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1.
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
Journal of Integrative Agriculture    2020, 19 (5): 1283-1291.   DOI: 10.1016/S2095-3119(19)62755-3
摘要134)      PDF    收藏
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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2. Occurrence and molecular characterization of Potato spindle tuber viroid (PSTVd) isolates from potato plants in North China
QIU Cai-ling, ZHANG Zhi-xiang, LI Shi-fang, BAI Yan-ju, LIU Shang-wu, FAN Guo-quan, GAO Yan-ling, ZHANG Wei, ZHANG Shu, Lü Wen-he, Lü Dian-qiu
Journal of Integrative Agriculture    2016, 15 (2): 349-363.   DOI: 10.1016/S2095-3119(15)61175-3
摘要2260)      PDF    收藏
China is the largest potato producing country worldwide, with this crop representing the fourth largest staple food crop in China. However, the steady presence of Potato spindle tuber viroid (PSTVd) over the past five decades has a significant economic impact on potato production. To determine why PSTVd control measures have been ineffective in China, more than 1 000 seed potatoes collected between 2009 and 2014 were subjected to PSTVd detection at the Supervision and Testing Center for Virus-free Seed Potatoes Quality, Ministry of Agriculture, China. A high PSTVd infection rate (6.5%) was detected among these commercial seed potatoes. Some breeding lines of potato collected from 2012 to 2015 were also tested for PSTVd infection, revealing a high rate of PSTVd contamination in these potato propagation materials. Furthermore, comparison of the full-length sequences of 71 different Chinese PSTVd isolates revealed a total of 74 predominant PSTVd variants, which represented 42 different sequence variants of PSTVd. Comparative sequence analysis revealed 30 novel PSTVd sequence variants specific to China. Comprehensive phylogenetic analysis uncovered a close relationship between the Chinese PSTVd sequence variants and those isolated from Russia. It is worth noting that three intermediate strains and six mild strains were identified among these variants. These results have important implications for explaining the ineffective control of PSTVd in China and thus could serve as a basic reference for designing more effective measures to eliminate PSTVd from China in the future.
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