极端嗜热多肽的纯化、性质初探及对苗瘟的防效  

Purification of the extreme thermophilic polypeptides,their characterization and control effect on Magnaporthe grisea at seedling stage

作　　者：周华强[1] 谭芙蓉[1] 周颖[1] 郑爱萍[1] 李平[1] 

机构地区：[1]四川省农业生物技术工程研究中心四川农业大学水稻研究所,四川温江611130

出　　处：《西南农业学报》2007年第6期1211-1216,共6页Southwest China Journal of Agricultural Sciences

基　　金：国家"863"高技术研究发展计划项目(2002AA212151)

摘　　要：本试验用加热法从多粘类芽孢杆菌（Paenibacillus polymyxa）LM-3菌株的发酵液中纯化得到2个极端嗜热多肽。平板拮抗实验表明，5μl纯化多肽对稻瘟病菌（Magnaporthe grisea）抑菌率达89．6％。SDS—PAGE电泳显示纯化多肽分子量介于6000～7000Da之间。多肽复性后，其中之一对稻瘟病菌表现出强的拮抗活性，命名为APPLM3（Antagonism Polypeptidefrom Paenibacilluspolymyxa LM-3）；另一个则无此活性，命名为PPLM3（Polypeptide from Paenibacillus polymyxa LM-3）。APPLM3经氨基酸测序，获得了其N-末端5个氨基酸序列（H2N—ANDPR）；以该序列为靶序列在NCBI上进行相似性检索，发现其可能与3个假设蛋白（或推导蛋白）相关。以APPLM3为主效成分配得5种微生物源生防乳剂，对水稻苗瘟进行了田间防效试验。试验结果表明，这5种乳剂的防效都优于LM-3拮抗液直接喷雾，其中AB4处理的防效最佳，达80．73％，其拮抗成分、吐温80、乳化剂OP、溶剂的质量比为50：6．5：1．75：41．75。APPLM3为首次报道的兼具极端嗜热性和稻瘟病菌拮抗活性的小分子多肽。Two extreme thermophilic polypeptides were isolated from fermentation liquor of strain LM-3 which belonged to Paenibacillus polymyxa. The inhibition capacity of 5 μl purified polypeptides against Magnaporthe grisea reached 89.6 % according to results on agar plates. These two polypeptides had a molecular weight between 6 000 and 7 000 Da by SDS-PAGE. They exhibited extreme thermophilic characteristic and were recovered from electro-lanes for detection of inhibition activity to M. grisea respectively. One polypeptide with strong inhibition activity to M. grisea ,was named APPLM3 （ Antagonism Polypeptide from Paenibacillus polymyxa LM-3）, and the other without such activity was named PPLM3 （ Polypeptide from Paenibacillus polymyxa LM-3 ）. N-terminal amino acid residues of APPLM3 were then sequenced and the sequence was H2N-ANDPR by which restriction similarity comparison of amino acid sequence was conducted on NCBI Website. Five microbial pesticides （emulsion） were confected with polypeptide APPLM3. Their control effect against rice blast disease was conducted in paddy field at seedling stage. The results indicated that the control effect of the 5 microbial pesticides was superior to the fermentation liquor of LM-3 bacterium. Of the 5 treatments in the present study,AB4 showed the strongest control effect which reached 80.73 % ,and the quality fraction of its components was 50： 6.5： 1.75： 41.75. APPLM3 polypeptide was the first one being reported ,which combined the extreme thermophilic characteristic and strong antifungal activity against M. grisea.

关 键 词：多粘类芽孢杆菌 稻瘟病菌 极端嗜热多肽 拮抗作用 防效 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]