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Sensitivity of Colletotrichum spp. Isolated from Grapes in Korea to Carbendazim and the Mixture of Carbendazim Plus Diethofencarb

Sookyeong Hwang,Hye-ryoung Kim,Joo-Hyung Kim,Jong-Han Park,Sang-Bum Lee,S. Cheong,Heungtae Kim

Published 2010 in Plant Pathology Journal

ABSTRACT

National Institute of Horticultural & Herbal Science, RDA, Suwon 441-440, Korea(Received on June 16, 2009; Accepted on November 10, 2009)Thirty-six isolates of Colletotrichum spp. were obtainedfrom infected grapes in two different locations of Korea;18 isolates from Cheonahn, where carbendazim (MBC)and the mixture of MBC and diethofencarb (NPC) hadbeen applied to control grape ripe rot, and 18 isolatesfrom Cheongju, where no fungicides had been used.Sequences analysis of the internal transcribed spacer(ITS) and the β-tubulin gene identified 34 of the 36isolates as Colletotrichum gloeosporioides. The remainingtwo isolates from Cheongju were identified as C. acutatum.Of the 18 isolates from Cheonahn, 12 were resistant toboth MBC and the mixture (MBC+NPC), and six weresensitive to them. All C. gloeosporioides isolates fromCheongju, but not the two C. acutatum isolates, weresensitive to these fungicides. Sequence analysis of the β-tubulin gene in all isolates revealed that C. gloeosporioidesresistant to MBC and MBC+NPC had a tyrosine insteadof phenylalanine at the amino acid position 200. The ap-pearance of resistance to MBC and the mixture in C.gloeosporioides correlated with the history of fungicideapplication in Korea. Keywords : fungicide resistance, mixture of carbendazimplus diethofencarb, grape ripe rot, Colletotrichum gloeo-sporioides, Colletotrichum acutatumGrape ripe rot, a serious Colletotrichum bunch rot diseasethat occurs on mature grapes as they ripen, was firstreported in the United States in 1891 and has been found inmost regions where grapes are grown, especially in warmand humid areas such as the southeastern USA (Pearsonand Goheen, 1988). Initially, C. gloeosporioides was consi-dered to be the causal organism of the disease. However,both C. gloeosporioides and C. acutatum were implicatedin ripe-rot disease outbreaks on muscadine grapes in theUnited States (Daykin and Milholland, 1984; Kummuanget al., 1996). More recently, C. acutatum was found to bethe causal organism for ripe rot in Korea along with C.gloeosporioides (Hong et al., 2008). In Japan, ripe rot hasalso been the most serious pre-harvest disease of grapeberries with the main causal pathogens being Glomerellacingulata and C. acutatum (Ozoe et al., 1972; Yamamoto etal., 1999). In Korea, several fungicides have been used to controlgrape ripe rot. Because unlike C. gloeosporioides, C.acutatum is resistant to certain fungicides such as benzimi-dazoles (BENs), correct taxonomic identification is there-fore important in selecting appropriate fungicides fordisease management (Chung et al., 2006; Kim et al., 2007;Peres et al., 2004). For approximately 30 years, BENsincluding carbendazim (MBC), benomyl, and thiabendazolehave been widely and successfully used to protect manycrops from phytopathogens (Russell, 1995). These agentsinhibit nuclear division via disruption of microtubuleassembly during mitosis by binding to the β-tubulin subunit(Davidse and Flach, 1977). However, The extended use ofthese compounds has resulted in the selection of resistantpathogen genotypes, which can remain dominant in apopulation for several years after discontinued BEN use(Moorman and Lease, 1992). The first cases of resistancewere reported in fungi with short life cycles, such asBotrytis cinerea, in grape vineyards (Leroux and Clerjeau,1985). Due to the emergence of many types of resistantpopulations of phytopathogenic fungi in the field, theefficacy of these chemicals to control plant diseases hasdecreased (Bonnen and Hopkins, 1997; Maymon et al.,2006; Washington et al., 1992). In most reported cases ofacquired resistance to BENs, point mutations in the β-tubulin gene were responsible (Koenraadt et al., 1992).Studies of the mutants generated in the laboratory haverevealed a small number of amino acid substitutions in theβ-tubulin gene that cause resistance (Fujimura et al., 1992).Numerous cases of resistance to BENs have beenreported for several species of Colletotrichum in variouscrops (Kim et al., 2007; Maymon et al., 2006; Peres et al.,2004; Sanders et al., 2000; Wong et al., 2008; Yang andTeBeest, 1995). Although BENs have been widely used to

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