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LncRNA在柑橘木虱与黄龙病病原菌互作中的差异表达分析及验证
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引用本文:陈梦瑶,曹征鸿,贺康,梅洋,魏圣飞,肖花美.LncRNA在柑橘木虱与黄龙病病原菌互作中的差异表达分析及验证.植物保护学报,2019,46(6):1252-1261
DOI:10.13802/j.cnki.zwbhxb.2019.2018184
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陈梦瑶 宜春学院生命科学与资源环境学院, 江西省作物生长发育调控重点实验室, 宜春 336000
浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058 
 
曹征鸿 浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058  
贺康 浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058  
梅洋 浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058  
魏圣飞 浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058  
肖花美 宜春学院生命科学与资源环境学院, 江西省作物生长发育调控重点实验室, 宜春 336000
浙江大学昆虫科学研究所, 农业农村部作物病虫分子生物学重点实验室, 杭州 310058 
xiaohuamei625@163.com 
中文摘要:为明确柑橘黄龙病的唯一自然传播媒介——柑橘木虱Diaphorina citri的长链非编码RNA(long non-coding RNA,lncRNA)是否参与调控黄龙病病原菌Candidatus Liberibacter asiaticus(CLas)的侵染及复制,采用生物信息学预测及PCR扩增方法进行lncRNA的预测、特征分析、验证及差异表达分析。结果显示,柑橘木虱的13个转录组RNA-Seq数据中共有10 192个lncRNA基因,对应15 747条lncRNA转录本;与蛋白质编码基因相比,柑橘木虱lncRNA具有更少的外显子数量和更短的转录本长度;随机选取的10条lncRNA基因中,有7条lncRNA基因在无菌柑橘木虱广州品系或赣州品系中有表达,其中1条lncRNA基因TCONS_00034665在无菌广州品系和无菌赣州品系中存在差异表达;带菌和无菌柑橘木虱成虫中预测获得2个差异表达的lncRNA基因TCONS_00096118和TCONS_00234564,实时荧光定量PCR验证发现TCONS_00234564与预测结果一致,在带菌柑橘木虱成虫中高表达。表明lncRNA参与了黄龙病病原菌与寄主柑橘木虱的互作。
中文关键词:长链非编码RNA  柑橘木虱  黄龙病病原菌  RNA-Seq  验证
 
Differential expression analysis and validation of lncRNA between Asian citrus psyllid and Candidatus Liberibacter asiaticus interaction
Author NameAffiliationE-mail
Chen Mengyao Key Laboratory of Crop Growth and Development Regulation of Jiangxi Province, College of Life Sciences and Resource Environment, Yichun University, Yichun 336000, Jiangxi Province, China
Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China 
 
Cao Zhenghong Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
He Kang Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
Mei Yang Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
Wei Shengfei Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
Xiao Huamei Key Laboratory of Crop Growth and Development Regulation of Jiangxi Province, College of Life Sciences and Resource Environment, Yichun University, Yichun 336000, Jiangxi Province, China
Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China 
xiaohuamei625@163.com 
Abstract:Asian citrus psyllid Diaphorina citri was the only natural vector of Huanglongbing (HLB). In order to clarify whether long non-coding RNA (lncRNA) engaged in regulating infection and replication of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of HLB in vivo, the bioinformatics, RTPCR and qPCR amplification technology were used to conduct lncRNA prediction, characteristic analysis, validation and differential expression analysis. The result showed that 10 192 lncRNA genes corresponding to 15 747 lncRNA transcripts were predicted from 13 Asian citrus psyllid RNA-Seq datasets. Asian citrus psyllid lncRNA genes had less exons and shorter transcripts comparing to protein-coding genes. Ten lncRNA genes were randomly selected for RT-PCR validation and seven lncRNAs expressed in both CLas-free Guangzhou strain and CLas-free Ganzhou strain except another lncRNA only expressedinCLas-freeGanzhoustrain.ThelncRNAdifferentialexpressionanalysispredictedtwolncRNAs:TCONS_00096118 and TCONS_00234564 were expressed differentially in CLas-infected and CLasfree adults. qPCR was used to validate the prediction result:TCONS_00234564 matched the result and highly expressed in CLas-infected adults. It indicated that lncRNA involved in the interaction of pathogen CLas and its host Asian citrus psyllid.
keywords:lncRNA  Diaphorina citri  Candidatus Liberibacter asiaticus  RNA-Seq  validation
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