转Bt cry1Ah基因抗虫玉米对亚洲玉米螟、棉铃虫和黏虫的抗性评价

利用农杆菌介导法将Bt cry1Ah基因转入玉米自交系综31,获得对玉米螟有显著抗性的转基因玉米HGK60,为了研究其对鳞翅目害虫的杀虫活性,在室内和田间分别用亚洲玉米螟、棉铃虫和黏虫幼虫对HGK60玉米的杀虫效果进行检测。室内生物活性检测结果表明,取食HGK60玉米心叶的亚洲玉米螟幼虫在3 d后死亡率达100%;HGK60玉米对棉铃虫幼虫有毒杀作用,玉米不同部位对棉铃虫幼虫的杀虫效果存在差异;与非转基因玉米对照相比,取食HGK60玉米叶片一周后的黏虫幼虫体重增长明显受到抑制。田间生物活性检测结果表明,HGK60玉米对于亚洲玉米螟和棉铃虫有很强的杀虫活性,达到高抗级别,对于黏虫有一定的杀虫活性,为抗性级别。     

植物次生物质对烟青虫和棉铃虫食物利用及中肠解毒酶活性的影响

用分别添加0-5%(干重比)棉酚、烟碱、番茄苷和辣椒素4种植物次生物质的人工饲料饲养烟青虫Helicoverpa assulta和棉铃虫H. armigera 5龄幼虫48 h,测定这些次生物质对烟青虫和棉铃虫的营养效应和中肠谷胱甘肽S-转移酶（GST）及羧酸酯酶（CarE）活性的影响。结果表明：在实验浓度下,棉酚可显著降低烟青虫的相对消化率,但对棉铃虫却有助食作用;番茄苷抑制烟青虫的取食和生长,对其近似消化率和食物利用率也有显著的抑制作用,但食物转化率有明显升高,对棉铃虫的各营养指标无显著影响;烟碱对烟青虫和棉铃虫的相对生长率均无影响;辣椒素使烟青虫的取食量有大幅度的提高,对棉铃虫的取食量无影响,但引起其相对消化率的提高。由此可见,棉铃虫对4种次生物质有普遍的适应性,而烟青虫只对寄主植物所含的烟碱和辣椒素有较好的适应性。烟青虫和棉铃虫幼虫中肠CarE活性不受4种次生物质的影响,烟碱和辣椒素对烟青虫GST有显著的诱导作用,番茄苷对烟青虫GST活性则有抑制作用,4种次生物质对棉铃虫GST均无显著影响。     

实夜蛾属二近缘种对寄主植物次生物质的反应:次生物质对幼虫生长和食物利用的影响

实夜蛾属(Heliothis)的棉铃虫(H. armigcra)和烟青虫(H. assulta)是近缘种,幼虫期取食多种相同的植物,其中含有不同的次生物质.本项工作是在人工饲料中分别加入0.5%的烟碱、番茄苷、棉子酚、丹宁酸等饲养早期六龄的幼虫,测定它们对生长和食物利用的影响.结果表明这些次生物质对两种幼虫有不同的作用:烟碱对棉铃虫没有明显影响,但对烟青虫的取食却有一定的刺激作用.丹宁酸、棉子酚、番茄苷可抑制两种幼虫的生长,而以番茄苷抑制烟青虫的生长最为显著.番茄苷主要通过抑制取食来影响幼虫的生长,而丹宁酸和棉子酚则具有降低消化率的作用.通过次生物质对这两种幼虫效应的比较可知,棉铃虫有较大的忍耐力.     

棉铃虫齿唇姬蜂的寄生对棉铃虫体内主要代谢物质的影响

寄生蜂是调节寄主害虫种群数量的一个重要因子.棉铃虫齿唇姬蜂Campoletis chorideae Uchida是棉铃虫Helicoverpa armigera Hübner和烟青虫Helicoverpa assulta Guenée的主要寄生性天敌,对两种害虫的田间自然控制起着非常重要的作用.本文选取棉铃虫齿唇姬蜂-棉铃虫系统为研究对象,从齿唇姬蜂的寄生对棉铃虫体内主要代谢物质的影响等方面进行了研究.试验结果显示,棉铃虫齿唇姬蜂的寄生使棉铃虫幼虫组织中糖原含量发生了明显的变化,与未被寄生的同龄棉铃虫幼虫相比,被寄生幼虫组织中的糖原含量下降了近一倍,被寄生的棉铃虫幼虫血淋巴和虫体组织中蛋白质、脂肪的含量均显著下降.     

玉米螟和棉铃虫卵壳结构特征的电镜观察

玉米螟Ostrinia nubilalis属鳞翅目螟蛾科,幼虫为害玉米、大麻、泽麻、甜菜、粟等,蛀食茎内,使作物受害甚烈。棉铃虫Heliothis armigera属鳞翅日夜蛾科,于幼虫期蛀害棉铃,是使棉花减产的重要害虫。如何消灭玉米螟和棉铃虫于卵期,即消灭于其危害之前,是人们关切的问题。近年我国许多省市利用赤眼蜂防治玉米螟、棉铃虫等,已取得显著成绩。在我国东北和华北等地区的许多省市日前已大面积推广应用赤眼蜂防治玉米螟,放蜂后对玉米螟卵块的寄生率可达96％,卵粒寄生率可达85％以上。在利用赤眼蜂防治棉铃虫卵方面也取得初步成效,据报导在吉林、湖北和山西等省利用赤眼蜂防治棉铃虫卵,寄生率达70—100％,放蜂后蕾铃被害率减少1—2倍,百株虫口也     

番茄是烟青虫的寄主植物吗？

长期以来,国内有关文献一直将番茄列为烟青虫Helicoverpa assulta的寄主植物,但田间调查结果往往与此矛盾,本研究的目的是阐明番茄是否为烟青虫的寄主植物。室内试验表明,虽然烟青虫成虫在盆栽番茄植株上产卵较多,但无论是用番茄离体嫩叶还是用盆栽植株饲养,初孵幼虫绝大部分在1龄死亡,平均存活时间很短,不会对番茄造成有经济意义的危害;即使用番茄青果饲养的3龄幼虫也不能活到6龄。根据人工饲料饲养结果估测,番茄苷对初孵幼虫的致死中浓度LC₅₀为0.0744%,叶和青果中报道的番茄苷含量已接近或超过此水平,这是幼虫不能存活的主要原因之一。田间调查也表明,烟青虫并不危害番茄。因此,番茄不是烟青虫的寄主植物。文献中的记载很可能是将棉铃虫误判为烟青虫。     

光对棉铃虫和烟青虫杂交的影响

【目的】力求建立一种准确鉴定室内棉铃虫Helicoverpa armigera(Hübner)、烟青虫H.assulta(Guenée)及其杂交种的分子技术,探讨棉铃虫和烟青虫杂交的可能。【方法】室内暗期设置0.5 lx黑光灯和白炽灯,测定不同配对模式下3日龄处女棉铃虫和烟青虫的杂交率;筛选可区分室内建立的棉铃虫、烟青虫及杂交种家族的微卫星位点;于架设有黑光灯的温室内释放棉铃虫和烟青虫混合种群,利用微卫星分子标记技术检测子代微卫星位点大小。【结果】两种蛾类在任何一种配对模式下均可杂交,混合种群处理杂交率为2.92%,且均为烟青虫雌蛾与棉铃虫雄蛾配对交配;黑光灯、白炽灯和黑暗条件下杂交率无显著性差异;筛选出的Har SSR1、Har SSR9和Har SSR10在两种蛾类上的等位基因片段大小不一样;两年温室实验共鉴定360头子代,混合种群中未检测到杂交后代。【结论】交配笼中棉铃虫和烟青虫能进行种间杂交,弱光不能提高二者杂交率;混合种群处理只发现一种杂交配对模式,说明了棉铃虫雄蛾的交配竞争能力更强;成功利用微卫星分子标记技术鉴定室内建立的棉铃虫、烟青虫及杂种家族,提供了一种简单准确的分子鉴定手段;两年温室实验未检测到杂交种,说明2种蛾类之间存在着交配前生殖隔离机制。     

雄性棉铃虫和烟青虫对雌性信息素的触角电生理反应

利用触角电位图(Electroantennogram,EAG)技术,比较研究了二近缘种棉铃虫、烟青虫对其性信息素主要成分Z—11—16：Ald、Z—9—16：Ald的触角电生理反应。剂量反应曲线表明,对Z—11—16：Ald,棉铃虫和烟青虫均有明显的EAG反应,且随浓度的增加而增强,但棉铃虫比烟青虫的反应较强;对Z—9—16：Ald,烟青虫有很强的EAG反应,棉铃虫的反应则很弱;对Z—11—16：Ald和Z—9—16：Ald以97：3和7：93比例形成的混合物,棉铃虫、烟青虫均有EAG反应,但二者之间没有显著差异[动物学报49(6)：795～799,2003]。     

棉铃虫和烟青虫幼虫下颚栓锥感器对某些化合物反应特性的比较

利用电生理学技术,以两个食性有差异的近缘种棉铃虫和烟青虫为研究对象,比较研究了6龄幼虫下颚上两对栓锥感器对刺激取食素和抑食素的感受机理.结果表明:(ⅰ)两种幼虫栓锥感器对蔗糖和印楝素敏感的细胞均主要分布在侧栓锥感器,对肌醇敏感的细胞主要分布于中栓锥感器;(ⅱ)烟青虫幼虫中栓锥感器对肌醇刺激的反应敏感性强于棉铃虫;(ⅲ)在测试的6种抑食素中,仅印楝素在两种昆虫的侧栓锥感器上可诱导高的神经脉冲;(ⅳ)抑食素对刺激取食素引发栓锥感器神经脉冲有抑制作用,其中对于棉铃虫,辣椒素对蔗糖引发脉冲的抑制率最大,番茄苷对肌醇引发脉冲的抑制率最大,棉酚对蔗糖和肌醇引发脉冲的抑制率均为最小;对于烟青虫,单宁、棉酚和番茄苷对蔗糖引发脉冲均有较大的抑制率,棉酚对肌醇引发脉冲的抑制率最大,烟碱对蔗糖和肌醇引发脉冲的抑制率均为最小.     

球孢白僵菌在棉铃虫表皮上萌发的研究

在扫描电子显微镜下,观察了球孢白僵菌三株不同毒力菌株的分生孢子在棉铃虫幼虫表皮表面的萌发,发现高、低毒力菌株的分生孢子都能在棉铃虫幼虫表皮上萌发和生长,但高毒力菌株的萌芽管伸长至分生孢子长径的1.5～2.5倍时能发生定向生长,垂直向里穿透表皮,低毒力菌株的萌芽管呈蔓延生长,极少有穿透表皮的发生,偶见极度伸长的菌丝穿透表皮。此外,还发现球孢白僵菌穿透菌丝穿透表皮的过程不是垂直向里一步完成的。     

Genetic differentiation of Helicoverpa armigera (Hübner) and H. assulta (Guenée) (Lepidoptera: Noctuidae) based on AFLP markers

Abstract Here we use amplified fragment length polymorphism (AFLP) to assess genetic differentiation of Helicoverpa armigera and H. assulta . The results indicated that both species-specific fingerprints and cluster analysis showed the ability of AFLP technique to discriminate the two sibling species; among a total 1963 AFLP markers amplified from nine primer combinations: 777 (39.6%) were H. armigera -specific, 602 (30.7%) were H. assulta -specific, and 584 (29.7%) were common bands. The mean number of H. armigera -specific bands was significantly more than that of H. assulta -specific bands for nine primer combinations ( P < 0.05); the intraspecific distance of H. armigera and H. assulta was 0.123 0 and 0.110 7 respectively, and the interspecific distance was 0.178 3. In addition, the percentage of polymorphic loci and estimated average heterozygosity were used to estimate genetic diversity of the two species. This study therefore demonstrates that AFLP analysis is a sensitive and reliable technique to study genetic differentiation and genetic relationships between species and provides sufficient molecular markers for future linkage map construction, location and eventual cloning of genes involved in traits differentiation.     

Comparative study on the responses of maxillary sensilla styloconica of cotton bollwormHelicoverpa armigera and Oriental tobacco budwormH. assulta larvae to phytochemicals

Using the electro-physiological technique, the sensory mechanisms of maxillary sensilla styloconica to stimulants and deterrents were explored on two closely related species, the generalist Helicoverpa armigera and the specialist H. assulta. The results showed that: (i) in both species, cells sensitive to sucrose and azadirachtin were mainly in the lateral sensillum styloconicum, and those to inositol were in the medial sensillum styloconicum; (ii) sensitivity of medial sensillum styloconicum in H. assulta to inositol was higher than that in H. armigera; (iii) among 6 tested deterrents, only azadirachtin evoked high impulse discharge from the lateral sensillum styloconicum in both insects; (iv) the deterrents could disturb stimulants evoking impulse discharge from maxillary sensilla styloconica of both species in different degrees: To sucrose evoking impulses on lateral sensillum styloconicum, for H. armigera capsaicin had a strong inhibition and gossypol had a weak inhibition, while for H. assulta tann     

棉铃虫不同寄主种群遗传分化的RAPD分析

应用RAPD技术对生活于辣椒,烟草和番茄3种不同寄主上的棉铃虫（Helicover pa armigera)种群,并以烟草烟青虫（H.assulta)为外群进行了种群遗传分化的研究,研究结果表明：4个处理内个体间最大的遗传距离均小于处理间最小的遗传距离,烟青虫的遗传距离为0．177－0．346,棉铃虫的辣椒种群为0．289－0．404,烟草种群为0．396－0．505,番茄种群为0．329－0．382,采用UPDGA法进行聚类,并构建系统发育树。各处理成聚的遗传距离为：烟青虫是0．334,棉铃虫的辣椒种群是0．372,烟草种群是0．463,番茄种群是0．360。各处理均首先成聚,棉铃虫与烟青虫成聚的遗传距离（0．703）,稍大于棉铃虫各种群的成聚值（0．639）。上述结果表明,棉铃虫不同寄主种群已经存在明显的遗传分化。     

斜纹夜蛾和烟青虫在烟草上的生态位

斜纹夜蛾和烟青虫同是烟草上食叶类的重要害虫,对烟叶产量和品质均可构成严重的影响.研究两者在烟草上的生态位和种间关系,结果表明斜纹夜蛾的空间生态位宽度和空间生态位重叠指数均略大于烟青虫,两者空间生态位的比例相似性为0.0973,说明两种害虫在空间资源上几乎不存在竞争.斜纹夜蛾和烟青虫的时间生态位及时间生态位重叠指数很接近,时间生态位的比例相似性高达0.9485.斜纹夜蛾和烟青虫在烟草植株上、中、下部叶片共存的比例分别为6.11%、3.23%和0.51%.由于对空间垂直资源的不同选择,使它们可以同时在同株烟草上为害而互不干扰.     

Using COI gene sequence to barcode two morphologically alike species: the cotton bollworm and the oriental tobacco budworm (Lepidoptera: Noctuidae)

Due to limited morphological difference, the two closely related sister species, the cotton bollworm, Helicoverpa armigera (Hübner) and the oriental tobacco budworm, H. assulta (Guenée) (Lepidoptera: Noctuidae), are very difficult to distinguish, especially at the larvae stage. Recently, DNA sequence has been widely used as a bio-barcode for species identification. In this study, we attempted to distinguish H. armigera and H. assulta using the mitochondrial cytochrome C oxidase subunit I gene (COI) gene sequence as the barcode. We determined a 658 bp segment of the COI gene for 28 individuals of H. armigera, 8 individuals of H. assulta, and 10 individuals of Mamestra brassicae (as the outgroup) in Yunnan Province, southwest of P. R. China, together with one H. assulta and two H. armigera reported sequences from GenBank. Twenty-three haplotypes were identified in all 49 samples. As expected, network analysis of the haplotypes of the three species presented a clustering pattern consistent with the respective species status. Haplotypes of the same species differed from each other by no more than three nucleotide substitutions. However, each haplotype of H. armigera differed from that of H. assulta by at least 22 nucleotide substitutions. Both species differed from M. brassicae by more than 50 nucleotide substitutions. 17 unique diagnostic nucleotides were also used to discriminate the two species. The finding of large COI sequence differences between H. armigera and H. assulta suggested that it could be used to distinguish the two morphologically alike species and be employed for quick species identification during pest control.     

Pheromone biosynthetic pathways in the moths Helicoverpa zea and Helicoverpa assulta

Sex pheromones of many Lepidopteran species have relatively simple structures consisting of a hydrocarbon chain with a functional group and usually one to several double bonds. The sex pheromones are usually derived from fatty acids through a specific biosynthetic pathway. We investigated the incorporation of deuterium-labeled palmitic and stearic acid precursors into pheromone components of Helicoverpa zea and Helicoverpa assulta. The major pheromone component for H. zea is (Z)11-hexadecenal (Z11-16:Ald) while H. assulta utilizes (Z)9-hexadecenal (Z9-16:Ald). We found that H. zea uses palmitic acid to form Z11-16:Ald via delta 11 desaturation and reduction, but also requires stearic acid to biosynthesize the minor pheromone components Z9-16:Ald and Z7-16:Ald. The Z9-16:Ald is produced by delta 11 desaturation of stearic acid followed by one round of chain-shortening and reduction to the aldehyde. The Z7-16:Ald is produced by delta 9 desaturation of stearic acid followed by one round of chain-shortening and reduction to the aldehyde. H. assulta uses palmitic acid as a substrate to form Z9-16:Ald, Z11-16:Ald and 16:Ald. The amount of labeling indicated that the delta 9 desaturase is the major desaturase present in the pheromone gland cells of H. assulta; whereas, the delta 11 desaturase is the major desaturase in pheromone glands of H. zea. It also appears that H. assulta lacks chain-shortening enzymes since stearic acid did not label any of the 16-carbon aldehydes.     

Genetic basis of sex pheromone blend difference between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

The two closely related moth species, Helicoverpa armigera and H. assulta, are sympatric in China. Both species use a mixture of (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as their sex pheromones but in widely different ratios. Hybridization and backcrossing experiments between H. armigera and H. assulta were conducted and sex pheromone compositions of the parent species, their F(1) hybrids and backcrosses were compared to study the genetic basis of the production of their sex pheromone blend composition. Results show that the difference in sex pheromone blend ratios of these Helicoverpa species is mainly controlled by an autosomal locus with two alleles, with the allele from H. armigera being almost completely dominant over that derived from H. assulta.     

Comparative study of the cuticular hydrocarbon composition of Melipona bicolor Lepeletier, 1836 (Hymenoptera, Meliponini) workers and queens

In social insects, cuticular hydrocarbons are involved in species, kin, caste and nestmate recognition. Gas chromatography and mass spectrometry were used to compare the cuticular hydrocarbon composition of workers, males and queens of Melipona bicolor. The cuticular hydrocarbon composition of this species was found to consist mainly of C23, C25:1, C25, C27:1, C27, C29:1 and C29, which are already present in imagoes that have not yet abandoned the brood cell. This composition varied quantitatively and qualitatively between and within the castes and sexes. The newly emerged workers and young queens (virgins) had similar cuticular hydrocarbon profiles, which were different from those of the males. When the females start executing their tasks in the colony, the cuticular hydrocarbon profile differences appear. The workers have less variety, while the queens conserve or increase the number of cuticular hydrocarbon compounds. The queens have more abdominal tegumentary glands than the workers, which apparently are the source of the new cuticular compounds.     

Evidence for no sexual isolation between Drosophila albomicans and D. nasuta

Sexual isolation, the reduced tendency to mate, is one of the reproductive barriers that prevent gene flow between different species. Various species‐specific signals during courtship contribute to sexual isolation between species. Drosophila albomicans and D. nasuta are closely related species of the nasuta subgroup within the Drosophila immigrans group and are distributed in allopatry. We analyzed mating behavior and courtship as well as cuticular hydrocarbon profiles within and between species. Here, we report that these two species randomly mated with each other. We did not observe any sexual isolation between species or between strains within species by multiple‐choice tests. Significant difference in the courtship index was detected between these two species, but males and females of both species showed no discrimination against heterospecific partners. Significant quantitative variations in cuticular hydrocarbons between these two species were also found, but the cuticular hydrocarbons appear to play a negligible role in both courtship and sexual isolation between these two species. In contrast to the evident postzygotic isolation, the lack of sexual isolation between these two species suggests that the evolution of premating isolation may lag behind that of the intergenomic incompatibility, which might be driven by intragenomic conflicts.     

湖北恩施烟夜蛾幼虫田间分布格局的地统计学分析

烟夜蛾以幼虫取食烟叶,给烟叶生产带来巨大损失.运用地统计学的方法,研究了湖北恩施烟区烟夜蛾幼虫的空间结构和发生动态.结果表明： 恩施烟区烟夜蛾幼虫在烟叶移栽后40 d开始为害,至烟叶成熟前期达到高峰;球形 指数套合模型为烟夜蛾幼虫田间分布的主要模型,属聚集〖JP2〗分布;空间变异度C/(C₀+C)均大于0.75,说明其结构性变异较大,随机性变异较小;烟夜蛾幼虫在烟叶旺长期进行了较大迁移,之后其数量趋于稳定,烟叶成熟期尤其稳定.