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Folia microbiologica
Apr, 2021;66 (2): 273-283.

Loop-mediated isothermal amplification (LAMP) assay for detection of sesame phyllody phytoplasmas in Vietnam.

Nguyen Bao Quoc, Nguyen Thi Thanh Xuan, Nguyen Mai Nghiep, Nguyen Doan Nguyen Phuong, Ton Bao Linh, Nguyen Ngoc Bao Chau, Nguyen Duc Xuan Chuong, Nguyen Chau Nien, Matthew Dickinson
Author Information
  1. Nguyen Bao Quoc: Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Vietnam. baoquoc@hcmuaf.edu.vn. ORCID
  2. Nguyen Thi Thanh Xuan: Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Vietnam.
  3. Nguyen Mai Nghiep: Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Vietnam.
  4. Nguyen Doan Nguyen Phuong: Research Institute for Biotechnology and Environment, Nong Lam University, Ho Chi Minh City, Vietnam.
  5. Ton Bao Linh: Department of Biotechnology, Nong Lam University, Ho Chi Minh City, Vietnam.
  6. Nguyen Ngoc Bao Chau: Faculty of Biotechnology, Ho Chi Minh City Open University, Ho Chi Minh City, Vietnam.
  7. Nguyen Duc Xuan Chuong: Faculty of Agronomy, Nong Lam University, Ho Chi Minh City, Vietnam.
  8. Nguyen Chau Nien: Faculty of Agronomy, Nong Lam University, Ho Chi Minh City, Vietnam.
  9. Matthew Dickinson: School of BioSciences, University of Nottingham, Nottingham, UK.
PMID: 33404955 DOI: 10.1007/s12223-020-00842-0

Abstract

Phloem-limiting phytoplasmas are known to be causal agents of phyllody, which is recognized by the abnormal development of floral structures resulting in serious yield losses in sesame plants. Currently, identification of the various groups of phytoplasmas that cause sesame phyllody (SP) is conducted by nested PCR, RFLP, and multiplex real-time qPCR assays. However, these methods require intensive labor and are costly and time-consuming so can only be undertaken in well-equipped labs. Here, diagnostic loop-mediated isothermal amplification (LAMP)-based assays allowing rapid detection of specific groups of phytoplasmas within 30 min were developed based on detection of the 16S rRNA sequence of phytoplasmas. Universal 16S rRNA phytoplasma primers and seven primer sets of different 16Sr group phytoplasmas (16SrI, 16SrII, 16SrIII, 16SrIV, 16SrV, 16SrX, 16SrXI) and universal plant cytochrome oxidase (cox) gene primers were used to detect 16S rRNA group phytoplasma sequences and the cox gene in sesame plants. The LAMP assays were carried out using a real-time fluorometer with amplification plots and annealing curves visualized directly. Results demonstrated that the 16SrI and 16SrII group phytoplasmas were causal agents of sesame phyllody in Vietnam. LAMP-based assays for in-field detection of sesame phyllody-causing phytoplasmas revealed advantages and potential applicability in comparison with conventional approaches. To the best of our knowledge, this is the first assessment of multiple phytoplasma infection associated with sesame phyllody disease in Vietnam using LAMP-based assays.

Keywords

LAMP-based assay Nested PCR Phytoplasmas Rapid detection Sesame phyllody

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Grants

  1. No. 21/FIRST/1a/HUAF/FIRST-MOST

MeSH Term

DNA, Bacterial
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques
Phytoplasma
Plant Diseases
RNA, Ribosomal, 16S
Real-Time Polymerase Chain Reaction
Sesamum
Vietnam

Chemicals

DNA, Bacterial
RNA, Ribosomal, 16S
Journal Article

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