Cloning and Sequencing of a Gene Encoding Aminopeptidase N in the Midgut of Helicoverpa armigera(Hubner)

A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACE technique in this paper. The full-length of APN-Harm is 3 043 bp. Open reading frame is 2 856 bp in length, encoding 951 amino acid residues. Its predicted molecular weight and isoelectric point are 108. 3 kDa and 5.29, respectively. This deduced amino acid sequence shares some common structural features with aminopeptidase N from several moth species, including the consensus zinc-binding motif HEXXHX18E and the GAMEN motif common to gluzincin aminopeptidases. The first 20 amino acid residues at N-termini is hydrophobic transmembrane helix. The sequence of APN-Harm was deposited in GenBank and the accession number is AY181026.

Effect of Selected Insecticides on Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) on Tomato (Lycopersicon esculentum Miller) and Their Successful Management

Helicoverpa armigera H. is a polyphagous notorious pest of a number of economic crops including tomato. In tomato, it can reduce its yield as high as 70% due to fruit boring. Although some predators and parasitoids have been recorded in the field for their natural control, their manipulation to a successful crop management is still awaited. To overcome this pest, insecticides play a significant role in its effective crop management program globally. Tomato is an absolutely perishable commodity and needs the specified period required between the application, of conventional pesticides of organophosphate group, and consumption which can hardly be afforded. Therefore, nine insecticides from the new chemistries, which were relatively safer to human and environment, i.e. Coragen (chlorantraniliprole), steward (indoxacarb), Belt (flubendamide), Delegate (spintoram), Volium Flexy (chlorantraniliprole + thiamethaxim), Fipronil (grafter), Proclaim (emamectin benzoate), Pirate (chlofenapyr) and Lufenuron (lufenuron), were tested on tomato crop heavily infested by this pest in the farmers field in districts Lodhran and Bahawalpur (Pakistan) in the month of April 2014. The mortality was compared after 4, 7 and 10 days and treatments were compared with control as well with one another. The average maximum % mortality, i.e. 89.36 and 85.09 of the pest, was observed with Volium Flexy, and Delegate also worked well even after 7 and 10 days. Similarly, the results on the basis of damaged fruits and percent loss of yield pointed out that Chlorantraniliprole, Flubendiamide and Indoxacarb had resulted better as compared with others, although the difference was statistically non significant. These new chemistry pesticides are suggested to be used at tomato crop against Helicoverpa armigera which are best fitted in the IPM program for the control of the pest.

Relationship between Temperature and Occurrence of Adult Helicoverpa armigera(Hübner)

[Objective] We aimed to investigate the relationship between temperature and occurrence of Helicoverpa armigera （H0bner） in Shihezi Zong Chang. [Method] The data of occurrence of adult Helicoverpa armigera （Hubner） and temperature in 2007-2010 was analyzed for building the prediction model. [Result] The amount of overwintering generation of Helicoverpa armigera （Hubner） increased with the in- crease of temperature and its population dynamics showed a positive correlation with temperature. The amount of first and second generation of Helicoverpa armigera （Hubner） decreased with the increase of temperature and its population dynamics showed a negative correlation with temperature. [Conclusion] This research would provide a basis for the forecast and prevention of Helicoverpa armigera （Hubner） in North Xinjiang.

不同光波及光强度下棉铃虫(Helicoverpa armigera)成虫的行为反应

运用室内行为实验方法研究了不同波长单色光和白光刺激的棉铃虫[Helicoverpaarmigera(H櫣ｂｎｅｒ）］成虫趋光、避光反应及其与光强度之间的相互关系?峁允?:(1)一定时间暗适应的棉铃虫蛾对单色光和白光刺激均具有趋光行为反应 ,依其趋光反应率大小 ,其光谱反应曲线在340 -605nm波谱范围内 ,表现为大小较复杂的几个峰 ,其中两个峰分别在蓝光区483nm、紫外340nm ,另在近紫外的400nm、绿黄光区的538nm有峰。(2)一定光强度范围内 ,棉铃虫蛾的趋光反应率随单色光和白光光强度的增强而增大 ,至一定光强度时增加变缓 ,呈现近似S型曲线式样。(3)性别、日龄对棉铃虫蛾的光谱、光强度反应曲线均有一定影响 ,雌性较雄性的趋光反应率和可适应的光强度高 ,1、3、5日龄蛾中以3日龄的趋光反应率和可适应的光强度高。 (4)棉铃虫蛾的趋光、避光反应行为具有相似的波长及光强度选择机制。

光因子对棉铃虫(Helicoverpa armigera)雌蛾性信息素产生及其求偶行为的影响

分为 3部分 :第 1部分为光因子对棉铃虫雌蛾求偶行为的影响 ;第 2部分为光因子对棉铃虫雌蛾性信息素 ( Z-1 1 -1 6:Ald)含量的影响 ;第 3部分为光因子对棉铃虫雌蛾脑因子活性的影响。研究结果表明 :用非正常光周期、间隔光照、不同光质处理棉铃虫雌蛾后 ,求偶峰值下降 ,求偶节律混乱 ,性信息素含量下降 ,脑因子活性受到抑制。主要研究目的是探讨一种不使用或少使用农药的新型生态防治方法。

棉铃虫(Helicoverpa armigera)幼虫性特征和精子的发生

以棉铃虫精巢为材料,气干法制片,Giemsa染色,研究了棉铃虫精巢的形态变化和精子发生的动态过程。结果表明:棉铃虫的精巢自预蛹期开始会发生融合现象;雄性棉铃虫染色体数目为n=31。还发现雌性幼虫的腹部有4个腺体,可以作为幼虫的性别特征。

棉铃虫[Helicoverpa armigera(Hübner)]蛾复眼视网膜电位研究

运用视网膜电位（Ｅｌｅｃｔｒｏｒｅｔｉｎｏｇｒａｍ ， ＥＲＧ） 技术研究了棉铃虫［ Ｈｅｌｉｃｏｖｅｒｐａ ａｒｍｉｇｅｒａ（Ｈüｂｎｅｒ）］ 成虫在暗适应过程中对单色光和复合光（ 即白光） 刺激的光感受性变化， 并初步分析了ＥＲＧ 的波形成分组成及影响因素。结果显示：（１） 依ＥＲＧ 振幅大小（ 峰－ 峰值） ，棉铃虫蛾的光谱敏感曲线在３４０ － ６０５ｎｍ 波谱内有三个大小不等的峰，其中第一峰位于绿－ 黄光区的５６２ｎｍ ，第二峰位于蓝光区的４８３ｎｍ ，第三个峰在近紫外区的４００ｎｍ ，显示其复眼至少具有三种感受器类型。（２） 一定光强度范围内， 一定日龄的棉铃虫蛾的ＥＲＧ 值随单色光和白光刺激光强度的增强而增大， 至一定光强度时增加变缓，呈近似Ｓ 型曲线式样。表明该蛾复眼具有较强的光强度自调节和适应机制。（３） 棉铃虫蛾复眼的ＥＲＧ 波形成分由４ 个部分组成：开光反应、正相电位、持续负电位和闭光反应，波长、光强度及光刺激时程对其各成分有不同影响。

棉铃虫Helicoverpa armigera的环境友好型诱杀技术研究

2003年和2004年，在山东惠民采用杀虫灯、性诱剂、杨树枝把等措施开展了诱杀棉铃虫Helicoverpa armigera成虫的防治试验。杀虫灯＋10支性诱剂，杀虫灯＋5支性诱剂＋5支杨树枝把，杀虫灯＋10支杨树枝把，杀虫灯4种措施的日均诱蛾量分别为11．4、10．4、8．6和7．0头，性诱剂和杨树枝把均未显著提高对棉铃虫的诱杀效果。不同诱捕方案对棉铃虫累积诱蛾量随日期的变化符合Gompertz方程。普通灯和时控灯对棉铃虫的日均诱蛾量分别为19．9头和17．4头，二者差异不显著；时控灯对异色瓢虫和龟纹瓢虫的日均诱捕量均少于普通灯；减少开灯时间可在不显著降低棉铃虫诱杀量的同时，有效保护捕食性瓢虫。与普通灯相比，设置时间合理的时控灯（夜间19：00—23：00和3：00—5：00开灯）未显著降低棉铃虫的日均诱蛾量，而对异色瓢虫和龟纹瓢虫的日均诱捕量分别为2．0头和4．9头，均显著低于普通灯的3．7头和9．2头，合理设置开关灯控制模式可在不显著降低对棉铃虫诱杀效果的同时，降低近50％的天敌杀伤率，节约用电40％。

棉铃虫Helicoverpaarmigera(Hübner)精母细胞减数分裂研究

采用压片法HSG显带技术,对棉铃虫精母细胞减数分裂进行了研究,描述了其减数分裂各时期的特征。同 时通过C-带处理表明棉铃虫染色体单倍数目为n=31,染色体长度从2.05μm至6.20μm不等。

棉铃虫(Helicoverpa armigera)FK506结合蛋白基因的克隆、表达与纯化

为了深入了解FK506结合蛋白基因的作用和探索调控棉铃虫CYP6B6的表达机制,基于单酵母杂杂交结果采用RT-PCR的方法从棉铃虫的中肠cDNA中扩增得到了FK506结合蛋白基因,将测序正确的目的片段克隆至原核表达载体pET32a中,在大肠杆菌BL21中用异丙基-β-D-硫代半乳糖(IPTG)诱导表达,通过镍柱离子亲和层析纯化目的蛋白。用SDS-PAGE检测目的蛋白的表达和纯化结果,并用Western blotting进行验证。结果表明,克隆得到的目的基因大小为327 bp,编码108个氨基酸残基,预测蛋白质分子量和等电点分别是11.78 kD和7.87。氨基酸序列分析表明该序列具有完整的开放阅读框,且没有信号肽。重组质粒pET32a-FKBP在大肠杆菌BL21中获得表达,主要以可溶性形式存在,经亲和层析柱纯化获得目的蛋白,Western blotting检测发现纯化的目的蛋白大小正确且纯度高。

土壤特性对棉铃虫(Helicoverpa armigera Hübner)发生的影响及发生程度分区

统计分析表明 ,土壤的多种理化性质与棉铃虫的发生程度间存在着密切的关系 ,其中起主要作用的是土壤容重、砂粒含量和粘粒含量 3个因子 ,其直接通径系数依次为 -0 .652 1、0 .673 5和 0 .3 4 56。这些因子使土壤表现出不同的粘性 ,决定了土壤的透气性、持水量和板结程度等 ,从而影响到棉铃虫幼虫入土后蛹的存活率。用上述 3个因子进行聚类分析 ,可将通州市划分为 3个棉铃虫发生区 ,各区域在测报和防治上应进行区别对待。

棉田与玉米田棉铃虫(Helicoverpa armigera Hübner)种群动态特征的比较

1998年在江苏丰县系统调查了棉田和玉米田中第三、四代棉铃虫的种群动态 ,用生命表方法比较了两种寄主田内棉铃虫种群的发育速率和存活率。结果表明 ,棉铃虫卵峰期至 4龄幼虫高峰期的历期在玉米田要比棉田快 4 d,并以产卵于玉米抽雄扬花期的发育最快。两类寄主田内平均存活率无显著差异 ,但产卵于玉米抽雄扬花期时 ,其卵至 6龄幼虫的存活率比产在其它阶段玉米上的存活率高。在江苏棉区目前的作物布局和耕作制度下 ,玉米田第三代棉铃虫是棉田第四代棉铃虫的重要虫源。

交配和黑光灯处理对棉铃虫Helicoverpa armigera生物钟基因cryptochromes mRNA表达的影响

[目的]明确交配和黑光灯处理对棉铃虫生物钟基因cryptochromes mRNA表达的影响。[方法]应用实时定量PCR(SYBR Green)技术检测不同条件下棉铃虫cryptochromes(cry1和cry2)基因的表达。提取棉铃虫头部总RNA,经DNase I消化后进行反转录合成cDNA,并采用特异性引物分别对cry1、cry2和EF-1α基因进行实时定量PCR扩增。[结果]黑光灯处理棉铃虫2h,其cry1mRNA的表达量明显降低;cry2mRNA的表达量小于对照,但两者之间差异性并不显著。交配对棉铃虫cry1和cry2mRNA表达均存在显著影响,并且雌、雄两性cry1和cry2mRNA表达量在交配后随时间延长呈下降趋势。[结论]该结果对进一步研究cry基因的功能以及棉铃虫防治具有重要意义。

Cloning and Sequence Analysis of Proteasome β5 Fragment in Helicoverpa armigera

[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).

石榴棉铃虫(Heliothis armigera(Hubner)发生规律及其防治研究

:石榴棉铃虫 (Heliothisarmigera (Hubner) ,属鳞翅目 (Lepidoptera) ,夜蛾科 (Noctuidae) ,是棉花的主要害虫 ,但也是云南蒙自石榴的主要害虫之一。但其发生危害规律尚不清楚 ,致使防治效果不佳。于 1 998～ 1 999年对该虫发生规模及综合防治进行了研究 。

Cryptochromes mRNA Expression Under Mating and Black light Treatment on Helicoverpa armigera

[ Objective ] The paper was to confirm the effect of mating and black light treatment on cryptochromes mRNA expression of Hclicoverpa armigera. [ Method ] Quantitative real-time PCR （ SYBR Green） technique was applied to detect the expression of cryptochromes gene （ cryl and cry2 ） of H. armigera under different conditions. Total RNA was extracted from the head of H. armigera, and carried out reverse transcription to synthesize cDNA after digested by DNase I. Specific primers were used to carry out quantitative real-time PCR on cryl, cry2 and EF-Ict gene, respectively. [ Result] The expression of cryl mRNA of H. ar- migera significantly decreased after exposure to black light for 2 h, the mRNA expression of cry2 was smaller than control, they had no significant difference with each other. Mating had significant effect on mRNA expression of cryl and cry2 of H. armigera, and the mRNA expression of cryl and cry2 of male and female adults showed decreasing trend with the prolongation of time after mating. [ Conclusion] The result had important significance for further studying on function of cry gene and the control of cotton bollwonn.

河北省棉铃虫(Helicoverpa armigera Hübner)种群数量的气象综合预报方法的研究

为了客观地反映棉铃虫种群数量变化与气候背景的关系规律 ,创建了棉铃虫气象多时段综合因子的因子组建方法 ;建立了贡献度权重修正的气象距离指标方法和模型 ;匹配以传统的多元回归模型 ,进行棉铃虫种群数量的气象监测和预报 ;将其自身生物潜能和气象条件影响相结合 ,建立了棉铃虫灾害的生物气象定量综合预报模型。在棉铃虫发生的气象条件评价和发生程度预测预报应用中 ,趋势准确率达到 93 %。

The Sublethal Effects of Chlorantraniliprole on Helicoverpa armigera (Lepidoptera: Noctuidae)

Sublethal effects of chlorantraniliprole to Helicoverpa armigera （Hiibner） larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed on the insecticide-containing diet for 7 d, the LC10, LC20, LC40, and LC50values were 3.790, 7.977, 21.577, and 33.121 μg active ingredient L^-1, respectively. Chlorantraniliprole at sublethal concentrations significantly reduced the larval body mass, emergence ratio, adult longevity and egg hatching rate in both the parental and offspring generations. The pupation and copulation rate in the parental generation and the pupal mass in the offspring also strongly decreased. Reproduction was seriously disturbed in both the parental and offspring groups even when only one of the partners was exposed to chlorantraniliprole at larval stages. However, at the lowest concentration of exposure （LC^0）, offspring fecundity was strongly reduced only when both partners were exposed. The net reproductive rate （R0）, intrinsic rate of increase （rm）, and gross reproduction rate （GRR） at LC20, LC40 concentrations were significantly lower than that of the control. Post-exposure effects also included an extended larval developmental time and increased male proportion in both generations. The doubling time （Dt） at LC20, LC40 concentrations as well as gross reproduction rate （GRR） at LC10 concentration were also significantly increased. Chlorantraniliprole might have significant effects on H. armigera population dynamics even at sublethal concentrations on both parental and offspring generations.

Structure,Binding Characteristics,and 3D Model Prediction of a Newly Identified Odorant-Binding Protein from the Cotton Bollworm,Helicoverpa armigera (Hübner)

The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,encoding a protein with 147 amino acids,namely HarmOBP5.HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography.SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics.Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles,including （E）-β-farnesene,ethyl butyrate,ethyl heptanoate,and acetic acid 2-methylbutyl ester.Based on three-dimensional （3D） model of AaegOBP1 from Aedes aegypti,a 3D model of HarmOBP5 was predicted.The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H.armigera.This study provides clues for better understanding physiological functions of OBPs in H.armigera and other insects.

Response of successive three generations of cotton,bollworm,Helicoverpa armigera (Hübner),fed on cotton bolls under elevated CO_2

The growth, development and consumption of successive three generations of cotton bollworm, Helicoverpa armigera （Htibner）, fed on cotton bolls grown under elevated CO2 （double-ambient vs. ambient） in open-top chambers were examined. Significant decreases in protein, total amino acid, water and nitrogen content and increases in free fatty acid were observed in cotton bolls. Changes in quality of cotton bolls affected the growth, development and food utilization of H. armigera. Significantly longer larval development duration in three successive generations and lower pupal weight of the second and third generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower fecundity was also found in successive three generations of H. armigera fed on cotton bolls grown under elevated CO2. The consumption per larva occurred significant increase in successive three generations and frass per larva were also significantly increased during the second and third generations under elevated CO2. Significantly lower relative growth rate, efficiency of conversion of ingested food and significant higher relative consumption rate in successive three generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower potential female fecundity, larval numbers and population consumption were found in the second and third generations of cotton bollworm fed on cotton bolls grown under elevated CO2. The integrative effect of higher larval mortality rate and lower adult fecundity resulted in significant decreases in potential population consumption in the latter two generations. The results show that elevated CO2 adversely affects cotton bolls quality, which indicates the potential population dynamics and potential population consumption of cotton bollworm will alleviate the harm to the plants in the future rising CO2 atmosphere.