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A universal probe for simultaneous detection of six pospiviroids and natural infection of potato spindle tuber viroid (PSTVd) in tomato in China
ZHANG Yu-hong, LI Zhi-xin, DU Ya-jie, LI Shi-fang, ZHANG Zhi-xiang
2023, 22 (3): 790-798.   DOI: 10.1016/j.jia.2022.08.119
Abstract332)      PDF in ScienceDirect      

Several viroids in the genus Pospiviroid can infect tomato (Solanum lycopersicum) and cause severe diseases, posing a serious threat to tomato production.  For simultaneous detection of six tomato-infecting pospiviroids - columnea latent viroid (CLVd), pepper chat fruit viroid (PCFVd), potato spindle tuber viroid (PSTVd), tomato apical stunt viroid (TASVd), tomato chlorotic dwarf viroid (TCDVd), and tomato planta macho viroid (TPMVd), we developed a universal probe based on a highly conserved 61 nt long sequence shared among them.  Compared with their specific probes, the universal probe has a similar, though slightly reduced, detection sensitivity and has the advantages of simple and cost-effective preparation and simultaneous detection of the six pospiviroids.  In addition, the universal probe was used in dot-blot hybridization assays for a large-scale survey of viroid(s) in tomato plantings in China.  Only PSTVd was detected in a few greenhouse-planted tomato plants.  Sequence analysis revealed that these tomato PSTVd isolates may have been introduced from tomato seeds imported from abroad. 

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Analysis of fig tree virus type and distribution in China
Mahmut Mijit HE Zhen, HONG Jian, LU Mei-guang, LI Shi-fang, ZHANG Zhi-xiang
2017, 16 (06): 1417-1421.   DOI: 10.1016/S2095-3119(16)61551-4
Abstract939)      PDF in ScienceDirect      
The common fig (Ficus carica L.) was one of the earliest horticultural crops to be domesticated.  A number of different viruses can infect fig trees including Fig mosaic virus (FMV) that has been detected in several commercial fig trees in Xinjiang, China.  However, the distribution of FMV and other fig-infecting viruses in China remains unknown.  In the present study, a sample from an ancient fig tree growing in Xinjiang was investigated by electron microscopy (EM) followed by PCR/RT-PCR, and FMV, Fig badnavirus 1 (FBV-1) and Fig leaf mottle-associated virus 1 (FLMaV-1) were detected.  Fig leaf samples (252) from commercial orchards across China were subjected to PCR/RT-PCR, and FMV, FBV-1 and Fig fleck-associated virus (FFkaV) were relatively abundant (44.4, 48.4 and 44%, respectively), while FLMaV-1 and Fig mild mottle-associated virus (FMMaV) were much scarcer (5.6 and 0.4%, respectively), and FLMaV-2, Fig cryptic virus (FCV), and Fig latent virus (FLV) were not detected.  The presence of disease-causing viruses in fig trees presents a significant challenge for fig producers in China.  This study may help to promote actions aimed at controlling fig viruses, especially FMV.
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Complete nucleotide sequences of two isolates of Cherry virus A from sweet cherry in China
GAO Rui, LI Shi-fang, LU Mei-guang
2016, 15 (7): 1667-1671.   DOI: 10.1016/S2095-3119(16)61343-6
Abstract2258)      PDF in ScienceDirect      
    Cherry virus A (CVA) is a member of the genus Capillovirus, in the family Betaflexiviridae. The infection rate of CVA was high in sweet cherry in China. We determined the complete nucleotide sequences of two isolates of CVA from Tai’an, Shandong Province, China using high fidelity PCR enzymes and specific primer pairs for amplifying long fragments in RT-PCR and RACE. The full-length sequences from isolates ChTA11 and ChTA12 are both 7 382 nucleotide (nt) long, excluding the poly(A) tail, encode two open reading frames (ORFs) and have similar genome organization to the two isolates in GenBank. The complete nucleotide sequence of ChTA11 is 98.2 and 81.2% nt identity to the isolates from Germany and India in GenBank, respectively, and the ChTA12 isolate is 98.2 and 81.0% similar. Analysis of the nucleotide and amino acid sequences showed that the domain of unknown function (DUF1717) is more variable compared with other domains. This is the first report of the complete nucleotide sequences of CVA isolates infecting sweet cherry in China.
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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
2016, 15 (2): 349-363.   DOI: 10.1016/S2095-3119(15)61175-3
Abstract2260)      PDF in ScienceDirect      
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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