The Plant Pathology Journal 2009;25(4):322-327.
Published online December 31, 2009.
Molecular Analysis of Botrytis cinerea Causing Ginseng Grey Mold Resistant to Carbendazim and the Mixture of Carbendazin Plus Diethofencarb
Joo Hyung Kim, Ji Young Min, Young Seok Bae, Heung Tae Kim
A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture (CarRMixR), the second one resistant to carbendazim and sensitive to the mixture (CarRMixS), and the last one sensitive to both of them (CarSMixS). Carbendazim resistance correlated with a single mutation in β- tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp45I and BstUI allowed differentiation of the PCR fragment of β-tubulin gene of CarSMixS isolates from that of CarRMixR and CarRMixS isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.
Key Words: Botrytis cinerea, beta-tubulin gene, carbendazim, fungicide resistance, mixture of carbendazim plus diethofencarb, PCR-RFLP

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