Development of Dignoses of Entomopathogenic Fungi and Other Organisms Using Molecular Biological Techniques

在植物防疫中以蟲生真菌, 例如綠殭菌( Nomuraea rileyi )或其他真菌可感染多種經濟作物害蟲.為評估這些微生物在田間防治害蟲之效果及偵測其在自然環境生態之分布, 開發一個快速診斷菌種及田間追蹤的有效方法為首要工作.利用分生技術及特定專一性核酸探針, 除了在流行病學的研究上可用來診斷及鑑定田間與外來菌種, 以及進行田間實際偵測特定菌株之效益之外, 對於菌種遺傳演化上的研究亦有重要的參考價值.本計畫擬調查台灣地區蟲生真菌的發生分布與菌種收集, 並從中篩選致病力高的菌株以供害蟲防治之應用.採用分生技術建立菌種DNA圖譜與專一的核酸探針開發, 為重要之研究項目, 此技術開發後尚可供作一個模式應用於其他外侵生物或害蟲種類之診斷用.The entomopathogenic fungi, for example, Nomuraea rileyi, and other pathogens are infectious to many economic crops and therefore are useful for plant protection.In order to assess and monitor these pathogenic microorganisms in nature and fields, it is important to develop rapid and precise diagnostic techniques for their identification as well as distribution in the field condition.Molecular techniques and specific nucleic acid probes are useful for microbial genetics and evolution in addition to studies on species diagnoses and identification in fields with regard to microbial epidemics.The present proposal is to investigate occurrence and distribution of the entomopathogenic fungi in Taiwan, and to screen highly virulent pathogens for pest control.Furthermore, one of the main research items will be to establish DNA profiles and to find out nucleic acid probes, which are specific to entomopathogenic fungi.In addition, the developed technology in this proposal could be a model for diagnosis of the invading pests or other insects

Development of Microbial Control of Insect Pests of Vegetables under Structure

本計畫擬用本省分離純化之病原微生物包括黑殭菌( Metarhizium anisopliae )、白殭菌( Beauveria bassiana )、綠殭菌( Nomuraea rileyi )、蜡蚧輪枝孢菌( Verticillium lecanii )及昆蟲寄生性線蟲( Steinernema abbasi ), 針對設施有機栽培常發生害蟲如: 小菜蛾( Plutella xylostella )、斜紋夜盜蛾( Spodoptera litura )、蚜蟲( Aphis gossypii )、粉蝨、黃條葉蚤( Phyllotreta striolata )等之幼蟲及成蟲, 藉由土壤之施用處理及葉表之施用處理進行防治技術之開發試驗, 及作物栽培防治流程之比較及效果評估.In Taiwan, crucifer our crops and other leafy vegetables are major vegetables cultivated under structure.The insects such as Phyllotreta striolata, Plutella xylostella, Spodoptera litura and Spodoptera exigua were considered economic important pests under structure.Integrated pests management ( IPM )by using multiple control measures is encouraged to adopt for solving the problems of insect pests.Microbial control plays an important role in the IPM system.The entomopathogens including Metarhizium anisopliae, Beauveria bassiana, Nomuraea rileyi, Verticillium lecanii and Steinernema abbasi are useful as microbial agents.In this project, an integrated control approach by combing soil treatments and foliage spraying with these microbial insecticides will be conduced to control insect pests under structure, and evaluation of the entomopathogens against those pests will be performed

Research and Development on Bioinsecticides Using Indigenous Entomopathogenic Nematodes (II)

本計畫主要研究目的在於利用本土產, 具有可發展為生物農藥製劑潛力的蟲生線蟲Steinernema abbasi, 以群體合作方式研發量產、配方與劑型之技術, 針對本省作物之重要害蟲, 進行蟲生線蟲適用之寄主範圍評估, 及感染溫度、溼度、長期儲存環境等對蟲生線蟲影響之生物特性加以測試.並利用農業加工生產之有機廢棄物, 開發本地產蟲生線蟲( S.abbasi )之大量生產技術, 及不同基質量產產品對斜紋夜盜( Spodoptera litura )幼蟲及其他害蟲殺蟲效力之生物檢定, 以確定產品管制.另外開發感染期三齡幼蟲( IJs )線蟲之分離方法, 回收保存技術.同時將進行土壤環境因子對線蟲之活動行為及對殺蟲效力之影響.並在設施栽培耕作環境下, 開發最適合用線蟲防治害蟲的操作方法, 提供有機栽培農民安全防治害蟲之用, 期能提高農產品之價值.The present proposal is to study the indigenous entomopathogenic nematode, Steinernema abbasi, to be a potential bioinsecticide.Technology for mass production and formulation of this nematode-based bioinsecticide will be developed under the collaboration between institution and industrial sector.Host range, influence of temperature, humidity, and storage on biological nature of the nematode will be conducted.Mass production of S.abbasi will be studied using organic wastes of agricultural production.Bioassays on efficacy of the products produced from different substrates against Spodoptera litura or other insect pests will be carried out for their quality control.In addition, techniques for isolating, recovering and storing the infective juveniles ( IJs )of nematodes will be developed.Meanwhile, effects of soil factors on nematode activity, behaviour, and insecticidal ability will be investigated.The optimal and effective techniques for applying the nematode-based bioinsecticides to control insects under the screen house or tunnel cultivation of highly economic crops will also be developed.This biological approach is expected to provide farmers with a safe and effective pest control measure as well as to raise the value of agro-products in the future

Research and Development on Bioinsecticides Using Indigenous Entopathogenic Nematodes

本計畫主要研究目的在於利用本土產,具有可發展為生物農藥製劑潛力的蟲生線蟲Steinernema abbasi,以群體合作方式研發量產、配方與劑型之技術,針對本省作物之重要害蟲,進行蟲生線蟲適用之寄主範圍評估,及感染溫度、溼度、長期儲存環境等對蟲生線蟲影響之生物特性加以測試.並利用統全公司在加工生產之有機廢棄物,開發本地產蟲生線蟲(S. abbasi)之大量生產技術,及不同基質量產產品對斜紋夜盜(Spodoptera litura)幼蟲及其他害蟲殺蟲效力之生物檢定,以確定產品管制.另外開發感染期三齡幼蟲(IJs)線蟲之分離方法,回收保存技術.同時將進行土壤環境因子對線蟲之活動行為及對殺蟲效力之影響.並在設施栽培耕作環境下,開發最適合用線蟲防治害蟲的操作方法,提供有機栽培農民安全防治害蟲之用,期能提高農產品之價值.The present proposal is to study the indigenous entomopathogenic nematode, Steinernema abbasi, to be a potential bioinsecticide. Technology for mass production and formulation of this nematode-based bioinsecticide will be developed under the collaboration between institution and industrial sector. Host range, influence of temperature, humidity, and storage on biological nature of the nematode will be conducted. Mass production of S. abbasi will be studied using organic wastes of the products from Tung-cheng Company. Bioassays on efficacy of the products produced from different substrates against Spodoptera litura or other insect pests will be carried out for their quality control. In addition, techniques for isolating, recovering and storing the infective juveniles (IJs) of nematodes will be developed. Meanwhile, effects of soil factors on nematode activity, behaviour, and insecticidal ability will be investigated. The optimal and effective techniques for applying the nematode-based bioinsecticides to control insects under the screen house or tunnel cultivation of highly economic crops will also be developed. This biological approach is expected to provide farmers with a safe and effective pest control measure as well as to raise the value of agro-products in the future