|本期目录/Table of Contents|

[1]宋起萱,浦艳飞,余蓉培,等.蝴蝶兰建兰花叶病毒和齿兰环斑病毒的实时荧光定量PCR检测[J].江苏农业科学,2023,51(15):21-28.
 Song Qixuan,et al.Real-time fluorescence quantitative PCR detection of cymbidium mosaic virus and odontoglossum ringspot virus in Phalaenopsis[J].Jiangsu Agricultural Sciences,2023,51(15):21-28.
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蝴蝶兰建兰花叶病毒和齿兰环斑病毒的实时荧光定量PCR检测(PDF)
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《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第51卷
期数:
2023年第15期
页码:
21-28
栏目:
生物技术
出版日期:
2023-08-05

文章信息/Info

Title:
Real-time fluorescence quantitative PCR detection of cymbidium mosaic virus and odontoglossum ringspot virus in Phalaenopsis
作者:
宋起萱1浦艳飞1余蓉培23瞿素萍23杨春梅23吴丽芳23汪国鲜23阮继伟23
1.云南大学资源植物研究院,云南昆明 650504; 2.云南省农业科学院花卉研究所/国家观赏园艺工程技术研究中心,云南昆明 650205;3.玉溪云星生物科技有限公司,云南玉溪 653100
Author(s):
Song Qixuanet al
关键词:
蝴蝶兰建兰花叶病毒齿兰环斑病毒实时荧光定量PCR病毒检测
Keywords:
-
分类号:
S436.8+1
DOI:
-
文献标志码:
A
摘要:
建兰花叶病毒(Cymbidium mosaic virus,简称CymMV)和齿兰环斑病毒(Odontoglossum ringspot virus,简称ORSV)是蝴蝶兰生产中的主要病毒病害,建立稳定、灵敏的检测体系是进行病毒早期检测和脱毒技术研究的前提。本研究首先对CymMV和ORSV 的外壳蛋白(coat protein,简称CP)基因片段进行克隆和序列比对,随后设计特异性引物,分别构建2种病毒的RT-qPCR(real time quantitative PCR,简称RT-qPC)检测体系,并对云南地区20份样品进行检测。研究结果显示,CymMV和ORSV CP基因序列与其他地区分离物中这2种病毒的核苷酸序列相似性分别为97.92%~98.66%和99.12%~99.56%,其CP基因序列具有保守性。CymMV和ORSV在RT-qPCR体系中最低检出浓度分别为30.9、35.0 copies/μL,灵敏度较RT-PCR(reverse transcription polymerase chain reaction,简称 RT-PCR)提高10倍。利用RT-qPCR体系对20份温室样品进行检测,CymMV阳性率为100%,ORSV阳性率为90%,阳性率均高于RT-PCR检测结果。本研究构建的CymMV和ORSV RT-qPCR检测体系将为蝴蝶兰病毒病早期监测和脱毒种苗的培育奠定基础。
Abstract:
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参考文献/References:

[1]梁芳,邓祖丽颖,许申平,等. 蝴蝶兰2种病毒的同步检测及其CP融合反义表达载体构建[J]. 西南农业学报,2018,31(8):1622-1627.
[2]Chien K W,Agrawal D C,Tsay H S,et al. Elimination of mixed ‘Odontoglossum ringspot’ and ‘Cymbidium mosaic’ viruses from Phalaenopsis hybrid ‘V3’ through shoot-tip culture and protocorm-like body selection[J]. Crop Protection,2015,67:1-6.
[3]Pai H,Jean W H,Lee Y S,et al. Genome-wide analysis of small RNAs from Odontoglossum ringspot virus and Cymbidium mosaic virus synergistically infecting Phalaenopsis[J]. Molecular Plant Pathology,2020,21(2):188-205.
[4]Lee C H,Zheng Y X,Jan F J. The orchid-infecting viruses found in the 21st century[M]//Orchid biotechnology Ⅲ. New Jersey:World Scientific,2017:145-164.
[5]Kim S M,Choi S H. Simultaneous detection of Cymbidium mosaic virus and Odontoglossum ringspot virus in orchids using multiplex RT-PCR[J]. Virus Genes,2015,51(3):417-422.
[6]Kondo H,Takemoto S,Maruyama K,et al. Cymbidium chlorotic mosaic virus,a new sobemovirus isolated from a spring orchid (Cymbidium goeringii) in Japan[J]. Archives of Virology,2015,160(8):2099-2104.
[7]Wong S M,Mahtani P H,Lee K C,et al. Cymbidium mosaic potexvirus RNA:complete nucleotide sequence and phylogenetic analysis[J]. Archives of Virology,1997,142(2):383-391.
[8]Voinnet O,Lederer C,Baulcombe D C. A viral movement protein prevents spread of the gene silencing signal in Nicotiana benthamiana[J]. Cell,2000,103(1):157-167.
[9]明艳林,郑国华. 齿兰环斑病毒的研究进展[C]//陈海如.第三次全国植物病毒和病毒防治学术研讨会论文集. 云南昆明:中国植物病理学会,2003:158-162.
[10]Ryu K H,Park W M. Nucleotide sequence analysis of a cDNA clone encoding the 34K movement protein gene of odontoglossum ringspot virus,ORSV-Cy,the Korean isolate[J]. Plant Molecular Biology,1994,26(3):995-999.
[11]刘沛然,王四清. 建兰花叶病毒及齿兰环斑病毒的检测方法[J]. 热带作物学报,2009,30(1):99-103.
[12]涂小云,董小艳,郭春梅,等. 多重RT-PCR检测蝴蝶兰3种病毒CymMV、ORSV和CMV[J]. 江苏农业科学,2017,45(5):91-93.
[13]Yin J L,Shackel N A,Zekry A,et al. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I[J]. Immunology and Cell Biology,2001,79(3):213-221.
[14]刘美,王毓洪,彭斌,等. 黄瓜绿斑驳花叶病毒实时荧光定量PCR检测体系的建立[J]. 植物保护学报,2022,49(4):1063-1070.
[15]王艳娇,崔甜甜,黄爱军,等. 柑橘脉突病毒实时荧光定量RT-PCR检测体系的建立与应用[J]. 园艺学报,2016,43(8):1613-1620.
[16]王丽,王振东,乔奇,等. 甘薯褪绿矮化病毒西非株系实时荧光定量PCR检测方法的建立及应用[J]. 植物病理学报,2014,44(5):461-468.
[17]高艳玲,范国权,程胜群,等. 苜蓿花叶病毒RT-PCR和RT-qPCR检测技术体系的建立与应用[J]. 植物保护学报,2022,49(2):515-527.
[18]Al Rwahnih M,Osman F,Sudarshana M,et al. Detection of Grapevine leafroll-associated virus 7 using real time qRT-PCR and conventional RT-PCR[J]. Journal of Virological Methods,2012,179(2):383-389.
[19]Shabrangy A,Roustan V,Reipert S,et al. Using RT-qPCR,proteomics,and microscopy to unravel the spatio-temporal expression and subcellular localization of hordoindolines across development in barley endosperm[J]. Frontiers in Plant Science,2018,9:775.
[20]Elmahdy E M,Shaheen M N F,Mahmoud L H I,et al. Detection of norovirus and hepatitis a virus in strawberry and green leafy vegetables by using RT-qPCR in Egypt[J]. Food and Environmental Virology,2022,14(2):178-189.
[21]Vazquez-Iglesias I,McGreig S,Pufal H,et al. A novel high-throughput sequencing approach reveals the presence of a new virus infecting Rosa:rosa ilarvirus-1 (RIV-1)[J]. Journal of Virological Methods,2022,300:114417.
[22]任锐,魏永路,朱根发,等. 广东省国兰病毒病害调查及CymMV和ORSV基于cp基因的系统进化分析[J]. 植物保护学报,2020,47(2):372-383.
[23]Rao X Q,Li Y,Sun J,et al. Genetic diversities of Cymbidium mosaic virus and Odontoglossum ringspot virus isolates based on the coat protein genes from orchids in Guangdong Province,China[J]. Journal of Phytopathology,2015,163(4):324-329.
[24]吴伟文,贺振,刘娴,等. 江苏省建兰齿兰环斑病毒的鉴定和遗传多样性分析[J]. 植物保护学报,2019,46(4):745-753.
[25]Pallás V,Más P,Sánchez-Navarro J A. Detection of plant RNA viruses by nonisotopic dot-blot hybridization[M]//Foster G D,Taylor S C.Plant Virology Protocds. New Jersey:Humana Press,1998:461-468.
[26]梁芳,张燕,王若斓,等. 建兰花叶病毒SYBR GreenⅠ实时荧光定量PCR检测方法的建立[J]. 江西农业大学学报,2017,39(3):572-580.
[27]宋蒙,徐雷锋,曹雨薇,等. 百合中车前草花叶病毒的实时荧光定量PCR检测[J]. 园艺学报,2021,48(12):2497-2505.
[28]Eun A J C,Seoh M L,Wong S M. Simultaneous quantitation of two orchid viruses by the TaqMan real-time RT-PCR[J]. Journal of Virological Methods,2000,87(1/2):151-160.
[29]Jensen D D. Virus diseases of orchids in the Netherlands[J]. Netherlands Journal of Plant Pathology,1970,76(3):135-139.
[30]Zettler F W,Ko N J,Wisler G C,et al. Viruses of orchids and their control[J]. Plant Disease,1990,74(9):621-626.
[31]柳爱春,刘超,赵芸,等. 利用ELISA检测两种兰花病毒的研究[J]. 浙江农业学报,2009,21(2):91-95.
[32]谢林娜,苏梦芸,朱明明,等. 不同品种蝴蝶兰2种病毒的ELISA检测及其症状表现[J]. 江苏农业科学,2017,45(3):80-83.
[33]周国辉,陈晓琴,李梅辉,等. 广东地区两种兰花病毒病害的分子鉴定及检测[J]. 中国病毒学,2004,19(2)149-152.

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备注/Memo

备注/Memo:
收稿日期:2022-11-06
基金项目:云南省重大科技专项计划(编号:202102AE090052);云南大学专业学位研究生实践创新基金(编号:ZC-22222419)。
作者简介:宋起萱(1999—),女,四川攀枝花人,硕士研究生,研究方向为观赏植物研究。E-mail:songqixuan0218@126.com。
通信作者:余蓉培,博士,副研究员,研究方向为观赏植物研究,E-mail:yrongpei@126.com;瞿素萍,硕士,研究员,研究方向为花卉组培繁育,E-mail:qsp@yaas.org.cn。
更新日期/Last Update: 2023-08-05