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巨桉EgrWRKY70基因克隆和初步表达分析     被引量:3

Cloning and Expression Analysis of Egr WRKY70 in Eucalyptus grandis

文献类型:期刊文献

中文题名:巨桉EgrWRKY70基因克隆和初步表达分析

英文题名:Cloning and Expression Analysis of Egr WRKY70 in Eucalyptus grandis

作者:姚海荣[1,2,3] 曾炳山[2] 范春节[2] 裘珍飞[2] 郭光生[2] 覃伟权[3] 阎伟[3]

第一作者:姚海荣

机构:[1]海南大学环境与植物保护学院;[2]中国林业科学研究院热带林业研究所;[3]中国热带农业科学院椰子研究所

年份:2016

卷号:37

期号:7

起止页码:1341-1348

中文期刊名:热带作物学报

外文期刊名:Chinese Journal of Tropical Crops

收录:CSTPCD;;北大核心:【北大核心2014】;CSCD:【CSCD_E2015_2016】;

基金:国家科技计划项目(No.2013AA102705);中央级公益性科研院所基本科研业务费专项资金项目(No.RITFYWZX201304)

语种:中文

中文关键词:巨桉;Egr;WRKY70;基因克隆;差异表达

外文关键词:Eucalyptus grandis; Egr WRKY70; Gene cloning; Gene expression

分类号:S792.39

摘要:WRKY家族为植物所特有,参与调控植物逆境胁迫、生长发育和衰老等生物学过程。为了获得WRKY70同源基因在桉树响应胁迫中的作用和功能,克隆巨桉(Eucalyptus grandis)中的Egr WRKY70基因,通过生物信息学分析和q RT-PCR进行功能的初步鉴定。结果表明,Egr WRKY70基因编码321个氨基酸,蛋白分子量为36.18 ku,只有一个WRKY结构域,属于WRKY转录因子III a类成员。同时,Egr WRKY70与麻风树的Jcu WRKY70的亲缘关系最近,其氨基酸序列相似性达到58.2%。半定量RT-PCR分析表明,Egr WRKY70主要在巨桉叶片、根和花中表达,而在茎部表达量较低。实时定量q RT-PCR分析显示:Egr WRKY70基因在低温和盐胁迫下,表达量快速升高然后下降;在盐胁迫下Egr WRKY70基因表达仍然维持在较高水平;在油菜素内酯和水杨酸处理时,Egr WRKY70基因能够快速的响应,使用茉莉酸甲酯处理时其表达量没有发生明显的变化。由此表明,Egr WRKY70能够通过不同的响应方式参与桉树生物和非生物胁迫的应答。
The plant-specific WRKY transcription factor family appears to be involved in the regulation of various biological processes including response to stresses, growth and development, and senescence. In order to identify the function response to stress of WRKY70 in Eucalyptus grandis, Egr WRKY70 was cloned and analyzed by using reverse transcription polymerase chain reaction(RT-PCR) and real time quantitative PCR(q RT-PCR). Egr WRKY70 encoded a predict protein of 321 amino acids. The theoretical molecular weight of Egr WRKY70 was 36.18 ku with p I of 5.28, unstability index of 63.67. Egr WRKY70 contained a conserved WRKY domain and belonged to group IIIa. The results of multiple alignments and phylogenetic analysis revealed that Egr WRKY70 closed to Jcu WRKY70 with 58.2% similarity. The expression of Egr WRKY70 was detected in flowers, leaves and roots by semi-quantitative reverse transcription and polymerase chain reaction, and Egr WRKY70 did not nearly expressed in stem including xylem and phloem. Expression of Egr WRKY70 increased firstly and then decreased with the duration of the treatment of salt and cold. However, the Egr WRKY70 expression maintained at a high level under salt stress. Besides, after the treatments of salicylic acid and brassinolide, the levels of Egr WRKY70 expression increased rapidly and then decreased. The expression of Egr WRKY70 almost did not changed after the treatment of methyl jasmonate. These results suggested that Egr WRKY70 may have relationship with biotic and abiotic stress responses in E. grandis.

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