华，李万里，马淑娟. 金龟子绿僵菌及其粗毒素对樟巢螟幼虫的致病性[J]. , 2014, 57(4): 418-427.
[13]	袁红霞, 褚峰, 秦粉菊, 刘佳, 何亚清, 刘航. 镉胁迫对家蚕脂肪体脂质过氧化物含量及抗氧化酶活性和mRNA表达的影响[J]. , 2014, 57(2): 168-175.
[14]	黄琼, 胡杰, 王勤. 两种色型黄粉虫酚氧化酶原的cDNA克隆、生物信息学分析及表达水平检测[J]. , 2013, 56(6): 594-604.
[15]	黄琼, 胡杰, 孙灵, 王勤. 黄粉虫热休克蛋白70基因的克隆、序列分析与表达（英文）[J]. , 2013, 56(5): 475-485.