A Queensland plant nursery wishes to import tissue culture plantlets of oriental

Question

A Queensland plant nursery wishes to import tissue culture plantlets of oriental lily varieties from Himachal Pradesh in northern India. Lilies from this region have been previously reported as suffering a phytoplasma disease. To meet Department of Agriculture quarantine requirements, imported lily plantlets must be grown out in an Australian post-quarantine entry glasshouse facility and test negative for phytoplasma disease before they can be released and taken into commercial production. You are responsible for testing these quarantine lilies and deciding if they are disease-free and able to be released in Australia. There is no commercial kit available. Explain the process you would undertake to develop a diagnostic test for phytoplasma in imported lilies. In your answer include: (1) a list of the resources you need to develop a reliable diagnostic test; and (2) list at least two examples of relevant diagnostic technology platforms which you might develop (these assays will need to be sensitive!)

Explanation / Answer

Answer:

1)Here are some References:
a) Paper by Dumonceaux et.al in NCBI -Molecular Diagnostic Tools for Detection and Differentiation of Phytoplasmas Based on Chaperonin-60 Reveal Differences in Host Plant Infection Patterns.
b) Paper by Yi Fang et. al in NCBI, on Current and Prospective Methods for Plant Disease Detection
c) A book -Recent Advances in the Diagnosis and Management of Plant Diseases- would also be helpful.

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Phytoplasmas are recalcitrant to cultivation and are often difficult to detect and identify due to their erratic distribution, low concentration, seasonal fluctuation and enzyme-inhibitory plant poly-saccharide and polyphenolic compounds especially in woody perennial plant hosts. Rapid, sensitive, accurate and early diagnosis of phytoplasma diseases is indispensable to reduce their economical impact. Several serological and molecular techniques have been developed for accurate and sensitive detection of phytoplasmas in both host plants and insect vectors.

Sensitive ELISA to detect Phytoplasma.
Restrction Fragment Length Ploymorphism Techniques(RFLPs) could also be used to detect Phytoplasma.
Quantitative PCR and bioimaging also helps in detecting Phyoplasma titers in plants.

Also Dectection kit which is available using loop-mediated isothermal amplification (Sensitive rapid diagnostic kit) could also be reffered.

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