CAJ | 학술논문

绿木霉(Trichoderma virens)是一种重要菌寄生真菌，该菌已广泛应用于多种病害的生物防治上。本研究构建了双元载体pCATPH-I(GenBank登录号为：KC252999)，并利用该载体成功实现了农杆菌介导的绿木霉遗传转化，转化效率可达385~460个转化子/106绿木霉分生孢子。转化子的PCR和遗传稳定性分析表明， T-DNA已整合进该菌的基因组中且在无选择压力的条件下能够稳定遗传。利用该转化体系成功建立了超过2000个转化子的转化子库，通过转化子与病原真菌立枯丝核菌(Rhizoctonia solani)对峙培养在该转化子库中成功筛选到两个重寄生缺陷突变体，D441和A281。本研究的完成可为绿木霉的功能基因组学及重寄生分子机制的研究打下基础。
록목매(Trichoderma virens)시일충중요균기생진균，해균이엄범응용우다충병해적생물방치상。본연구구건료쌍원재체pCATPH-I(GenBank등록호위：KC252999)，병이용해재체성공실현료농간균개도적록목매유전전화，전화효솔가체385~460개전화자/106록목매분생포자。전화자적PCR화유전은정성분석표명， T-DNA이정합진해균적기인조중차재무선택압력적조건하능구은정유전。이용해전화체계성공건립료초과2000개전화자적전화자고，통과전화자여병원진균립고사핵균(Rhizoctonia solani)대치배양재해전화자고중성공사선도량개중기생결함돌변체，D441화A281。본연구적완성가위록목매적공능기인조학급중기생분자궤제적연구타하기출。
Trichoderma virens, which has been extensively used as a biocontrol agent against lots of plant diseases for many years, was an important mycoparasite. In this research, a binary vector, pCATPH-I, was constructed and submitted in GenBank under accession number KC252999. Trichoderma virens was successfully transformed for random integration of transforming DNA (T-DNA) with a high transformation efficiency of 385~460 transformants/106 conidia by co-culturing with Agrobacterium tumefaciens AGL-1 which carries pCATPH-1. PCR and genetic stability analysis of the transformants showed that T-DNA integrated into the genome and can be stably inherited without selective pressure. A T-DNA insertion library of more than 2 000 transformants was constructed by this transformation system. By confronting cultivation with Rhizoctonia solani, two mutants defective in hyperpara-sitism, designated as D441 and A281, were screened out from that T-DNA insertion library. This work can lay a foundation for researching on the functional genome and hyperparasitism mechanism of Trichoderma virens.