内蒙古赤峰地区辣椒病毒种类鉴定及其遗传进化特征

周艳芳; 王秀芝; 张晓梅; 罗香文; 刘泽红; 崔聪聪; 孟令强; 孙军玲

doi:10.19303/j.issn.1008-0384.2024.07.009

内蒙古赤峰地区辣椒病毒种类鉴定及其遗传进化特征

doi: 10.19303/j.issn.1008-0384.2024.07.009

1.
赤峰市农牧科学研究所，内蒙古　赤峰　024031
2.
湖南省农业科学院，湖南　长沙　410125
3.
湖南省慈利县高峰土家族乡农业综合服务中心，湖南　张家界　427205

基金项目: 国家大宗蔬菜产业技术体系赤峰综合试验站（CARS-23-G21）；湖南省自然科学基金（2021JJ30415）；国家自然科学基金（32072383）；湖南省科技创新项目（2022CX08、2023CX39）

详细信息

作者简介:
周艳芳（1986 — ），女，硕士，副研究员，主要从事植物病虫害研究，E-mail: yecaode@126.com

通讯作者:
王秀芝（1964 — ），女，研究员，主要从事辣椒新品种选育研究，E-mail: yzhdx@163.com

中图分类号: S436.418.1+2
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- 被引次数: 0
出版历程
- 收稿日期: 2023-10-28
- 修回日期: 2024-01-11
- 网络出版日期: 2024-08-15
- 刊出日期: 2024-07-01

Identification and Genetic Evolution of Viruses Infecting Chili Peppers at Chifeng, Inner Mongolia

1.
Chifeng Academy of Agricultural and Animal Science of Inner Mongolia, Chifeng, Inner Mongolia　024031, China
2.
Hunan Academy of Agricultural Science, Changsha, Hunan　410125, China
3.
Rural integrated Service Center of Gaofeng Tujia Village of Cili County of Hunan Province, Zhangjiajie, Hunan　427205, China

摘要

摘要: 目的系统检测内蒙古赤峰市辣椒主产区病毒发生种类，并分析病毒的遗传分化情况，以期为该地区辣椒病毒病精准防控提供理论依据。方法从内蒙古赤峰地区4个辣椒主产区采集疑似病毒侵染的辣椒样本47份，采用sRNA高通量测序技术结合生物信息学分析，以及常规RT-PCR检测，明确侵染辣椒的病毒种类及复合侵染情况；采用常规Sanger测序测定常规RT-PCR扩增片段序列，进行遗传进化分析。结果内蒙古赤峰地区4个辣椒主产区辣椒混合病样的sRNA高通量测序，共检测到6种病毒，常规RT-PCR共验证到其中4种病毒；4个地区病毒种类存在明显差异，松山区城子乡优势种类为蚕豆萎蔫病毒2（Broad bean wilt virus 2，BBWV2）和马铃薯Y病毒（Potato virus Y, PVY），喀喇沁旗锦山镇为PVY和辣椒轻斑驳病毒（Pepper mild mottle virus, PMMoV）；宁城县和喀喇沁旗西桥镇为BBWV2；复合侵染类型以2种病毒复合侵染为主，复合侵染率最高达61.54%。对测定的序列系统发育分析表明，赤峰地区4个辣椒产区的PMMoV和BBWV2存在遗传分化，PVY可能出现了新的遗传型。结论内蒙古赤峰地区不同辣椒主产区发生的病毒种类、检出率和病毒复合侵染种类均存在明显差异。
- 内蒙古赤峰地区 /
- 辣椒病毒病 /
- sRNA高通量测序 /
- RT-PCR /
- 复合侵染 /
- 系统发育分析
Abstract: Objective Viruses that infected chili pepper crops in Chifeng City, Inner Mongolia were identified, and their genetic relationship analyzed. Methods The 47 infected chili pepper plants at 4 major producing areas in Chifeng City were diagnosed, and viruses identified by sRNA high-throughput sequencing with bioinformatic analysis and RT-PCR. Fragments of RT-PCR were sequenced using the Sanger method to conduct a phylogenetic analysis with the MEGA software. Results Six viruses were identified by sRNA high-throughput sequencing. Of which, 4 were re-verified by RT-PCR. The dominant viruses infecting the 4 regions differed significantly. They were Broad bean wilt virus 2(BBWV2) and Potato virus Y(PVY) in Chengzi County of Songshang District, PVY and Pepper mild mottle virus(PMMoV) in Jinshan County of Kalaqin District, and BBWV2 in Ningcheng County and Xiqiao County of kalaqin District. When the infection was caused by multiple viruses, it was mostly by two major pathogens and had the highest rate of occurrence at 61.54%. The phylogenetic analysis on the sequenced nucleotides of the viruses showed distinctive genetic differences between PMMoV and BBWV2 and, possibly, PVY of a novel genetic type. Conclusion There are significant differences in the types of viruses, detection rates, and types of virus co-infections that occur in different chili production areas of Chifeng City in Inner Mongolia .
- Chifeng City of Inner Mongolia /
- chili pepper viral disease /
- sRNA sequencing /
- RT-PCR /
- multiple species infection /
- phylogenetic analysis

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图 1 sRNA长度分布

Figure 1. Distribution of sRNA length

下载: 全尺寸图片幻灯片

图 2 BBWV2 RNA依赖的RNA聚合酶基因部分核苷酸序列系统发育分析

Figure 2. Phylogenetic analysis on partial nucleotide sequence of RNA dependent RNA polymerase gene of BBWV2

下载: 全尺寸图片幻灯片

图 3 PMMoV CP基因部分核苷酸序列系统发育分析

Figure 3. Phylogenetic analysis on partial nucleotide sequence of CP gene of PMMoV

下载: 全尺寸图片幻灯片

图 4 PVY CP基因部分核苷酸序列系统发育分析

Figure 4. Phylogenetic analysis on partial nucleotide sequence of CP gene of PVY

下载: 全尺寸图片幻灯片

表 1 sRNA测序数据质量

Table 1. Quality of data obtained by sRNA

样本编号 Sample No.	原始读数量 Reads/M	原始序列量 Bases /M	GC含量 GC content/ %	质量值Q30/%	片段数量 Contig number	片段长度中值 Contig N50/bp
城子乡CZ	12.14	607	52.34	97.45	284	157
锦山镇KQ	13.34	667	48.28	97.46	394	182
宁城县NC	13.43	671	51.06	97.37	276	168
西桥镇XQ	12.80	640	51.64	97.60	258	146
Q30：Phred 数值大于 30 的碱基占总体碱基的百分比。 Q30: ratio of bases of Phred number over 30 in total bases.

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表 2 辣椒病毒检出率

Table 2. Positive viral detection rate on chili pepper plants

采样地点 Sampling site	辣椒样本病毒检出率 Rates of positive detection of viruses in pepper plants/%
采样地点 Sampling site	SPVG	PMMoV	BBWV2	PVY	INLV	SCMV
城子乡CZ	0	61.54	92.31	0	23.08	0
锦山镇KQ	0	58.33	41.67	83.33	0	0
宁城县NC	0	25.00	58.33	0	0	0
西桥镇XQ	0	20.00	30.00	0	0	0

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表 3 辣椒病毒复合侵染率

Table 3. Occurrence rate of multiple viruses infection on chili pepper plants

采样地点 Sampling site	病毒复合侵染种类 Viruses of fused infection	病毒检出率 Rates of positive detection of viruses/%
城子乡CZ	BBWV2+PMMoV	61.54
	BBWV2+INLV	23.08
	BBWV2+INLV+PMMoV	23.08
锦山镇KQ	PMMoV+BBWV2	41.67
	PMMoV+PVY	58.33
	BBWV2+PVY	41.67
	PMMoV+BBWV2+PVY	25.00
宁城县NC	PMMoV+BBWV2	25.00
西桥镇XQ	PMMoV+BBWV2	15.38

下载: 导出CSV

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