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蝽类昆虫酯酶同工酶、消化酶及体内微生物作为分类依据的可行性研究
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摘要
本文采用酯酶同工酶电泳、镜检与分离培养及消化酶分析对蝽类昆虫酯酶同工酶、体内微生物及主要消化酶类进行了初步研究,结果如下:
     1 酯酶同工酶电泳表明:蝽类昆虫酯酶同工酶谱存在明显的差异,特征酶谱重复性和稳定性较好。蝽类昆虫科之间酶谱差异大于各属间的差异;各属间的酶谱也有显著差异;而属内各种间的酶谱比较接近,但每种都有自己的特征酶谱,彼此易于区别。在鉴别蝽类昆虫中,酯酶同工酶作为生化分类指标有一定的可靠性。
     2 利用镜检、分离培养等方法确定了蝽类体内存在细菌。从菌落形态上看,姬猎蝽和黑食蚜盲蝽之间有相同的菌落存在,即圆形的乳白色菌落和圆形的桔黄色菌落;斑须蝽和小长蝽的相同菌落是圆形的白色菌落。植食性的蝽类和肉食性的蝽类之间存在明显的菌落差异。斑须蝽(Dolycorisbaccars (Linuacus))成虫、若虫、产出体外的卵与虫体内未产出的卵内均含有细菌,对本次试验中蝽类昆虫体内细菌进行功能测定表明,这些细菌都有分解蛋白的能力,但都没有分解淀粉的能力,肉食性蝽类体内细菌分解蛋白形成的透明环带较宽;植食性蝽类体内细菌分解蛋白形成的透明环带较窄。本试验中透明圈宽度可以作为所选试虫植食性肉食性分类的依据。
     3 试验采用α-淀粉酶活力/蛋白酶活力比值作为指标,对蝽类昆虫进行了食性鉴定,结果表明:肉食性蝽类的α-淀粉酶/蛋白酶比值范围在0.3~0.8之间,而植食性蝽类的α-淀粉酶/蛋白酶比值范围在2.0~6.0之间,食性差异明显。植食性蝽的α-淀粉酶活力/蛋白酶活力比值显著大于捕食性蝽。
The availability for classification of Hemiptera insects by using the esterase isoenzynes, microorganisms and digestive enzymes characters has been studied. The results are as follows.
    1.There were differences in the electrophoretic pattern of the bugs. Differences of zymograms between families were more significant than those between genera. Although zymograms within genera were rather similar, all the species could be unequivocally distinguished from each other. The feasibility about using isoenzymes in systematics was also discussed.
    2.The microorganisms in the bugs' body were identified through bacterioscopy; isolation and culture.The result showed that microorganisms were bacterium. The flesh-eating bugs, Nabis stenoferus Hsiao and Deraeocoris punctulastus Fall, had identical colony character; plant-eating bugs, Dolycoris baccarus (Linuacus) and Nysius ericae (Schilling), also had the same ones. However the significant differences of colony exited between flesh-eating bugs and plant-eating bugs.Bacterium were found in ova; nymphs and adults of Dolycoris baccarus (Linuacus). All the bacterium in the bugs' body could decompose protein, but could not break down starch. The transparent rings formed by bacterium decomposing protein between flesh-eating bugs and plant-eating bugs were different. The former were wider than latter.
    3.The ratio of amylase to protease enzyme activies(A/P) was used as one of the criteria for identifying the feeding habits of bug.The A/P for plant-eating bugs was about 2.0-6.0, for the flesh-eating bugs was 0.3-0.8. The rate of plant-eating bugs was fairly higher than that of flesh-eating bugs.
引文
[1]姚涌.RAPD技术在医学昆虫分类中的应用[J].疾病控制杂志,2000,4(4):讲座与综述.
    [2]彩万志,庞雄飞,花保祯,等.普通昆虫学[M].北京:中国农业大学出版社,2001:271.
    [3]王心丽.试论昆虫种类鉴定的准确性[J].昆虫知识,1999,36(3):174~173.
    [4]Imai, H T, R H Crozier&R W Taylor. 1977. Karyotype evolution in Australian ants[J]. Chromosoma 59: 341~393.
    [5]Imal, H T, R W Taylor, R H Crozler. Chromosomal Polymorphism in the ant Myrmecta (pilosula) n=1[J].Ann, Report Nat. Inst (Jap.) , 1988, 38: 82~84.
    [6]Muramoto N. A chromosome study of thirteen species of heteropteran Insects Heteroptera) [J].La Kromosomo Ⅱ1981, 23: 668~675.
    [7]彩万志.昆虫细胞分类学的基本问题及染色体系统发育的重建方法[J].昆虫分类学报,1994,16(1):4~14.
    [8]吴福桢,郭予元.中国小蠊属蜚蠊种类及分布、生活习性和经济意义[J].昆虫学报,1984,27(4):439~442.
    [9]姚世鸿,王景佑,谷晓民,等.贵州两种黄脊蝗核型和C—带的比较研究[J].昆虫学报,1995,38(3):312~316.
    [10]李本文.长沙地区德国小蠊的染色体核型[J].湖南医学院学报,1984,9(4):358~360.
    [11]张惠如,李本文.长沙地区美洲大蠊的染色体的研究[J].湖南医学院学报,1985,10(2):137~139.
    [12]陈天华,徐进,赖焕林.等.黑松、马尾松及其杂种的核型比较[J].植物资源与环境学报,1994,3(1):9~14.
    [13]龚鹏,张孝羲,杨效文.用微卫星引物pcr分析棉蚜不同蚜型的DNA多态性[J].昆虫学报,2001,44(4):416~421.
    [14]王桂荣.rapd技术及其在昆虫学研究中的应用[J].昆虫知识,1999,36(3):184~188.
    
    
    [15]王继恒,曲传智.随即扩增多态性DNA技术及其在蚊虫研究中的应用[J].中国媒介生物学及控制杂志.2000,11(2):156~158.
    [16]顾耘,王思芳,张迎春.应用RAPD技术时引物数量和统计方法的研究[J].莱阳农学院学报,2000,17(4):239~242.
    [17]张晓俊,李三暑,黄玉清.近几年我国褐飞虱迁飞的研究概况及RAPD技术的应用[J].江西农业大学学报,200l,23(2):253~258.
    [18]桂慕燕,左正宏,王学民.RAPD分析在绢丝昆虫亲缘关系研究中的应用□.柞蚕品种间的遗传差异[J].遗传,2001,23(5):452~454.
    [19]万由衷,曲志才,曹清玉.不同种群灰飞虱(Laodelphax striatellus)的RAPD分析[J].复旦学报(自然科学版),2001,40(5):535~543.
    [20]郑哲民,黄刚.螽斯总科昆虫基因组DNA的提取和种内雌雄及体色表现型不同个体间RAPD带型变异的研究[J].陕西师范大学学报(自然科学版),2002,30(1):99~107.
    [21]田英芳,郑哲民.5种蟋蟀的RAPD研究初报(直翅目:蟋蟀总科)[J].陕西师范大学学报(自然科学版),2002,30(2):91~93.
    [22]蒋国芳,陆敢,黄琨.用RAPD标记研究蚱属五个种间的亲缘关系[J].昆虫学报,2002,45(9):499~502.
    [23]王思芳,顾耘,顾颂东.四个齿爪鳃金龟类群分类地位的RAPD分析[J].华东昆虫学报,2002,11(2):26~29.
    [24]高明嫒,王心丽,李重九.表皮碳氢化合物分析用于棉铃虫与烟青虫幼虫分类鉴别[J].昆虫知识,1999,36(5):166~269.
    [25]韩雅莉,谭竹均,郑哲民.网翅蝗科九种蝗虫的酯酶同工酶研究(直翅目:蝗总科)[J].昆虫分类学报,1999,21(2):87~92.
    [26]彩万志,李淑娟,米青山.昆虫拟态的多样性[J].昆虫知识,2002,39(5):390~397.
    [27]梁铬球,郑哲民.中国蝗总科(Acridoidea)昆虫分类研究综述[J].昆虫知识,1994,31(2):119~121.
    [28]王歆,徐如宏,张立异.小麦外源基因检测研究进展[J].山地农业生物学报
    
    2001,20 (1):70~75.
    [29]Saul S H, M J Sinsko, P R Grimstad. Identification of sibling species, Aedes triseriatus and Ae. hendersoni, by electrophoresis[J]. J med Ent, 1977, 13(6): 705~708.
    [30]Mahon, R J C A Green, R H Hunt. Diagnostic allozymes for routine identification of adults of the Anopheles gambiae complex (Diptera, Culicidae)[J]. Bull ent Res, 1976, 66(1): 25~31.
    [31]Miles S J. A biochemical key to adult members of theAnopheles gambiae group of species (Diptera: Culicidae)[J]. J Med Entomol, 1979, 15: 297~299.
    [32]Wagner R P. and R K Selander, Isoenzymes in insects and their significance[J]. Ann Rev Entomol, 1974, 19: 117~138.
    [33]李绍文,孟玉萍,张宗炳,等膜翅目酯酶同工酶的比较研究[J].昆虫学报,1987,30(3):266~270.
    [34]Berlocher S H. Insect molecular systematics[J]. Ann. Rev. Entomol. 1984, 29:403~433.
    [35]杨国海,梁广勤.同工酶电泳技术在植物检疫害虫鉴定中的应用[J].植物检疫,1992,6(5):335~338.
    [36]童富淡,胡家恕.东方蜜蜂与西方蜜蜂酯酶同工酶的比较研究[J].浙江大学学报,2001,27(1):113.
    [37]徐萍莉,谢爱林,谷晓明.2种负蝗酯酶同工酶的比较研究[J].贵州师范大学学报(自然科学版),2001,19(3):17~19.
    [38]郭晓霞,郑哲民.菜粉蝶不同发育期酯酶同工酶的比较研究[J].昆虫学报,2002,45(3):401~403.
    [39]唐桦,郑哲民.光肩星天牛与黄斑星天牛酯酶同工酶的比较研究[J].北京林业大学学报,2002,24(1):66~68.
    [40]王学忠,李菊升,王丕玉.微小按蚊种团酯酶同工酶比较研究[J].实用寄生虫病杂志,2000,8(1):9~11.
    [41]李鹤,施宗伟,沈佐锐.三种瘤背豆象幼虫酯酶同工酶的比较研究[J].昆虫知
    
    识,2000,38(1):62~64.
    [42]Miles S J. A biochemical key to adult members of the Anophezes gambiae group of species (Diptera: Culicidae) [J]. Entomol, 1979, 15(3): 297~299.
    [43]缪建吾,等.中国六中蚊虫的酯酶同工酶的比较研究[C].昆虫学研究集刊,第一集,1980,89~92.
    [44]将成山,等.中华按蚊和雷氏按蚊嗜人亚种八中同工酶的遗传分析及其亲缘关系的研究[J].遗传学报,1983,10(5):390~397.
    [45]Berlocher S H. Insect molecular systematics Ann[J]. Rev Entomol, 1984, 29:403~433.
    [46]梁广勤,杨国海,梁帆,等.利用染色体和同工酶技术鉴定寡鬃实蝇幼虫[J].植物检疫,1994,8(1):4~9.
    [47]邓晓峰,王俊,屈娟,等.台湾省斑潜蝇种类及其防治研究进展[J].植物检疫,1997,11(5):284~289.
    [48]S 拉杰 乌尔森,N B克里希纳穆尔蒂,S N 黑格德.家蚕个体发育中酯酶同工酶的研究(英文)[J].蚕业科学,1997,23(2):109~113.
    [49]沈文飚,徐朗莱,黄丽琴,等.棉铃虫各龄期酯酶同工酶的比较[J].昆虫知识,2001,35(1):15~16.
    [50]谢寿安,吕淑杰,袁锋,等.华山松大小蠹不同龄期幼虫酯酶同工酶的比较研究[J].湖北农学院学报,2002,22(5):388~393.
    [51]薛增召,张满让,霍绍棠,等.杨陵、太白两地麦二叉蚜种群酯酶同工酶比较研[J].西北农业学报,1996,5(1):93~94.
    [52]沈文飚,徐朗莱,黄丽琴,等.棉铃虫各龄期酯酶同工酶的比较[J].昆虫知识,1998,35(1):15~16.
    [53]汪桂玲,蒋葵,黄原.5种瓢虫酯酶同工酶种间及种内变异的比较研究[J].陕西师范大学学报(自然科学版),1998,26(3):91~94.
    [54]张迎春,郑哲民,杨建雄,等.五种瓢虫酯酶同工酶的比较研究及其在分类上的应用(鞘翅日:瓢虫科)[J].昆虫分类学报,1999,21(2):123~127.
    [55]唐桦,郑哲民.光肩星天牛与黄斑星天牛酯酶同工酶的比较研究[J].北京林业
    
    大学学报,2002,24(1):66~68.
    [56]http://www.google.com/search?h1=zh-CN&inlang=zh-CN&ie=GB2312&q=Markert+group+gene++structure+enzyme+isozymes++decide&btnG=Google%CB%D1%CB%F7&1r=, 2003-05-17.
    [57]http://agri-history.net/history/mulu16.htm,2003-5-17.
    [58]徐萍莉,谢爱林,谷晓明.2种负蝗酯酶同工酶的比较研究[J].贵州师范大学学报(自然科学版),2001,19(3):17~19.
    [59]雷朝亮,宗良炳.两种小花蝽酯酶同工酶的比较研究[J].华中农业大学学报,1989,8(4):342~345.
    [60]Pereira J, Dujardin J P, Salvatella R. Enzymatic variability and phylogenetic relatedness among Triatoma infestans, T platensis, T delpontei and T rubrovaria[J].Heredity, 1996, 77(1): 47~54.
    [61]Costa J, Maria G R F S, Veronica M S, etal. Isoenzymes detect variation in populations of Triatoma.braziliensis.Neiva,. 1911(Hemiptera,.Reduviidae, Triatominae) [J]. Memoriasdo Instituto Oswaldo Cruz, 1997, 92(4): 459~464.
    [62]NorieauF, GutierrezT, ZegarraM.etal. Cryptic speciation in Triatoma sordida (Hemiptera: Reduviidae) from the bolivian Chaco[J]. Trop Med. Int Health, 1998,3(5): 364~372.
    [63]Harry M, Moreno G, Goyffon M. Genetic vari-ability in populations of Rhodnius prolixusvector of Chagas disease in Venezuela[J]. Evol Biol (Bogota), 1992, 6 (10): 175~194.
    [64]Harry M, Galindez I, Carlou M L. Isozymic data question the specific status of some blood-sucking[J]. Med Vet Entomol, 1993, 6(1): 37~43.
    [65]Harry M. Isozymic data question the specific status of blood-sucking bugs of the genus Rhodnius, vectors of Chagas' disease[J]. Trans R Soc Trop Med Hrg, 1993, 87(4): 492.
    [66]马春燕,郑哲民.四种蝽科昆虫酯酶同工酶的比较研究(半翅目:蝽科)[J].昆虫分类学报,2000,22(4):257~261.
    
    
    [67]http://science.scu.edu.tw/micro/1024/micro_encyc/01/001 .htm
    [68]T D.布洛克.微生物学[M].北京:人民出版社,2000:119~120.
    [69]徐孝华,普通微生物学[M].北京:中国农业大学出版社.1991:227
    [70]The Principles of Insect Physiology[M].V B Wigglesworth London Chapman and Hall, 1972, 412: 510.
    [71]上海植物生理学会.植物生理学实验手册[M].上海:上海科学技术出社,1985,167~169.
    [72]牟吉元,普通微生物学[M].北京:中国农业大学出版社,1998,71~72.
    [73]Buchner P. Endosymbiosis and apvsymbiosis in arthropods[J]. Symp Soc Gen Mic robid B, 1965, 297~332.
    [74]Douglas A.E. Sulphate utilisaation in anaphid symbiosis[J]. Insect Biochem. 1988,18,599~605.
    [75]Koyama K. Nutritional physiology of the brown rice planthopper, Nilaparvata lugens stal (Homoptera:Delphacidae) [J]. Ⅱ Essential amino acids fer nymphal development. APPL. Entomol Zool, 1985, 20: 424~430.
    [76]Koyama K, Arttificial reariny and nutritional physiology of the planthoppers and leaf hoppers on a holidic diet. JARQ[J]. 1988, 22: 20~27.
    [77]]傅强,等.高温处理后褐飞虱体内共生酵母菌和氨基酸需求的变化[J].昆虫学报,2001,44(4):534~539.
    [78]Pant N C, G Praenkel. studies on the symbiotic yeasts of two insect species Lasioderma serricome F. and stegobium paniceum L[J]. Biol, 1954, 107: 420~432.
    [79]Dowd P F in the cigarette Beetle (Coleoptera :Anobiidae)[J]. Econ Entomol, 1989,82: 396~400.
    [80]Shen S K, P F Dowd. Detoxification spectrum of the cigarette bettle syinbiont symbiotophrina kochit in culture[J]. Entomol, 1991, 60: 51~59.
    [81]Potrikus C J, Breznak J A. Nigtrogen-fixing Enterobacter agglomerans isolated from guts of wood-eating termites[J]. Appl Envir Microbiol, 1977, 33: 392~399.
    [82]Cochran D G. Nitrogen excretion in cockroaches[J]. A Rev Ent, 1985, 30: 29~49.
    
    
    [83]Ishilawa H. and Yamaji M. Symbionim, an aphid endosymbiont spectfic protein-I.Production of insect deficient in symbiont[J]. Insect Biochem 1985, 15: 155~163
    [84]Ohtake C. and Ishikawa H. Effects of heat treatment on the symbiotic system of an aphid mycetocyte[J]. Symbiosis, 1991, 11: 19~30.
    [85]Amiressaami M.Investigation of the light microscopical and ultraastructure of the Derneton-S-methyt resistance aphids urder constderation of the mycetome symbionts of pjorodon humuli Schrank[J]. Endocytobiology, 1982,425.
    [86]徐红星,郑许松,童中华,杀虫剂对褐飞虱体内共生菌的影响[J].浙江农业学报,2000,12(3):126~128.
    [87]吕仲贤,俞晓平,陈建明,共生菌在褐飞虱致害性变化中的作用[J].昆虫学报,2001,44(2):197~204.
    [88]吕仲贤,俞晓,陈建明,不同虫源和致害性的褐飞虱体内共生菌的种群动态[J].华东昆虫学报,2001,(1):115~118.
    [89]龚鹏,沈佐锐,李志红.国麦蚜体内的沃尔巴克氏体(Wolbachia)的检测[J].昆虫知识,2002,39(3):188~190.
    [90]崔晓峰,吴云锋,林林.桃蚜体内与病毒结合的共生菌Buchnera groEL基因的克隆和序列分析[J].中国病毒学,2002,17(1):69~72.
    [91]郑乐怡,归鸿,昆虫分类[M].上册,南京:南京师范大学出版社,1999,457.
    [92]Wigglesworth, VB. The Principles of Insect Physiology[M]. Chapman & Hall,1972, 412: 510.
    [93]寄主植物对棉蚜羧酸酯酶活性的影响[J].高希武,昆虫学报,1992,35(3):267~271.
    [94]伦宁格 A L.生物化学[M].北京:科学出版社,1990,234~238.
    [95]Hofer R, Schiemer F. Proteolytic activity in the digestive tract of several of fish with different feeding habits[J]. Oecologia, 1981, 48: 342~345.
    [96]Agrawal V P. Digestive enzymes of three teleost fishes[J]. Acta Physiol Hung, 1975, 46:93~98.
    [97]Prejs A, Blaszezyk M. Relationship between food and cellulase activity in
    
    frcshwater fishes[J]. Fish Biol, 1997, 11:447~452.
    [98]Kawai S, Ikeda S. Effects of dietary changes on the activities of digestive enzymes in carp intestine[J]. Bull Japan Soc science Fish, 1972, 38(3): 265~269.
    [99]Kenji Takil, Sadao Shimeno. The effect of feeding stimulant in diet on digestive enzyme activities of eel[J]. Bull Japan Soc science Fish, 1986, 52(8): 1449~1454.
    [100]薛元海.棉铃虫消化酶活性水平可作为评价棉花抗虫性的一个指标[J].中国棉花,1994,21(2):30.
    [101]刘强,郑乐怡.干旱区重要农业害虫——麦蝽属的中国新纪录种[J].干旱区资源与环境,1994,8(4):91~92.
    [102]柴冠卿.斑须蝽对泡桐的为害及防治[J].植保技术与推广,1995,3:22.
    [103]李国阳,刘从明,张卫标.扶沟棉田盲椿象猖獗为害[J].植保技术与推广,2002,22,(3):47.
    [104]孙黎明.用毒饵诱杀茶翅蝽效果佳[J].北京农业,1997,8:20.
    [105]崔素贞,小花蝽生物学特性及其对主要棉虫控制作用的研究[J].棉花学报,1994,6(增刊):78~83.
    [106]刘万学,万方浩,张帆.棉田优势天敌异须盲蝽的研究[J].中国生物防治,2000,(164):148~151.
    [107]秦玉洁,吴伟坚,梁广文.节瓜蓟马的新天敌—中华微刺盲蝽[J].昆虫天敌,2001,23(3):115~118.
    [108]白世红,朱承美,徐元春.山东林木药用昆虫概述[J].山东林业科学,2000,增刊:41~42.
    [109]http://www. sxrb.com/mag6/20010829/colart45825.htm. 2003-5-12.
    [110]郭尧君,蛋白质电泳实验技术[M].北京:科学出版社,2001,72.
    [111]莽克强,.徐乃正,方荣祥,等.聚丙烯凝胶电泳[M].北京:科学出版社,1975,34~39.
    [112]蒋成山.中华按蚊和雷氏按蚊嗜人亚种八种同工酶的遗传学分析及其亲缘关系的研究[J].遗传学报,1983,10(5):390~397.
    [113]郑乐怡,动物分类原理与方法[M].北京:高等教育出版社,1987,74.
    
    
    [114]吕仲贤,俞晓平,陈建明,等.褐飞虱体内共生菌种群及其与抗性水稻品种的关系[J].生物防治,2001,44(2):197~204.
    [115]龚明霞.几种革兰氏染色方法的比较与体会[J].卫生职业教育,2002,5:39.
    [116]河北农业大学《微生物学试验》编写组,微生物学试验[M].保定:河北农业大学出版社,115~122.
    [117]郭勇.酶工程[M].北京:中国轻工业出版社,2000,308~311.
    [118]王中仁.植物等位酶分析[M].北京:科学技术出版社,1998,108~110.
    [119]王琛柱,钦俊德.棉铃虫幼虫中肠主要蛋白酶活性的鉴定[J].昆虫学报,1996,39(1):7~13.

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