樟树NBS-LRR类抗病基因家族分析与CcRNL基因克隆

doi:10.13560/j.cnki.biotech.bull.1985.2017-0570

生物技术通报 ›› 2018, Vol. 34 ›› Issue (2): 142-149.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0570

樟树NBS-LRR类抗病基因家族分析与CcRNL基因克隆

郑永杰, 伍艳芳, 李江, 章挺, 汪信东

江西省林业科学院国家林业局樟树工程技术研究中心,南昌 330032;

收稿日期:2017-07-10 出版日期:2018-02-26 发布日期:2018-03-12
作者简介:郑永杰,男,硕士,研究实习员,研究方向：林木遗传育种;E-mail：zyj_bio2015@163.com
基金资助:
江西省科技计划项目（20143ACF60014）,江西省林业科学院博士启动项目

Identification of NBS-LRR-like Disease-resistant Genes in Cinnamomum camphora and Clone of Two CcRNL Genes

ZHENG Yong-jie, WU Yan-fang, LI Jiang, ZHANG Ting, WANG Xin-dong

Camphor Engineering Technology Research Center for State Forestry Administration,Jiangxi Academy of Forestry,Nanchang 330032;

Received:2017-07-10 Published:2018-02-26 Online:2018-03-12

摘要/Abstract

摘要： 樟树（Cinnamomum camphora L. Presl.）是自然界中较少受病原体侵害的优良抗性种质资源。利用生物信息的方法对樟树叶组织转录组数据中NBS类抗病基因进行了鉴定,对其编码的假定蛋白的理化性质、保守基序及系统发生关系等进行分析。结果表明,在樟树叶组织转录组中共找到28个具有完整编码框的NBS类抗病基因,大小在2 118-3 378 bp之间,编码蛋白等电点在5.35-9.03之间;在NBS区域内鉴定获得8个保守motifs,系统发育树将28个成员分为5个亚组。通过PCR法克隆获得2个RNL类抗病基因全长,分别命名为CcRNL1和CcRNL2,在氨基酸序列水平二者相似性为71%,在多肽N端都具有RPW8-like保守序列,暗示它们可能具有拟南芥RPW8基因所具有的广谱抗白粉病功能。采集感白粉病和正常樟树幼叶进行表达分析,结果显示CcRNL1可被樟树白粉病菌诱导表达,推测其可能参与了响应白粉病菌胁迫反应过程。

关键词: 樟树, NBS类抗病基因, 樟树白粉病, 诱导表达

Abstract: Cinnamomum camphora is an germplasm resource with excellent resistance against pathogens in nature. Based on transcriptome data of C. camphora leaf tissue,the disease-resistant genes（R）of NBS（nucleotide-binding site）family were identified,and the physic-chemical properties,motifs and phylogenetic relationship of encoded putative proteins were analyzed through bioinformatics analysis. As results showed,28 NBS-like disease-resistant genes with complete encoding frame were identified from leaf transcriptome of C. camphora leaf tissue. The lengths of ORF ranged in 2 118-3 378 bp,and the estimated isoelectric point of putative proteins were in 5.35-9.03. The 8 motifs were identified in the NBS domain. The 28 R genes were divided into 5 subgroups based on the phylogenetic analysis. Additionally,2 RNL genes were cloned by PCR and designated as CcRNL1 and CcRNL2. The two genes shared 71% similarity in amino acid sequence,and there was RPW8-like conservative motif in the N-terminal of the polypeptides,indicating that they might have broad-spectrum resistance to powdery mildew as gene RPW8 in Arabidopsis thaliana has. Analyzing the young leaves from powdery mildew-infected C. camphora and normal one,the results revealed that CcRNL1 expressed by the induction of powdery mildew in C. camphora,therefore it was inferred to be involved in the response to stress process of powdery mildew pathogens.

Key words: Cinnamomum camphora, NBS-like disease-resistant gene, powdery mildew, expression analysis

郑永杰, 伍艳芳, 李江, 章挺, 汪信东. 樟树NBS-LRR类抗病基因家族分析与CcRNL基因克隆[J]. 生物技术通报, 2018, 34(2): 142-149.

ZHENG Yong-jie, WU Yan-fang, LI Jiang, ZHANG Ting, WANG Xin-dong. Identification of NBS-LRR-like Disease-resistant Genes in Cinnamomum camphora and Clone of Two CcRNL Genes[J]. Biotechnology Bulletin, 2018, 34(2): 142-149.

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樟树NBS-LRR类抗病基因家族分析与CcRNL基因克隆

Identification of NBS-LRR-like Disease-resistant Genes in Cinnamomum camphora and Clone of Two CcRNL Genes

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