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A Rapid Identification Method of Bactrocera cilifera (Hendel) with species-specific primers(SS-COI)

Plant Diseases and Pests 2021年第2期12卷 33-37页

作者： Zhen Huang Qiongxia Guo College of Plant Protection Fujian Agriculture and Forestry UniversityFuzhou 350002China Fuzhou Changle Airport Customs Fuzhou 50209China Fuzhou Customs Technical Center Fuzhou 350001China

[Objective]The paper was to establish a rapid identification method of Bactrocera cilifera(Hendel)with species-specific primers(SS-COI).[Method]Using ***(Hendel)as the positive control,and 19 species of fruit flies such as ***(Coquillett)and ***(Hendel)as the negative controls,a pair of species-specific primers,YF290 and YR511,were designed and screened for accurate identification of ***,based on mitochondrial DNA COI sequence.[Result]The PCR products were amplified and detected by *** a clear and single band was observed at about 222 bp in the positive control,while no bands were found in the other negative controls.[Conclusion]The established rapid identification method with species-specific primers(SS-COI)is of great practical significance for rapid identification of fruit flies intercepted from import and export fruits and vegetables at ports,and for rapid clearance and early warning of import fruits and vegetables at ports.

关键词： Bactrocera cilifera(Hendel) species-specific primers species-specific PCR(SS-PCR) mt DNA COI Rapid identification

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Development of species-specific PCR primers and Sensitive Detection of the Tylenchulus semipenetrans in China

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Agricultural Sciences in China 2011年第2期10卷 252-258页

作者： LIU Guo-kun CHEN Juan XIAO Shun ZHANG Shao-sheng PAN Dong-ming College of Plant Protection Fujian Agriculture and Forestry UniversityFuzhou 350002P.R.China College of Horticulture Fujian Agriculture and Forestry UniversityFuzhou 350002P.R.China

Tylenchulus semipenetrans is the most economically important and widespread nematode pest of citrus in ***-ITS of 14 populations of *** which were collected from different citrus groves or Chinese fir（Cunninghamia lanceolata） plantations in China were amplified and *** species-specific primers were designed for the first time to diagnosis *** based on the sequences of rDNA-ITS regions of geographic population *** primers were sensitive to amplify the expected band size（297 bp） from DNA template of a single second-stage juvenile（J2） or different life stages of *** specific band was amplified from 15 non-target nematode species which were commonly found in citrus *** and reliability of the primers were validated by further PCR amplification of 16 extra populations of *** collected from 4 provinces of *** primers successfully detected a single J2 of *** within a whole nematode community comprising a large numbers of non-target *** developed diagnostic technique can be used for accurate identification of *** and also as a decision tool for nematode management for citrus or Chinese fir in China.

关键词： Tylenchulus semipenetrans rDNA-ITS species-specific primers diagnosis detection

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Molecular authentication of the traditional Chinese medicine Tongren Dahuoluo Wan and its alternative formulation

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Frontiers of Agricultural Science and Engineering 2017年第3期4卷 353-357页

作者： Jikun WANG Jing DU Meng CAO Lu YAO Suhua XIE Jiafu CHEN Xingbo ZHAO National Engineering Laboratory for Animal Breeding Key Laboratory of Animal Genetics Breeding and ReproductionMinistry of Agriculture College of Animal Science and Technology China Agricultural University Research Institute of Beijing Tongrentang Co. Ltd.

Tongren Dahuoluo Wan has been a popular traditional Chinese medicine in international pharmaceutical markets for hundreds of years. Leopard bone powder is the key element in its formulation. However, the leopard has been listed for wildlife conservation, which limits the use of the leopard bone supplies. Therefore, an alternative formulation which substitutes leopard bone with zokor bone in the formula of Tongren Dahuoluo Wan is now manufactured. To develop a simple and reliable molecular method for authenticating the two patent medicines,mitochondrial nucleotide polymorphic sites of 12 S rRNA,COI and Cytb genes were screened in leopard and zokor bones, and nine pairs of species-specific primers were verified for discriminating the two species. For the patent medicine authentication, we set up a molecular diagnostic assay to resolve the difficulties of low concentration of target DNAs and presence of PCR-inhibitory substances in this complex medicine, and successfully confirmed leopard or zokor content using the nine pairs of species-specific primers. We recommend a common technical strategy for authentication of species origins in traditional Chinese medicine, and discuss the experimental solutions for technical problems of molecular diagnostic assays.

关键词： Tongren Dahuoluo Wan molecular diagnostic assay Eospalax baileyi Panthera pardus species-specific primers

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Shoot rot of Zizania latifolia and the first record of its pathogen Pantoea ananatis in China

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2022年第4期23卷 328-338页

作者： Zilan XIAO Jianping DENG Xiaojun ZHOU Liyan ZHU Xiaochan HE Jingwu ZHENG Deping GUO Jingze ZHANG Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects Institute of BiotechnologyZhejiang UniversityHangzhou 310058China Plant Protection and Plant Inspection Station of Jinyun County Jinyun 321401China Jinhua Academy of Agricultural Sciences Jinhua 321017China Department of Horticulture College of Agriculture and BiotechnologyZhejiang UniversityHangzhou 310058China

The aquatic grass Zizania latifolia grows symbiotically with the fungus Ustilago esculenta producing swollen structures called Jiaobai,widely cultivated in China.A new disease of *** was found in Zhejiang Province,*** lesions appeared on the leaf sheaths or sometimes on the leaves near the leaf *** lesions extended along the axis of the leaf shoots and formed long brown to dark brown streaks from the leaf sheath to the leaf,causing sheath rot and death of entire leaves on young *** pathogen was isolated and identified as the bacterium Pantoea ananatis,based on 16 S ribosomal RNA(r RNA)gene sequencing,multilocus sequence analysis(atp D(β-subunit of ATP synthase F1),gyr B(DNA gyrase subunit B),inf B(translation initiation factor 2),and rpo B(β-subunit of RNA polymerase)genes),and pathogenicity *** observations using scanning electron microscopy revealed that the bacterial cells colonized the vascular tissues in leaf sheaths,forming biofilms on the inner surface of vessel walls,and extended between vessel elements via the perforated *** achieve efficient detection and diagnosis of ***,species-specific primer pairs were designed and validated by testing closely related and unrelated species and diseased tissues of *** is the first report of bacterial sheath rot disease of *** caused by *** in China.

关键词： Zizania latifolia Phylogeny Pantoea ananatis Multilocus analysis Scanning electron microscopy species-specific primers

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