功能性微生物製劑應用於甘藍栽培與黑腐病防治之評估

The Evaluated of Functional Microbial Agents Application for the Cabbage Cultivation and Black Rot Disease Control

郭建志(Chien-Chih Kuo) ； 莊天心(Tian-Hsin Chuang) ； 陳俊位(Chien-Wei Chen) ； 廖君達(Chung-Ta Liao)

《臺中區農業改良場研究彙報》 137期 (2017/12) Pp. 37-57

摘要

本研究篩選促進生長與對病害發生具保護效果之微生物菌株，並導入甘藍苗期與田間施用方法，結合病蟲害發生時機資料，建立甘藍病害綜合管理策略。本試驗自南投縣魚池鄉及霧社鄉的土壤樣本中，篩選出30株具耐熱性之微生物菌株，經由溶血測試後，選取17株不溶血的微生物菌株，進行後續的溶磷活性分析、4種分解酵素能力測定，結果選擇溶磷活性達500 (μg/ml/day)以上的5株菌株進行5種甘藍病原菌的抗菌圖譜測定，其結果以TCB237對甘藍黃葉病、甘藍苗立枯病Rhizoctonia solani、甘藍黑腐病與甘藍軟腐病的拮抗測試最為優異，其次為TCB207，而TCB203、TCB263與TCB298則無明顯的拮抗效果。在菌種鑑定部分，TCB203為巨大芽孢桿菌Bacillus megaterium、TCB207為Bacillus velezensis、TCB237與TCB263為Bacillus sp. (尚未確認其種名)，TCB298則是鑑定為類芽孢桿菌(臺中種) Paenibacillus taichungensis。經由搖瓶培養的試驗結果，所有菌株以LB broth培養菌量均可達10^9 cfu/ml以上，選定LB broth為基礎發酵培養基。應用此5菌株經LB broth發酵72 hr後，進行甘藍台中2號盆栽與溫室之根部澆灌試驗，盆栽處理的結果，經6次澆灌後，以TCB237處理最佳，其葉片數、株高及根長相較對照組具有顯著差異。此外，在溫室栽培的處理中，仍以TCB237處理之單球重最高。其後應用本場原有的液化澱粉芽孢桿菌微生物製劑Tcba05、TCB102-B7與TCB9407，進行甘藍台中2號黑腐病帶菌種子處理，檢測處理後之黑腐病帶菌率，結果以Tcba05、TCB102-B7處理後的甘藍種子無檢出黑腐病產生，顯示種子預先浸泡於微生物製劑中，可以降低種子帶菌風險及苗期病害的發生。此3株微生物製劑，施用於夏季甘藍台中2號的試驗田中，經根部澆灌與葉部噴施共7次，以TCB102-B7處理之黑腐病罹病度最低，僅6.25%，而Tcba05處理9.17%，對照組則達45%，顯示預先澆灌與噴施可降低甘藍黑腐病的發生。本研究已篩選具有耐熱、溶磷與抗黑腐病之微生物菌株TCB237，具發展微生物農藥之潛力，可融入甘藍整合性病害管理中，成為友善耕作的植物保護資材。

關鍵字

甘藍 ； 功能性微生物 ； 芽孢桿菌

並列摘要

The aim of this study are to screen and introduce growth-promoting, disease protecting microbial agents for cabbage integrated crop and diseases management. In this study, we isolated and screened about 30 microbial strains from the two soil samples in Lianhuachih and Wushe area. According to the blood agar test, phosphate-solubilizing analysis and 4 kinds of enzyme decomposition analysis, five strains were selected for further study. The five strains were selected based on the phosphate solubilizing activity that was 500 μg/ml/day at least, the antagonistic spectrum against 5 types of pathogen fungi and bacteria of cabbage. The result show that TCB237 could inhibit 4 kinds of pathogens. Based on the sequences of 16S rRNA and gyrB genes comparison with GenBank database. Strain TCB203 was identified as Bacillus megaterium, TCB207 as Bacillus velezensis, TCB237 and TCB263 as Bacillus spp. TCB298 as Paenibacillus taichungensis. The cell density of each strain cultured in LB medium could reach 10^9 cfu/ml, thus LB medium was selected for microbials fermentation.The five strains were applied on cabbage for six times in field trials and the leaves number, plant height and root length were recorded. We founded that the growth of cabbage with treatment of strain TCB237 was better than other treatments. In addition, we applied the 3 Bacillus sp. microbial agents Tcba05, TCB102-B7 and TCB9407 on seeds inoculated with X. campestris pv. campestris, the results showed that no black rot pathogen was detected with the treatment of strain Tcba05 and TCB102-B7, suggesting that soaking the seeds with microbial agents can reduce the black rot disease occurance in seedling stage. Application 3 Bacillus sp. microbial agents Tcba05, TCB102-B7 and TCB9407 on cabbage Taichung No.2 in the field trial in summer. The disease severity of black rot with treatment of strain TCB102-B7 by root irrigation and leaves spraying was 6.25%, while the treatment with Tcba05 and water as the control were 9.17% and 45%, respective. It showed pre-irrigation and spraying could inhibited the black rot disease severity effectually. TCB237 which had phosphate-solubilizing characteristic had the potential to development the microbial pesticides, which would integrated into cabbage cultivation system, to be the plant protection materials which environment friendly.

並列關鍵字

cabbage ； microbial agent ； Bacillus sp