Inheritance of resistance and cross resistance pattern in indoxacarb-resistant diamondback moth Plutella xylostella L.

Leaf-dip assay of Plutella xylostella against indoxacarb showed that the concentration that produced 50% mortality (LC₅₀) of indoxacarb ranged from 20.1 to 11.9 ppm, with highest in Nasik and lowest levels in Coimbatore strains. In selection studies, the LC₅₀ of indoxacarb was 18.5 ppm at generation 1 (G1), which increased to 31.3-fold (167.8 ppm) resistance after ten exposed generations (G10) as compared to unexposed. The LC₅₀ of quinalphos was 74.4 ppm, which increased to 10.0-fold (631.5 ppm) resistance after G10. The LC₅₀ of cypermethrin resistant strain resulted in an 11.5-fold increase in resistance after G10. In P. xylostella , heritability (h²) after ten generations of selection was estimated at 0.4. The number of generations required for tenfold increase in LC₅₀ (1/R) were 6.7. The response to indoxacarb selection in P. xylostella was 0.2 and the selection differential was estimated as 0.4. The phenotypic standard deviation was 0.2. Reciprocal crosses between indoxacarb-resistant and susceptible strains showed that the inheritance of indoxacarb resistance was autosomal. The degree of heritability (D_LC) (0.4, 0.4) indicated incomplete recessive inheritance of indoxacarb resistance.     

Equivalent intensity but differential dominance of sodium channel blocker insecticide resistance conferred by F1845Y and V1848I mutations of the voltage-gated sodium channel in Plutella xylostella

Two point mutations (F1845Y and V1848I) in the voltage-gated sodium channel gene of Plutella xylostella are involved in the target-site resistance to sodium channel blocker insecticides (SCBIs). The contribution of the individual mutations to the SCBI resistance and the associated inheritance modes is as yet unclear. Through 2 rounds of single-pair crossing and marker-assisted selection, 2 P. xylostella strains (1845Y and 1848I) bearing homozygous F1845Y or V1848I mutant alleles were successfully established from a field-collected population, and the contribution of each mutation to SCBI resistance, as well as associated inheritance patterns, was determined. When compared with the susceptible SZPS strain, each of the mutations individually conferred equally high-level resistance to indoxacarb (378 and 313 fold) and metaflumizone (734 and 674 fold), respectively. However, dominance levels of resistance to SCBIs were significantly different between the 2 resistant strains. Resistance of the 1845Y strain to indoxacarb and metaflumizone was inherited as an autosomal and incompletely dominant trait (D values ranged from 0.43 to 0.76). In contrast, that of the 1848I strain followed an autosomal but incompletely recessive to semidominant mode (D values: −0.24 to 0.09). Our findings enriched the current understanding of inheritance and mechanisms of SCBI resistance in P. xylostella, and will help develop resistance management programs for P. xylostella and other economic pests.     

氰氟虫腙对小菜蛾阿维菌素抗性和敏感种群的亚致死效应(英文)

在实验室条件下采用生命表技术研究了氰氟虫腙亚致死剂量（LC₂₅）对小菜蛾Plutella xylostella阿维菌素抗性（AV-R）和敏感（AV-S）种群的亚致死效应, 旨在为小菜蛾对阿维菌素的抗性治理提供理论基础。结果表明： 氰氟虫腙对小菜蛾3龄幼虫抗性种群的LC₅₀和LC₂₅分别为0.24 mg/L和0.09 mg/L; 对敏感种群的LC₅₀和LC₂₅分别为0.20 mg/L和0.07 mg/L。氰氟虫腙亚致死剂量0.09 mg/L 处理小菜蛾后, 对处理代的影响表现为显著降低处理种群的化蛹率、 蛹重、 羽化率、 繁殖力; 明显延长蛹期, 缩短成虫产卵期和寿命; 对子代种群的影响表现为显著降低卵的孵化率、 幼虫各龄期的存活率, 延长发育历期。处理种群的内禀增长率（r_m）、 周限增长率（λ）和净增值率（R₀）显著低于对照种群（P<0.0001）。亚致死剂量的氰氟虫腙对小菜蛾抗性种群的影响大于敏感种群, 对处理代种群的影响大于子代种群。氰氟虫腙亚致死剂量可以极大地影响小菜蛾尤其是阿维菌素抗性种群的种群动态, 因此氰氟虫腙对于小菜蛾的抗性治理具有积极的作用。     

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。     

小菜蛾对杀虫双和杀螟丹抗性的现实遗传力

利用室内选育119代的小菜蛾Plutella xylostella抗性品系,分析了小菜蛾对杀虫双和杀螟丹抗药性的现实遗传力,并对遗传力和杀虫剂的杀死率对抗性发展速率的影响进行了预测,结果表明,室内继代药剂选择119代后,小菜蛾对杀虫双和杀螟丹抗性的遗传力很低（h^2=0.03)。因此,小菜蛾对这两种药剂产生更高水平抗性的潜力不大,在选择的前半段（F30）,杀虫双和杀螟丹的抗性遗传力（分别为0.14t 0.11)高于选择后半段（F80)的遗传力（均为0.05),选择前、后半段的选择差数没有明显差别。即使在室内选择条件下,假如斜率=2.0(δp=0.5),h^2=0.05,群体中70%个体致死时要获得10倍的抗性,约需34代,田间条件下,由于等位基因频率的变化、环境变异等因素的影响,抗性增长10倍则需要更长的时间,因此田间条件下小菜蛾在短期内不易对杀虫双和杀螟丹产生高水平抗性。     

不同小菜蛾田间种群对氟啶脲的抗性动态

【目的】了解不同种植模式的小菜蛾Plutella xylostella(L.)田间种群对氟啶脲(Chlorfluazuron)的抗性动态,旨在为该药剂的田间抗性管理提供参考。【方法】参照中华人民共和国农业行业标准"十字花科蔬菜小菜蛾抗性监测技术规程"(NY/T 2360-2013)进行。【结果】与敏感种群相比,不同监测点小菜蛾种群由于地理位置、种植模式、用药习惯等的不同,对氟啶脲的抗性表现出地区差异和年度间差异。小户型连年种植模式的广东广州地区小菜蛾田间种群对该药剂抗性相对最高,以中等及以上抗性水平为主,最高抗性比(RR值)为437.58;而规模化连年种植模式的广东惠州地区小菜蛾种群对该药剂的抗性仅次于广东广州地区,也以中等及以上抗性水平为主,最高RR值为337.18;小户型连年种植为主的连州地区小菜蛾种群对氟啶脲的抗性动态与广东惠州地区的非常相似,抗性水平比广东广州地区和广东惠州地区稍轻;山区小户型多样化种植模式的广西柳州地区小菜蛾田间种群对该药剂的抗性相对最低,以低水平抗性为主,最高RR值仅为23.24。但4个抗性监测点也存在抗性发展大趋势基本一致的共同点。具体表现在4个监测点小菜蛾种群对该药剂的抗性在2008年均为中等以上抗性水平,随后通过调整了用药策略,多数地区2009—2011年抗性下降至低水平(RR值小于10)。2012年随着田间防治压力的上升,氟啶脲用药量和用药频次再次增加,造成多数地区抗性再次上升至中等及以上水平,并持续到2014年。【结论】小菜蛾对氟啶脲的抗性田间稳定性不高,停用或过度依赖该药剂一段时间,抗性即出现下降或上升,基本与用药情况、频次和强度等呈正相关。     

Evidence for behavioural resistance by the diamondback moth, Plutella xylostella (L.)

Abstract:  The diamondback moth (DBM), Plutella xylostella (L.) (Lep., Plutellidae), is an important pest of brassicaceous crops worldwide. It has developed resistance to almost every synthetic insecticide applied in the field and consequently is often difficult to control in crucifer-growing areas. DBM oviposition behaviour was investigated in greenhouse and growth chamber conditions with seven cruciferous and one non-cruciferous plant varieties. In free choice tests, females deposited more eggs on the stem near the soil-stem interface than on leaves. Our findings suggest that DBM is capable of developing behavioural resistance through oviposition site selection to avoid lethal doses of foliar-applied insecticides in the field.     

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.     

湖南不同地区小菜蛾对药剂敏感性比较

采用浸叶法在室内测定了湖南长沙和怀化地区小菜蛾Plutella xylostella(L.)田间种群对10种药剂的敏感性。结果表明:湖南长沙和怀化地区田间小菜蛾除对丁醚脲和BT制剂仍处于敏感(抗性倍数<3)状态外,对其它8种药剂产生了不同程度的抗药性,其中以长沙地区小菜蛾田间种群对高效氯氰菊酯抗性倍数最高(抗性倍数为33.58)。长沙和怀化种群对药剂的相对毒力倍数以巴丹最低(1.20)而以多杀菌素最高(2.59)。     

小菜蛾抗药性及其治理对策的研究进展

小菜蛾（Plutella xylostella L.）的抗药性研究是一项全球性的重要课题。本从其抗药性发展、抗药性机理研究进展、抗药性检测及研究手段的状况、抗药性治理对策等方面,对小菜蛾抗药性的研究进展进行综述。目前,对小菜蛾抗药性的分子水平上的探讨甚少。     

小菜蛾对几种常用药剂的敏感性测定

采用叶片浸溃法,在2001年10月和2002年10月分别测定了福州上街蔬菜基地的小菜蛾,对10种杀虫剂的敏感性。结果表明,上街菜区小菜蛾对灭多威、杀虫双、辛硫磷、毒死蜱、杀灭菊酯和功夫菊酯等农药极不敏感,已不能有效控制小菜蛾的危害。而小菜蛾对氟啶脲、阿维菌素、氟虫腈和虫螨腈比较敏感。通过两次毒力测定比较(2002年10月的LC50;2001年10月的LC50),结果表明,大多数杀虫剂敏感性变化较小。但小菜蛾对阿维菌素敏感性明显下降,毒力倍数达6．65倍,对氟虫腈毒力倍数相差1．48倍。此外,2002年10月小菜蛾对功夫菊酯的LC50为2001年10月的2．02倍。用浸叶法和浸溃法测定多杀菌素、氟虫腈与虫螨腈推荐浓度对小菜蛾幼虫的毒杀作用。结果表明3种药剂浸叶法处理效果均较理想。     

小菜蛾芳基酰胺酶表达及其与呋喃虫酰肼抗性的关系

以对硝基乙酰苯胺为底物,采用紫外分光光度计比色法,研究了小菜蛾Plutella xylostella(L.)体内芳基酰胺酶酶促反应的最适反应条件、各发育阶段和不同组织中芳基酰胺酶的活性及亚细胞分布及其与小菜蛾对呋喃虫酰肼抗性的关系。结果表明,小菜蛾芳基酰胺酶最适反应条件为35℃,pH7.5,底物浓度为1.2×10-3mol/L,反应时间30min。利用非变性聚丙烯凝胶电泳活性染色及SDS-PAGE法测得小菜蛾芳基酰胺酶的分子量为61.1ku。小菜蛾4龄幼虫的芳基酰胺酶活性最高。不同亚细胞层中,芳基酰胺酶活力大小依次为线粒体层>微粒体层>细胞核和细胞碎片层>可溶性部分。芳基酰胺酶主要分布在小菜蛾4龄幼虫的腹部,其余依次为表皮、头部和胸部。小菜蛾呋喃虫酰肼抗性品系芳基酰胺酶的活性是敏感品系的3.89倍,表明该酶参与了小菜蛾对呋喃虫酰肼的抗性形成。     

小菜蛾酪氨酸酶生物化学性质研究

采用时间动力学法对小菜蛾Plutella xylostella酪氨酸酶的最适反应条件、对抑制剂的敏感度以及发育期变化规律进行了研究。结果表明：小菜蛾酪氨酸酶反应的最适pH值为6.4,反应线性时间为0～2 min,在22℃和4℃下其活性分别可保持12 h 和72 h,在-20℃下至少可保持7 天。不同发育阶段小菜蛾酪氨酸酶活性的大小依次为预蛹＞4龄幼虫＞蛹＞1龄、2龄、3龄幼虫＞成虫。苯基硫脲（phenyl thiourea,PTU）对小菜蛾阿维菌素抗性品系和敏感品系酪氨酸酶活性的抑制中浓度（I₅₀）值分别为1.1796 μmol/L和1.2795 μmol/L,二者无明显差异。     

小菜蛾在温带地区越冬研究进展

小菜蛾是世界性重要害虫。在热带和亚热带地区冬季的十字花科植物上能正常发育繁殖,可见到各种虫态。但在温带冬季十字花科植物不能生长的地区,小菜蛾的越冬成为一个重要生态学问题。综述了亚洲、北美洲和欧洲小菜蛾越冬的研究进展。小菜蛾在日本的北海道、本州岛的北陆和东北大部分地区不能越冬,越冬北限相当于冬季积雪覆盖时间为60d的区域;在中国,小菜蛾不能在寒冷的东北地区越冬,在长江中下游以南地区冬季可见各虫态,但越冬北限尚不清楚;小菜蛾在冬天气候温和的韩国以及澳大利亚东南部继续发生;北美洲的加拿大西部和安大略地区大量的试验证明小菜蛾不能成功越冬,在美国南部小菜蛾冬季可正常发生,北部小菜蛾的越冬尚未见系统研究报道,但确认春季从南部运输的受小菜蛾感染的甘蓝等种苗是美国北部的重要虫源。小菜蛾在欧洲各地越冬的系统研究未见报道,没有证据表明在英国小菜蛾会发生有显著意义的越冬。目前小菜蛾越冬研究主要采用冬季直接试验观察和基于耐寒性试验的越冬预测两种方法。冬季直接试验观察法包括:(1)利用人工饲养的小菜蛾在田间各种潜在的越冬场所的越冬试验;(2)在秋播、野生或残留的十字花科植物上进行冬季种群的系统抽样调查;(3)越冬前后在前茬为十字花科植物的田块广泛搜寻普查小菜蛾的存活个体。基于耐寒性试验的越冬预测法:在获取小菜蛾越冬场所温度的基础上设计低温处理模式,试验低温处理后小菜蛾的存活率及后续发育和生殖。将试验数据和各地气温或小气候相结合,对小菜蛾在的越冬可能性进行推断。     

印楝素乳油对小菜蛾种群的控制作用

通过室内外系统试验探讨了印楝素乳油对小菜蛾Plutella xylostella种群的控制作用。结果表明,其控制作用主要表现在对成虫产卵的显忌避作用。对幼虫的拒食作用以及对生长发育的影响和减少幼虫危害为等方面,印楝素乳油0.005mL/59.0%,使小菜蛾种群趋势指数（I值）降为2.4干扰作用控制指数（IIPC）为0.1079,采用状态空间分析法就印楝素乳油施用后小菜蛾种群数量动态的模拟分析亦表明,其显的控制作用表现在种群发展初期对成虫的忌避作用。     

浙江临海小菜蛾成虫数量季节消长规律

在浙江临海相隔60km的2个十字花科蔬菜种植区,于2002~2005年利用性诱剂诱捕器每日对小菜蛾Plutella xylostella(L.)雄成虫进行诱集并计数。对两地成虫数量季节消长与种群基数、气温、雨量和日照等因子的相关性做了统计分析,建立了5个预测模型。应用这些模型可预测全年36个旬期的雄成虫数量变化,从而可预测春季高峰期并对全年的发生量和为害程度进行预警,这对提高当地小菜蛾监测预警与综合防治水平具有实践意义。     

小菜蛾对溴氰菊酯抗性相关的DNA片段的克隆与分析

采用代表性差异分析（representational difference analysis, RDA ）方法,以小菜蛾Plutella xylostella (L.)的敏感品系作为对照组,溴氰菊酯抗性近等基因系（near isogenic line）作为测试组,通过三轮消减杂交得到大小为 150～300 bp的差异扩增带,经亚克隆测序并与GenBank等数据库同源比较,发现差异片段与已知抗性相关基因无同源性;以随机选取的差异片段作探针进行Southern blot分析,在测试组扩增子和三轮差异产物中都获得了阳性结果,而在对照组扩增子中的结果是阴性的。这些结果表明测序片段可能是与抗性相关的新基因序列或调控序列。     

萜类化合物对小菜蛾幼虫的拒食活性

采用叶碟浸液法测试从松节油合成的26个不同结构萜类化合物样品对小菜蛾Plutella xylostella(L.)4龄幼虫的拒食活性。结果显示,编号为2,4,8,13,27,31的6个化合物,即诺卜醇、诺卜丙基醚、内型异莰烷基甲醇丙酸酯、4-(1-甲基乙烯基)-1-环己烯-1-乙醇丙酸酯、羟基香茅醛丙酸酯、羟基香茅醛1,2-丙二醇缩醛具有较高的拒食活性。处理有效成分为0.01g/mL时,这6个化合物24h拒食率为70%～100%。其中2,4,8号3个化合物拒食活性最高,处理有效成分为0.01g/mL和0.001g/mL时,24h拒食率分别为98.33%,99.80%,100.00%和85.60%,83.90%,66.97%;并且持效性较好,72h拒食率分别为92.67%,89.97%,98.73%和63.20%,63.30%,45.93%。     

颗粒体病毒对不同蔬菜上小菜蛾实验种群的控制作用

通过组建施用颗粒体病毒Plutella xylostellagranulosis virus(PxGV)后4种十字花科蔬菜上的小菜蛾Plutella xylostellaL.实验种群生命表发现:小菜蛾颗粒体病毒除了能引起幼虫感病死亡外,还能显著地降低成虫的产卵量,推迟产卵高蜂1~2 d,但对化蛹及雌蛾寿命无显著影响。在种群水平上,病毒对小菜蛾的控制作用综合表现为压低了小菜蛾种群增长趋势指数,在菜心、芥蓝、芥菜和小白菜上小菜蛾实验种群增长趋势指数在分别为13.7,12.3,10.7和22.7,病毒对小菜蛾种群的干扰作用控制指数(IIPC)分别为0.334,0.177,0.280和0.433,表明病毒对小菜蛾种群的控制能力受到寄主植物的影响,在芥蓝上控制能力最强,其次为芥菜和菜心,而小白菜最次。     

Management of diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae) by mating disruption

Abstract Field trials were conducted in China in 2008 and 2009 to evaluate the efficacy of mating disruption (MD) on diamondback moth, Plutella xylostella, in cabbage, Brassica oleracea var. capitata. Effectiveness was positively correlated with the MD dispenser density in the field. A density of 167 MD dispensers per ha produced an average population decrease of about 50% compared to the conventional‐practice field. Significant fewer males were captured in pheromone‐treated and conventional‐practice fields than in the blank control field, but the difference was not significant between the pheromone‐treated and conventional‐practice fields. In addition, fewer eggs and larvae were observed in pheromone‐treated fields. Our results suggest mating disruption coupled with minimal insecticidal supplements is a promising solution for resistance management and control of diamondback moth infestation.