Dataset Information

Molecular detection of watermelon mosaic virus associated with a serious mosaic disease on Cucurbita pepo L. in Shanxi, China.

ABSTRACT: With continued expansion of Cucurbita pepo L. cultivation, viral diseases affecting the crop have become more serious in recent years, causing enormous losses in yield and quality. A virus sample was obtained from Wenshui in Shanxi province, China. Double-stranded RNA technology and sequence-independent amplification (SIA) were used to identify the virus that induced C. pepo L. mosaic disease. SIA and sequencing results showed the presence of watermelon mosaic virus (WMV) in diseased C. pepo L. leaves. The complete sequence of WMV from the Shanxi isolate (i.e., WMV-WS) was cloned and analyzed for further characterization. The genomic RNA of WMV-WS is 10,040 nucleotides in length and encodes a putative polyprotein of 3218 amino acids. Phylogenetic analysis indicate that all WMV isolates were divided into four groups and WMV-WS isolate belong to Group 4. Further analysis showed that these WMV isolates were not only to a certain degree related to the host, but also related to geographical origin of isolates. Our results provide information for a better understanding of the genetic diversity of WMV isolates infecting C. pepo L. in Shanxi, China.

SUBMITTER: Niu Y

PROVIDER: S-EPMC5684996 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature

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Molecular detection of watermelon mosaic virus associated with a serious mosaic disease on Cucurbita pepo L. in Shanxi, China.

Niu Yanbing Y Pang Xiaojing X Cheng Yuanyuan Y Wang Defu D Guo Shang S Liu Yong Y

Virusdisease 20170629 3

With continued expansion of Cucurbita pepo L. cultivation, viral diseases affecting the crop have become more serious in recent years, causing enormous losses in yield and quality. A virus sample was obtained from Wenshui in Shanxi province, China. Double-stranded RNA technology and sequence-independent amplification (SIA) were used to identify the virus that induced C. pepo L. mosaic disease. SIA and sequencing results showed the presence of watermelon mosaic virus (WMV) in diseas ...[more]

PMID: 29291219

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Project description:The genus Cucurbita comprises many popular vegetable and ornamental plants, including pumpkins, squashes, and gourds, that are highly valued in China as well as in many other countries. During a survey conducted in Zhejiang province, Southeast China in 2016, severe symptoms of viral infection were observed on Cucurbita maxima Duch. ex Lam. Diseased plants showed symptoms such as stunting, mosaicking, Shoe string, blistering, yellowing, leaf deformation, and fruit distortion. Approximately, 50% of Cucurbita crops produced in Jinhua were diseased, causing an estimated yield loss of 35%. In this study, we developed a method using all known virus genomes from the NCBI database as a reference to map small RNAs to develop a diagnostic tool that could be used to diagnose virus diseases of C. maxima. 25 leaf samples from different symptomatic plants and 25 leaf samples from non-symptomatic plants were collected from the experimental field of Jihua National Agricultural Technology Garden for pathogen identification. Small RNAs from each set of three symptomatic and non-symptomatic samples were extracted and sequenced by Illumina sequencing. Twenty-four different viruses were detected in total. However, the majority of the small RNAs were from Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), and Cucumber mosaic virus (CMV). Mixed infections of these three viruses were diagnosed in leaf samples from diseased plants and confirmed by reverse transcription PCR (RT-PCR) using primers specific to these three viruses. Crude sap extract from symptomatic leaf samples was mechanically inoculated back into healthy C. maxima plants growing under greenhouse conditions. Inoculated plants developed the same disease symptoms as those observed in the diseased plants and a mixed infection of ZYMV, WMV, and CMV was detected again by RT-PCR, thus fulfilling Koch's postulates. The diagnostic method developed in this study involves fewer bioinformatics processes than other diagnostic methods, does not require complex settings for bioinformatics parameters, provides a high level of sensitivity to rapidly diagnose plant samples with symptoms of virus diseases and can be performed cheaply. This method therefore has the potential to be widely applied as a diagnostic tool for viruses that have genome information in the NCBI database.

Dataset Information

Molecular detection of watermelon mosaic virus associated with a serious mosaic disease on Cucurbita pepo L. in Shanxi, China.

Publications

Molecular detection of watermelon mosaic virus associated with a serious mosaic disease on <i>Cucurbita pepo</i> L. in Shanxi, China.

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