Inheritance of resistance and cross-resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from Pakistan

A field population of Plutella xylostella (L) from Pakistan was found to be highly resistant to deltamethrin (>500-fold) but had little or no resistance to spinosad, fipronil, indoxacarb, abamectin, Bacillus thuringiensis (Bt) var kurstaki, Bt var aizawai or Cry1Ac when compared with a susceptible laboratory population, Lab-UK. A sub-population was selected for six generations (laboratory G3-G8) with deltamethrin (delta-SEL), while a second sub-population was left unselected (UNSEL). Bioassays at G9 found that selection with deltamethrin gave resistance ratios of >230 compared with UNSEL (>6730 compared with Lab-UK). The delta-selected population showed no apparent cross-resistance to spinosad, fipronil or indoxacarb. Logit regression analysis of F1 reciprocal crosses between delta-SEL and Lab-UK indicated that resistance to deltamethrin in the delta-SEL population was inherited as an autosomal, incompletely dominant (D(LC) = 0.67) trait. A direct test of monogenic inheritance based on a back-cross of F1 progeny with delta-SEL suggested that resistance to deltamethrin was controlled by more than one factor.     

抗Bt棉铃虫种群的交互抗性研究

分别用对CyrlAc抗性达11．6倍的Bt棉叶和Bt粉剂（Btk)汰选棉铃虫（Heliocoverpa armigera)种群（对Btk的抗性指数为5．2倍）,测定了其对不同Bt制剂,Bt毒素,化学农药的交互抗性,结果表明：停止汰选后,Bt棉叶汰选种群对CrylAc的抗性指数为5．9倍,对CrylAc 1C的交互抗性为3．7倍,对Btk的敏感性降低,对Bta,Cry2A无交互抗性,Btk汰选种群对Btk的抗性指数为4．1倍,对Bta,CrylAc,CrylAc 1C的交互抗性为2．3,3．1,2．3倍,对Cry2A无交互抗性,两汰选种群对化学农药灭多威,辛硫磷,氯氰菊酯的敏感性均有所增加。     

Genetics and evidence for an esterase-associated mechanism of resistance to indoxacarb in a field population of diamondback moth (Lepidoptera: Plutellidae)

Bioassays (at generation G2) with a newly collected field population (designated CH3) of Plutella xylostella L. from farmers' fields in the Cameron Highlands, Malaysia, indicated resistance ratios of 813-, 79-, 171-, 498- and 1285-fold for indoxacarb, fipronil, spinosad, deltamethrin and Bacillus thuringiensis toxin Cry1Ac respectively compared with a laboratory susceptible population (Lab-UK). At G2 the field-derived population was divided into two subpopulations: one was selected (G2 to G7) with indoxacarb (indoxa-SEL), while the second was left unselected (UNSEL). A significant reduction in the resistance ratio for each compound was observed in UNSEL at G8. For indoxa-SEL, bioassays at G8 found that selection with indoxacarb gave a resistance ratio of 2594 compared with Lab-UK and of 90 compared with UNSEL. The toxicity of fipronil, spinosad and deltamethrin was not significantly different in indoxa-SEL at G8 compared with G2 but was significantly greater than UNSEL at G8. The toxicity of Cry1Ac was significantly reduced in indoxa-SEL at G8 compared with G2 but was also significantly greater than UNSEL at G8. This suggests that indoxacarb selection maintained resistance to these compounds in the indoxa-SEL population. Synergist studies indicated that resistance to indoxacarb in indoxa-SEL was esterase associated. Logit regression analysis of F1 reciprocal crosses between indoxa-SEL and Lab-UK indicated that resistance to indoxacarb was inherited as an autosomal, incompletely recessive (D(LC) = 0.35) trait. Tests of monogenic inheritance suggested that resistance to indoxacarb was controlled by a single locus.     

Cross-resistance and inheritance of resistance to Bacillus thuringiensis toxin Cry1Ac in diamondback moth (Plutella xylostella L) from lowland Malaysia

A field population of Plutella xylostella from Malaysia (SERD4) was divided into five sub-populations and four were selected (G2-G5) with the Bacillus thuringiensis insecticidal crystal (Cry) toxins Cry1Ac, Cry1Ab, Cry1Ca and Cry1Da. Bioassay at G6 gave resistance ratios of 88, 5, 2 and 3 for Cry1Ac, Cry1Ab, Cry1Ca and Cry1Da respectively compared with the unselected sub-population (UNSEL-SERD4). The Cry1Ac-selected population showed little cross-resistance to Cry1Ab, Cry1Ca and Cry1Da, (3-, 2- and 3-fold compared with UNSEL-SERD4), whereas the Cry1Ab-SEL sub-population showed marked cross-resistance to Cry1Ac (40-fold), much greater than Cry1Ab itself. In contrast, the Cry1Ca- and Cry1Da-SEL sub-population showed little if any cross-resistance to Cry1Ac and Cry1Ab. The mode of inheritance of resistance to Cry1Ac was examined in Cry1Ac-selected SERD4 by standard reciprocal crosses and back-crosses using a laboratory insecticide-susceptible population (ROTH). Logit regression analysis of F1 reciprocal crosses indicated that resistance to Cry1Ac was inherited as an incompletely dominant trait. At the highest dose of Cry1Ac tested, resistance was recessive, while at the lowest dose it was almost completely dominant. The F2 progeny from a back-cross of F1 progeny with ROTH were tested with a concentration of Cry1Ac that would kill 100% of ROTH. The mortality ranged between 50 and 95% in seven families of back-cross progeny, which indicated that more than one allele on separate loci were responsible for resistance to Cry1Ac.     

Selection and heritability of resistance to Bacillus thuringiensis subsp kurstaki and transgenic cotton in Helicoverpa armigera (Lepidoptera: Noctuidae)

Compared with an unselected susceptible population, a cotton bollworm, Helicoverpa armigera (Hübner), population selected for 22 generations with transgenic cotton leaves (modified Cry1A) in the laboratory developed 11.0-fold resistance to Cry1Ac (one single-protein product MVPII). Resistance to Bacillus thuringiensis Berliner subsp kurstaki (Btk) was selected for 22 generations with a 5.2-fold increase in LC50. The estimated realized heritabilities (h2) of resistance for transgenic-cotton- and Btk-selected populations were 0.1008 and 0.2341, respectively. This reflects the higher phenotypic variation in response to Cry1Ac in the transgenic-cotton-selected population. This variation may have been caused by differences in protein toxin levels expressed in different growth stages of the transgenic cotton. Because of the different slopes of the probit regression lines between Cry1Ac and Btk, the estimated realized h2 cannot be used visually to compare resistance development to Cry1Ac and Btk in H armigera. Thus, the response quotient (Q) of resistance was also estimated. The Q values of resistance for transgenic-cotton- and Btk-selected populations were 0.0763 and 0.0836, respectively. This showed that the rate of resistance development would be similar in both selection populations. This result indicates that the selection of resistance using transgenic cotton is different from that selected using the single toxin. Resistance risk to transgenic cotton and Btk in field populations was assessed assuming different pressures of selection by using the estimated h2. Assuming the h2 of resistance in a field population was half of the estimated h2, and the population received prolonged and uniform exposure to transgenic cotton or Btk causing >70% mortality in each generation, we predicted that resistance would increase 10-fold after <23 generations for Cry1Ac in transgenic cotton-selected-populations and after <21 generations for Btk in Btk-selected populations. Cross-resistance would be expected after <48 generations for Btk in transgenic-cotton-selected populations and after <21 generations for Cry1Ac in Btk-selected population. The results show that the potential to evolve resistance is similar in both transgenic-cotton- and Btk-selected populations, but that cross-resistance development to Btk is slower in transgenic-cotton-selected populations than cross-resistance development to Cry1Ac in Btk-selected populations.     

小菜蛾Btk抗性品系对四种Bt杀虫晶体蛋白抗性发展的研究

使用苏云金芽胞杆菌库斯塔克亚种（Btk）可湿性粉剂对小菜蛾进行继代汰选获得F80代和F100代抗性品系。通过分别测定Cry1Ab、Cry1Ac、Cry1Ah和Cry1Ca四种杀虫晶体蛋白对小菜蛾Btk抗性品系F80代和F100代的室内毒力,明确了小菜蛾Btk抗性品系对四种Bt杀虫晶体蛋白抗性发展规律。研究结果表明,汰选至F80代时,Cry1Ca对小菜蛾的毒力最高,LC50约12.1mg/L,其次为Cry1Ac,LC50约47.7mg/L;而汰选至F100代时,仍以Cry1Ca对小菜蛾的毒力最高,LC50约21.4 mg/L,其次为Cry1Ab,LC50约72.2 mg/L。与相对敏感品系相比,小菜蛾抗性品系对Cry1Ac的抗性发展较快（抗性倍数高达67.3～106.8倍）,对Cry1Ab的次之（抗性倍数高达60.0～66.1倍）,而对Cry1Ca和Cry1Ah的抗性发展较慢（抗性倍数分别为3.4～6.0倍和1.6～2.5倍）。以上结果说明,在主效杀虫基因为Cry1Ac的Btk药剂选择压力下,小菜蛾对四种Bt杀虫晶体蛋白的抗性发展速度差异较大,且Cry1Ac和Cry1Ab存在交互抗性风险...     

Characterisation of abamectin resistance in a field‐evolved multiresistant population of Plutella xylostella

BACKGROUND: Plutella xylostella (L.) has evolved resistance to various kinds of insecticide in the field. Reversion and selection, cross‐resistance, inheritance and mechanisms of abamectin resistance were characterised in a field‐derived multiresistant population of P. xylostella from China. RESULTS: Compared with a susceptible Roth strain, the field‐derived TH population showed ～5000‐fold resistance to abamectin. Rapid reversion of abamectin resistance was observed in the TH population when kept without insecticide selection. The TH‐Abm strain, selected from the TH population with abamectin, developed 23 670‐fold resistance to abamectin, a high level of cross‐resistance to emamectin benzoate and low levels of cross‐resistance to spinosad and fipronil. Genetic analyses indicated that abamectin resistance in the TH‐Abm strain was autosomal, incompletely dominant and polygenic. P450 monooxygenase activities in the TH‐Abm strain were significantly elevated compared with the TH strain. Piperonyl butoxide (PBO) inhibited a small part of abamectin resistance in the TH‐Abm strain. CONCLUSION: Field‐evolved high‐level resistance to abamectin in the TH population was not stable. Selection of the TH population with abamectin resulted in an extremely high level of cross‐resistance to emamectin benzoate and low levels of cross‐resistance to spinosad and fipronil. Enhanced oxidative metabolism was involved in, but may not be the major mechanism of, polygenic abamectin resistance in the TH‐Abm strain. Copyright © 2009 Society of Chemical Industry     

Mode of inheritance and stability of resistance to Bacillus thuringiensis var kurstaki in a diamondback moth (Plutella xylostella) population from Malaysia

Genetic inheritance of resistance to Bacillus thuringiensis var kurstaki (BTK) was examined in a diamondback moth (Plutella xylostella) population collected from the Melaka region of Malaysia. A BTK‐selected sub‐population (BTK‐SEL) which was more than 100‐fold resistant to BTK compared with a susceptible (ROTH) population of P xylostella was used with standard reciprocal crosses and back‐crosses between ROTH and BTK‐SEL. Logit regression analysis of F ₁ reciprocal crosses indicated that BTK resistance was inherited as an incompletely recessive autosomal trait and controlled by a single locus. In contrast, other studies have shown that resistance to Cry1Ac is inherited as an incompletely dominant autosomal trait in a Cry1Ac‐selected sub‐population of the same Melaka population. The frequency of the allele responsible for resistance decreased without exposure to insecticide in the laboratory. © 2000 Society of Chemical Industry     

Cross-resistance between a Bacillus thuringiensis Cry toxin and non-Bt insecticides in the diamondback moth

BACKGROUND: Bacillus thuringiensis Berliner (Bt) crystal (Cry) toxins are expressed in various transgenic crops and are also used as sprays in integrated pest management and organic agricultural systems. The diamondback moth (Plutella xylostella L.) is a major worldwide pest of crucifer crops and one that has readily acquired field resistance to a broad range of insecticides.RESULTS: Selection of a subpopulation of the P. xylostella SERD4 population with the pyrethroid deltamethrin increased resistance to both deltamethrin and Cry1Ac relative to an unselected subpopulation. Selection of a second subpopulation with the Bt toxin Cry1Ac also increased resistance to both Cry1Ac and deltamethrin. A complementation test between the Cry1Ac-selected and deltamethrin-selected subpopulations suggested the presence of a common genetic locus or loci that control resistance to both insecticides. A piperonyl butoxide analogue with potent inhibitory activity against insect esterases significantly increased the toxicity of Cry1Ac and deltamethrin against the respective resistant subpopulations, but showed no such synergism with the unselected subpopulation of SERD4.CONCLUSION: Selection of one resistance phenotype resulted in the simultaneous selection of the other. This phenomenon could be due to a single mechanism acting against both classes of insecticide or to genetically linked, but separate, mechanisms.     

转Bt 基因抗虫玉米对棉铃虫杀虫活性的初步评价

国内外在转 Bt基因抗虫作物的研究和应用上已取得重大进展 ,其中 Bt抗虫棉已在美国、澳大利亚、中国等进入商品化应用阶段 ,近年来种植面积增长很快 ,并取得了显著的经济、生态和社会效益。 1 997年下半年 ,我国农业部分别批准了孟山都公司申报的保铃棉○R在河北省商业化生产和 Bt玉米在我国多省份的田间种植。其中保铃棉 ○R已由河北省定名为新棉 33B,1 998年在全省实际种植了近 1 0万 hm2 ,1 999年的种植面积已达 1 5万 hm2 。我国自主研制的 Bt抗虫棉也于 1 998年开始商品化应用 ,当年种植面积即达 1 0万 hm2以上 ,1 999年推广总面积…     

Cry1Ba3、Cry1Ia8蛋白对Cry1Ac抗性小菜蛾的杀虫活性研究

利用2种苏云金芽胞杆菌原毒素Cry1Ba3、Cry1Ia8及其组合,分别对Cry1Ac抗性种群小菜蛾幼虫进行生物活性测定。结果表明Cry1Ba3、Cry1Ia8对2种目标试虫均有高毒力,LC50分别为0.2175、0.6706μg/mL;Cry1Ba3毒力3倍于Cry1Ia8。2种蛋白混配的结果也表现出高毒力,LC50为0.4375μg/mL,没有显著的协同增效作用,也不存在拮抗。敏感与抗性小菜蛾种群生测结果统计分析比较,结果表明这2种蛋白及其组合与Cry1Ac并无交互抗性。     

入侵云南草地贪夜蛾种群对5种常用Bt蛋白的敏感性评价

草地贪夜蛾Spodoptera frugiperda(J. E. Smith)是原分布于美洲大陆热带和亚热带地区的一种重要玉米害虫。在当地,种植抗虫转基因玉米是防控草地贪夜蛾危害的主要手段。该虫于2019年1月入侵我国云南省,为明确入侵我国云南的草地贪夜蛾种群对常用Bt蛋白的敏感性水平,本文通过饲料表面涂抹法测定了瑞丽草地贪夜蛾幼虫对Cry1Ab、Cry1Ac、Cry1F、Cry2Ab以及Vip3A等5种Bt蛋白的敏感性。结果表明:几种常用Bt蛋白对瑞丽草地贪夜蛾致死作用顺序为Vip3A>Cry1Ab>Cry1F>Cry2Ab>Cry1Ac,对草地贪夜蛾抑制生长发育的顺序为Cry1Ab>Cry1F>Vip3A>Cry1Ac>Cry2Ab。此外,与美国相对敏感种群比较,云南瑞丽草地贪夜蛾种群对Cry1Ab、Cry1Ac、Cry1F、Cry2Ab和Vip3A的敏感性指标在0.28～3.76之间,表明该入侵种群对此5种Bt蛋白均未产生抗性。此研究可为将来建立以转Bt基因玉米作为防控草地贪夜蛾的技术体系提供依据。     

V-ATPase H表达量下调增强棉铃虫对Cry1Ac的抗性

棉铃虫Helicoverpa armigera是世界性重要农业害虫。目前防治棉铃虫的主要手段是种植转苏云金芽胞杆菌Bacillus thuringiensis(Bt)杀虫蛋白的转基因作物。本文旨在研究棉铃虫V-ATPase H在Cry1Ac蛋白毒力和抗性中的作用。利用实时荧光定量qRT-PCR技术分析V-ATPase H在Cry1Ac抗、感品系棉铃虫幼虫中肠及敏感品系棉铃虫幼虫受Cry1Ac诱导后的表达情况;在昆虫Sf9细胞中过表达V-ATPase H对其进行细胞定位,通过细胞毒力试验验证其对Cry1Ac毒力的影响。结果发现棉铃虫V-ATPase H基因在抗性品系中低表达,并且V-ATPase H在受到Cry1Ac诱导时也低表达;在Sf9细胞内表达V-ATPase H蛋白发现其在整个细胞中都有分布,过表达该蛋白后增强了细胞对Cry1Ac蛋白的敏感性。结果表明V-ATPase H参与Cry1Ac蛋白的毒力。     

Long-term selection for resistance to transgenic cotton expressing Bacillus thuringiensis toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Selection experiments for resistance to transgenic Bt cotton expressing Cry1Ac toxin in Helicoverpa armigera (Hübner) were conducted using a leaf-feeding method with 42 selection episodes over 45 generations. The cotton bollworm developed resistance to transgenic Bt cotton after 12 generations (F12) of selection. The survival rate of F12 neonates feeding on leaves of seedling stage for 4 days and boll-opening stage for 5 days of R19 line were ca 34 and 72%, respectively, compared with ca 0 and 40% for a non-selected sister strain (NYCS), but lower than or similar to that of F12 feeding on leaves of non-Bt cotton, Sumian 12 (ca 87 or 76%). Resistance to B thuringiensis HD-1 Dipel in neonates (F12) was ca 6-fold. After 42 generations of selection, the strain developed a very high level of resistance to Cry1Ac protoxin, 210 g kg(-1) MVPII wettable powder and 200 g litre(-1) MVPII liquid formulation, the resistance ratios being 1680-, 1780- and ca 1200-fold, respectively, compared with a laboratory susceptible strain (HZS). When compared with the non-selected NYCS, the resistance ratios to the above toxins were ca 540-, 580- and 510-fold, respectively, but to Dipel only ca 16-fold. The results indicated that it is very important to develop and implement effective resistance-management strategies and to detect early resistance to Bt cotton in field populations.     

Field-evolved resistance to Bt toxin Cry1Ac in the pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), from India

BACKGROUND: The pink bollworm is one of the most destructive pests of cotton. Transgenic cotton producing Bt toxin Cry1Ac or a combination of Cry1Ac and Cry2Ab2 has been used effectively against this pest. However, some other insects have evolved resistance to Bt toxins in the field. During the 2007–2008 and 2008–2009 seasons, pink bollworm populations in India were surveyed to evaluate their responses to Cry1Ac and seed powder containing Cry1Ac and Cry2Ab2. RESULTS: The results provide evidence that resistance to Cry1Ac had evolved by 2008 in a population sampled from non‐Bt cotton in the Amreli district of Gujarat in western India. The median lethal concentration of Cry1Ac for five‐day‐old larvae (LC₅₀) was significantly higher for insects derived in 2008 from Amreli than for any of the other field populations tested from four locations in India. For Cry1Ac, the mean LC₅₀ for the strain derived from Amreli in 2008 was 44 times higher than for the most susceptible population. However, for seed powder of Bollgard II containing primarily Cry2Ab2, the 2008 Amreli population was only slightly less susceptible than the most susceptible population. CONCLUSIONS: The data reported here constitute the first evidence of field‐evolved resistance of pink bollworm to Cry1Ac. This initial evidence spurred more extensive evaluations during the 2009–2010 growing season, which confirmed field‐evolved resistance to Cry1Ac in Amreli. The lack of cross‐resistance to Cry2Ab2 suggests that plants producing this toxin are likely to be more effective against resistant populations than plants producing only Cry1Ac. Copyright © 2011 Society of Chemical Industry     

Evidence for Resistance to Bacillus thuringiensis (Bt) subsp. kurstaki HD-1, Bt subsp. aizawai and Abamectin in Field Populations of Plutella xylostella from Malaysia

The efficacy of Bacillus thuringiensis (Bt) subsp. kurstaki HD-1 (‘Dipel’™; Btk; CryIA & CryII) and Bt. subsp. aizawai (‘Florbac’™; Bta; CryIA & CryIC) was assessed against larvae from various field populations of Plutella xylostella (F2 generation) collected in the Cameron Highlands, Malaysia in April 1994 and a lowland population (SERD 2; F10 generation) collected in December 1993. Evidence of resistance to Btk and to a lesser extent Bta is reported in these populations (LC₅₀ Toxicity Ratios [TR]=3–14 and 2–8 respectively), most notably in SERD 2. The first recorded evidence of resistance to abamectin (TR=17–195-fold) in field populations of P. xylostella is also reported. In an unselected sub-population of SERD 2, the TR values for Btk, Bta and abamectin declined 2- to 3-fold (P<0·01) over six generations in the laboratory (F10–F16) while in sub-populations of SERD 2 selected with these products (F11–F15) there was a significant (P<0·01) increase in the TR (15-, 3- and 2·5-fold respectively) when compared with the F10 generation. This suggests the presence of marked resistance to Btk and some resistance to Bta and abamectin. There is also evidence of slight cross-resistance to Btk in the Bta-selected sub-population but no evidence for the reverse selection of resistance or for cross-resistance between Btk and abamectin. Concurrent selection studies (F11–F15) with another sub-population of SERD 2 demonstrated resistance to the acylurea insect growth regulator, teflubenzuron (‘Nomolt’™) (29-fold increase in TR). Based on the selection experiments with SERD 2, estimates of realised heritability (h²) of resistance gave very high values for teflubenzuron and Btk (c.0·7) and moderate values for abamectin and Bta (c.0·3). The results are discussed in relation to integrated pest management (IPM) and insecticide resistance management (IRM) strategies for P. xylostella.     

Cross-resistance between acylurea insect growth regulators in a strain of Plutella xylostella L. (Lepidoptera: Yponomeutidae) from Malaysia

The activities of the acylurea insect growth regulators, chlorfluazuron, teflubenzuron and difubenzuron, and the neurotoxic macrocyclic lactone, abamectin were assessed against a laboratory susceptible (FS) strain and a field (Cameron Highlands, Malaysia (CH)) strain of the diamondback moth, Plutella xylostella L. using a leaf-dip bioassay at 20°C. The time taken to achieve end-point mortality was found to vary considerably (9–17 days), being fastest with abamectin against the FS strain and slowest with difubenzuron against the CH strain. The order of activity (LC₅₀ at F6/7) against second-instar larvae of both strains was: abamectin > chlorfluazuron = teflubenzuron ? difubenzuron. Subsequent assays (F14) with the acylureas, flufenoxuron and hexaflumuron against the FS strain suggested that the former was slightly more active than chlorfluazuron or teflubenzuron, the latter slightly less active. The CH population was found to be 12.6-, 6.7-, 6.4- and 2.3-fold less sensitive to difubenzuron, teflubenzuron, chlorfluazuron and abamectin respectively than the FS strain. Selection of sub-populations of the CH strain with chlorfluazuron (CHL-SEL) and teflubenzuron (TEF-SEL) for six generations (F6-11), resulted in LC₅₀ resistance ratios of 109- and 315-fold respectively when compared with the FS strain, equivalent to an 18- and a 46-fold increase in resistance compared with the unselected CH strain. Marked cross-resistance was also demonstrated between chlorfluazuron and teflubenzuron in both sub-populations. However, there was no evidence of cross-resistance to dijlubenzuron and abamectin and little or no cross-resistance to flufenoxuron and hexaflumuron. Resistance to chlorfluazuron and teflubenzuron appeared to be relatively unstable in the TEF-SEL compared with the CHL-SEL sub-population (over 6–9 generations). However, reselection of the TEF-SEL population with chlorfluazuron (F18–20) led to a very rapid increase in resistance to chlorfluazuron and particularly teflubenzuron. For the latter compound, resistance factors of about 1000000 were obtained (F19, 21). Such values are probably only semi-quantitative, as above a certain level of resistance feeding bioassays with acylureas (compounds which are active to a significant extent by ingestion) are likely to become rate-limiting.     

Bt杀虫蛋白对棉铃虫的抗虫毒力和增效作用

用 Bt杀虫蛋白 (ICP) Cry1A汰选的棉铃虫 (H elicoverpa armigera)抗性种群和同源对照种群分别测定了 4种不同类型 Bt ICP的抗虫毒力。结果表明 :4种杀虫蛋白对对照种群的毒力顺序为 :Cry1Ac>Cry1Ac+ 1C>Cry2 A>Cry1C;对抗性种群的毒力顺序为 :Cry1Ac+ 1C>Cry1Ac>Cry2 A>Cry1C,其中Cry1Ac+ 1C表现出对抗性种群有显著的协同增效作用。采用从 Bt液体发酵上清液中提取的增效粉 ,与Cry1Ac以 1∶ 1和 1∶ 2的比例混合 ,对 Cry1Ac有显著的增效作用 ,用抗性种群测定的增效比值明显大于对照种群 ,处理 5d的增效比值大于 14d的     

棉铃虫V-ATPase亚基B是Cry1Ac的功能受体

目前对转苏云金芽胞杆菌Bacillus thuringiensis（Bt）的研究发现,液泡型ATP酶（Vacuolar-type proton ATPase,V-ATPase）可能是Bt的一类新型受体。我们前期通过构建棉铃虫的中肠酵母文库筛选Cry1Ac的结合蛋白发现棉铃虫V-ATPase亚基B（V-ATPase B）可以与Cry1Ac结合。为明确V-ATPase B在Cry1Ac毒力和昆虫对Cry1Ac抗性机制中的作用,本研究首先采用实时荧光定量PCR技术分析了该基因在抗感Cry1Ac棉铃虫幼虫中及其受到Cry1Ac诱导时的基因表达情况;通过Ligand blot进一步地证实了其与Cry1Ac的结合特性;并通过在Sf9细胞中表达V-ATPase B的试验验证了其功能。结果表明V-ATPase B在抗性品系及受到Cry1Ac诱导时均下调表达,Ligand blot证实了V-ATPase B与Cry1Ac特异性结合;并且在昆虫细胞内过表达该基因,会增强Cry1Ac的细胞毒力。研究结果表明棉铃虫V-ATPase B是Cry1Ac的功能受体,并有可能通过降低基因表达来参与棉铃虫对Cry1Ac的抗性形成。     

Effects of Cry1Ac toxin of Bacillus thuringiensis and nuclear polyhedrosis virus of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on larval mortality and pupation

In the laboratory, the percentage mortality and pupation of Helicoverpa armigera (Hübner) were investigated when larvae were exposed to Cry1Ac of Bacillus thuringiensis Berliner, nuclear polyhedrosis virus of H. armigera (HaNPV) or Cry1Ac and HaNPV together. The results revealed that interactions between Cry1Ac and HaNPV varied with bioassay method and concentration of the suspension. When larvae were infected using a suspension containing both HaNPV and Cry1Ac, most combinations of Cry1Ac (62.5, 125 and 250 microg mL(-1)) and HaNPV (1.2 x 10(6), 6.0 x 10(6) and 3.0 x 10(7) PIB mL(-1)) showed an antagonistic effect. In the bioassay procedure where larvae were force fed diet containing Cry1Ac 48 h after being infected by HaNPV, interaction between Cry1Ac (0.5, 1, 2, 4 and 8 microg mL(-1)) and HaNPV (6.0 x 10(6) and 3.0 x 10(7) PIB mL(-1)) showed an additive effect, while combinations of Cry1Ac (0.5, 1, 2 and 4 microg mL(-1)) and HaNPV (1.2 x 10(6) PIB mL(-1)) showed an antagonistic effect. In the bioassay procedure where larvae being infected by HaNPV were fed on Cry1Ac diet from neonate to death or pupation, the results suggested that Cry1Ac and HaNPV showed an additive interaction. The percentage mortality was lower in the treatment of larvae infected by transgenic Bt cotton leaf discs containing HaNPV suspension than in the treatment of larvae by conventional cotton leaf discs containing HaNPV, while the pupation rate was higher. The combination of Bt cotton and HaNPV showed antagonism. The present results showed that a combination of Cry1Ac and HaNPV usually resulted in mortality levels greater than in the case of Cry1Ac but not greater than with the virus alone.