抗高效氯氰聚酯家蝇品系的交互抗性及其相关酶的活性研究

发布时间：2018-01-16 06:42

本文关键词：抗高效氯氰聚酯家蝇品系的交互抗性及其相关酶的活性研究　出处：《山东师范大学》2011年硕士论文　论文类型：学位论文

【摘要】：本文以家蝇为实验对象,通过实验的前期准备,熟悉了家蝇的饲养方法及抗性培育方法,并采用点滴法性作为抗性培育及生物测试的方法。实验所用的家蝇敏感品系是山东疾病控制预防中心昆虫饲养室长期饲养的,抗性培育是从实验室内饲养的敏感品系(S)开始,所使用的杀虫剂为高效氯氰菊酯(beta-cypermethrin)。对抗性培育家蝇的采用点滴法进行抗药性倍数的测定后,并选择不同抗性水平的家蝇对其他三种杀虫剂进行抗性倍数的测定,以观察抗高效氯氰菊酯抗性家蝇对其他杀虫剂的交互抗性。实验观察记录了家蝇抗药性培育的结果;统计了不同抗性水平家蝇对其他三种杀虫剂的交互抗性的变化;并测量了不同抗性水平家蝇体内GSH-PX、AChE、SOD活性的变化,并对相关酶的同工酶的电泳谱带进行分析;观察了不同抗性水平家蝇的羽化率的变化;最后观察了不同杂交条件下对家蝇抗性衰减的影响。通过对家蝇品系26代的抗性培育,家蝇的抗性倍数从敏感品系(S),到F_(26)代时抗性倍数达到152.9079,抗性家蝇抗性水平已经达到高抗水平,这对家蝇的防制产生了较大的困难。在家蝇的抗性培育过程中,分别对抗性选育的F_5、F_(10)、F_(16)、F_(22)和F_(26)家蝇品系对溴氰菊酯(deltamethrin)、敌敌畏(DDVP)和残杀威(propoxur)3种杀虫剂的抗性倍数的测定。结果显示,抗高效氯氰菊酯家蝇在F_(26)时抗性倍数增长了44.8196倍,存在明显的交互抗性;对DDVP的抗性倍数到F_(26)时增长了8.7899倍,有交互抗性;对残杀威,抗性倍数保持在1倍左右,在F_(26)时,抗性倍数最低,为0.6074倍,存在负交互抗性。由此可以得出,拟除虫菊酯类抗性家蝇对同类的药剂存在明显的交互抗性,对有机磷杀虫剂有一定的交互抗性,对氨基甲酸酯类杀虫剂存在负交互抗性。这些结果对于选用具有负交互抗性杀虫剂防制家蝇有指导作用。为检测抗性选育的家蝇品系对家蝇体内酶的作用,分别对不同抗性水平家蝇的GSH-PX、AChE和SOD三种酶进行了酶活的测定,结果显示,家蝇体内的三种酶的酶活均发生了较大的变化。GSH-PX随着家蝇抗性的增长,活性逐渐增强,在F_(26)时达到最高,相对活性达到236.55%,其POD同工酶酶谱带颜色均有加深趋势,特别是PX_3和PX_4条带在F_(18)达到最深,说明酶表达量增加,活性加强,因此可以看出GSH-PX作为重要的解毒酶系,活性增强与抗性的增长有关;AChE作为重要的靶标酶系,随着家蝇抗性的增高,AChE活性降低,因此表现为靶标不敏感,有利于降低杀虫剂的危害,在酯酶同工酶酶谱中,E_1和E_3颜色逐渐加深,而E_5条带颜色变浅,甚至消失,说明酯酶同工酶结构性质有所改变;SOD作为昆虫体内重要保护酶系,家蝇的抗性增长的同时,SOD活性逐渐增强,到F_(26)时到达最强,相对活性达到297.35%,活性增加了近3倍,SOD活性增强有利于消除家蝇体内氧自由基,达到保护作用,在SOD同工酶酶谱中,4条谱带颜色逐渐加深,与SOD活性变化相符。在本研究过程中,为观察家蝇的抗性增长对家蝇羽化率的影响,选取不同抗性世代家蝇测定其羽化率情况。结果显示,随着抗性的增高,家蝇的羽化率随之降低,羽化率从敏感品系的为91.33%,到F_(26)时降到44.67%,下降率达到51.10%。说明家蝇抗性的增长是以羽化率降低为代价的,这也是抗性适合度的一个重要方面。此外,在抗性家蝇培育通过对第26代家蝇连续培养7代,观察每代家蝇在不同的杂交情况下抗性衰退情况。结果显示,敏感品系雌性家蝇与抗性品系的雄性家蝇杂交后抗性衰减最快,经过7代后抗性倍数降到2.2989,衰减率达到98.50%,基本接近敏感水平;抗性雌性家蝇与敏感雄性家蝇的杂交后衰减也较快,衰减率达到91.85%;在自然衰退状况下,抗性衰退稍慢,其衰减率也达到83.66%。结果说明,实验室培育的单抗品系,在无杀虫剂的选择压力下,抗性的衰退较快,因此在对防制家蝇的过程中,为防止抗性的产生,同一种杀虫剂的停用或换用其他杀虫剂有利于抗性的衰减,增加家蝇防制效果。
[Abstract]:In this paper, housefly as the experimental object, through the preparation of the experiment, familiar with housefly breeding methods and resistance breeding, and the drop method as the method of resistance breeding and biological testing. Housefly sensitive strains used in the experiment is the Shandong Center for Disease Control and prevention insectaria long-term breeding, resistance training is susceptible rearing in the laboratory (S) from the start, the use of pesticides for cypermethrin (beta-cypermethrin). Against the cultivation using topicalapplication method for determination of housefly resistance ratio, determination and select resistance levels of different housefly resistance ratio of the other three kinds of insecticides, in order to observe the effect of anti cross resistance to cypermethrin resistance of housefly to other insecticides. The observed resistance of housefly breeding results; the statistics of different resistance levels of housefly to other three kinds of insecticides to Change of mutual resistance; and measured the different resistance levels of housefly in vivo GSH-PX, AChE, SOD activity, and the related enzyme electrophoresis isozyme bands were analyzed; observed changes in different resistance level housefly emergence rate; finally the effect on attenuation of housefly resistance under different hybridization.
Through resistance to the 26 generation of housefly strain, the resistance multiple of Musca domestica from the susceptible strain (S) to the F_ (26) generation reached 152.9079, and the resistance level of the resistant Musca domestica has reached a high level, which caused great difficulties for the control of housefly.
In the cultivation of the resistance of housefly resistance breeding process, respectively F_5, F_ (10), F_ (16), F_ (22) and F_ (26) strains of housefly to deltamethrin (deltamethrin), dichlorvos (DDVP) and propoxur (propoxur) determination of resistance ratio of 3 insecticides. Results display, anti cypermethrin of housefly in F_ (26) when the resistance ratio increased by 44.8196 times, there are obvious cross resistance; the resistance ratio of DDVP to F_ (26) an increase of 8.7899 times, have cross resistance to propoxur;, the resistance ratio maintained at about 1 times in F_ (26), the resistance index was lowest, 0.6074 times, has negativecross resistance. It can be concluded that the pyrethroid resistance of Musca domestica to the same insecticide exist obvious cross resistance, some cross resistance to organophosphorus insecticides, has negativecross resistance to carbamate pesticides. These results have negative cross resistance to insecticides selection It has a guiding role in the control of housefly.
For the detection of strains of housefly resistance breeding of housefly enzymes, respectively on Housefly with different resistance levels of GSH-PX, AChE and SOD of three enzymes were determined. The enzyme showed that three enzymes in vivo housefly enzyme activity were changed greatly with.GSH-PX of Musca domestica to growth activity gradually, in the F_ (26) reached the highest, relative activity reached 236.55%, bands of color are the deepening trend of POD isozyme, especially PX_3 and PX_4 bands in F_ (18) reached the deepest, it is suggested that the enzyme expression increase, strengthen the activity, therefore GSH-PX can be seen as an importantdetoxification enzyme. Enhanced activity associated with resistance to growth; AChE as an important target enzymes, with the increase of housefly resistance, AChE activity decreased, so the performance is not sensitive to the target, to reduce the harm of pesticides, the esterase isozyme, E_1 and E_3 color gradually increase Deep, E_5 band color shallow, or even disappear, illustrate the structural properties of esterase isozyme change; SOD as an important protective enzyme in insects, the resistance of housefly increased at the same time, SOD activity gradually increased to F_ (26) to reach the strongest, the relative activity reached 297.35%, the activity increased by nearly 3 times, SOD enhanced activity of Musca domestica is conducive to the elimination of free radicals, achieve the protective effect in the SOD isozymes, 4 bands of color gradually deepened, consistent with the change of SOD activity.
In the course of study, to observe the growth of housefly resistance effect of housefly emergence rate, selection of different generations to determine the resistance of housefly emergence rate. The results showed that with the increase of resistance, the housefly emergence rate decreased the emergence rate from 91.33% for the susceptible strain, F_ (26) dropped to 44.67% and the rate of decline of 51.10%. shows that the growth of housefly resistance is on the cost of reduced emergence rate, an important aspect of the suitability of this is resistance.
In addition, in the cultivation of the twenty-sixth generation of resistance of housefly Musca domestica cultured for 7 generations, and observed each generation of housefly resistance in hybrid condition of recession. The results showed that male hybrid housefly with female resistant strains of sensitive strains after resistance decay most rapidly, after 7 generations the resistance ratio down to 2.2989, decay rate reached 98.50%, close to the sensitive level; hybrid resistance and sensitive male female housefly Musca domestica after attenuation fast decay rate reached 91.85%; recession in natural conditions, the resistance slowerdecay the decay rate has reached 83.66%.. The results show that the monoclonal antibodylaboratory strains cultivated in the choice of pressure free of pesticides, resistance rapid decline, so in the process of house fly control in order to prevent the generation of resistance to disable the same insecticides or replaced by other pesticides in favor decay resistance, increase the control effect. The housefly

【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R184

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