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Identification of a novel emaravirus infecting lilac through next-generation sequencing
WANG Ya-qin, SONG Yu, CAO Meng-ji, CHENG Qi, WU Jian-xiang, HU Tao
2020, 19 (8): 2064-2071.   DOI: 10.1016/S2095-3119(19)62806-6
Abstract143)      PDF in ScienceDirect      
Emaraviruses are multipartite negative-sense single-stranded viruses that usually infect flowering plants.  Using unbiased next-generation sequencing (NGS) approach, a novel virus from lilac leaves with yellow mottle symptom was identified.  Five viral genomic RNA segments were detected and completely sequenced.  The molecular characteristics of the viral RNAs and proteins resemble those of members of the genus Emaravirus.  The virus was found to be mechanically transmissible to Nicotiana benthamiana. Sequence alignment and phylogenetic analyses of the virus support the proposal of creating a new species in the genus Emaravirus for which the name lilac chlorotic ringspot-associated virus (LiCRaV) is proposed. 
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Monoclonal antibody-based serological detection of potato virus M in potato plants and tubers
ZHANG Yu, GAO Yan-ling, HE Wan-qin, WANG Ya-qin, QIAN Ya-juan, ZHOU Xue-ping, WU Jian-xiang
2020, 19 (5): 1283-1291.   DOI: 10.1016/S2095-3119(19)62755-3
Abstract134)      PDF in ScienceDirect      
Potato virus M (PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide.  To investigate and control this viral disease, efficient and specific detection techniques are needed.  In this study, PVM virions were purified from infected potato plants and used as the immunogen to produce hybridomas secreting PVM-specific monoclonal antibodies (MAbs).  Four highly specific and sensitive murine MAbs, i.e., 1E1, 2A5, 8A1 and 17G8 were prepared through a conventional hybridoma technology.  Using these four MAbs, we have developed an antigen-coated plate (ACP)-ELISA, a dot-ELISA and a Tissue print-ELISA for detecting PVM infection in potato plants and tubers.  PVM could be detected in infected potato plant tissue crude extracts diluted at 1:10 240 (w/v, g mL–1) by the dot-ELISA or at 1:163 840 (w/v, g mL–1) by the ACP-ELISA.  The Tissue print-ELISA is the quickest and easiest assay among the three established serological assays and is more suitable for onsite large-scale sample detection.  Detection results of the field-collected samples showed that PVM is currently widespread in the Yunnan and the Heilongjiang provinces in China.  The field sample test results of the developed serological assays were supported by the results from RT-PCR and DNA sequencing.  We consider that the newly established ACP-ELISA, dot-ELISA and Tissue print-ELISA can benefit PVM detection in potato plant and tuber samples and field epidemiological studies of PVM.  These assays can also facilitate the production of virus-free seed potatoes and breeding for PVM-resistant potato cultivars, leading to the successful prevention of this potato viral disease.
 
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Genome-wide identification, characterization, and expression analysis of the SWEET gene family in cucumber
HU Li-ping, ZHANG Feng, SONG Shu-hui, TANG Xiao-wei, XU Hui, LIU Guang-min, WANG Ya-qin, HE Hong-ju
2017, 16 (07): 1486-1501.   DOI: 10.1016/S2095-3119(16)61501-0
Abstract969)      PDF in ScienceDirect      
    SWEETs (sugars will eventually be exported transporters) are a novel class of recently identified sugar transporters that play important roles in diverse physiological processes. However, only a few species of the plant SWEET gene family have been functionally identified. Up till now, there has been no systematic analysis of the SWEET gene family in Cucurbitaceae crops. Here, a genome-wide characterization of this family was conducted in cucumber (Cucumis sativus L.). A total of 17 CsSWEET genes were identified, which are not evenly distributed over the seven cucumber chromosomes. Cucumber SWEET protein sequences possess seven conserved domains and two putative serine phosphorylation sites. The phylogenetic tree of the SWEET genes in cucumber, Arabidopsis thaliana, and Oryza sativa was constructed, and all the SWEET genes were divided into four clades. In addition, a number of putative cis-elements were identified in the promoter regions of these CsSWEET genes: nine types involved in phytohormone responses and eight types involved in stress responses. Moreover, the transcript levels of CsSWEET genes were analyzed in various tissues using quantitative real-time polymerase chain reaction. A majority (70.58%) of the CsSWEET genes were confined to reproductive tissue development. Finally, 18 putative watermelon ClaSWEET genes and 18 melon CmSWEET genes were identified that showed a high degree of similarity with CsSWEET genes. The results from this study provided a basic understanding of the CsSWEET genes and may also facilitate future research to elucidate the function of SWEET genes in cucumber and other Cucurbitaceae crops.
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