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Pathogen and Biological Contamination Management

The Road Ahead

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Part of the Methods in Molecular Biology™ book series (MIMB,volume 318)

Abstract

Multiplication of certified pathogen-free stock plants in vitro makes an important contribution to the production of disease-free planting material for vegetatively propagated crops. Meristem culture is extensively used to eliminate pathogens and contaminants from microbially contaminated plants prior to micropropagation. The approach to pathogen and contamination management differs. It is essential to avoid the release of pathogen-contaminated microplants and to ensure this the plant pathogen-testing guidelines and protocols issued by the Food and Agriculture Organisation (FAO) and its regional representative organizations should be followed. Where in vitro methods are used to eliminate pathogens, the progeny plants should be established in vivo under quarantine conditions and tested under FAO guidelines before being used as stock plants for in vitro multiplication. At establishment of microplants in vitro (stage 1), cultures should be culture-indexed for the presence of microbial contaminants. If pathogenand contaminant-free cultures are established, then the risk is that of managing laboratory contamination by common environmental microorganisms based on Hazard Analysis Critical Control Points (HACCP) principles.

International plant health certification organizations are conservative and rely on established pathogen indexing protocols. They are reluctant to accept DNA-based tests and do not accept testing of in vitro cultures. Given that in time both of these restrictions may be relaxed, micropropagators may look forward to availing more of diagnostic service providers using polymerase chain reaction-based multiplex assays for pathogen-indexing and advances in diagnostic kits for environmental microorganisms in support of laboratory contamination management; with the caveat that molecular tests for pathogens may continue to require confirmation by inoculation of indicator plant species.

Key Words

  • Environmental microorganisms
  • HACCP
  • meristem culture
  • molecular beacons
  • multiplex assays
  • PCR

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Cassells, A.C., Doyle, B.M. (2006). Pathogen and Biological Contamination Management. In: Loyola-Vargas, V.M., Vázquez-Flota, F. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology™, vol 318. Humana Press. https://doi.org/10.1385/1-59259-959-1:035

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  • DOI: https://doi.org/10.1385/1-59259-959-1:035

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-547-7

  • Online ISBN: 978-1-59259-959-2

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