| 摘要： 紅龍果為國內近年來新興的果樹產業，栽培面積逐年增加，易衍生病蟲害的問題。 影響紅龍果的產量及果品經濟價值的病害包含紅龍果炭疽病(Anthracnose)、紅龍果莖潰 瘍病(Stem canker)，以及近年新發生的紅龍果濕腐病(Wet rot)等，但目前僅只有炭疽病 有推薦化學藥劑可供使用，其餘病害仍無有效的藥劑使用。因此，本場致力於開發有益微生物製劑，期望可融入紅龍果栽培體系中，藉以降低病害的威脅與延長果實的貯存時間。本場自轄內紅龍果栽培田土壤中，篩選數十株微生物菌種，經由分解酵素測試與抗菌活性分析，其中3株微生物菌株具有澱粉、纖維素、脂肪及蛋白質分解酵素活性，對於紅龍果炭疽病、莖潰瘍病與濕腐病之真菌病原具有抑制其菌絲生長之功效。此外，利用PCR技術可以自3菌株之核酸中，增幅出伊枯草菌素A(Iturin-A)、表面活性素(Surfactin)之專一性DNA片段，顯示此3菌株具有產生抗生物質之能力。經由Biolog system細菌自動鑑定系統、16S rRNA與gyrB之序列分析後，鑑定為液化澱粉芽孢桿菌Bacillus amyloliquefaciens。此3菌株經由少量400ml搖瓶、10公升醱酵量產試驗，所產製的液態製劑之菌量可穩定達到108 109cfu/ml。經由果實浸泡3菌株醱酵液10分鐘，後續接種炭疽病及濕腐病，Tcb45處理之發病率為最低，相較對照組具有延緩發病之效果。紅龍果濕腐病先期田間試驗中，於開花期、花瓣摘除期與果實套袋前，各噴施1次微生物製劑，結果以Tcb45菌株處理之果實腐敗病率為最低，僅2.38%，對照組不摘花處理則已達13.34%，具有顯著差異。因此，Tcb45菌株可作為後續開發微生物製劑的潛力菌株。Pitaya was the emerging fruit tree industry recently years in Taiwan, the cultivated area expensive was increasing recently, the pest would be accompanied. Among them, the Pitaya anthracnose, stem canker and wet rot diseases would cause the Dragon fruit yield loss and low quality. There are only some fungicides for control anthracnose. Therefore, we devoted to develop and research the beneficial microorganisms agent to reduce losses caused by the diseases and extension the fruit storage time, it is expect to integrate the dragon fruit cultivation system. We isolated and screen dozens microorganism strains from soil in dragon fruit field. The 3 strains all have the enzyme activities of amylase, protease, cellulase and lipase. In addition, examination the antagonistic efficacy of the mycelium growth of Colletotrichum gloeosporioides, Neoscytalidium dimidiatum and Gibertella persicaria in vitro in dual cultures on PDA. The Iturin-A, Surfactin antibiotic specific DNA fragment could be amplified by PCR technique. The result showed that 3 stranis can produce the antibiotics. Use the Biolog automatic identification system and 16s rRNA and gyrB gene identify 3 strains as Bacillus amyloliquefaciens. Through the 400ml shake flask, 10L fermentation broths, the total endospores count could reach to 108 109 cfu/ml. We soaking the dragon fruit into the 3 Bacillus spp. fermentation broths 100 times for 10 minutes, follow-up inoculation the mycelium blocks of anthracnose and wet rot. The resulted showed that Tcb45 treatment have the lowest disease incidence. Compared to control treatment, the Tcb45 strains would have the efficacy to delay the pathogenesis. Early field trial of Pitaya wet rot, we spray the 3 Bacillus agents 3 times in flowering stage, petals removal and before bagging time, respectively. The resulted show that the lowest fruit decay ratio were 2.38% by used Tcb45 treatment, the control without remove petals were reach 13.34% with a significant difference. Therefore, the Tcb45 strain would have potential to develop microorganism agent. |
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