地衣芽孢杆菌C和L-3菌株生物活性的初步研究
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摘要
通过常规微生物形态学观察和生理生化特性鉴定,结果表明菌株C和L-3在分类学上均属于地衣芽孢杆菌。经对两菌株摇瓶发酵液抑菌活性初步测定,表明其对玉米小斑病菌(Bipolaris maydis)、小麦赤霉病菌(Fusarium graminearum)、莴苣菌核病菌(Sclerotinia sclerotiorum)等11种植物原真菌具有较强的抑制作用,而对根癌土杆菌(Agrobacterium tumefaciens)、花生青枯病菌(Pseudomonassolanacearum)等6种病原细菌基本无抑制活性;以玉米小斑病菌为指示菌,对两菌株发酵液的除菌上清和菌体,分别进行了活性检测试验,结果表明,活菌体有抑菌活性,且抑菌活性物质均在发酵培养的过程中产生,该活性物质可被60%固体(NH4)_2 SO_4沉淀,初步确定此类活性物质为蛋白质。
以小麦、玉米、白菜的种子为试验材料,在室内测定两个菌株发酵液对三种作物种子萌发的影响试验,结果表明:两菌株的除菌上清液中含有可刺激作物种子萌发的活性物质,且对所试的禾本科作物种子促进萌发的效果较好,而对白菜种子则不明显。试验确定了此活性物质并非培养基中的营养组分,而是在发酵培养的过程中,由菌体分泌产生的代谢产物。通过尾穗苋黄化苗子叶苋红合成法测定,此类活性物质可能是细胞分裂素。
通过小麦盆栽和玉米田间试验,分别测定了菌株C和L-3的发酵液对作物生长的刺激作用,在所设置的三个处理,即:喷叶、灌根和喷叶+灌根处理中,喷叶+灌根处理的效果最好,可以在一定程度上提高小麦和玉米的产量。
According to the morphological,physiological and biochemical features, bacterium C and L-3 are identified as Bacillus Iicheniformis.The two strains can control 11 plant diseases which caused funguns,such as Fusarium graminearum,Bipolaris maydis and Sclerotinia sclerotiorum,by mensurating the antimicrobial activity of zymotic fluid,but there is no obvious activty to the 6 bacterium in the test.Trough the determination of biological activity,supernatant and thalli of two strains restrain Bipolaris maydis which is as a direction.It shows that antimicrobial substance is prouduced at fermenting,and the substance is confirmed as a protein primarily,because it can be deposited by 60%(NH4)_2 SO_4.
Determining the hormesis of zymotic fluid of the two stains to three plant seeds, which indicate activity substance in supernatant to stimulate plant growth,and the effect of wheat and corn is much better than that of cabbage.The results show that the substance is not component of culture medium,but the metabolite excreted by the stains. Trough synthesizing Amaranthin by yellow seedling of Amaranthus Gandatus L.,it may be a Cytomin.
According to potted plant of wheat and field trial of corn with different treatments, the result of spraying zymotic fluid over leaves and irrigating it in root synchronously demonstrated that the output of wheat and corn can be improved.
以小麦、玉米、白菜的种子为试验材料,在室内测定两个菌株发酵液对三种作物种子萌发的影响试验,结果表明:两菌株的除菌上清液中含有可刺激作物种子萌发的活性物质,且对所试的禾本科作物种子促进萌发的效果较好,而对白菜种子则不明显。试验确定了此活性物质并非培养基中的营养组分,而是在发酵培养的过程中,由菌体分泌产生的代谢产物。通过尾穗苋黄化苗子叶苋红合成法测定,此类活性物质可能是细胞分裂素。
通过小麦盆栽和玉米田间试验,分别测定了菌株C和L-3的发酵液对作物生长的刺激作用,在所设置的三个处理,即:喷叶、灌根和喷叶+灌根处理中,喷叶+灌根处理的效果最好,可以在一定程度上提高小麦和玉米的产量。
According to the morphological,physiological and biochemical features, bacterium C and L-3 are identified as Bacillus Iicheniformis.The two strains can control 11 plant diseases which caused funguns,such as Fusarium graminearum,Bipolaris maydis and Sclerotinia sclerotiorum,by mensurating the antimicrobial activity of zymotic fluid,but there is no obvious activty to the 6 bacterium in the test.Trough the determination of biological activity,supernatant and thalli of two strains restrain Bipolaris maydis which is as a direction.It shows that antimicrobial substance is prouduced at fermenting,and the substance is confirmed as a protein primarily,because it can be deposited by 60%(NH4)_2 SO_4.
Determining the hormesis of zymotic fluid of the two stains to three plant seeds, which indicate activity substance in supernatant to stimulate plant growth,and the effect of wheat and corn is much better than that of cabbage.The results show that the substance is not component of culture medium,but the metabolite excreted by the stains. Trough synthesizing Amaranthin by yellow seedling of Amaranthus Gandatus L.,it may be a Cytomin.
According to potted plant of wheat and field trial of corn with different treatments, the result of spraying zymotic fluid over leaves and irrigating it in root synchronously demonstrated that the output of wheat and corn can be improved.
引文
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18.刘波,刘文斌,王恬.地衣芽孢杆菌在异育银鲫日粮中的应用.湛江海洋大学学报,2005,25(6):31-35
19.刘文斌等.3种益生素配伍对异育银鲫生长、消化及肠道菌群组成的影响.海洋与湖沼,2007,38(1):29-35
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2.曹煜成,李卓佳,冯娟,陈永青,吴灶和.地衣芽孢杆菌胞外产物消化活性的研究.热带海洋学报,2005,24(6):6-11
3.陈中义,张杰,黄大昉.植物病害生防芽孢杆菌抗菌机制与遗传改良研究.植物病理学报,2003,33(2):97-103
4.陈志谊.拮抗细菌B.916防治水稻纹枯病作用机制的研究.[硕士学位论文].南京:南京农业大学,1998
5.丁海涛,李顺鹏,沈标,崔中利.拟除虫菊酯类农药残留降解菌的筛选及其生理特性研究.土壤学报,2003,40(1):123-129
6.东秀珠,蔡妙英等.常见细菌系统鉴定手册.北京:2001,
7.方敦煌,李天飞,沐应祥,周黎,杨硕媛,陆庆华.拮抗细菌GP13防治烟草黑胫病的田间应用.云南农业大学学报,2003,18(1):49-52
8.范青,田世平,刘海波.两种拮抗菌α-1,3-葡萄糖酶和几丁质酶的产生及其抑菌的可能机理.科学通报,2001,46(20):1713-1717
9.郭贵海,王崇文.肠道菌群调节剂的研究进展.临床内科杂志,2002,19(2):88-90
10.顾真荣,马承铸,韩长安.产几丁质酶芽孢杆菌对病原真菌的抑制作用.上海农业学报,2001,17(4):88-92
11.郝华昆,贾洁,韩俊华,牛天贵.产抗菌蛋白芽孢杆菌R21—4的鉴定及其抗菌谱研究.食品与发酵工业,2006,32(10):54-58
12.何李黎.地衣芽孢杆菌ZJU12所产细菌素的分离纯化及其理化性质和基因克隆的研究.[硕士学位论文].杭州:浙江大学图书馆,2006
13.胡洪波,梁洁,刘月英,傅锦坤.微生物吸附贵金属的研究与应用.微生物学通报,2002,29(3):94-97
14.纪兆林,童蕴惠,凌筝,陈夕军,徐敬友.拮抗细菌对部分水果产后病原真菌的抑制及防腐作用.南京农专学报,2003,19(4):23-27
15.纪兆林,唐丽娟,张清霞,徐敬友,陈夕军,童蕴惠.地衣芽孢杆菌W10抗菌蛋白的分离纯化及其理化性质研究.植物病理学报,2007,37(3):260-264
16.李孱,白景华.细胞分裂素及其产生菌研究概况.东北农业大学学报,1995,26(3):305-310
17.林东,徐庆,刘忆舟,魏军明,瞿礼嘉,顾红雅,陈章良.枯草芽孢杆菌SO113分泌蛋白的抑菌作用及抗菌蛋白的分离纯化.农业生物技术学报,2001,9(1):77-80
18.刘波,刘文斌,王恬.地衣芽孢杆菌在异育银鲫日粮中的应用.湛江海洋大学学报,2005,25(6):31-35
19.刘文斌等.3种益生素配伍对异育银鲫生长、消化及肠道菌群组成的影响.海洋与湖沼,2007,38(1):29-35
20.蒙显英,黎起秦,冯家勋,叶云峰,段承杰.芽孢杆菌产生的抗菌物质的研究进展.中国植保导刊,2004,12:13-15
21.梅汝鸿.增产菌.技术与市场,1997,7(1):15-16
22.卜祥斌,陈洁,刘红艳,罗有文,周岩民.沸石粉、寡糖及益生素在黄鸡饲料中的应用效果研究,家畜生态学报.2006,27(1):37-40
23.彭化贤,刘波微,陈小娟,刘海艳.水稻稻瘟病拮抗细菌的筛选与防治初探.中国生物防治,2002,18(1):25-27
24.齐东梅,梁启美,惠明,牛天贵.棉花枯萎、黄萎病拮抗芽孢杆菌的抗菌蛋白特性.微生物学通报,2005,32(4):42-46
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