Molecular Identification of Four Viruses that Cause Mixed Infection of Pumpkin in Chengdu,Sichuan Province of China

doi:10.16420/j.issn.0513-353x.2020-0770

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (11): 2299-2310.doi: 10.16420/j.issn.0513-353x.2020-0770

• Original article • Previous Articles Next Articles

Molecular Identification of Four Viruses that Cause Mixed Infection of Pumpkin in Chengdu,Sichuan Province of China

LIN Xiaomin, PENG Meifang, FAN Xiaoli, CHEN Kegui()

Institute of Biotechnology and Nuclear Technology,Sichuan Academy of Agricultural Sciences,Chengdu 610061,China

Received:2021-03-11 Revised:2021-06-10 Published:2021-12-02
Contact: CHEN Kegui E-mail:keguichen@yahoo.com

Abstract

Abstract:

In recent years the disease caused by virus has become more and more serious issue in pumpkin field production. In this study,samples from the field in Chengdu,Sichuan Province of China were collected in May,2020. Deep sequencing of small RNA（sRNA）from the pumpkin sample revealed that there were four viruses of cucurbit aphid-borne yellows virus（CABYV）,watermelon mosaic virus（WMV）,papaya ringspot virus（PRSV）and zucchini yellow mosaic virus（ZYMV）. All the viruses were further confirmed by reverse transcriptional PCR（RT-PCR）and Sanger sequencing of the RT-PCR fragments. Alignments and phylogenetic analyses with sequences of the viruses in GenBank showed that CABYV and ZYMV were more similar with the viruses recently reported in China,whereas WMV and PRSV were similar with the other viruses in China and abroad,as well. Besides,the sequence alignments and phylogenetics further suggested that two isolates for both CABYV and PRSV might be currently infecting pumpkin in Sichuan Province. Discovery of the complicated mixed virus-infection in pumpkin might be of potential significance for the virus disease control in pumpkin production.

Key words: pumpkin, virus, mixed infection, CABYV, PRSV, WMV, ZYMV

CLC Number:

S642.1

LIN Xiaomin, PENG Meifang, FAN Xiaoli, CHEN Kegui. Molecular Identification of Four Viruses that Cause Mixed Infection of Pumpkin in Chengdu,Sichuan Province of China[J]. Acta Horticulturae Sinica, 2021, 48(11): 2299-2310.

Figures/Tables 7

Table 1 PCR primers used in this study

扩增的病毒 Virus	引物名称 Primer name	引物序列（5′-3′） Sequence
CABYV	CABYV-F	ACATTATTAGCGCAGAGAGTCCC
CABYV	CABYV-R	CCACTCGCAATATTTCTCCGTGTA
ZYMV	ZYMV-F	GATAGTTCTTTAACTCCAGCCTTACTG
ZYMV	ZYMV-R	TCCGTCCATCATGAACCACA
WMMV	WMV-F	TACACAAGTTAGCCATACCACATCA
WMMV	WMV-R	CACTCGCTCTTTAGCTCTTGG
PRSV	PRSV-F	CTCTCATTCCAAGAGGCTCGA
PRSV	PRSV-R	GTGTCTGGGTTATGATGGATGG

Fig. 1 Symptoms of the pumpkin plants with virus disease A:Plant leaves;B:Young leaves;C:Aphids on the abaxial surface;D:Whiteflies on the abaxial surface.

Fig. 2 RT-PCR amplification from the sample RNA M:Marker;-:Negative control without template.

Fig. 3 Neighbor-joining phylogram of CABYV obtained from alignment of nucleotide sequence（MW002437）of this isolate with others from GenBank

Fig. 4 Neighbor-joining phylogram of PRSV obtained from alignment of nucleotide sequence（MW002438）of this isolate with others from GenBank

Fig. 5 Neighbor-joining phylogram of WMV obtained from alignment of nucleotide sequence（MV002439）of this isolate with others from GenBank

Fig. 6 Neighbor-joining phylogram of ZYMV obtained from alignment of nucleotide sequence（MW002440）of this isolate with others from GenBank

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Molecular Identification of Four Viruses that Cause Mixed Infection of Pumpkin in Chengdu,Sichuan Province of China

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