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Characterization of HrpWAaca protein of Acidovorax avenae subsp. cattleyae OAC1 and improvement of hrp gene inducing medium for Acidovorax spp.
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Acidovorax avenae subsp. cattleyae (Aaca) OAC1 causes a brown spot disease on Cattleyae, while A. a. subsp. citrulli (Aac) causes Bacterial fruit blotch disease (BFB) on cucurbitaceae. These bacteria causing diseases in host plants or eliciting hypersensitive response (HR) on nonhost plants depend on type III secretion system (T3SS), encoded by hrp/hrc gene cluster. In this study, we analyzed the biological function of hrpWAaca of Aaca by loss-of-function strategy. The hrpWAaca mutant elicited a delayed HR on Nicotiana tabacum, but still maintained the same virulence as its wild type on its host Oncidium and Phalaenopsis. Hence, HrpWAaca might be not play an essential role in pathogenicity. The purified or crude extracted HrpWAaca-His6 proteins could elicit HR with or without heat treatment. Moreover, HrpWAaca-His6 protein contains 17.58% glycine but no cysteine, and HrpWAaca could be secreted into bacterial milieu under T3SS inducing condition. Therefore, HrpWAaca is a harpin-like protein. The Pel domain in C-terminus of HrpWAaca has the pectate lyase activity based on the pel assay with polyglacturonic acid as a substrate. Besides, Gel filtration assay showed the purified HrpWAaca-His6 could be detected as dimer (120 kDa)、hexamer (400 kDa) and oligomer (>2000 kDa) in size. To control the tomato bacterial spot caused by Xanthomonas vesicatoria, HrpWAaca-His6 solution was sprayed onto tomato leaves 1 or 4 days before inoculated with X. vesicatoria. The result showed that HrpWAaca-His6 could reduce bacterial spot severity after 4 days harpin treatment. To improve the T3SS induction medium for Aac, extracts of watermelon suspension cell and loofah were total using gfp as a reporter gene under the control of hrcC promoter. The results indicated that both extracts significantly improved induction efficiency of the XVM2 medium.
農業試驗所 - 蝴蝶蘭細菌性褐斑病菌Acidovorax avenae subsp. cattleyae專一性引子之開發 / 許秀惠;宋嘉仁;申屠萱; Hseu, S.H.;Sung, C.J.

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