[1]朱雪梅,郭广君,潘宝贵,等.辣椒抗PMMoV基因L4连锁标记的验证分析[J].江苏农业学报,2022,38(06):1627-1636.[doi:doi:10.3969/j.issn.1000-4440.2022.06.022]
　ZHU Xue-mei,GUO Guang-jun,PAN Bao-gui,et al.Test analysis of L4-linked markers for PMMoV resistant gene in pepper[J].,2022,38(06):1627-1636.[doi:doi:10.3969/j.issn.1000-4440.2022.06.022]
点击复制

辣椒抗PMMoV基因L4连锁标记的验证分析()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年06期
页码:
1627-1636
栏目:
园艺
出版日期:
2022-12-31

文章信息/Info

Title:
Test analysis of L4-linked markers for PMMoV resistant gene in pepper
作者:
朱雪梅1; 2; 郭广君2; 潘宝贵2; 刁卫平2; 刘金兵2; 高长洲2; 王述彬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/江苏省高效园艺作物遗传改良重点实验室,江苏南京210014）
Author(s):
ZHU Xue-mei1; 2; GUO Guang-jun2; PAN Bao-gui2; DIAO Wei-ping2; LIU Jin-bing2; GAO Chang-zhou2; WANG Shu-bin2
(1.College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;2.Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Efficient Horticultural Crop Genetic Improvement，Nanjing 210014，China)
关键词:
辣椒; 辣椒轻斑驳病毒; 分子标记辅助筛选
Keywords:
pepper; pepper mild mottle virus; molecular marker assisted selection
分类号:
S641.3
DOI:
doi:10.3969/j.issn.1000-4440.2022.06.022
文献标志码:
A
摘要:
近年来，辣椒轻斑驳病毒（Pepper mild mottle virus，PMMoV）病的发生日趋严重，已严重危害中国辣椒生产。L系列等位基因（L1、L2、L3、L4）是抗烟草花叶病毒属的主要抗性基因，其中L4基因抗性最强且具有广谱性。为加速L4基因的转育应用，本研究对前人报道的3对与L4基因紧密连锁的分子标记进行检验分析。结果显示，标记L4SC340退火温度不稳定，极易造成假阳性，不适用于L4抗性基因的辅助筛选。标记087H3T7和L4-SCAR可以筛选出携带L3和L4抗性基因的辣椒种质，但无法区分携带L3和L4基因的抗性材料，同时087H3T7存在杂合基因型过高的问题。二者比较来说，L4-SCAR标记的筛选准确度高于087H3T7，但是L4-SCAR为显性标记，无法区分杂合基因型。因此上述3对标记均不能用于筛选携带L4抗性基因的种质材料，但是在明确抗性材料基因型的情况下，L4-SCAR标记结合087H3T7可用于L3和L4抗性基因转育后代抗性单株的辅助筛选。本研究结果可为加速辣椒抗PMMoV育种提供更实用的分子标记。
Abstract:
In recent years, disease caused by pepper mild mottle virus (PMMoV) has become more and more serious and has harmed pepper production in China severely. L-series allelic genes (L1, L2, L3, L4) are the main resistant genes against tobamovirus, and L4 gene was reported to show the strongest resistant to tobamovirus with a broad spectrum. In order to accelerate the application of transgenic breeding of the L4 gene, three pairs of reported molecular markers closely linked to L4 gene were examined and analyzed in this study. The results showed that, the annealing temperature of marker L4SC340 was unstable and could easily cause false positives, so L4SC340 was unsuitable for the assisted screening of L4 resistant gene. Markers 087H3T7 and L4-SCAR could be used to screen pepper germplasms carrying L3 and L4 resistant genes, but they could not be used to distinguish resistant materials carrying L3 and L4 genes. Meanwhile, 087H3T7 had the problem of excessive heterozygous genotypes. In comparison, the screening accuracy of L4-SCAR marker was higher than that of 087H3T7, but L4-SCAR was a dominant marker that could not distinguish heterozygous genotype. Therefore, none of the three pairs of markers could be used to screen the germplasm material carrying L4 resistant gene. However, through clarifying the genotype of the resistant materials, L4-SCAR marker combined with 087H3T7 could be used for assisted screening of resistant individuals in the breeding of transgenic progeny with L3 and L4 resistant genes. Results of the study can provide more practical molecular markers for accelerated breeding of PMMoV resistant pepper.

参考文献/References:

[1]王立浩,张正海,曹亚从,等. “十二五”我国辣椒遗传育种研究进展及其展望 [J]. 中国蔬菜, 2016(1):1-7.
[2]张宝玺,王立浩,张正海,等. 甜椒L3应对辣椒轻斑驳病毒及中椒系列新品种的选育 [J]. 中国农业科学, 2020, 53(18): 3846-3855.
[3]刘勇，李凡，李月月，等. 侵染我国主要蔬菜作物的病毒种类、分布与发生趋势 [J]. 中国农业科学, 2019, 52(2): 55-77.
[4]GREENLEAF W H, COOK A A, HEYN A. Resistance to tobacco mosaic virus in Capsicum, with reference to the Samsun latent strain [J]. Phytopathology, 1964, 54: 1367-1371.
[5]向本春，谢浩，崔星明，等. 新疆辣椒轻微斑驳病毒的分离鉴定 [J]. 病毒学报, 1994, 10(3): 240-245.
[6]杨中周,邓竹根,李曼,等. 辣椒轻斑驳病毒研究的回顾与展望 [J]. 中国瓜菜, 2021, 34(9): 1-6.
[7]DEMSKI J W. Tobacco mosaic virus is seedborne in pimiento peppers [J]. Plant Disease, 1981，65: 723-724.
[8]BROADBENT L, FLETCHER J T. The epidemiology of tomato mosaic: sources of TMV in commercial tomato crops under glass [J]. Annals of Applied Biology, 2010, 57(1): 113-120.
[9]BOUKEMA I W. Allelism of genes controlling resistance to TMV in Capsicum L [J]. Euphytica, 1980, 29(2): 433-439.
[10]BOUKEMA I W. Resistance to a new strain of TMV in Capsicum chacoense Hunz [J]. Capsicum Newslett, 1982, 1: 49-51.
[11]BOUKEMA I W. Resistance to TMV in Capsicum chacoense Hunz. is governed by an allele of the L-locus [J]. Capsicum Newsl, 1984,3:47-78.
[12]TOMITA R, SEKINE K T, MIZUMOTO H, et al. Genetic basis for the hierarchical interaction between Tobamovirus spp. and L resistance gene alleles from different pepper species [J]. Mol Plant Microbe Interact, 2011, 24(1): 108-117.
[13]于海龙,王立浩,张宝玺,等. 分子标记辅助选育聚合烟草花叶病毒属病毒抗性基因(L4)及马铃薯Y病毒抗性基因(Pvr4)的甜椒自交系 [J]. 中国蔬菜, 2021(1): 41-47.
[14]MATSUNAGA H, SAITO T, HIRAI M, et al. DNA Markers linked to pepper mild mottle virus (PMMoV) resistant locus (L4) in Capsicum [J]. Engei Gakkai Zasshi, 2003, 72(3): 218-220.
[15]KIM J H, JUNG-HEON H, HYOUNG Y J, et al. Development of a sequence characteristic amplified region marker linked to the L4 locus conferring broad spectrum resistance to tobamoviruses in pepper plants [J]. Molecules and Cells, 2008, 25(2):205-210.
[16]YANG H B, LIU W Y, KANG W H, et al. Development of SNP markers linked to the L locus in Capsicum spp. by a comparative genetic analysis [J]. Molecular Breeding, 2009, 24(4): 433-446.
[17]YANG H B, LIU W Y, KANG W H, et al. Development and validation of L allele-specific markers in Capsicum [J]. Molecular Breeding, 2012, 30(2): 819-829.
[18]LEE J D, HAN J H, YOON J B. A Set of Allele-specific markers linked to L locus resistant to tobamovirus in Capsicum spp.[J]. Korean Journal of Horticultural Science & Technology, 2012, 30(3): 286-293.
[19] LORAINE A E, BLAKLEY I C, JAGADEESAN S, et al. Analysis and visualization of RNA-Seq expression data using RStudio, bioconductor, and integrated genome browser [M]. USA: Humana Press, 2015.
[20]李宁,宫丽媛,高升华,等. 辣椒种质的抗性基因分子标记检测 [J]. 中国蔬菜, 2020(8):19-32.

相似文献/References:

[1]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,(06):1273.[doi:doi:10.3969/j.issn.1000-4440.2016.06.013]
[2]郭广君,孙茜,刘金兵,等.基于辣椒基因组重测序的InDel标记开发及应用[J].江苏农业学报,2015,(06):1400.[doi:doi:10.3969/j.issn.1000-4440.2015.06.032]
　GUO Guang-jun,SUN Qian,LIU Jin-bing,et al.Development and application of pepper InDel markers based on genome re-sequencing[J].,2015,(06):1400.[doi:doi:10.3969/j.issn.1000-4440.2015.06.032]
[3]李廷芳,吴淑华,赵文浩,等.青海海东设施辣椒轻斑驳病毒的分子检测[J].江苏农业学报,2017,(04):958.[doi:doi:10.3969/j.issn.1000-4440.2017.04.036]
　LI Ting-fang,WU Shu-hua,ZHAO Wen-hao,et al.Molecular detection of mild mottle virus isolated from pepper in Haidong, Qinghai province[J].,2017,(06):958.[doi:doi:10.3969/j.issn.1000-4440.2017.04.036]
[4]刘潮,韩利红,宋培兵,等.辣椒类甜蛋白基因家族鉴定及表达分析[J].江苏农业学报,2018,(01):122.[doi:doi:10.3969/j.issn.1000-4440.2018.01.018]
　LIU Chao,HAN Li-hong,SONG Pei-bing,et al.Identification and expression analysis of thaumatin-like protein gene in pepper[J].,2018,(06):122.[doi:doi:10.3969/j.issn.1000-4440.2018.01.018]
[5]王运儒,秦玉燕,杨秀娟,等.40％氯虫·噻虫嗪水分散粒剂在辣椒及土壤中的残留消解动态[J].江苏农业学报,2018,(01):207.[doi:doi:10.3969/j.issn.1000-4440.2018.01.030]
　WANG Yun-ru,QIN Yu-yan,YANG Xiu-juan,et al.Dissipation of chlorantraniliprole and thiamethoxam in pepper and soil after field application in the form of 40% water dispersible granules[J].,2018,(06):207.[doi:doi:10.3969/j.issn.1000-4440.2018.01.030]
[6]潘宝贵,钱恒彦,戈伟,等.辣椒应答冷信号转导机制研究进展[J].江苏农业学报,2019,(03):743.[doi:doi:10.3969/j.issn.1000-4440.2019.03.034]
　PAN Bao-gui,QIAN Heng-yan,GE Wei,et al.Research progress of cold signal transduction mechanisms in pepper[J].,2019,(06):743.[doi:doi:10.3969/j.issn.1000-4440.2019.03.034]
[7]高晶霞,吴雪梅,牛勇琴,等.辣根素水乳剂对连作辣椒生长及土壤酶活性的影响[J].江苏农业学报,2021,(01):116.[doi:doi:10.3969/j.issn.1000-4440.2021.01.015]
　GAO Jing-xia,WU Xue-mei,NIU Yong-qin,et al.Effect of athomin water emulsion on growth and soil enzyme activities of continuous cropping pepper[J].,2021,(06):116.[doi:doi:10.3969/j.issn.1000-4440.2021.01.015]
[8]郭广君,朱雪梅,潘宝贵,等.利用InDel分子标记辅助选育辣椒抗黄瓜花叶病毒病种质[J].江苏农业学报,2021,(05):1251.[doi:doi:10.3969/j.issn.1000-4440.2021.05.021]
　GUO Guang-jun,ZHU Xue-mei,PAN Bao-gui,et al.Innovation of pepper germplasm resource with resistance to cucumber mosaic virus by InDel molecular marker assisted selection[J].,2021,(06):1251.[doi:doi:10.3969/j.issn.1000-4440.2021.05.021]
[9]刘雪梅,王悦,李玮琦,等.辣椒miR169及其靶基因的生物信息学分析[J].江苏农业学报,2021,(06):1510.[doi:doi:10.3969/j.issn.1000-4440.2021.05.020]
　LIU Xue-mei,WANG Yue,LI Wei-qi,et al.Bioinformatics analysis of miR169 and its target genes in pepper[J].,2021,(06):1510.[doi:doi:10.3969/j.issn.1000-4440.2021.05.020]
[10]赵小慧,刘冲,郁凯,等.利用小RNA深度测序技术鉴定江苏盐城辣椒病毒种类[J].江苏农业学报,2023,(01):37.[doi:doi:10.3969/j.issn.1000-4440.2023.01.005]
　ZHAO Xiao-hui,LIU Chong,YU Kai,et al.Identification of viruses from peppers in Yancheng, Jiangsu province by small RNA deep sequencing[J].,2023,(06):37.[doi:doi:10.3969/j.issn.1000-4440.2023.01.005]
[11]吴淑华,赵文浩,李廷芳,等.南京辣椒上一种斑驳类型病毒病的分子鉴定[J].江苏农业学报,2015,(06):1284.[doi:doi:10.3969/j.issn.1000-4440.2015.06.014]
　WU Shu-hua,ZHAO Wen-hao,LI Ting-fang,et al.Molecular identification of a virus causing mottle symptoms in pepper leaves in Nanjing[J].,2015,(06):1284.[doi:doi:10.3969/j.issn.1000-4440.2015.06.014]

备注/Memo

备注/Memo:
收稿日期：2022-04-07基金项目：国家自然科学基金面上项目（32072597）；江苏省种业振兴揭榜挂帅项目[JBGS（2021）065]；国家现代农业产业技术体系建设专项（CARS-23-G42）作者简介：朱雪梅 （1996-），女，山东滨州人，硕士研究生，主要研究方向为辣椒遗传育种和分子生物学。（E-mail）1668178637@qq.com。郭广君为共同第一作者。通讯作者：王述彬，（E-mail）wangsbpep@163.com
更新日期/Last Update: 2023-01-13