小菜蛾对杀虫双、杀螟丹抗性品系选育及其抗性消长规律的研究(英语)

用改进的蛭石萝卜苗法饲养小菜蛾,以点滴法处理对杀虫双、杀螟丹两种药剂淘汰选育的小菜蛾抗性品系和不以药剂处理的敏感品系的各项生物学特性与同期田间种群比较,各品系间差异不显著,均没有生长发育和繁殖上的不利情况.5年来药剂淘汰选育,杀虫双品系至F_(85)代其抗性增长178倍;杀螟丹品系F_(80)代其抗性增长87倍,均已形成高抗品系.两个抗性品系经停止药剂淘汰选育后5代,抗性倍数分别从167倍降至57倍和74倍降至16倍,说明这两个抗性品系的抗性是不稳定的,在没有药剂选择压力的情况下,开始几代抗性下降较快,当下降到一定水平后抗性便比较稳定,似乎很难恢复原有的敏感性.     

不同品系小菜蛾成虫脑突触体 蛋白质磷酸化的研究

对小菜蛾Plutella xylostella(L.)敏感品系、抗溴氰菊酯品系、抗杀虫双品系和抗杀螟丹品系的成虫脑突触体蛋白质磷酸化进行了研究比较。结果表明：蛋白质磷酸化在各个品系中的表现是不一样的。cAMP和钙加钙调蛋白对不同品系小菜蛾脑蛋白质磷酸化都有不同程度的刺激作用;3种杀虫剂均对各品系小菜蛾的磷酸化反应有影响,杀虫双、杀螟丹表现为抑制,溴氰菊酯表现为加强。这种影响在敏感品系中表现得比抗性品系中要强烈。     

小菜蛾对杀虫双和杀螟丹抗药性遗传的DNA随机扩增多态性研究

利用室内选育119代的抗杀虫和抗杀螟丹小菜蛾Plutella xylostella品系和敏感品系,通过反复回交建立近等基因系。用Operon公司全盛的80个引物对抗杀虫比近等基因系和抗杀螟丹近等基因系小菜蛾基因组DNA进行PCR扩增;在抗杀虫双近等基因系中3个引物分别产生了1条特异扩增带,2个引物分别产生了2条特异扩增带,1个引物产生3条特异拉增带;在抗杀螟丹近等基因系中,有4个引物分别产生了1条特异扩增带,1个引物产生了2条特异扩增带,在培育近等基因系的多次回交过程中,与抗性有关的遗传因子被逐步换到敏感品系的基因组中,因此可以认为这些特异带与小菜蛾对杀虫双和杀螟丹的敏感性有关。     

小菜蛾对杀螟丹抗性遗传的研究

利用室内选育的抗杀螟丹小菜蛾Plutella xylostella (L.)品系和敏感品系研究了该品系的抗性遗传形式,结果表明,小菜蛾对杀螟丹的抗性形式为常染色体多基因遗传,并呈不完全显性。该品系对６种常用杀虫剂的抗性谱测定结果表明,对杀虫双有较严重的正交互抗性;对敌敌畏、杀扑磷有低度交互抗性;对溴氰菊酯、灭多威和叶蝉散等药剂无交互抗性。还发现该品系对杀螟丹的抗性与乙酰胆碱酯酶和羧酸酯酶活性无关。     

杀虫双和杀螟丹选育对小菜蛾抗药性的形成及其抗性机制

用杀虫双和杀螟丹在实验室以点滴法处理小菜蛾Plutella xylostella L.四龄幼虫,以连续继代药剂淘汰选育其抗药性。至35代,药剂汰选的小菜蛾对杀虫双和杀螟丹的抗药性较选育前正常品系分别提高了51倍和25倍。其抗药性的形成发展均呈S形,可认为已成为抗性品系。以有机磷、氨基甲酸酯、拟除虫菊酯及有机氮等11种杀虫剂测试抗杀虫双小菜蛾品系和抗杀螟丹小菜蛾品系对常用药剂的敏感度结果表明：对杀虫双、杀螟丹和杀虫环之间有较严重的正交互抗性;对敌敌艮、马拉硫磷和杀螟松有轻微交互抗性产生;对溴氰菊酯、氯氰菊酯、氯菊酯和灭多威、久效威等药剂更加敏感,呈负交互抗性。用聚丙烯酰胺凝胶电泳(PAGE)法测定表明,抗药性产生与特异性酯酶的形成有一定关系。用比色法和酸度法测定,抗性品系的乙酰胆碱酯酶(AchE)活性降低,羧酸酯酶(CarE)活性无差异。加增效剂Pb和SV₁：于四龄幼虫表皮,对抗杀虫双小菜蛾晶系分别有6.28及1.45倍的增效作用;对抗杀螟丹小菜蛾品系分别有4.85及1.39倍的增效作用,可见多功能氧化酶(MFO)为小菜蛾抗杀虫双和抗杀螟丹的重要因子。     

小菜蛾对苦皮藤素抗性选育及交互抗性测定

敏感小菜蛾Plutella xylostella (L.)经苦皮藤素20代的抗性选育,其抗性增长21.57倍。选育的小菜蛾抗性品系对杀虫双、杀螟丹和叶蝉散分别有4.63、4.11和3.71倍的交互抗性;对溴氰菊酯、氯菊酯、氯氰菊酯分别有0.22、0.01和0.26倍的负交互抗性。高抗杀螟丹、杀虫双小菜蛾品系对苦皮藤素无明显交互抗性,而高抗溴氰菊酯小菜蛾品系对苦皮藤素有3.61倍的交互抗性。     

小菜蛾对阿维菌素的抗性遗传方式 和相对适合度研究

就小菜蛾Plutella xylostella对阿维菌素的抗性遗传方式和抗性品系的相对适合度进行了研究。室内选育的阿维菌素抗性品系与同源的敏感品系杂交、F₁代自交、F₁代与亲本回交,结果表明：杂交后的显性度（D）分别为-0.64和-0.52,说明小菜蛾对阿维菌素的抗性是常染色体、不完全隐性遗传;χ²检验证实,可能是多基因控制的抗性遗传。杂交F₁代乙酰胆碱酯酶（AChE）、羧酸酯酶（CarE）和谷胱甘肽转移酶（GST）活性比抗性亲本有所降低,F2代及回交后代三种酶的活性继续降低。种群适合度研究表明,抗性品系相对于敏感品系有0.84的适合度。     

小菜蛾对定虫隆抗性种群的选育及交互抗性研究

用昆虫生长调节剂(insect growth regulator,简称IGR)定虫隆(chlorfluazuron)对源自深圳田间的小菜蛾(SZ-S)Plutella xylostella(L.)在室内进行抗性种群选育,经过8代饲养和6次药剂汰选,获得抗性种群(CH-R),与相对敏感种群Sz—S比较,抗性指数(R1)为23.78倍。CH-R种群在去除选择压力条件下饲养5代,抗性逐渐下降。抗性汰选前后分别测定了10种药剂的剂量—死亡毒力回归线,发现CH—R抗性种群对三氟氯氰菊酯、氯氰菊酯、辛硫磷、喹硫磷、灭多威、磺胺脲类衍生物—杀螨隆、微生物杀虫剂Bt和齐墩螨素无明显交互抗性,抗性倍数为0.4-1.7;对两种沙蚕毒素类杀虫剂杀螟丹和杀虫丹的敏感性却有所上升,有负交互抗性趋势。活体增效剂试验表明,增效醚(PBO)和三苯基磷酸酯(TPP)对定虫隆均有一定的增效活性,PBO的增效比最高为29倍,能够完全抑制对定虫隆的抗性,说明多功能氧化酶可能是主导抗性机制之一。     

Functional and genetic characteristics of Chlorantraniliprole resistance in the diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae)

The diamondback moth (Plutella xylostella) is a globally distributed and important economic pest, and it has developed resistance to all conventional insecticide classes used in the field. Chlorantraniliprole is a new chemical class of insecticide that acts as a conformation‐sensitive activator of the insect ryanodine receptor (RyR). In the present study, a field strain (16.3‐fold resistance to chlorantraniliprole) was collected in Korea and lab‐selected with chlorantraniliprole for more than one year. The resulting strain presented 2,157‐fold resistance to chlorantraniliprole. A point mutation (G4946E) in the RyR gene was observed at a high frequency in the resistant strain. Enzyme assays indicated that glutathione S‐transferase (GST) and P450 activity in the resistant strain were 2.4‐ and 1.96‐times higher than that of the susceptible strain, respectively. The expression of the RyR, GST (sigma, omega, and zeta) and CYP321E1 gene was higher in the resistant strain than in the susceptible strain. The F₁ progeny resulting from reciprocal crosses did not reveal maternal effects or a diamide‐susceptible phenotype, which suggests an autosomal nearly recessive mode of inheritance. In addition, we surveyed the susceptibility to 13 insecticides (3 diamides, 2 synthetic pyrethroids, 2 spinosyns, 1 organophosphate, 1 oxadiazine, 1 avermectin, and 3 others) in the chlorantraniliprole‐resistant strain. The resistant strain exhibited high cross‐resistance to flubendiamide (5,910 fold) and showed no cross‐resistance to spinetoram, spinosad, indoxacarb, and metaflumizone. These results can serve as an important basis for guiding the use of insecticides in the field.     

Abamectin resistance in strains of vegetable leafminer,Liriomyza sativae (Diptera: Agromyzidae) is linked to elevated glutathione S‐transferase activity

Abamectin resistance was selected in the vegetable leafminer, Liriomyza sativae (Blanchard) (Diptera: Agromyzidae) under laboratory conditions, and cross‐resistance patterns and possible resistance mechanisms in the abamectin‐resistant strains (AL‐R, AF‐R) were investigated. Compared with the susceptible strain (SS), strain AL‐R displayed 39‐fold resistance to abamectin after 20 selection cycles during 25 generations, and strain AF‐R exhibited 59‐fold resistance to abamectin after 16 selection cycles during 22 generations. No cross‐resistance to cyromazine was found in both abamectin‐resistant strains. However, we failed to select for cyromazine resistance in L. sativae under laboratory conditions by conducting 17 selection cycles during 22 generations. However, moderate levels of cross‐resistance to abamectin (6–9 fold) were observed in strains which received cyromazine treatments. Biochemical analysis showed that glutathione S‐transferase (GST) activity in both abamectin‐resistant strains (AL‐R, AF‐R) was significantly higher than in the susceptible strain (SS), suggesting metabolically driven resistance to abamectinin L. sativae. Recommendations of mixtures or rotation of cyromazine and abamectin should be considered carefully, as consecutive cyromazine treatments may select for low‐level cross‐resistance to abamectin.     

小菜蛾对九种杀虫剂的抗药性

采用叶片药膜法测定了9种杀虫剂对海南、山东、广东、湖北武汉和襄樊等5个地区小菜蛾Plutella xylostella(L.)田间种群的毒力,和室内相对敏感品系比较。结果显示,5个地区的小菜蛾种群都对氯氰菊酯产生了10倍以上的抗性,广东和山东种群达到30倍以上的抗性。对阿维菌素的抗性山东小菜蛾种群达到135倍,广东种群达到30倍,其他地区均在10倍以下。但是,作用机制与阿维菌素类似的药剂,氟虫腈的抗性5个地区均在5倍或5倍以下。对昆虫生长调节剂定虫隆和氟虫脲的抗性除广东种群分别在10倍和20倍以上外,其他地区均在5倍以下。广东种群对多杀菌素的抗性为7.8倍,其他地区的种群没有产生抗性,多杀菌素和杀虫双均为作用于乙酰胆碱受体的药剂,但是作用的位点不同。杀虫双对5个种群的LC50值均是比较大的(>800 mg/L),尽管没有明显的抗性,不适宜用于小菜蛾田间防治。对呼吸链电子转移抑制剂溴虫腈的抗性均在10倍以下。对辛硫磷的抗性在2~5倍。     

STABILITY OF ORGANOPHOSPHATE RESISTANCE IN CULEX PIPIENS PALLENS COQUILLETT UNDER RELAXATION OF INSECITICDE SELECTION*

Abstract The stability of resistance in the organophosphate resistant strain of Culex pipiens pallens with amplification of single gene coding for esterase B₂ was determined under relaxation of insecticide selection. Insecticide resistance and amplification of esterase were both lost when strains were not homozygous for the presence of amplified gene, probably due to biological disadvantage of the esterase gene. LC₅₀ and LC₉₅ of chlorpyrifos for this strain were 0. 2099 mg/L and 0.9036 mg/L in the F₀ generation respectively, but 0. 0207 mg/L and 2. 8027 mg/L respectively in the F₁₆ generation. In the homozygous strain, insecticide resistance was still stable and amplification of esterase remained after 16 generations under relaxation of insecticide selection, which indicated that copy of esterase gene was not lost in the offspring of this strain. The evolution of insecticide resistance in mosquitoes is discussed.     

Transgenic broccoli expressing aBacillus thuringiensis insecticidal crystal protein: Implications for pest resistance management strategies

We usedAgrobacterium tumefaciens to transform flowering stalk explants of five genotypes of broccoli with a construct containing the neomycin phosphotransferase gene and aBacillus thuringiensis (Bt) gene [CryIA(c) type] optimized for plant expression. Overall transformation efficiency was 6.4%; 181 kanamycin-resistant plants were recovered. Of the 162 kanamycin-resistant plants tested, 112 (69%) caused 100% morality of 1st-instar larvae of aBt-susceptible diamondback moth strain. Southern blots of some resistant transformants confirmed presence of theBt gene. Selected plants that gave 100% mortality of susceptible larvae allowed survival of a strain of diamondback moth that had evolved resistance toBt in the field. F₁ hybrids between resistant and susceptible insects did not survive. Analysis of progeny from 26 resistant transgenic lines showed 16 that gave segregation ratios consistent with a single T-DNA integration. Southern analysis was used to verify those plants possessing a single T-DNA integration. Because these transgenic plants kill susceptible larvae and F₁ larvae, but serve as a suitable host for resistant ones, they provide an excellent model for tests ofBt resistance management strategies.     

Pleiotropy of insecticide resistance in the codling moth, Cydia pomonella

The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), has developed resistance to various insecticides. Relative fitness of one susceptible strain (Sv) and two strains selected for resistance to diflubenzuron (Rt) and deltamethrin (Rv), respectively, was measured in the absence of insecticide selection pressure. Mating rate, fecundity, fertility, developmental time, fifth instar weight, and adult longevity were compared. Both resistant strains were less fecund and fertile, developed more slowly, weighed less, and had shorter life-spans than the susceptible strain. These results indicate that biological constraints are associated with insecticide resistance in the codling moth. We also found that fitness estimates of the Rv strain did not differ statistically from those of the Rt strain. Enhanced mixed-function oxidase and glutathione-S-transferase activities have been shown to be involved in insecticide resistance in both Rt and Rv strains. This suggests that the fitness cost described in both resistant strains was mainly associated to metabolic resistance. The impact of such deleterious pleiotropy of insecticide resistance in C. pomonella in terms of resistance management in the field is discussed.     

Selection for resistance to acetamiprid and various other insecticides in the diamondback moth, Plutella xylostella (L.) (Lep., Plutellidae)

Abstract:  A diamondback moth (DBM) colony (KOBII) originally collected from cabbage fields of Iwaoka-cho in Kobe City, Japan, where acetamiprid is the primary insecticide for DBM control was selected for studies on resistance to acetamiprid, phenthoate, esfenvalerate and cartap, using the leaf-dipping method. Resistance to acetamiprid was slow to develop in KOBII compared with the other insecticides. Three selection experiments each with phenthoate and esfenvalerate yielded a resistance ratio (RR) of 140 and 222, respectively, in the selected strains compared with a part of KOBII that was not exposed to any insecticide (KOBII-NS). Three selection experiments with cartap also gave an RR of 15. However, six selection experiments with acetamiprid yielded an RR of just 9.5 in the selected strain. When cross-resistance to acetamiprid was evaluated, the cartap-resistant strain showed a moderate level of cross-resistance to acetamiprid. In the case of the phenthoate-resistant strain, a very low cross-resistance to acetamiprid was observed. There was however no discernible cross-resistance to acetamiprid in the esfenvalerate-resistant strain.     

Mode of Inheritance of Amitraz Resistance in a Brazilian Strain of the Southern Cattle Tick, Boophilus microplus (Acari: Ixodidae)*

The southern cattle tick, Boophilus  microplus (Canestrini), has developed resistance to amitraz in several countries in recent years. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the mode of inheritance of amitraz resistance with cross-mating experiments. The Muñoz strain, a laboratory reared acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. A modified Food and Agriculture Organization Larval Packet Test was used to measure the levels of susceptibility of larvae of the parental strains, F₁, backcross, F₂, and F₃ generations. Results of reciprocal crossing experiments suggested that amitraz resistance was inherited as an incomplete recessive trait. There was a strong maternal effect on larval progeny’s susceptibility to amitraz in both the F₁ and the subsequent generations. The values of the degree of dominance were estimated at ?0.156 and ?0.500 for the F₁ larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F₁ backcrossed with the resistant parent strain and that of the F₂ generations suggested that more than one gene was responsible for amitraz resistance in the Santa Luiza strain. Comparisons of biological parameters (engorged female weight, egg mass weight, and female-to-egg weight conversion efficiency index) indicated significant differences between different genotypes. The differences appeared to be heritable, but not related to amitraz resistance. Results from this study may have significant implications for the management of amitraz resistance.     

Tebufenozide resistance is associated with sex‐linked inheritance in Plutella xylostella

The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of cruciferous crops. Tebufenozide, a novel nonsteroidal ecdysone agonist, exhibits good efficacy and has played an increasingly important role in the control of Lepidopteran pests in China. For its resistance management, the genetic basis of tebufenozide resistance was studied using a laboratory selected resistant strain of DBM (resistant ratio, RR = 268). A series of crosses with laboratory susceptible and resistant strains revealed that tebufenozide resistance in this pest was partially biased toward female heredity, with a large difference in RR for F₁ (RR = 29) and rF₁ progeny (RR = 147). The dominance calculated for these 2 cross progeny was ?0.788 and 0.09, respectively. Further analysis showed that the susceptible male and female larvae were similar in their sensitivity to tebufenozide, but the resistant female larvae showed significantly higher resistance than the resistant male larvae. The heredity of tebufenozide resistance in DBM might be linked with the W sex chromosome, which suggested that DBM has the ability to develop high levels of resistance to tebufenozide. This is the first report of sex‐linked inheritance of tebufenozide resistance in P. xylostella (L.).     

Rapid selection and characterization of Cry1F resistance in a Brazilian strain of fall armyworm

Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thuringiensis Berliner, has been used for management of the fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), in Brazil since 2009. A strain of S. frugiperda, obtained from field collections of larvae in TC1507 maize in Minas Gerais state in 2010, was selected in the laboratory for resistance to Cry1F using leaves of TC1507 maize in two selection regimes. Continuous exposure of larvae to Cry1F was more effective than exposure for 6, 8, and 10 days in the selection of resistant S. frugiperda individuals. With only four generations of laboratory selection, a strain with high levels of resistance to Cry1F was obtained, as indicated by the survival of insects reared on leaves of TC1507 maize plants and by the more than 300‐fold resistance level measured in bioassays with the purified Cry1F protein. Importantly, reciprocal crosses between control and the Cry1F‐selected strains revealed that the resistance is autosomal and incompletely recessive, and the response obtained in the backcross of the F₁ generation with the resistant strain was consistent with simple monogenic inheritance. Additionally, there were no apparent fitness costs associated with resistance either for survival or larval growth on non‐Bt maize leaves. Our findings provide experimental evidence for rapid evolution of Cry1F resistance in S. frugiperda in the laboratory and further reinforce the potential of this species to evolve field resistance to the TC1507 maize as previously reported. The resistant strain isolated in this study provides an opportunity to estimate the resistance allele frequency in the field and to determine the biochemical and molecular basis of the resistance, which should provide further information to assist in the resistance management of S. frugiperda on transgenic maize producing B. thuringiensis proteins.     

艾氏剂环氧化酶及细胞色素P-450对小菜蛾抗药性发展的影响

本文对室内长期饲养的小菜蛾(Plutella xylostella L.)敏感品系和田间采集的抗性种群体内的艾氏剂环氧化酶及细胞色素P-450进行了比较研究。结果证明,艾氏剂环氧化酶在感性和抗性小菜蛾间存在着量及质的差异。 抗性种群的艾氏剂环氧化酶的Vmax和Km值分别为感性品系的5.4倍和6.5倍。抗性种群的细胞色素P-450的含量是感性品系的1.1-1.3倍。艾氏剂环氧化酶在量上及质上的差异及细胞色素P-450含量的提高是导致小菜蛾抗药性发生与发展的重要机制之一。而且质的差异较之量的差异可能起着更为重要的作用,     

Pas1 is a common lung cancer susceptibility locus in three mouse strains

Inherited predisposition to lung cancer is a phenotypic trait shared by different mouse inbred strains that show either a high or an intermediate predisposition. Other strains are instead genetically resistant. The Pas1 locus is the major determinant of lung cancer predisposition in the A/J strain (Gariboldi et al. 1993). To define the determinants of susceptibility to lung tumorigenesis in the highly susceptible SWR/J and in the intermediately susceptible BALB/c mice, we analyzed (BALB/c × SWR/J)F₂ and (BALB/c × C3H/He)F₂ crosses by genetic linkage experiments. The present results provide unequivocal evidence that the same Pas1/+ allele that leads to lung cancer predisposition is shared by A/J, SWR/J, and BALB/c strains. The intermediate susceptibility of the BALB/c strain would result by interaction of Pas1 locus with lung cancer resistance loci. Received: 18 April 1997 / Accepted: 15 June 1997