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標題: 辣椒炭疽病菌高度分枝侵入構造(HBPS)的誘導與調控
Induction and regulation of highly branched penetration structure of Colletotrichum acutatum
作者: 陳美雅
Chen, Mei-Ya
Contributors: 李敏惠
Miin-Huey Lee
植物病理學系所
關鍵字: 辣椒炭疽病菌;高度分枝侵入構造;訊號傳遞路徑;外在環境因子;光照
Colletotrichum acutatum;highly branched penetration structure (HBPS);signal transduction pathway;environmental factors;light
日期: 2012
Issue Date: 2013-11-19 12:38:50 (UTC+8)
Publisher: 植物病理學系所
摘要: 辣椒是台灣重要的熱帶栽培作物之一,在台灣 C. acutatum 是造成辣椒炭疽病的的重要病原菌之一,當 C. acutatum 嚴重感染辣椒果實時,可造成辣椒嚴重損失。本實驗室前人的研究中首度發現 C. acutatum 可在辣椒果實的角質層形成一個高度分枝的侵入構造稱 “Highly branched penetration structure”。本研究主要是探討辣椒炭疽病菌高度分枝侵入構造的誘導與調控。首先使用有機溶劑萃取四層辣椒角質層 PC1、PC2、PC3 與 PC4, 並探討 C. acutatum Coll-153 菌株在這四種角質層的 HBPS 形成情形,結果 Coll-153 可在 PC1 與 PC2 中形成高比例的 HBPS ,但無法在 PC3 與 PC4 中形成。由於 Coll-153 可在 PC2 形成最高比例的 HBPS,因此以 PC2 進行 HBPS 的誘導與調控因子之分析。實驗結果顯示外在環境因子之光照及滲透壓皆會影響 HBPS 的形成,其中以光照之影響最為顯著,當光照時間愈長時, HBPS 形成比例愈低。溫度及殺菌劑處理並不影響 HBPS 的形成。若早期處理產生氧化的藥劑則可抑制 HBPS 的情形,但於後期處理則無影響。以作用於細胞之不同訊息傳遞途徑之化學藥劑處理以探討 HBPS 形成的調控,結果顯示 HBPS 的形成受到 cAMP dependent protein kinase A (PKA) 、PIP3 及 MAP kinase 所媒介之訊息傳遞路徑所調控,HBPS 的形成亦受到細胞內的鈣離子濃度所調控,但 HBPS 的形成卻不受鈣離子幫浦 A23187、 鈣離子通道抑制劑 nicardipine hydrochloride 及第二型鈣離子/鈣調素依存型蛋白質激酶抑制劑 KN93 所調控。將光照、neomycin sulfate 及 cAMP 處理的條件應用在群香綠果接種試驗,結果三種處理條件對 HBPS 在辣椒果實之角質層的形成皆沒有影響, 但特定濃度的 neomycin sulfate 會抑制病斑的擴展。
Chili pepper (Capsicum spp.) anthracnose caused by Colletotrichum acutatum results in severe losses in yield and fruit quality in Taiwan. Our previous study has shown that C. acutatum penetrates the cuticle layer of chili pepper fruits by forming a highly branched penetration structure (HBPS) which was not previously characterized in any Colletotrichum-plant interactions. The purpose of the study is to investigate the induction and regulation of HBPS formation. Cuticle layer PC1, PC2, PC3 and PC4 from pepper fruit were isolated (Capsicum annuum, GroupZest) C. acuttaum strain Coll-153 forms high proportion of HBPS in PC1 and PC2, but hardly forms HBPS in PC3 and PC4. Overall C. acutatum forms the highest frequency of HBPS in PC2. Therefore, PC2 was used to study factors involved in HBPS induction and regulation. The results show that HBPS formation can be regulated by light and osmotic stress. Light has highly significant impact on HBPS formation, the longer exposure time the lower proportion of HBPS formation. The formation of HBPS doesn’t seem to be regulated by stress caused by temperature and fungicides. Oxidative stress, when treated at early stage, was able to inhibit HBPS formation, but the inhibition was not found when treated at late stage. Pharmacological effectors of signal transduction pathways were used to study the regulation of HBPS formation. HBPS formation was found to be inhibited by the stimulators of cAMP dependent protein kinase A and by the inhibitors of phospholipase C and MAP kinase. The imbalance of cellular calcium level also seems to have a role on HBPS formation. However, HBPS was not inhibited by calcium ionophore A23187, calcium channel blocker nicardipine hydrochloride and calcium/calmodulin kinase II inhibitor. Light, neomycin sulfate and cAMP did not inhibit HBPS formation in chili pepper fruits when treated on 18 h post-inoculated fruits. However, certain amounts of neomycin sulfate could reduce disease of chili pepper fruits caused by C. acutatum Coll-153.
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