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Application of molecular detection on Botrytis elliptica and establishment of identification for intra-species of Botrytis spp.
灰黴病菌可在許多植物上造成病害，並適合在低溫高濕的環境下發病。在 PDA 上可形成白色菌落，其後可產生分生孢子及黑色不規則的菌核。灰黴病菌在酸性環境下生長情形較在鹼性環境中良好。不同培養基不影響其產胞量，但培養於 125 ml 三角瓶之灰黴病菌其孢子產量較培養於 9 cm 之培養皿者為高。觀察菌核之發芽率與發芽方式和溫度之間的關係時發現，發芽率隨著溫度的上昇而升高，至 28 ℃ 後開始下降，而菌核僅在 20 及 24 ℃ 時發芽產生分生孢子，其餘處理皆發芽形成菌絲。在神岡鄉調查灰黴病之田間生態時發現灰黴病菌之病勢進展與風向有關，推測此乃因分生孢子為其主要二次感染源之故。利用選擇性培養基檢測灰黴病菌時可在空氣中測得 Botrytis cinerea 及 B. elliptica 的存在，但在百合種球上卻並未檢測到灰黴病菌。利用專一性引子對 66B03R/66B03F 對所收集的百合灰黴病菌菌株進行專一性測試，雖然可以在植物葉片上測得濃度為 103 conidia/ml的孢子，但並無法對於所有的 B. elliptica 測試菌株產生反應。利用 ITS 區域及 PCR-RFLP 可將 B. cinerea、B. elliptica、B. gladiolorum 及 B. tulipae 加以區分，且在以 PCR 增幅後可在 1000 bp 及 1200 bp 左右發現另一條帶的產生，但其意義及與 ITS 區域是否有關仍須進一步的研究。而分析其蛋白質電泳可發現種間的差異性，較種內的差異性為高且可在 26 kDa 處發現僅 B. elliptica 具有之蛋白質。然其功能及意義仍須進一步研究。
Many plants could cause disease by Botrytis sp. At low temperature and high humidity. It usually formed white colony, black irregular sclerotia and gray conidia spores on PDA plate. Botrytis sp. had better growth rate when cultured at acidic medium than at basic medium. When cultured at 2% PDA plate and 10% V8 juice medium there was no relationship for sporulation, but more conidia spores could be get when cultured at 125 ml flask than cultured at 9 cm petri dish ether on 2% PDA plate or 10% V8 juice medium. Sclerotia formed conidia spores after germination at 20 to 24 ℃, and the germination rate increased till 28 ℃ then decreased. The wind direction was highly related with the disease progress of B. elliptica when investigated the lily field at Shengan. The conidia spores might be the main organism for secondary inoculums was suggested. Both conidia spores of B. cinerea and B. elliptica in air could be detected by selective medium, but did not have any reaction on bulb form Netherlands. The concentration of 103 conidia/ml spores could be detected on the lily leaf by the specific primer 66B03R/66B03F when test collected isolate of B. elliptica, but the specific band did not amplified by all tested isolates. B. cinerea, B. elliptica, B. gladiolorum and B. tulipae could be identical by using PCR-RFLP in ITS region. Two bands about 1000 bp and 1200 bp also cloud be found after PCR amplification, but the meaning and relationship between ITS region was unknown. The protein pattern was found little difference on interspecies than intraspecies. A protein pattern at 26 kDa was found only in B. elliptica, but the function and meaning need for further research.
|Appears in Collections:||植物病理學系|
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