Identification of Viruses Infecting Passiflora edulis in Guizhou Province by Small RNA Sequencing

doi:10.3969/j.issn.1000-2561.2019.08.018

Abstract

Abstract:

Virus disease is one of the main diseases of Passiflora edulis, and the identification of virus species can provide a theoretical basis for the disease prevention. In this study, six suspected virus-infected leaf samples were mixed and analyzed by small RNA deep sequencing. Cucumber mosaic virus (CMV) and Telosma mosaic virus (TeMV) were detected from the mixed sample. Viruses identified by small RNA sequencing were validated by virus coat protein gene (CP) specific RT-PCR from the above six samples. CMV and TeMV was detected from all six samples, respectively. The open reading frame of CP gene of CMV and TeMV obtained by RT-PCR was 657 bp and 816 bp, correspondingly. The nucleotide sequence identity of CMV-GZ (accession number: MH623077), TeMV-GZ1 (accession number: MH623078) and TeMV-GZ2 (accession number: MH623079) was 82.3%-95.9%, 87.8%-99.3% and 87.8%-98.3%, respectively, with the reported CMV and TeMV isolates. CMV and TeMV in P. edulis was first reported Guizhou. The combined infection of the two viruses may be a potential threat to P. edulis.

Key words: Passiflora edulis, virus, small RNA sequencing, identification

CLC Number:

S436.67

YAN Jiawen,YUAN Qifeng,XIE Pu,WANG Lijuan,MA Yuhua,WANG Zhengyuan. Identification of Viruses Infecting Passiflora edulis in Guizhou Province by Small RNA Sequencing[J]. Chinese Journal of Tropical Crops, 2019, 40(8): 1577-1584.

Figures/Tables 10

References 23

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品种 Cultivar	样品编号 Sample name	采集地点 Source	田间症状 Symptoms
‘紫香1号’	ZF	贞丰县鲁容乡	褪绿、皱缩、矮化
‘紫香1号’	ZN	镇宁县良田镇	斑驳、皱缩、曲叶、矮化
‘紫香1号’	WM	望谟县郊纳镇	褪绿、皱缩
‘紫香1号’	LD	罗甸县逢亭镇	褪绿、皱缩、矮化
‘平塘1号’	PT	平塘县克度镇	斑驳、皱缩
‘紫香1号’	CJ	从江县下江镇	斑驳、皱缩、曲叶

品种 Cultivar	样品编号 Sample name	采集地点 Source	田间症状 Symptoms
‘紫香1号’	ZF	贞丰县鲁容乡	褪绿、皱缩、矮化
‘紫香1号’	ZN	镇宁县良田镇	斑驳、皱缩、曲叶、矮化
‘紫香1号’	WM	望谟县郊纳镇	褪绿、皱缩
‘紫香1号’	LD	罗甸县逢亭镇	褪绿、皱缩、矮化
‘平塘1号’	PT	平塘县克度镇	斑驳、皱缩
‘紫香1号’	CJ	从江县下江镇	斑驳、皱缩、曲叶

引物名称 Primer name	引物序列 Sequence (5′-3′)	退火温度 T_m/℃	PCR产物大小 Amplification length/bp	引物用途 Primer type
CMV-F1	CCGCTCCGCTTCCTCCTC	62.7
CMV-R1	TTGAATACACGAGCACGGCG	62.0	506	CMV检测
TeMV-F1	ATGCAGGCAACACAGGCAAAG	62.3
TeMV-R1	GTCCATCATCACCCACACGC	61.4	398	TeMV检测
CMV-F2	ATGGACAAATCCGAATCAACC	60.0
CMV-R2	TCAAACTGGGAGCACACCA	58.0	657	CP基因克隆
TeMV-F2	TCAAGTAAGGTGGATGATGT	56.0
TeMV-R2	CTGCACAGAACCTACCCCA	58.0	816	CP基因克隆

引物名称 Primer name	引物序列 Sequence (5′-3′)	退火温度 T_m/℃	PCR产物大小 Amplification length/bp	引物用途 Primer type
CMV-F1	CCGCTCCGCTTCCTCCTC	62.7
CMV-R1	TTGAATACACGAGCACGGCG	62.0	506	CMV检测
TeMV-F1	ATGCAGGCAACACAGGCAAAG	62.3
TeMV-R1	GTCCATCATCACCCACACGC	61.4	398	TeMV检测
CMV-F2	ATGGACAAATCCGAATCAACC	60.0
CMV-R2	TCAAACTGGGAGCACACCA	58.0	657	CP基因克隆
TeMV-F2	TCAAGTAAGGTGGATGATGT	56.0
TeMV-R2	CTGCACAGAACCTACCCCA	58.0	816	CP基因克隆

病毒名称 Virus name	参考序列长度 Reference length/bp	匹配序列长度 Consensus length/bp	Contig数量 Contig count/个	覆盖率 Rate/%
CMV isolate DS segment	3 360	2 875	20	85.6
RNA1-KU255787 mapping	3 360	2 875	20	85.6
CMV isolate ZMBJ segment	3 054	2 629	16	86.1
RNA2-KT350981 mapping	3 054	2 629	16	86.1
CMV isolate DS segment	2 220	1 610	33	72.5
RNA3-KU255789 mapping	2 220	1 610	33	72.5
TeMV isolate Hanio complete	9 689	6 130	114	63.3
genome DQ851493 mapping	9 689	6 130	114	63.3