本文關(guān)鍵詞：抗高效氯氰聚酯家蠅品系的交互抗性及其相關(guān)酶的活性研究　出處：《山東師范大學(xué)》2011年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 家蠅 抗性培育 交互抗性 酶活 同工酶 羽化率 抗性衰退

【摘要】：本文以家蠅為實(shí)驗(yàn)對(duì)象,通過(guò)實(shí)驗(yàn)的前期準(zhǔn)備,熟悉了家蠅的飼養(yǎng)方法及抗性培育方法,并采用點(diǎn)滴法性作為抗性培育及生物測(cè)試的方法。實(shí)驗(yàn)所用的家蠅敏感品系是山東疾病控制預(yù)防中心昆蟲(chóng)飼養(yǎng)室長(zhǎng)期飼養(yǎng)的,抗性培育是從實(shí)驗(yàn)室內(nèi)飼養(yǎng)的敏感品系(S)開(kāi)始,所使用的殺蟲(chóng)劑為高效氯氰菊酯(beta-cypermethrin)。對(duì)抗性培育家蠅的采用點(diǎn)滴法進(jìn)行抗藥性倍數(shù)的測(cè)定后,并選擇不同抗性水平的家蠅對(duì)其他三種殺蟲(chóng)劑進(jìn)行抗性倍數(shù)的測(cè)定,以觀察抗高效氯氰菊酯抗性家蠅對(duì)其他殺蟲(chóng)劑的交互抗性。實(shí)驗(yàn)觀察記錄了家蠅抗藥性培育的結(jié)果;統(tǒng)計(jì)了不同抗性水平家蠅對(duì)其他三種殺蟲(chóng)劑的交互抗性的變化;并測(cè)量了不同抗性水平家蠅體內(nèi)GSH-PX、AChE、SOD活性的變化,并對(duì)相關(guān)酶的同工酶的電泳譜帶進(jìn)行分析;觀察了不同抗性水平家蠅的羽化率的變化;最后觀察了不同雜交條件下對(duì)家蠅抗性衰減的影響。 通過(guò)對(duì)家蠅品系26代的抗性培育,家蠅的抗性倍數(shù)從敏感品系(S),到F_(26)代時(shí)抗性倍數(shù)達(dá)到152.9079,抗性家蠅抗性水平已經(jīng)達(dá)到高抗水平,這對(duì)家蠅的防制產(chǎn)生了較大的困難。 在家蠅的抗性培育過(guò)程中,分別對(duì)抗性選育的F_5、F_(10)、F_(16)、F_(22)和F_(26)家蠅品系對(duì)溴氰菊酯(deltamethrin)、敵敵畏(DDVP)和殘殺威(propoxur)3種殺蟲(chóng)劑的抗性倍數(shù)的測(cè)定。結(jié)果顯示,抗高效氯氰菊酯家蠅在F_(26)時(shí)抗性倍數(shù)增長(zhǎng)了44.8196倍,存在明顯的交互抗性;對(duì)DDVP的抗性倍數(shù)到F_(26)時(shí)增長(zhǎng)了8.7899倍,有交互抗性;對(duì)殘殺威,抗性倍數(shù)保持在1倍左右,在F_(26)時(shí),抗性倍數(shù)最低,為0.6074倍,存在負(fù)交互抗性。由此可以得出,擬除蟲(chóng)菊酯類(lèi)抗性家蠅對(duì)同類(lèi)的藥劑存在明顯的交互抗性,對(duì)有機(jī)磷殺蟲(chóng)劑有一定的交互抗性,對(duì)氨基甲酸酯類(lèi)殺蟲(chóng)劑存在負(fù)交互抗性。這些結(jié)果對(duì)于選用具有負(fù)交互抗性殺蟲(chóng)劑防制家蠅有指導(dǎo)作用。 為檢測(cè)抗性選育的家蠅品系對(duì)家蠅體內(nèi)酶的作用,分別對(duì)不同抗性水平家蠅的GSH-PX、AChE和SOD三種酶進(jìn)行了酶活的測(cè)定,結(jié)果顯示,家蠅體內(nèi)的三種酶的酶活均發(fā)生了較大的變化。GSH-PX隨著家蠅抗性的增長(zhǎng),活性逐漸增強(qiáng),在F_(26)時(shí)達(dá)到最高,相對(duì)活性達(dá)到236.55%,其POD同工酶酶譜帶顏色均有加深趨勢(shì),特別是PX_3和PX_4條帶在F_(18)達(dá)到最深,說(shuō)明酶表達(dá)量增加,活性加強(qiáng),因此可以看出GSH-PX作為重要的解毒酶系,活性增強(qiáng)與抗性的增長(zhǎng)有關(guān);AChE作為重要的靶標(biāo)酶系,隨著家蠅抗性的增高,AChE活性降低,因此表現(xiàn)為靶標(biāo)不敏感,有利于降低殺蟲(chóng)劑的危害,在酯酶同工酶酶譜中,E_1和E_3顏色逐漸加深,而E_5條帶顏色變淺,甚至消失,說(shuō)明酯酶同工酶結(jié)構(gòu)性質(zhì)有所改變;SOD作為昆蟲(chóng)體內(nèi)重要保護(hù)酶系,家蠅的抗性增長(zhǎng)的同時(shí),SOD活性逐漸增強(qiáng),到F_(26)時(shí)到達(dá)最強(qiáng),相對(duì)活性達(dá)到297.35%,活性增加了近3倍,SOD活性增強(qiáng)有利于消除家蠅體內(nèi)氧自由基,達(dá)到保護(hù)作用,在SOD同工酶酶譜中,4條譜帶顏色逐漸加深,與SOD活性變化相符。 在本研究過(guò)程中,為觀察家蠅的抗性增長(zhǎng)對(duì)家蠅羽化率的影響,選取不同抗性世代家蠅測(cè)定其羽化率情況。結(jié)果顯示,隨著抗性的增高,家蠅的羽化率隨之降低,羽化率從敏感品系的為91.33%,到F_(26)時(shí)降到44.67%,下降率達(dá)到51.10%。說(shuō)明家蠅抗性的增長(zhǎng)是以羽化率降低為代價(jià)的,這也是抗性適合度的一個(gè)重要方面。 此外,在抗性家蠅培育通過(guò)對(duì)第26代家蠅連續(xù)培養(yǎng)7代,觀察每代家蠅在不同的雜交情況下抗性衰退情況。結(jié)果顯示,敏感品系雌性家蠅與抗性品系的雄性家蠅雜交后抗性衰減最快,經(jīng)過(guò)7代后抗性倍數(shù)降到2.2989,衰減率達(dá)到98.50%,基本接近敏感水平;抗性雌性家蠅與敏感雄性家蠅的雜交后衰減也較快,衰減率達(dá)到91.85%;在自然衰退狀況下,抗性衰退稍慢,其衰減率也達(dá)到83.66%。結(jié)果說(shuō)明,實(shí)驗(yàn)室培育的單抗品系,在無(wú)殺蟲(chóng)劑的選擇壓力下,抗性的衰退較快,因此在對(duì)防制家蠅的過(guò)程中,為防止抗性的產(chǎn)生,同一種殺蟲(chóng)劑的停用或換用其他殺蟲(chóng)劑有利于抗性的衰減,增加家蠅防制效果。
[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

【學(xué)位授予單位】：山東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R184

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本文編號(hào)：1431977