TMV 和 PVX 胁迫下云烟和番茄实时荧光定量内参基因筛选

张宁,魏周玲,杜利娟,青玲,孙现超

doi:10.13718/j.cnki.xdzk.2016.07.002

西南大学学报（自然科学版）

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TMV 和 PVX 胁迫下云烟和番茄实时荧光定量内参基因筛选

张宁,魏周玲,杜利娟,青玲,孙现超. TMV 和 PVX 胁迫下云烟和番茄实时荧光定量内参基因筛选[J]. 西南大学学报（自然科学版）, 2016, 38(7). doi: 10.13718/j.cnki.xdzk.2016.07.002

引用本文:

ZHANG Ning,WEI Zhou-ling,DU Li-juan,QING Ling,SUN Xian-chao. Selection of Reference Genes in Tobacco and Tomato Stressed by TMV or PVX[J]. Journal of Southwest University Natural Science Edition, 2016, 38(7). doi: 10.13718/j.cnki.xdzk.2016.07.002

Citation:

ZHANG Ning,WEI Zhou-ling,DU Li-juan,QING Ling,SUN Xian-chao. Selection of Reference Genes in Tobacco and Tomato Stressed by TMV or PVX[J]. Journal of Southwest University Natural Science Edition, 2016, 38(7). doi: 10.13718/j.cnki.xdzk.2016.07.002

TMV 和 PVX 胁迫下云烟和番茄实时荧光定量内参基因筛选

张宁,魏周玲,杜利娟,青玲,孙现超

西南大学植物保护学院/植物病害生物学重庆市高校级重点实验室,重庆,400716

Selection of Reference Genes in Tobacco and Tomato Stressed by TMV or PVX

ZHANG Ning,WEI Zhou-ling,DU Li-juan,QING Ling,SUN Xian-chao

摘要: 合适的内参基因是采用实时荧光定量PCR研究基因表达获得可信数据的基础。为了获得合适的内参基因，更好地研究烟草花叶病毒（Tobacco mosaic virus ，TMV）和马铃薯X病毒（Potato virus X ，PVX）与烟草和番茄2种寄主的互作，本项研究根据已报道的云烟和番茄看家基因设计引物，使用 geNormPlus软件分别对其在 TMV 和PVX侵染下的云烟97和番茄中表达量的稳定性进行了评估。结果表明：2种寄主植物中供试看家基因中各有4对引物满足qPCR的要求，分别是云烟 tub ，CYP ，GAPDH ，β‐tub ，番茄18S ，UK ，ACT ，EFl‐α。在2种病毒侵染后的云烟内参基因中表达最稳定的为CYP ，番茄中内参基因为ACT 。最终确定这2个看家基因分别作为实时荧光定量PCR研究烟草和番茄在TMV和PVX胁迫下寄主体内基因表达变化的内参基因。该结果为进一步探讨2种病毒共同侵染时，它们与烟草和番茄2种寄主互作生物学效应研究奠定了基础。
- 烟草花叶病毒、马铃薯X病毒、寄主、实时荧光定量PCR、内参基因
Abstract: It is the base to select proper reference genes for getting valid data to analyze gene expression by quantitative real‐time PCR(RT‐qPCR) .In order to get proper reference genes of tobacco and tomato infec‐ted by TMV or PVX for studying the interaction between the viruses and their hosts ,we designed the primer pairs of the housekeeping genes of tobacco and tomato based on the previous studies ,and evaluated their expression stability in TMV or PVX infected tobacco and tomato with geNormPlus software .The re‐sults showed that the primer pairs of tub ,CY P ,GA PDH and β‐tub in tobacco and the primer pairs of 18S ,UK ,ACT and EFl‐αin tomato matched the request of RT‐qPCR .CY P was the most stable house‐keeping gene in tobacco infected by TMV or PVX ,and ACT was the most stable housekeeping gene in to‐mato .So ,these two genes were determined to be the reference genes normalization of qRT‐PCR data in to‐bacco or tomato in virus‐host interaction system ,which provides a base for studying the biological effect of the interaction between TMV or PVX and its hosts .
- Tobacco mosaic virus、Potato virus X、host、quantitative real time PCR、reference gene .

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TMV 和 PVX 胁迫下云烟和番茄实时荧光定量内参基因筛选

张宁,魏周玲,杜利娟,青玲,孙现超

西南大学植物保护学院/植物病害生物学重庆市高校级重点实验室,重庆,400716

关键词:

烟草花叶病毒、马铃薯X病毒、寄主、实时荧光定量PCR、内参基因

摘要: 合适的内参基因是采用实时荧光定量PCR研究基因表达获得可信数据的基础。为了获得合适的内参基因，更好地研究烟草花叶病毒（Tobacco mosaic virus ，TMV）和马铃薯X病毒（Potato virus X ，PVX）与烟草和番茄2种寄主的互作，本项研究根据已报道的云烟和番茄看家基因设计引物，使用 geNormPlus软件分别对其在 TMV 和PVX侵染下的云烟97和番茄中表达量的稳定性进行了评估。结果表明：2种寄主植物中供试看家基因中各有4对引物满足qPCR的要求，分别是云烟 tub ，CYP ，GAPDH ，β‐tub ，番茄18S ，UK ，ACT ，EFl‐α。在2种病毒侵染后的云烟内参基因中表达最稳定的为CYP ，番茄中内参基因为ACT 。最终确定这2个看家基因分别作为实时荧光定量PCR研究烟草和番茄在TMV和PVX胁迫下寄主体内基因表达变化的内参基因。该结果为进一步探讨2种病毒共同侵染时，它们与烟草和番茄2种寄主互作生物学效应研究奠定了基础。

English Abstract

Selection of Reference Genes in Tobacco and Tomato Stressed by TMV or PVX

西南大学植物保护学院/植物病害生物学重庆市高校级重点实验室,重庆,400716

Keywords:

烟草花叶病毒、马铃薯X病毒、寄主、实时荧光定量PCR、内参基因

Abstract: It is the base to select proper reference genes for getting valid data to analyze gene expression by quantitative real‐time PCR(RT‐qPCR) .In order to get proper reference genes of tobacco and tomato infec‐ted by TMV or PVX for studying the interaction between the viruses and their hosts ,we designed the primer pairs of the housekeeping genes of tobacco and tomato based on the previous studies ,and evaluated their expression stability in TMV or PVX infected tobacco and tomato with geNormPlus software .The re‐sults showed that the primer pairs of tub ,CY P ,GA PDH and β‐tub in tobacco and the primer pairs of 18S ,UK ,ACT and EFl‐αin tomato matched the request of RT‐qPCR .CY P was the most stable house‐keeping gene in tobacco infected by TMV or PVX ,and ACT was the most stable housekeeping gene in to‐mato .So ,these two genes were determined to be the reference genes normalization of qRT‐PCR data in to‐bacco or tomato in virus‐host interaction system ,which provides a base for studying the biological effect of the interaction between TMV or PVX and its hosts .