阿维链霉菌NRRL8165的摇瓶发酵及其与苏云金芽胞杆菌制剂的协同作用
|
摘要
本论文取得了以下几方面成果：
1．筛选了薄层层析法分离阿维菌素的展层剂体系。当展层剂体系为：二氯甲烷：乙酸乙酯：甲醇=30：3：1时，能有效分离阿维菌素B1a和B1b组份。通过正交设计，对阿维链霉菌(Streptomyces avermitilis)菌株NRRL8165的摇瓶培养基进行了筛选，改良培养基的发酵效价是对照培养基的1.87倍。同时，研究了菌株NRRL8165发酵过程中的代谢变化。
2．建立了用高效液相色谱法(二极管阵列检测器)分离、测定阿维菌素的方法。定量测定中，回收率达到98.8％和99.4％，相对标准偏差为0.1％和0.3％。结果表明：该方法具有良好的精确度和重现性。
3．以小菜蛾、棉铃虫和甜菜夜蛾为供测昆虫，测定了阿维菌素与苏云金芽胞杆菌的协同作用。当阿维菌素与苏云金芽胞杆菌的质量比分别为1：500～1：1500、1：500～1：1500和1：10000～1：20000时，对小菜蛾、棉铃虫和甜菜夜蛾都表现出增效作用。
4．将阿维菌素与苏云金芽胞杆菌混合进行喷雾干燥，喷雾干燥产品对小菜蛾，棉铃虫和甜菜夜蛾均表现出相加作用，说明喷雾干燥产品保持了对上述昆虫的毒力。通过初筛、复筛分别测定了AU-a、AU-b、AU-C、AU-d、AU-e五种抗紫外剂对阿维菌素B1组份的保护作用，其中抗紫外剂AU-b效果最好。将抗紫外剂AU-b以1：50(抗紫外剂与阿维菌素的质量比)加入到喷雾干燥产品中。在紫外灯下照射2小时后，与不含抗紫外剂的喷雾干燥产品相比，含抗紫外剂AU-b的喷雾干燥产品中阿维菌素B1组份的损失降低了13.9％；含抗紫外剂AU-b的喷雾干燥产品的毒力损失降低了12.6％。
The solvent system of thin-layer chromatography was screened. Avermectins can be isolated in the developing solvent chloroform: ethylacetate: methanol (30:3:1). By orthogonal test, the fermentation media of Streptomyces avermitilis NRRL8165 were screened, and the yield of Avermectin Bl component of improved media was 1.87 times as high as that of the control medium. Furthermore, we studied the metabolism of NRRL8165 during the fermentation.
The method of isolation and determination of Avermectins by High Performance Liquid Chromatography with Photodiode Array Detector was examined. The Relative Standard Deviation values were 0.2percent and 0.5percent; the rates of recovery were 98.8% and 100.0%. These showed that the method was precise and reliable.
The synergism of Avermectins and Bacillus thuringiensis to Plutella xylostella and Heliothis armigera in 1:500~1:1500(w/w) were tested. When the proportions were 1:10000~ 1:20000, the mixtures were synergism to Spodoptera exigua.
The mixtures of Avermectins and Bacillus thuringiensis were dried by Spray-Dryer. The powder was available. The results of bioassay showed that the toxicity of this powder remained. Furthermore, effects of five kinds of anti-ultraviolet material (AU-a, AU-b, AU-c, AU-d, and AU-e) on protecting Avermectin Bl components were tested. We found that the material AU-b was superior to the others. We added the material AU-b to the powder in the proportion of 1:50(material AU-b: Avermectin, w/w). After 2 hours' exposure under the ultraviolet lamp, the loss of Avermectins in the powder with material AU-b reduced 13.9% compared with that in the powder without anti-ultraviolet material. In addition, the loss of toxicity in the powder with material AU-b reduced 12.6% in bioassay.
1．筛选了薄层层析法分离阿维菌素的展层剂体系。当展层剂体系为：二氯甲烷：乙酸乙酯：甲醇=30：3：1时，能有效分离阿维菌素B1a和B1b组份。通过正交设计，对阿维链霉菌(Streptomyces avermitilis)菌株NRRL8165的摇瓶培养基进行了筛选，改良培养基的发酵效价是对照培养基的1.87倍。同时，研究了菌株NRRL8165发酵过程中的代谢变化。
2．建立了用高效液相色谱法(二极管阵列检测器)分离、测定阿维菌素的方法。定量测定中，回收率达到98.8％和99.4％，相对标准偏差为0.1％和0.3％。结果表明：该方法具有良好的精确度和重现性。
3．以小菜蛾、棉铃虫和甜菜夜蛾为供测昆虫，测定了阿维菌素与苏云金芽胞杆菌的协同作用。当阿维菌素与苏云金芽胞杆菌的质量比分别为1：500～1：1500、1：500～1：1500和1：10000～1：20000时，对小菜蛾、棉铃虫和甜菜夜蛾都表现出增效作用。
4．将阿维菌素与苏云金芽胞杆菌混合进行喷雾干燥，喷雾干燥产品对小菜蛾，棉铃虫和甜菜夜蛾均表现出相加作用，说明喷雾干燥产品保持了对上述昆虫的毒力。通过初筛、复筛分别测定了AU-a、AU-b、AU-C、AU-d、AU-e五种抗紫外剂对阿维菌素B1组份的保护作用，其中抗紫外剂AU-b效果最好。将抗紫外剂AU-b以1：50(抗紫外剂与阿维菌素的质量比)加入到喷雾干燥产品中。在紫外灯下照射2小时后，与不含抗紫外剂的喷雾干燥产品相比，含抗紫外剂AU-b的喷雾干燥产品中阿维菌素B1组份的损失降低了13.9％；含抗紫外剂AU-b的喷雾干燥产品的毒力损失降低了12.6％。
The solvent system of thin-layer chromatography was screened. Avermectins can be isolated in the developing solvent chloroform: ethylacetate: methanol (30:3:1). By orthogonal test, the fermentation media of Streptomyces avermitilis NRRL8165 were screened, and the yield of Avermectin Bl component of improved media was 1.87 times as high as that of the control medium. Furthermore, we studied the metabolism of NRRL8165 during the fermentation.
The method of isolation and determination of Avermectins by High Performance Liquid Chromatography with Photodiode Array Detector was examined. The Relative Standard Deviation values were 0.2percent and 0.5percent; the rates of recovery were 98.8% and 100.0%. These showed that the method was precise and reliable.
The synergism of Avermectins and Bacillus thuringiensis to Plutella xylostella and Heliothis armigera in 1:500~1:1500(w/w) were tested. When the proportions were 1:10000~ 1:20000, the mixtures were synergism to Spodoptera exigua.
The mixtures of Avermectins and Bacillus thuringiensis were dried by Spray-Dryer. The powder was available. The results of bioassay showed that the toxicity of this powder remained. Furthermore, effects of five kinds of anti-ultraviolet material (AU-a, AU-b, AU-c, AU-d, and AU-e) on protecting Avermectin Bl components were tested. We found that the material AU-b was superior to the others. We added the material AU-b to the powder in the proportion of 1:50(material AU-b: Avermectin, w/w). After 2 hours' exposure under the ultraviolet lamp, the loss of Avermectins in the powder with material AU-b reduced 13.9% compared with that in the powder without anti-ultraviolet material. In addition, the loss of toxicity in the powder with material AU-b reduced 12.6% in bioassay.
引文
1．上海农药所。井冈霉素。上海人民出版社，1977
2．天津轻工业学院，无锡轻工业学院，大连轻工业学院。工业发酵分析。北京轻工业出版社，1980
3．刘明秋。苏云金芽胞杆菌杀虫剂油剂的研究。[硕士论文]。武汉，华中农业大学，1999
4．刘波，林国宪，朱育菁，林抗美，陈皓，苑宝玲。BtA高效生物杀虫剂研制关键技术的研究。福建农业学报，1998，13(增刊1)：90-105
5．刘波，苑宝玲，葛慈斌。高效生物杀虫剂BtA的化学藕合技术的研究。农业科技推广通讯，1999，6：3—11
6．刘勇。苏云金杆菌发酵培养基和可湿性粉剂的研究。【硕士论文】。武汉，华中农业大学，1995
7．刘毅荣。阿维菌素片剂研制报告。青海畜牧兽医杂志，1996，26(4)：17—18
8．华汝麒，高峰，刘向文。液相色谱法光电二极管阵列检测器对双基药中硝化甘油和中定剂定性定量方法的研究。色谱，1988，6(5)：293-295
9．孙宇晖，邓子新。阿弗菌素生物合成基因簇的研究进展。中国抗生素杂志，(在印刷中)
10．朱育菁，林抗美，刘波，刘浩官。BtA高效生物杀虫剂对桃蚜防治效果的研究。福建农业学报，1998，13(增刊1)：140-144
11．何绍江。克虫星对安哥拉改良山羊的驱虫试验。四川畜牧兽医，1995，3：11-12
12．吴泽宜。农药词汇。科学出版社，1981
13．宋渊。一种高效杀虫大环内酯类抗生素一阿佛米丁(Avermectins)的研究。【硕士论文】。北京，北京农业大学，1993
14．张邵狮。除虫链霉菌DNA降解基因簇add的定位和异源表达。[硕士论文]。武汉，华中农业大学，1997
15．张宗炳。杀虫药剂的毒力测定。科学出版社。1988
16．张敏恒。国内农药开发的一些新特点。农药，1999，38(10)：27-30
17．张敏恒。新型高效广谱杀虫杀螨剂阿维菌素。农药，1998，37(3)；36-37
18．张瑞亭等。农药的混用与混剂。化学工业出版社，1987
19．李开国，王龙祥。生化分析仪器理论及应用。上海科学技术文献出版社，1996
20．李光伟，李建明，高永。0.3％阿维菌素微乳剂的研究。农药，1999，38(7)：7-8
21．李林，王兆守，喻子牛。对小菜蛾较高毒力的阿弗曼链霉菌突变株及发酵培养基的筛选。湖北农业科学，1999(1)：31-34
22．杜荣骞。生物统计学。高等教育出版社，1985
23．杨持。生物统计学。内蒙古大学出版社，1987
24．芮昌辉，孟香清，范贤林，赵建国。三种农药新制剂防治抗药性棉铃虫和小菜蛾试验。农药，1999，38(5)：33-34
25．邵赟．胡凤祖，师治贤。高效液相色谱法测定阿弗菌素含量。色谱，1998，16(1)：87-88
26．陈文君。植物化学保护实验技术导论。陕西科学技术出版社，1988
27．陈钧，吴性良，吉建国。二极管陈列检测高效液相色谱重叠峰的双波长分析法。色谱，1997，15(1)：5-7
28．周启，王道本。农用抗生素和微生物杀虫剂。中国农业出版社，1995
29．林抗美，刘波，林国宪，朱育菁，苏若秋，陈皓。BtA高效生物杀虫剂对棉铃虫防治效果的研究。福建农业学报，1998，13(增刊1)：135—139
30．林单坤。色谱法。科学出版社，1982
31．欧阳立明。小菜蛾对苏云金杆菌的抗性调查、抗性筛选及增效因子筛选。[硕士论文]。武汉，华中农业大学，1998
32．河北省三河市外专局，河北省三河市蔬菜局，福建省农科院生物技术中心农业环保技术研究室。生物农药BtA(天霸)防治蔬菜主要害虫田间药效试验。农业科技推广通讯，1999，6：1-2
33．金志华，金一平，宋友礼。Avermectin产生菌异亮氨酸诱导变种的选育。中国抗生素杂志。1997，22(2)；84-286
34．姚汝华。微生物工程工艺原理。华南理工大学出版社，1996
35．姜竹茂。食品添加剂的微胶囊方法及应用。食品工业科技，1998，5：73-75
36．胡润茂。一族新的广谱驱虫抗生素——除虫菌素(Avermectin)及其衍生物双氧除虫菌素。应用微生物，1986，5：1-8
37．凌世海。我国农药加工工业现状和发展建议。农药，1999，38(10)：19-26
38．原昭三，森定雄，花井俊彦。现代色谱分析法—原理和实际应用。邱宗萌，孙琢琏译。科学技术文献出版社，重庆分社，1988
39．唐根源，吴红京。高效液相色谱法光电二极管阵列检测器对苯乙烯羧化反应液的分析和鉴定。色谱，1990，8(1)：62-63
40．徐俊。阿维菌素产生菌DNA新型位点特异性修饰的分子遗传学研究。[硕士论文]。武汉，华中农业大学，1997
41．徐慧斌，沈杰，叶明忠，曹杰，张仓，王权，周金林，何国声。埃维菌素长效控释丸在棉羊体内的药物动力学研究。中国兽医寄生虫病，1996，4(2)：4-7
42．崔云龙，田明，邵宗泽。紫外线使苏云金芽胞杆菌伴胞晶体失活机理的研究。微生物学通报，1993，20(4)：193-195
43．扈洪波。Ivemaectin的制剂研究。中国兽药杂志，1998，32(2)：51-53
44．扈洪波。阿维菌素口服液体药剂的稳定性研究。畜牧兽医学报，1998，29(2)：185-188
45．曹贵明。阿维菌素(Avermectins)高产菌株的诱变育种和发酵条件的研究。[硕士论文】。北京，北京农业大学，1995
46．龚炳永，周亦昌。抗生素的分离与测定。色谱，1992，10(5)：269-277
47．喻子牛。苏云金杆芽胞菌制剂生产及应用。农业出版社，1993
48．喻子牛。苏云金杆菌。北京，科学出版社，1990
49．蒋志坚，马毓龙合编。农药加工从书可湿性粉剂。化学工业出版社，1991
50．慕立义。植物化学保护研究方法。中国农业出版社，1994
51. Chabalo J C, Mrozik H, Tolman R L, Eskola P, Lusi A, Peterson L H, Woods M F, Fisher M H, Campbell W C, Egerton J R and Ostlind D. Ivermectin, a new broad-spectrum antiparasitic agent. J Med. Chem, 1980,23:1134-1136
52. Christopher J D, Stephen P G, Alexander C G, Kelvin S H, Michael S P and John C R. Novel Avermectins produced by mutational biosynthesis. J Antibiot, 1991, 44(3): 357-365
53. Cimburkova E, Zima J, Novak J and Vanek Z. Nitrogen regulation of Avermectins biosynthesis in Streptornyces arvermitilis in a chemically defined medium. JBasic Microbiol, 1988, 28(8): 491-499
54. Dunkle R L and Shasha B S. Response of starch encapsulated Bacillus thuringiensis containing ultraviolet screens to sunlight. Environ Entomol, 1989, 18(6): 1035-1041
55. Edmund W H, Breland W H, Kelvin S, Hordom S, Edward L, Richard G W, Donald B, Hamish A I, Mcasthur and William W J. Branched-chain fatty acid requirement for Avermectin production by a mutant of Steptomyces avermitilis lacking branched-chain 2-oxo acid dehydrogenase activity. J Antibiot, 1991, 44(3): 349-356
56. Eva C,Vendvlka J, Jiri Z and Zdenko V. The effect of inorganic phosphate on the production of Avermectin in Streptomyces avermitilis. J Basic Microbiol, 1989, 29(6), 341-346
57．Fisher M H．Avermectin研究最新进展。农药译丛，1991，3(3)：19-26
58. Goudie A C, Evans N A, Gration K A F, Bishop B F, Gibson S P, Holdom K S, Kaye B, Wicks S R, Lewis D, Weatherley A J, Bruce C
I, Herbert A and Seymour D J. Doramectin-a potent novel endectocide. Veter Para, 1993, 49:5-15
59. Griego V M and Spence K D. Inactivation of Bacillus thuringiensis spores by ultraviolet and the visible light. Apple Environ Entomol, 1978, 35:906-910
60. Hafner, Edmund W H, Kelvin S, Lee, Edward S. Process for production of Avermectins. U S Patent 323, 198, 1994, Oct 14
61. Herbert B S, Marius R and Kuo S L. Microencapsulation of pesticides by interfacial polymerization utilizing isocyanate or aminoplast chemistry. Pestic Sci, 1998. 54:394-400
62. Hopwood D A. Understanding genetic control of antibiotic biosynthesis and sporulation in Streptomyces. Pro R Soc, B235:121-138
63. Ikeda H and Omura S. Avermectin biosynthesis. Chem Rev, 1997, 97:2591-2609
64. Ikeda H and Omura S. Control of Avermectin biosynthesis in Streptomyces avermitilis for the selective production of a useful component. JAntibiot, 1995, 48(7):549-562
65. Ikeda H and Omura S. Microorganism for selective production of a specific compound of Avermectin and a method for selective production. U S Patent 1998, 861. 1992, Dec. 30
66. Ikeda H, Kotaki H and Satoshi O. Genetic studies of Avermectin biosynthesis in Streptomyces avermitilis. J Bacteriol, 1987, 169:5613-5621
67. Ikeda H, Pang C H, Endo H, Ohta T, Tanaka H and Omura S. Construction of a single component producer from the wild type Avermectins producer Streptomyces avermitilis. J Antibiot, 1995, L08: 532-534
68. Ikeda H, Omura S and Tanaka H. Selective production of specific components of Avermectins in Streptomyces avermitilis. J Antibiot, 1991,
44: 560-563
69. Jan N, Jan K and Zdenko V. Nitrogen source regulates expression of alanine dehydrogenase isoenzymes in Streptomyces avermitilis in a chemically defined medium. Can J Microbiol, 1997. 43(2): 189-193
70. Louis S C, William F F, Byron H A, William J A V, Lawrence F C, Ralph A S, Walter F K and Peter G W. Photodegradtion of Avermectin Bla thin film on Glass. J Agric Food Chem, 1991, 39, 1310-1319
71. Lucille J C. Process for the Separation of Antibiotic Macrolides. United States patent, 4, 160, 083.1979, Jul 3
72. Marvin D S, Delia V, Otto D H, Deborah Z, Mary N, Kaplan L and Ostlind D A. Demethylavermctins Biosynthesis, Isolation and characterization. J Antibiot, 1985, Nov, 1494-1498
73. Marvin D S, Susan L A, Delia L V and Byron H A. purification and identification of dTDP-olean drose, the precursor of the oleandrose units of the Avermectins. J Bio Chem, 1990. 265 (28): 16965-16970
74. Matthews and Frank J. Process for isolation of Avermectin B1 components with improved purity and subsequent isolation of B2 components. U S Patent 931, 839, 1986 Nov. 18
75. Morris O N. Protection of Bacillus thuringiensis from inactivation by sunlight. Can Entomol, 1993, 115: 1215-1217
76. Toms C and Edward S I. Studies on the biosynthesis of Avermectins. Arch Biochem and Biophys, 1989, 270(2); 521-525
77. Toms C, Byron H A, Vincent P G and Edward S I. Further studies on the biosynthesis of Avermectins. J Ind Microbiol, 1989a, 4, 231-238
78. Toms C, Edward S I, Otto D H, Deorah Z and Ostlind D A. Directed biosynthesis of Avermectins. Arch Biochem and Biophys, 1989b, 269(2): 544-547
2．天津轻工业学院，无锡轻工业学院，大连轻工业学院。工业发酵分析。北京轻工业出版社，1980
3．刘明秋。苏云金芽胞杆菌杀虫剂油剂的研究。[硕士论文]。武汉，华中农业大学，1999
4．刘波，林国宪，朱育菁，林抗美，陈皓，苑宝玲。BtA高效生物杀虫剂研制关键技术的研究。福建农业学报，1998，13(增刊1)：90-105
5．刘波，苑宝玲，葛慈斌。高效生物杀虫剂BtA的化学藕合技术的研究。农业科技推广通讯，1999，6：3—11
6．刘勇。苏云金杆菌发酵培养基和可湿性粉剂的研究。【硕士论文】。武汉，华中农业大学，1995
7．刘毅荣。阿维菌素片剂研制报告。青海畜牧兽医杂志，1996，26(4)：17—18
8．华汝麒，高峰，刘向文。液相色谱法光电二极管阵列检测器对双基药中硝化甘油和中定剂定性定量方法的研究。色谱，1988，6(5)：293-295
9．孙宇晖，邓子新。阿弗菌素生物合成基因簇的研究进展。中国抗生素杂志，(在印刷中)
10．朱育菁，林抗美，刘波，刘浩官。BtA高效生物杀虫剂对桃蚜防治效果的研究。福建农业学报，1998，13(增刊1)：140-144
11．何绍江。克虫星对安哥拉改良山羊的驱虫试验。四川畜牧兽医，1995，3：11-12
12．吴泽宜。农药词汇。科学出版社，1981
13．宋渊。一种高效杀虫大环内酯类抗生素一阿佛米丁(Avermectins)的研究。【硕士论文】。北京，北京农业大学，1993
14．张邵狮。除虫链霉菌DNA降解基因簇add的定位和异源表达。[硕士论文]。武汉，华中农业大学，1997
15．张宗炳。杀虫药剂的毒力测定。科学出版社。1988
16．张敏恒。国内农药开发的一些新特点。农药，1999，38(10)：27-30
17．张敏恒。新型高效广谱杀虫杀螨剂阿维菌素。农药，1998，37(3)；36-37
18．张瑞亭等。农药的混用与混剂。化学工业出版社，1987
19．李开国，王龙祥。生化分析仪器理论及应用。上海科学技术文献出版社，1996
20．李光伟，李建明，高永。0.3％阿维菌素微乳剂的研究。农药，1999，38(7)：7-8
21．李林，王兆守，喻子牛。对小菜蛾较高毒力的阿弗曼链霉菌突变株及发酵培养基的筛选。湖北农业科学，1999(1)：31-34
22．杜荣骞。生物统计学。高等教育出版社，1985
23．杨持。生物统计学。内蒙古大学出版社，1987
24．芮昌辉，孟香清，范贤林，赵建国。三种农药新制剂防治抗药性棉铃虫和小菜蛾试验。农药，1999，38(5)：33-34
25．邵赟．胡凤祖，师治贤。高效液相色谱法测定阿弗菌素含量。色谱，1998，16(1)：87-88
26．陈文君。植物化学保护实验技术导论。陕西科学技术出版社，1988
27．陈钧，吴性良，吉建国。二极管陈列检测高效液相色谱重叠峰的双波长分析法。色谱，1997，15(1)：5-7
28．周启，王道本。农用抗生素和微生物杀虫剂。中国农业出版社，1995
29．林抗美，刘波，林国宪，朱育菁，苏若秋，陈皓。BtA高效生物杀虫剂对棉铃虫防治效果的研究。福建农业学报，1998，13(增刊1)：135—139
30．林单坤。色谱法。科学出版社，1982
31．欧阳立明。小菜蛾对苏云金杆菌的抗性调查、抗性筛选及增效因子筛选。[硕士论文]。武汉，华中农业大学，1998
32．河北省三河市外专局，河北省三河市蔬菜局，福建省农科院生物技术中心农业环保技术研究室。生物农药BtA(天霸)防治蔬菜主要害虫田间药效试验。农业科技推广通讯，1999，6：1-2
33．金志华，金一平，宋友礼。Avermectin产生菌异亮氨酸诱导变种的选育。中国抗生素杂志。1997，22(2)；84-286
34．姚汝华。微生物工程工艺原理。华南理工大学出版社，1996
35．姜竹茂。食品添加剂的微胶囊方法及应用。食品工业科技，1998，5：73-75
36．胡润茂。一族新的广谱驱虫抗生素——除虫菌素(Avermectin)及其衍生物双氧除虫菌素。应用微生物，1986，5：1-8
37．凌世海。我国农药加工工业现状和发展建议。农药，1999，38(10)：19-26
38．原昭三，森定雄，花井俊彦。现代色谱分析法—原理和实际应用。邱宗萌，孙琢琏译。科学技术文献出版社，重庆分社，1988
39．唐根源，吴红京。高效液相色谱法光电二极管阵列检测器对苯乙烯羧化反应液的分析和鉴定。色谱，1990，8(1)：62-63
40．徐俊。阿维菌素产生菌DNA新型位点特异性修饰的分子遗传学研究。[硕士论文]。武汉，华中农业大学，1997
41．徐慧斌，沈杰，叶明忠，曹杰，张仓，王权，周金林，何国声。埃维菌素长效控释丸在棉羊体内的药物动力学研究。中国兽医寄生虫病，1996，4(2)：4-7
42．崔云龙，田明，邵宗泽。紫外线使苏云金芽胞杆菌伴胞晶体失活机理的研究。微生物学通报，1993，20(4)：193-195
43．扈洪波。Ivemaectin的制剂研究。中国兽药杂志，1998，32(2)：51-53
44．扈洪波。阿维菌素口服液体药剂的稳定性研究。畜牧兽医学报，1998，29(2)：185-188
45．曹贵明。阿维菌素(Avermectins)高产菌株的诱变育种和发酵条件的研究。[硕士论文】。北京，北京农业大学，1995
46．龚炳永，周亦昌。抗生素的分离与测定。色谱，1992，10(5)：269-277
47．喻子牛。苏云金杆芽胞菌制剂生产及应用。农业出版社，1993
48．喻子牛。苏云金杆菌。北京，科学出版社，1990
49．蒋志坚，马毓龙合编。农药加工从书可湿性粉剂。化学工业出版社，1991
50．慕立义。植物化学保护研究方法。中国农业出版社，1994
51. Chabalo J C, Mrozik H, Tolman R L, Eskola P, Lusi A, Peterson L H, Woods M F, Fisher M H, Campbell W C, Egerton J R and Ostlind D. Ivermectin, a new broad-spectrum antiparasitic agent. J Med. Chem, 1980,23:1134-1136
52. Christopher J D, Stephen P G, Alexander C G, Kelvin S H, Michael S P and John C R. Novel Avermectins produced by mutational biosynthesis. J Antibiot, 1991, 44(3): 357-365
53. Cimburkova E, Zima J, Novak J and Vanek Z. Nitrogen regulation of Avermectins biosynthesis in Streptornyces arvermitilis in a chemically defined medium. JBasic Microbiol, 1988, 28(8): 491-499
54. Dunkle R L and Shasha B S. Response of starch encapsulated Bacillus thuringiensis containing ultraviolet screens to sunlight. Environ Entomol, 1989, 18(6): 1035-1041
55. Edmund W H, Breland W H, Kelvin S, Hordom S, Edward L, Richard G W, Donald B, Hamish A I, Mcasthur and William W J. Branched-chain fatty acid requirement for Avermectin production by a mutant of Steptomyces avermitilis lacking branched-chain 2-oxo acid dehydrogenase activity. J Antibiot, 1991, 44(3): 349-356
56. Eva C,Vendvlka J, Jiri Z and Zdenko V. The effect of inorganic phosphate on the production of Avermectin in Streptomyces avermitilis. J Basic Microbiol, 1989, 29(6), 341-346
57．Fisher M H．Avermectin研究最新进展。农药译丛，1991，3(3)：19-26
58. Goudie A C, Evans N A, Gration K A F, Bishop B F, Gibson S P, Holdom K S, Kaye B, Wicks S R, Lewis D, Weatherley A J, Bruce C
I, Herbert A and Seymour D J. Doramectin-a potent novel endectocide. Veter Para, 1993, 49:5-15
59. Griego V M and Spence K D. Inactivation of Bacillus thuringiensis spores by ultraviolet and the visible light. Apple Environ Entomol, 1978, 35:906-910
60. Hafner, Edmund W H, Kelvin S, Lee, Edward S. Process for production of Avermectins. U S Patent 323, 198, 1994, Oct 14
61. Herbert B S, Marius R and Kuo S L. Microencapsulation of pesticides by interfacial polymerization utilizing isocyanate or aminoplast chemistry. Pestic Sci, 1998. 54:394-400
62. Hopwood D A. Understanding genetic control of antibiotic biosynthesis and sporulation in Streptomyces. Pro R Soc, B235:121-138
63. Ikeda H and Omura S. Avermectin biosynthesis. Chem Rev, 1997, 97:2591-2609
64. Ikeda H and Omura S. Control of Avermectin biosynthesis in Streptomyces avermitilis for the selective production of a useful component. JAntibiot, 1995, 48(7):549-562
65. Ikeda H and Omura S. Microorganism for selective production of a specific compound of Avermectin and a method for selective production. U S Patent 1998, 861. 1992, Dec. 30
66. Ikeda H, Kotaki H and Satoshi O. Genetic studies of Avermectin biosynthesis in Streptomyces avermitilis. J Bacteriol, 1987, 169:5613-5621
67. Ikeda H, Pang C H, Endo H, Ohta T, Tanaka H and Omura S. Construction of a single component producer from the wild type Avermectins producer Streptomyces avermitilis. J Antibiot, 1995, L08: 532-534
68. Ikeda H, Omura S and Tanaka H. Selective production of specific components of Avermectins in Streptomyces avermitilis. J Antibiot, 1991,
44: 560-563
69. Jan N, Jan K and Zdenko V. Nitrogen source regulates expression of alanine dehydrogenase isoenzymes in Streptomyces avermitilis in a chemically defined medium. Can J Microbiol, 1997. 43(2): 189-193
70. Louis S C, William F F, Byron H A, William J A V, Lawrence F C, Ralph A S, Walter F K and Peter G W. Photodegradtion of Avermectin Bla thin film on Glass. J Agric Food Chem, 1991, 39, 1310-1319
71. Lucille J C. Process for the Separation of Antibiotic Macrolides. United States patent, 4, 160, 083.1979, Jul 3
72. Marvin D S, Delia V, Otto D H, Deborah Z, Mary N, Kaplan L and Ostlind D A. Demethylavermctins Biosynthesis, Isolation and characterization. J Antibiot, 1985, Nov, 1494-1498
73. Marvin D S, Susan L A, Delia L V and Byron H A. purification and identification of dTDP-olean drose, the precursor of the oleandrose units of the Avermectins. J Bio Chem, 1990. 265 (28): 16965-16970
74. Matthews and Frank J. Process for isolation of Avermectin B1 components with improved purity and subsequent isolation of B2 components. U S Patent 931, 839, 1986 Nov. 18
75. Morris O N. Protection of Bacillus thuringiensis from inactivation by sunlight. Can Entomol, 1993, 115: 1215-1217
76. Toms C and Edward S I. Studies on the biosynthesis of Avermectins. Arch Biochem and Biophys, 1989, 270(2); 521-525
77. Toms C, Byron H A, Vincent P G and Edward S I. Further studies on the biosynthesis of Avermectins. J Ind Microbiol, 1989a, 4, 231-238
78. Toms C, Edward S I, Otto D H, Deorah Z and Ostlind D A. Directed biosynthesis of Avermectins. Arch Biochem and Biophys, 1989b, 269(2): 544-547