[1]李亚茹,王银磊,赵丽萍,等.番茄ty-5和 Mi-1基因多重 PCR 体系的建立[J].江苏农业学报,2016,(04):869-873.[doi:10.3969/j.issn.100-4440.2016.04.024]
　LI Ya-ru,WANG Yin-lei,ZHAO Li-ping,et al.Identification of ty-5 gene and Mi-1 gene by multiplex PCR in tomato[J].,2016,(04):869-873.[doi:10.3969/j.issn.100-4440.2016.04.024]
点击复制

番茄ty-5和 Mi-1基因多重 PCR 体系的建立()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年04期
页码:
869-873
栏目:
园艺
出版日期:
2016-10-31

文章信息/Info

Title:
Identification of ty-5 gene and Mi-1 gene by multiplex PCR in tomato
作者:
李亚茹1; 2; 王银磊2; 赵丽萍2; 杨玛丽2; 姜静1; 2; 赵统敏2; 余文贵1; 2
1.南京农业大学园艺学院，江苏 南京 210095；2.江苏省农业科学院蔬菜研究所，江苏 南京 210014
Author(s):
LI Ya-ru1; 2; WANG Yin-lei2; ZHAO Li-ping2; YANG Ma-li2; JIANG Jing1; 2; ZHAO Tong-min 2;  YU Wen-gui2
1. College of Horticulture，Nanjing Agricultural University，Nanjing 210095，China; 2.Institute of Vegetable Crops，Jiangsu Academy of Agricultural Sciences，Nanjing 210014，China
关键词:
番茄; 番茄黄化曲叶病毒; 根结线虫; 抗性基因; 多重PCR
Keywords:
tomato;  tomato yellow leaf curl virus;  root-knot nematode;  resistance gene;  multiplex PCR
分类号:
S641.2
DOI:
10.3969/j.issn.100-4440.2016.04.024
文献标志码:
A
摘要:
为了用分子标记辅助选择法进行番茄育种时能快速鉴定抗番茄黄化曲叶病毒病基因ty-5和抗根结线虫病基因Mi-1，利用番茄材料1227（含有ty-5基因）、VFNT（含有Mi-1基因）和杂交获得的F2分离群体，通过分析扩增片段多态性差异，重新设计分子标记，建立了ty-5和Mi-1基因的多重PCR反应体系。通过对这2个基因的分离群体进行基因型和抗病性鉴定，证实标记检测结果与田间接种鉴定结果吻合率为95.%，可以将该PCR体系用于对这2个基因的同时鉴定。
Abstract:
To realize the rapid identification of gene ty-5 resistant to tomato yellow leaf curl virus (TYLCV) and gene Mi-1 resistant to root-knot nematode (RKN) disease for tomato breeding through marker-assisted selection, a multiplex PCR reaction system was developed with tomato lines 1227 (with ty-5 gene), VFNT (with Mi-1 gene) and segregation population F2 by analyzing the polymorphism of amplified fragment with redesigned molecular markers. The identification of the genotype and disease resistance in F2 population by multiplex PCR detection was 95% similar to that by field inoculation evaluation. The multiplex PCR system developed in this study is capable of simutaneous identification of ty-5 and Mi-1 in tomato.

参考文献/References:

[1]李敏. 番茄抗花叶病毒（ToMV）的鉴定及其类似序列分离[D]. 雅安:四川农业大学，2005.
[2]姚金晓. 番茄抗黄化曲叶病毒病基因的分子标记研究[D].金华:浙江师范大学，2010.
[3]裴华丽，刘永光，乔宁，等． 番茄黄化曲叶病毒Rep基因植物表达载体的构建及对番茄的遗传转化[J]． 江苏农业科学，2015，43(2): 41-45．
[4]黄狮，纠敏，汪伦记，等． 洛阳地区番茄黄化曲叶病的病原鉴定及防治策略[J]． 江苏农业科学，2014，42(11): 153-155 
[5]王银磊，赵统敏，杨文才，等． 温度对番茄根结线虫抗性相关基因表达的影响[J]． 江苏农业学报，2014，30(6): 1417-1422．
[6]TAHIRI A，SEKKAT A，BENNANI A. Distribution of tomato infecting begomoviruses and Bemisia tabaci biotypes in Morocco[J].Annals of Applied Biology，2006，149(2): 175-186.
[7]KOKLU G，ROJAS A. Molecular identification and the complete nucleotide sequence of a tomato yellow leaf curl virus isolate from Turkey [J]. Journal of Plant Pathology，2006，88(1): 61-66.
[8]TSUNAKI A, YAMAGUCHI A，KAWANO S，et al. Ageratum yellow vein virus isolated from tomato plants with leaf curl on Ishigaki Island，Okinawa，Japan[J]. Journal of General Plant Pathology，2010，76(4): 287-291.
[9]ANFOKA G H，ABHARY M，NAKHLA M K. Molecular identification of species of the tomato yellow leaf curl virus complex in Jordan[J]. Journal of Plant Pathology，2005，87 (1):65-70.
[10] SAWANGJIT S，CHATCHA W，PHANICH O. Molecular characterization of tomato infecting begomoviruses in Thailand[J]. Virus Research，2005，109 (1): 1-8.
[11] ANBINDER I，REUVENI M，AZARI R，et al． Molecular dissection of tomato leaf curl virus resistance in tomato line TY172 derived from Solanum peruvianum[J].Theoretical and Applied Genetics，2009，119(3): 519-530．
[12] HUTTON S F，SCOTT J W，SCHUSTER D J． Recessive resistance to tomato yellow leaf curl virus from the tomato cμltivar Ty-king is located in the same region as Ty-5 on chromosome 4[J].HortScience，2012，47(3): 324-327．
[13] SASSER J N． Root-knot nematodes: a global menace to crop production[J]. Plant Disease，1980，64(1): 36-41．
[14] JAOUANNET M，PERFUS-BARBEOCH L，DELEURY E，et al．A root-knot nematode-secreted protein is injected into giant cells and targeted to the nuclei[J]. New Phytologist，2012，194(4):924-931．
[15] EISENBACK J D，TRIANTAPHYLLOU H H． Root-knot nematodes:Meloidogyne species and races[M]. New York: Marcel Decker，1991: 191-274．
[16] SASSER J N． Root-knot nematodes: a global menace to crop production[J]. Plant Disease，1980，64(1): 36-41．
[17] GILBERT J C，MCGUIRE D C． Inheritance of resistance to severe root knot from Meloidogyne incognita in commercial type tomatoes[J]. Proc Amer Soc Hort Sci，1956，68: 437-442．
[18] 于力，朱为民，薛林宝，等. 番茄根结线虫病的研究进展[J]. 中国蔬菜，2006（11）:35-38.
[19] 陈明洁，方倜，柯涛，等. 多重PCR—一种高效快速的分子生物学技术[J]. 武汉理工大学学报，2005，10(27):37-40.
[20] WILLIAMSON V M， HO J Y，WU F F，et al. A PCR-based marker tightly linked to the nematode resistance gene, Mi, in tomato[J].Theor Appl Genet,1994，87：757-763.
[21] 卢圣栋．现代分子生物学实验技术[M]．北京：高等教育出版社,1993：98-103．
[22] 束永俊，李勇，朱振雷，等．大豆CAPS标记快速开发方法的建立于优化[J]．东北农业大学学报，2009，40(12): 62-65.

相似文献/References:

[1]赵丽萍,李永灿,赵统敏,等.灰霉菌毒素诱导番茄抗性突变体及相关防御酶活性[J].江苏农业学报,2016,(03):631.[doi:10.3969/j.issn.1000-4440.2016.03.023]
　ZHAO Li-ping,LI Yong-can,ZHAO Tong-min,et al.Resistant mutant induced by toxin of Botrytis cinerea in tomato and the activities of related defensive enzymes[J].,2016,(04):631.[doi:10.3969/j.issn.1000-4440.2016.03.023]
[2]吕敏,苏建坤,白和盛,等.桃蚜取食和机械损伤对番茄和辣椒 PAL、LOX 和 PPO 活性的诱导作用[J].江苏农业学报,2016,(06):1273.[doi:doi:10.3969/j.issn.1000-4440.2016.06.013]
　Lv?Min,SU Jian-kun,BAI He-sheng,et al.The activities of PAL, LOX and PPO in tomato and pepper plants induced by aphid herbivory and mechanical damage[J].,2016,(04):1273.[doi:doi:10.3969/j.issn.1000-4440.2016.06.013]
[3]姜静,王银磊,赵丽萍,等.番茄qRT-PCR内参基因的筛选[J].江苏农业学报,2017,(02):389.[doi:doi:10.3969/j.issn.1000-4440.2017.02.024]
　JIANG Jing,WANG Yin-lei,ZHAO Li-ping,et al.Selection of tomato reference genes for qRT-PCR[J].,2017,(04):389.[doi:doi:10.3969/j.issn.1000-4440.2017.02.024]
[4]罗伟君,唐琳,周佳丽,等.纳米锌肥对番茄果实锌含量与品质的强化[J].江苏农业学报,2016,(01):184.[doi:10.3969/j.issn.1000-4440.2016.01.028 ]
　LUO Wei-jun,TANG Lin,ZHOU Jia-li,et al.Improvement of zinc concentration and quality of tomato fruit by nano zinc fertilizer[J].,2016,(04):184.[doi:10.3969/j.issn.1000-4440.2016.01.028 ]
[5]乔俊卿,陈志谊,梁雪杰,等.枯草芽孢杆菌Bs916在番茄根部的定殖[J].江苏农业学报,2015,(06):1278.[doi:doi:10.3969/j.issn.1000-4440.2015.06.013]
　QIAO Jun-qing,CHEN Zhi-yi,LIANG Xue-jie,et al.Colonization of Bacillus subtilis Bs916 on tomato root[J].,2015,(04):1278.[doi:doi:10.3969/j.issn.1000-4440.2015.06.013]
[6]翟亚明,魏丽萍,杨倩.不同调控方式对设施盐渍化土壤特性和番茄产量及品质的影响[J].江苏农业学报,2015,(04):871.[doi:10.3969/j.issn.1000-4440.2015.04.025]
　ZHAI Ya-ming,WEI Li-ping,YANG Qian.Effects of regulatory measures on characters of greenhouse saline soil and tomato yield and quality[J].,2015,(04):871.[doi:10.3969/j.issn.1000-4440.2015.04.025]
[7]杨玛丽,赵统敏,余文贵,等.转基因 RNAi 技术在番茄研究中的应用[J].江苏农业学报,2015,(01):217.[doi:10.3969/j.issn.1000-4440.2015.01.034]
　YANG Ma-li,ZHAO Tong-min,YU Wen-gui,et al.Progress in RNA interference technique applied in tomato[J].,2015,(04):217.[doi:10.3969/j.issn.1000-4440.2015.01.034]
[8]姜静,王银磊,赵丽萍,等.抗番茄黄化曲叶病基因ty-5序列变异及表达分析[J].江苏农业学报,2017,(05):1105.[doi:doi:10.3969/j.issn.1000-4440.2017.05.022]
　JIANG Jing,WANG Yin-lei,ZHAO Li-ping,et al.Sequence variation and expression analysis of tomato yellow leaf curl virus resistance gene ty-5[J].,2017,(04):1105.[doi:doi:10.3969/j.issn.1000-4440.2017.05.022]
[9]唐玉新,曲萍,陆岱鹏,等.适合机械化移栽的番茄穴盘育苗基质配方筛选[J].江苏农业学报,2017,(06):1342.[doi:doi:10.3969/j.issn.1000-4440.2017.06.021]
　TANG Yu-xin,QU Ping,LU Dai-peng,et al.Screening of tomato plug seedling substrates proportion suitable for mechanized transplanting[J].,2017,(04):1342.[doi:doi:10.3969/j.issn.1000-4440.2017.06.021]
[10]周蓉,赵统敏,赵丽萍,等.高光泽番茄育种及果实表面光泽度[J].江苏农业学报,2018,(06):1437.[doi:doi:10.3969/j.issn.1000-4440.2018.06.033]
　ZHOU Rong,ZHAO Tong-min,ZHAO Li-ping,et al.High-glossiness tomato breeding and fruit surface glossiness[J].,2018,(04):1437.[doi:doi:10.3969/j.issn.1000-4440.2018.06.033]
[11]姜静,王银磊,李亚茹,等.江苏省及其他地区番茄黄化曲叶病毒的分子鉴定及序列分析[J].江苏农业学报,2018,(01):238.[doi:doi:10.3969/j.issn.1000-4440.2018.01.035]
　JIANG Jing,WANG Yin-lei,LI Ya-ru,et al.Molecular identification and sequence analysis of tomato yellow leaf curl virus in Jiangsu province and its peripheral area[J].,2018,(04):238.[doi:doi:10.3969/j.issn.1000-4440.2018.01.035]

备注/Memo

备注/Memo:
收稿日期：2015-11-26基金项目：江苏省自然科学青年基金项目（BK20140739）；江苏省农业自主创新基金项目[CX(14)5012]；江苏省农业科学院基本科研业务专项[ZX(15)2003、ZX(15)4022]；国家自然科学青年基金项目（31401884）作者简介：李亚茹(199-),女，河北石家庄人，硕士研究生,从事蔬菜遗传育种研究。(E-mail)yrlisecrect@163. com通讯作者：余文贵，(E-mail)wenguiyu@jass.ac.cn；赵统敏，（E-mail）tmzhaomail@163.com
更新日期/Last Update: 2016-11-01