害虫对氟虫腈的抗药性研究进展

氟虫腈是第一个商品化的苯基吡唑类杀虫剂,其作用机理是抑制γ-氨基丁酸(GABA)受体氯离子通道。氟虫腈为防治一些世界性的重大害虫包括对以前使用的杀虫剂具有严重抗性的害虫作出了重要贡献。然而,随着氟虫腈的大量使用,害虫对该药的抗药性问题受到日益关注。至今,重要的农业害虫如小菜蛾,烟粉虱、褐飞虱、白背飞虱,二化螟及斜纹夜蛾等已经对氟虫腈产生抗药性。文章就害虫对氟虫腈的抗药性发生概况、交互抗性及抗药性机理策略进行综述,以期为氟虫腈的合理使用提供理论基础。     

三唑磷、氟虫腈及其复配剂对四种miRNAs在斑马鱼组织中表达的影响

MicroRNAs(miRNAs)是一类长约22 nt的非编码小RNA,在基因表达中起重要调控作用.已有研究表明,农药三唑磷和氟虫腈能影响斑马鱼全鱼组织中部分miRNAs的正常表达,但未见对miRNA表达的组织特异性的研究.本研究采用荧光定量PCR技术研究了经三唑磷微乳剂、氟虫腈微乳剂及其复配剂处理后,四种miRNA(...     

氟虫腈对小菜蛾幼虫体内解毒酶的影响

本文从解毒酶系的角度研究了小菜蛾Plutella xylostella(L.)对氟虫腈的短期生理生化响应。用氟虫腈LC50剂量处理小菜蛾3龄幼虫一定时间后,测定小菜蛾幼虫体内多功能氧化酶(MFO)、酯酶(EST)和谷胱甘肽S-转移酶(GST)活性的动态变化。研究结果表明:经氟虫腈处理后小菜蛾体内解毒酶的活性均显著高于未经处理的对照,并且随着处理时间的延长,解毒酶活性逐渐提高。这说明氟虫腈对小菜蛾敏感品系的解毒酶具有一定的诱导作用,反之,解毒酶加快了小菜蛾体内对氟虫腈的代谢,降低了小菜蛾对氟虫腈的敏感性。     

五种植物精油对氟虫腈的增效与渗透促进作用

采用点滴法测定了花椒Zanthoxylum bungeanum Maxim,艾蒿Artemisia grgyi Levl. Et Vant,马尾松针Pinus massoniana Lamb,侧柏Platycladus orientalis(L.)Franco,黄蒿Artemisia annua L. 5种植物精油与氟虫腈联用对小菜蛾2龄幼虫的触杀毒力.研究结果表明,当植物精油浓度为1μL·mL-1时对氟虫腈的LC50均无显著增效作用,当上述精油浓度含量为10μL·mL-1时,艾蒿与侧柏氟虫腈能显著提高氟虫腈对小菜蛾幼虫的毒杀效果;致死中时(LT50)测定结果表明,当植物精油浓度为lpLL·mL-1时,仅艾蒿精油能缩短氟虫腈对小菜蛾幼虫的LT50,但当植物精油浓度为10μL·mL-1时,所有精油均能缩短氟虫腈对小菜蛾幼虫LT50;松针精油和侧柏精油能提高氟虫腈透皮吸收量,其余精油对氟虫腈没有显著促进渗透作用.     

土荆芥精油预处理后小菜蛾幼虫对氟虫腈敏感度的变化

采用微量点滴法测定了土荆芥Chenopodium ambrosioides精油预处理前后氟虫腈对小菜蛾Plutella xylostella 2龄幼虫的触杀作用,采用生化分析方法研究了土荆芥精油预处理前后小菜蛾幼虫体内乙酰胆碱酯酶（AChE）、羧酸酯酶（CarE）和谷胱甘肽-S-转移酶（GSTs）的比活力。研究结果表明,1μL·mL^-1和10μL·mL^-1的土荆芥精油预处理可增加小菜蛾幼虫对氟虫腈的敏感度,LC50分别为氟虫腈单用时的0.66倍和0.44倍;当用1μL·mL^-1和10μL·mL^-1土荆芥精油预处理小菜蛾2龄幼虫2h后,再用氟虫腈致死中量点滴处理,小菜蛾存活幼虫体内AChE、CarE和GSTs比活力均低于氟虫腈单独处理组。研究结果说明土荆芥精油预处理对氟虫腈的增效作用可能与主要抗性相关酶系活性变化有关。     

QuEChERS-GCMS测定液态乳中氟虫腈及其代谢物残留量

采用QuEChERS净化模式,通过气相色谱质谱仪（GCMS）建立了液态乳中氟虫腈及其代谢物残留量检测方法,并对两种不同净化材料的净化效果进行比较。样品由乙腈提取,QuEChERS净化,浓缩复溶后,通过GCMS进行检测,外标法定量。结果表明：氟虫腈及其代谢物残留量在2.00~200ng/mL内呈良好线性,线性相关系数（R2）均大于0.999,检出限为 0.400~1.00μg/kg;氟虫腈及其代谢物残留量在4 个添加水平（2.50、5.00、20.0、80.0μg/kg）下的加标回收率为 74.4%~120.2%,相对标准偏差（RSD,n=3）为1.0%~4.9%,小于10%。该方法准确、简单、快速,可适用于液态乳中氟虫腈及其代谢物残留量的同时测定。     

氟虫腈、吡虫啉作为黑翅土白蚁诱杀药剂的效果

毒力测定结果表明,0.025~0.4μg/mL氟虫腈和吡虫啉分别在药后3 d和5 d对黑翅土白蚁Odontotermes formosanus表现出明显的毒杀效果,氟虫腈和吡虫啉药后1 d的LC50分别为药后5 d的509倍和63.8倍,2种药剂对黑翅土白蚁的毒杀效果均比较缓慢。毒性传递试验表明,0.5μg/g毒沙处理白蚁1 h后,氟虫腈和吡虫啉的致死毒性均可被传毒白蚁传递给受毒白蚁。驱避作用试验表明,50μg/mL氟虫腈对黑翅土白蚁无明显的驱避作用,而50μg/mL吡虫啉对黑翅土白蚁表现出了明显的驱避作用。可见,2种供试药剂中,氟虫腈是较理想的白蚁诱杀药剂。     

湖北省褐飞虱对吡虫啉、噻嗪酮及氟虫腈的抗药性监测

采用稻茎浸渍法监测湖北省武穴、天门、监利、通城、孝感、公安及枣阳七地褐飞虱Nilaparvata lugens(Sta1)田间种群对吡虫啉、噻嗪酮、氟虫腈的抗药性。结果表明:2006、2007和2008年3年7地褐飞虱种群对吡虫啉的抗性分别为98.89～389.19倍、69.00～153.33倍和56.32～116.89倍,达高水平至极高水平抗性,但3年期间总体抗性水平呈现一定程度的下降趋势;2007和2008年2年7地褐飞虱对噻嗪酮的抗性分别为13.39～41.06倍和6.94～20.44倍,达低水平至高水平抗性;2007和2008年2年7地褐飞虱对氟虫腈抗性分别为0.97～2.64倍和0.63～1.42倍,为敏感阶段。此外,还对褐飞虱的治理进行讨论。     

miRNA-155对氟虫腈作用下斑马鱼ZF4细胞存活的影响

microRNA（miRNA）是一类长约20～24 nt的非编码单链小分子RNA. 环境毒理学研究表明,当生物暴露于环境化学物质时,会引起相关miRNA表达发生变化,进而调节基因的表达. 农药氟虫腈是一种苯基吡唑类杀虫剂,能影响斑马鱼中miR-155的正常表达,但尚未见异位表达的miR-155对斑马鱼细胞影响的研究. 本研究明确了异位表达的miR-155对氟虫腈作用下斑马鱼胚胎细胞ZF4存活的影响. 结果显示,氟虫腈处理ZF4细胞72 h的IC50值为39.05 μmol/L,瞬时转染miR-155 mimic,显著降低了ZF4细胞氟虫腈暴露的存活率.同时real-time PCR结果显示,其潜在靶基因cyb561d2表达显著降低;而转染miR-155 inhibitor,抑制细胞内源性的miR-155则可显著增强ZF4细胞抵抗氟虫腈细胞毒性的能力,同时cyb561d2表达则显著增高.以上结果表明,miR-155参与氟虫腈毒理学效应,其表达量的高低可以影响氟虫腈作用下ZF4细胞的存活率,而其可能的途径之一是通过潜在靶基因cyb561d2的表达调控来实现. 因此,miR-155可作为氟虫腈环境毒性的潜在的生物标志物,有望成为致癌环境化学物质对生物体作用机制研究的新途径.     

氟啶虫胺腈对意大利蜜蜂解毒、免疫和记忆相关基因表达的亚致死效应

【目的】新型杀虫剂氟啶虫胺腈广泛用于农业生产中的害虫防治,但其对蜜蜂健康影响的研究较少。本文探究了亚致死剂量的噻虫啉对意大利蜜蜂Apis mellifera ligustica 6种解毒相关基因(CYP9Q1、CYP9Q2、CYP9Q3、CYP9S1、CYP6AS5和CYP4G11)、4种免疫相关基因(Abaecin、Defensin、Hymenoptaeci和Apidaecin)和3种记忆相关基因(PKA、GluRA和NMDAR1)表达的影响。【方法】采用饲喂法确定氟啶虫胺腈对蜜蜂的经口急性毒性;并用1/3 LD_(50)和1/10 LD_(50)氟啶虫胺腈饲喂蜜蜂后48 h,运用荧光定量PCR技术分别检测了蜜蜂脑部解毒、免疫和记忆相关基因的表达变化,并统计各组蜜蜂的死亡率。【结果】氟啶虫胺腈对蜜蜂经口性半数致死剂量LD_(50)为0.099μg/蜂;饲喂1/3 LD_(50)氟啶虫胺腈能够显著抑制蜜蜂Abaecin和Defensin的表达(P 0.05),而PKA、NMDAR1、GluRA、CYP9Q2和CYP9Q3能够被其诱导上调表达(P 0.05);饲喂1/3 LD_(50)和1/10 LD_(50)氟啶虫胺腈均能显著诱导蜜蜂Hymenoptaecin、Apidaecin、CYP6AS5和CYP4G11上调表达(P 0.05)。但饲喂1/3 LD_(50)和1/10 LD_(50)氟啶虫胺腈的处理均不影响蜜蜂CYP9Q1和CYP9S1表达。【结论】氟啶虫胺腈的曝露可能会对意大利蜜蜂的免疫、记忆和解毒系统都有一定的影响,这对深入探究氟啶虫胺腈与蜜蜂之间互作的分子机制有一定生物学意义。     

抗二化螟的水稻品种筛选

【目的】筛选抗二化螟Chilo suppressalis水稻品种资源,明确其抗虫机理,是农业防治的基础。【方法】采用室内人工接虫为害和大田自然受害相结合的方法,鉴定了不同的水稻品种抗性;同时通过目测不同水稻植株外部形态特征、在网罩内供试品种上接入成虫后2 d的产卵量、幼虫孵化后2 d观察稻株内二化螟的存活率,探讨抗虫品种的抗虫机理。【结果】从344个水稻品种资源和当家品种中筛选出"碑田倒"和"银间杂糯"2个抗虫品种,以及"农飞"和"三香糯"等65个中抗品种,进一步对鉴定出的2个抗虫品种和65个中抗品种进行大田验证,发现符合率达88.06%;其中,抗虫品种"碑田倒"和"银间杂糯"稻株叶舌、叶耳紧贴,叶鞘紧包,脉间距较窄,其植株上的产卵量和幼虫存活率显著少于感虫品种"香稻"、"赣南早"、"三粒寸"、"赣优晚"及中抗品种"赛湖早"、"TKM6"。【结论】在籼稻、粳稻和糯稻中均可筛选出抗虫和中抗品种。其中,糯稻中的抗虫品种和中抗品种较多,占糯稻品种的33.3%;粳稻品种次之,中抗品种占20%,籼稻品种较少,为18.83%。其抗虫机理主要是稻株外部形态特征不利于二化螟蚁螟侵入、成虫产卵量少和幼虫生长发育不良、存活率低。     

二化螟人工饲料关键因子的优化及其优化配方的饲养效果

采用5因子4水平正交试验,将作者在以前筛选出的二化螟Chilosuppressalis(Walker)人工饲料的5个关键因子进行了进一步的优化,并得出了二化螟人工饲料的优化配方,其成分如下:稻茎粉18.75g、大豆粉15.00g、麦芽粉15.00g、稻糠粉6.25g、茭白茎粉7.50g、干酪素10.00g、酵母粉18.75g、纤维素7.50g、蔗糖15.00g、葡萄糖7.50g、维生素C7.50g、复合维生素B3.00g、胆固醇0.375g、氯化胆碱0.25g、Beck氏盐2.50g、山梨酸1.35g、琼脂13.49g和自来水809.25g。通过连续3代继代饲养试验,比较了该优化配方和二化螟天然饲料水稻茎的饲养效果。结果表明,在一些关键指标上(如存活率、化蛹率和羽化率等),所优化的二化螟人工饲料配方的饲养效果明显的好于其天然饲料水稻茎的;而在许多其它的指标上(如蛹期、成虫期、产卵量、卵期和孵化率等),该优化配方对二化螟的饲养效果与水稻茎的相当;但该配方饲养的二化螟在第3代时幼虫历期明显的比水稻茎的有所延长。     

用正交试验法优选二化螟人工饲料配方

采用15因子2水平正交试验,优选出1种二化螟Chilo suppressalis(Walker)人工饲料配方,筛选出影响二化螟发育的5个关键因子,分别是稻茎粉、麦芽粉、稻糠粉、酵母粉和蔗糖。其中稻茎粉、麦芽粉、酵母粉和稻糠粉影响二化螟的幼虫成活率,蔗糖和麦芽粉影响二化螟的幼虫历期,稻茎粉影响二化螟的蛹重。     

杀虫剂不同处理对二化螟为害、防治效果及水稻产量的影响

近年来,二化螟发生出现各代均偏重或大发生的新特点。为探索杀虫剂防治二化螟的新策略,采用田间小区试验在早晚稻田中分别研究5%氟虫腈加25%灭.水胺的不同施药时期和次数对二化螟幼虫的为害和防治效果及对水稻产量的影响。在试验地,二化螟第1和第2代发生在早稻田,第3和第4代发生在晚稻田。试验结果表明,在第1和第3代二化螟幼虫孵化高峰施药1次、隔10d再施药1次对早稻田和晚稻田的幼虫防效显著,分别达到了92.4%和92.6%。在早、晚稻田分别发生的第1和第3代2龄幼虫高峰期施药1次、同时在次代孵化高峰期再施药1次,均可显著降低虫伤株率和白穗株率,提高保穗效果,并显著提高单位面积上的水稻产量(3.98%~4.20%)。因此,对第1至第4代二化螟幼虫均进行1次药剂防治是二化螟各代均大发生的新形势下的防治策略。     

Molecular characterization and functional analysis of the vitellogenin receptor in the rice stem borer,Chilo suppressalis

As a member of the low-density lipoprotein receptor (LDLR) superfamily, vitellogenin (Vg) receptor (VgR) is responsible for the uptake of Vg into developing oocytes and is a potential target for pest control. Here, a full-length VgR complementary DNA (named as CsVgR) was isolated and characterized in the rice stem borer, Chilo suppressalis. The composite CsVgR gene contained an open reading frame of 5,484 bp encoding a protein of 1,827 amino acid residues. Structural analysis revealed that CsVgR contained two ligand-binding domains (LBDs) with four Class A (LDLR_A) repeats in LBD1 and seven in LBD2, which was structurally different from most non-Lepidopteran insect VgRs having five repeats in LBD1 and eight in LBD2. The developmental expression analysis showed that CsVgR messenger RNA expression was first detectable in 3-day-old pupae, sharply increased in newly emerged female adults, and reached a peak in 2-day-old female adults. Consistent with most other insects VgRs, CsVgR was exclusively expressed in the ovary. Notably, injection of dsCsVgR into late pupae resulted in fewer follicles in the ovarioles as well as reduced fecundity, suggesting a critical role of CsVgR in female reproduction. These results may contribute to the development of RNA interference-mediated disruption of reproduction as a control strategy of C. suppressalis.     

湖北江汉稻区优质香稻上二化螟的为害特性

研究了二化螟Chilo suppressalis(Walker)对优质香稻鄂香1号和武香988及优质常规中稻鄂中5号的田间为害特性。结果显示,1代二化螟危害造成的枯心苗数优质香稻略低于优质常规中稻。2代二化螟危害造成的枯孕穗和白穗优质香稻鄂香1号低于常规中稻,平均每15丛7.91株。田间剥查发现优质香稻中4~5龄2代二化螟所占百分比高于常规中稻,武香988中2代二化螟蛹所占比例高于常规中稻,达46.51%。3代二化螟量优质香稻低于常规中稻。结果说明香稻受二化螟的危害集中在2代,因而香稻二化螟的防治应注意消灭1代老熟幼虫和2代幼虫。     

Twelve nuclear microsatellite loci for rice stem borer (Chilo suppressalis W.)

We developed 12 polymorphic microsatellite markers for the rice stem borer Chilo suppressalis (Walker). These loci were screened for 96 individuals from eight populations across China. The total number of alleles ranged from 3 to 33 and the expected heterozygosity at these loci ranged from 0.131 to 0.671. These molecular markers will be useful for fully investigating the population structure and host preference of C. suppressalis at fine spatial scales.     

二化螟与稻纵卷叶螟幼虫取食诱导的水稻挥发物比较

前期研究结果表明,二化螟和稻纵卷叶螟幼虫为害所诱导的水稻挥发物能对二化螟绒茧蜂Apanteles chilonis Munakata、二化螟Chilo suppressalis(Walker)和稻纵卷叶螟Cnaphalocrocis medinalis(Guene)雌成虫的行为产生影响,并且两者的影响效应存在明显差异,但至今仍不清楚2种害虫危害后水稻挥发物的变化情况。利用动态吸附法对健康水稻苗、二化螟或稻纵卷叶螟幼虫为害苗的挥发物进行捕集与鉴定。在3种处理水稻苗中,共捕集到28种挥发物组分,包括萜类化合物、烷烃类化合物、酮类、醇类、水杨酸甲酯和一些未知化合物。二化螟和稻纵卷叶螟为害均能导致水稻挥发物总量和大多挥发物组分释放量的上升;同时,二化螟幼虫危害能诱导水稻新释放2-heptanonol,(E)-linalooloxide和2种未知挥发物组分,稻纵卷叶螟为害则可诱导水稻产生2-heptanonol,(E)-linalooloxide,MeSA和β-caryophyllene4种新化合物。2种虫害稻株间的比较表明,β-caryophyllene和MeSA是稻纵卷叶螟危害水稻后诱导产生的特有挥发物组分,并且linalool,α-copaene,(+)-cedrol,n-heptadecane和n-nonadecane在二化螟为害苗与稻纵卷叶螟为害苗之间差异显著。这一结果也许可以解释2种害虫诱导水稻挥发物对天敌和害虫行为影响不同的原因。     

Gene cloning and expression of aminopeptidase N and cadherin from midgut of the rice stem borer,Chilo suppressalis

Abstract The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here (CsCad) showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region (CR7), which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin Cry1Ab. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.     

Identification of tissues showing the lowest tolerance to freezing in larvae of the rice stem borer, Chilo suppressalis

Abstract.  Even though overwintering larvae of the rice stem borer, Chilo suppressalis , are freeze-tolerant, they cannot survive below −30 °C. Furthermore, nondiapausing larvae cannot survive freezing. However, the cause of death due to freezing is unclear. To identify the cause of death by freezing in larvae, those tissues most injured by low temperatures are identified using the vital stain trypan blue. In overwintering larvae, the midgut of dead larvae stains blue, and remarkable colour density differences between dead and surviving larvae are observed in the midgut. In nondiapausing larvae incubated at −10 °C for several hours, the fat body of dead larvae is strongly stained. Furthermore, increases in mortality with treatment time correspond with increases in the area of the fat body stained. Sterile nondiapausing larvae with lower supercooling points, below −20 °C, do not freeze at −10 °C and survive the treatment. However, all the larvae die when subjected to inoculative freezing at −10 °C, and the fat body stains blue. These results suggest that the midgut in overwintering larvae and the fat body in nondiapausing larvae have the lowest tolerance to freezing.