番茄碱对棉铃虫毒性分析

本文主要研究了番茄碱对棉铃虫的毒性作用．实验结果表明：（1）用不同浓度的番茄碱（tomatine）浸泡的棉花叶饲喂棉铃虫时，随浓度的增加及时间的延长，对棉铃虫的杀伤力增强．结果显示：饲喂第三天时，1mmol／L的浓度引起48％死亡，2mmol／L时60％死亡，4mmol／L时76％死亡，其IC50为1．78mmol／L．（2）用紫外光谱分析测得虫体内钙调蛋白的含量，结果显示：tomatine对其体内钙调蛋白的影响较大，随tomatine浓度的增加，钙调蛋白的含量逐渐降低．（3）紫外光谱图谱显示：番茄碱与钙调蛋白形成了一种新的物质（其特征吸收峰在190nm-220nm）．说明tomatine对棉铃虫的杀伤作用与钙调蛋白有关，此项研究为进一步探讨tomatine的作用分子机制提供了科学参数．     

棉铃虫幼虫蜕皮过程及RH-5992对其蜕皮的影响

用透射电镜技术研究了棉铃虫4龄幼虫在蜕皮过程中表皮层及皮细胞的变化，同时用含蜕皮激素类杀虫剂RH－5992的饲料饲喂4龄幼虫，导致其产生早熟，异常的致死蜕皮，在超微结构水平比较了正常蜕皮和RH－5992对皮细胞和新表皮形成的影响。     

含果蝇肌动蛋白基因的重组棉铃虫核型多角体病毒的构建

本实验利用昆虫杆状病毒表达载体，将果蝇肌动蛋白基因5Cactin克隆入杆状病毒表达载体，以棉铃虫单核衣壳核多角体病毒为亲本病毒，在脂质体介导下共转染棉铃虫细胞，空斑纯化得到重组病毒，以深入研究棉铃虫病毒在入侵、复制及装配过程中，宿主肌动蛋白的动态变化对病毒复制的作用与影响。     

Synthesis dynamic and developmental profile of prothoracicotropic hormone between diapause-and nondiapause-destined individuals in Helicoverpa armigera

Biosynthesis and secretion of prothoracicotropic hormone (PTTH) of diapause-and nondiapause-destined individuals in Helicoverpa armigera were studied using whole-mount immunocytochemistry and enzyme-linked immunosorbent assay (ELISA). The immunocytochemistry revealed that PTTH is expressed in two pairs of lateral neurosecretory cells of the brain. The presence of immunoreactivity has not significant difference between the brains of the diapause-and nondiapause-destined 6th instar larvae. However,the obvious differences of expressional pattern from day 4 pupae were observed be-tween the two types. PTTH titers in hemolymph from the 6th instar larvae to pharate adults were measured by the ELISA. Although there were similar titer changes between the two types of individuals at the larval stage,a significant difference from developmental expression was detected at the pupal stage,suggesting that the expression and secretion of PTTH does play a crucial role in regulation of pupal diapause of H. armigera.     

Host selection of Helicoverpa armigera and H. assulta and its inheritance

The difference in host selection between the polyphagous Helicoverpa armigera (Hübner) and the oligophagous H. assulta Quenée was examined, with cotton (Gossypium hirsutum), tomato (Lycopersicon esculentum) (hosts of H. armigera), tobacco (Nicotiana tobaccum) (host of both H. armigera and H. assulta), and bush redpepper (Capsicum frutescens) (host of H. assulta) as testing plants. A multiple-choice test was used with caged plant cuttings for adult oviposition and with leaf discs for larval feeding. A no-choice test was run for evaluating larval growth rate. The results indicated that the relationship between larval performance and adult preference of H. assulta was more conspicuous than that of H. armigera. Reciprocal hybridization between H. armigera and H. assulta followed by backcrossing of the hybrids (F1) with H. armigera was also carried out for genetic study on host selection of these two insect species. A two-choice test with cotton and bush redpepper leaf discs showed that H. armigera larvae preferred to feed on cotton, and H. assulta larvae to bush redpepper; feeding preferences of the two F1 lines were intermediate between those of their parents, but close to that of their female parent; preference indexes of backcross lines also showed that both maternal factor and chromosomal inheritance were involved in feeding selection of two Helicoverpa species.     

棉铃虫组织蛋白酶B在杆状病毒表达系统中的表达及鉴定

以棉铃虫（Helicoverpa armigera）组织蛋白酶B作外源基因重组构建克隆载体,自重组菌株HCB-DH10Bac、Histag-HCB-DH10Bac中提取穿梭质粒,脂质体法转染草地贪夜蛾卵巢细胞系Sf21细胞,转染液再感染细胞,收集感染3～4d的上清液,提取芽生病毒的DNA,PCR法鉴定外源HCB基因,感染上清进行SDS-PAGE、Western-blotting、蛋白酶活性检测．结果：感染上清中的芽生病毒的DNA作模板扩增出预期的1700bp片段,SDS-PAGE、westem-blotcing检测均在28ku处有明显表达产物,且表达产物有蛋白水解活性．     

中国绿刺蛾(Parasa sinica Moore)质型多角体病毒的研究

<正>本文对中国绿刺蛾质型多角体病毒(Ps CPV)进行了初步观察研究。该病毒的包含体呈六角形、四方形或不规则形,有强烈的折光性,大小为0.4～2.8微米。病毒粒子近球形,表面具管状突起,大小为48.2～55.3毫微米。这种病毒对中国绿刺蛾3～4龄幼虫的LC_(so)为1.21×10~5多角体/毫升,具有应用于生物防治的潜力。     

Degeneration of host prothoracic glands caused by Campoletis chlorideae polydnavirus

The development of last instar Helicoverpa armigera prothoracic glands was investigated first; then the effects on the prothoracic glands of Helicoverpa armigera was studied by injection with calyx fluid and polydnavirus (PDV) from the endoparasitoid Campoletis chlorideae. Results showed that 3 female equivalents of calyx fluid or 4 female equivalents of PDV induced degeneration of host prothoracic glands. 24 h after calyx fluid or PDV injection the ultrastructure of the gland cells showed a significant decrease in intercellular channel system, while an increase in the number of round mitochondria, lysosomes and whorled figures, the organelle of some cells has degenerated and only organelle debris remained in the cells. These results suggest that calyx fluid or PDV cause the inactivation and degeneration of host prothoracic glands.     

Synthesis dynamic and developmental profile of prothoracicotropic hormone between diapause- and nondiapause-destined individuals in Helicoverpa arrnigera

Biosynthesis and secretion of prothoracicotropic hormone （PTTH） of diapause- and nondiapausedestined individuals in Helicoverpa armigera were studied using whole-mount immunocytochemistry and enzyme-linked immunosorbent assay （ELISA）. The immunocytochemistry revealed that PTTH is expressed in two pairs of lateral neurosecretory cells of the brain. The presence of immunoreactivity has not significant difference between the brains of the diapause- and nondiapause-destined 6th instar larvae. However, the obvious differences of expressional pattern from day 4 pupae were observed between the two types. PTTH titers in hemolymph from the 6th instar larvae to pharate adults were measured by the ELISA. Although there were similar titer changes between the two types of individuals at the larval stage, a significant difference from developmental expression was detected at the pupal stage, suggesting that the expression and secretion of PTTH does play a crucial role in regulation of pupal diapause of H. armigera.     

棉铃虫泛素基因的克隆及序列分析

泛素介导的泛素-蛋白酶体通路(Ubiquitin-proteasome pathway,UPP)是真核细胞内依赖ATP的非溶酶体蛋白质降解途径,该途径对细胞内蛋白的选择性降解起着重要作用.设计一对简并引物,从棉铃虫Helicoverpa armigera卵巢细胞Ha831中克隆了泛素基因的编码区,GenBank登录号AY456195.序列分析表明,该编码区的长度为228bp,编码76个氨基酸、其编码蛋白的相对分子质量为8 560,等电点为6.56.同源性比较发现,棉铃虫泛素基因与其他真核生物泛素基因在氨基酸水平上具有96%以上的相似性,而与棉铃虫核多角体病毒泛素的相似性为76%,所有已知的泛素关键功能位点在该泛素蛋白中均保守存在.     

Interpretation of the biological species concept from interspecific hybridization of two Helicoverpa species

The biological species concept defines species in terms of interbreeding. Interbreeding between spe-cies is prevented by reproductive isolation mechanisms. Based on our results of interspecific hybridi-zation between Helicoverpa armigera and Helicoverpa assulta, reproductive isolation mechanisms of the two species are analyzed. A combination of prezygotic factors (absent sex attraction and physical incompatibility of the genitalia) and postzygotic factors (female absence and partial sterility in F1 hy-brids) causes reproductive isolation of the two species. In addition, the role of interspecific hybridiza-tion in speciation is discussed.     

沿江棉区棉田棉铃虫卵的空间格局

对安徽沿江棉田棉铃虫卵的空间分布动态规律进行了调查研究,结果表明棉铃虫卵在棉田的聚集程度随发生代次而逐渐增强,发现不同代次虫口密度相同对理论抽样数明显不同,提出了沿江各代估值调查中的理论抽样数,并对几种空间格局模型拟合棉铃虫的空间分布情况作了比较。     

Testes and chromosomes in interspecific hybrids betweenHelicoverpa armigera (Hübner) andHelicoverpa assulta (Guenée)

The interspecific hybridization betweenHelicoverpa armigera females andHelicoverpa assulta males yieldedF ₁ hybrids (RS), fertile males and sterile individuals with abnormal genitals. The reverse hybridization betweenH. assulta females andH. armigera males yielded F₁ hybrids (SR)-fertile males and fertile females. The morphology of testes and the karyotype of chromosomes of larvae in the hybrids were investigated. Among the 2d old fifth-instar SR larvae, individuals without testes were fertile females and those with testes were fertile males. The length and breadth of testes between SR and parental species were not significantly different (p>0.05). Among the 2d old fifth-instar RS larvae, the testes were observed in all the individuals, but it could be classified into two types. The length and the breadth of testes in Type 1 larvae were not significantly different from those of their parental species (p>0.05), while those in Type 2 were significantly less than those of their parental species (p<0.01). Mitotic metaphase I of brain cells showed the diploid chromosomes number of both reciprocal hybrids was 2n=62, as many as their parents. The haploid number of 31 was confirmed by counts from spermatocytes at meiotic metaphase from SR male larvae and Type 1 larvae of RS. Meiosis was not observed in spermatocytes of Type 2 larvae of RS. Considering the characteristics of adult hybrids of RS, it was concluded that Type 1 individuals in RS were fertile and those of Type 2 were sterile. The sterility of Type 2 individuals in RS is attributed to the abnormity in development of testes and the failing meiosis of spermatocytes. As a result, the normal spermatozoon could not been produced.     

棉铃虫性信息素合成激活肽基因的检测

性信息素合成激活肽是调节昆虫性信息素合成的神经肽．根据昆虫性信息素合成激活肽的氨基酸序列保守性,合成若干引物,以棉铃虫基因组ＤＮＡ为模板,用ＰＣＲ方法得到单一的扩增条带．ＲＴＰＣＲ方法从棉铃虫咽下神经结中检测出性信息素合成激活肽基因在棉铃虫基因组中不仅存在,而且表达．     

An analysis of structure fitting and bioactivity between sex pheromone of cotton bollworm, Helicoverpa armigera (Hübner) and its fluorinated analogs

A study on the structure-activty relationship between （Z）-hexadec-9-enal （Z-9-16：Ald） and its analog was conducted by comparing the structures of the sex pheromone of cotton bollworm, Helicoverpa armigera （Huebner） with its fluorinated analog using computer molecular fitting. It is demonstrated that the structure of analog substituting for hydrogen atom on the terminal carbon atom is similar to Z-9-16：Ald. The EAG result showed that there is no significant difference in activities between Z-9-16：Ald and its fluorinated analog synthesized.     

棉铃虫滞育解除的相关基因鉴定

运用差异显示PCR(DD-PCR)方法分析棉铃虫滞育解除蛹差异表达的基因,结果得到了56个差异片段.通过RT-PCR和Real-time PCR的方法,鉴定了滞育解除过程中有3个基因表达量在下调,有4个基因表达量在上调.这7个差异表达基因中有3个和已知的基因有较高的同源性:FKBP12,esr16和NADH dehydrogenase 1 alpha subcomplex subunit 6.这些差异基因为今后研究滞育解除的分子机制提供了新的线索.     

Interspecific hybridization ofHelicoverpa armigera andH. assulta (Lepidoptera: Noctuidae)

Reciprocal hybridization between Helicoverpa armigera (Hübner) and H. assulta Guenee followed by backcrossing of the hybrids (F₁) with H. armigera produced backcross (BC) lines consisting of fertile females and males. The F₁ of H. armigera female and H. assulta male had only male, no female sex. In this case Haldane’s rule applies, and therefore it is proved that the sex chromosomes of Helicoverpa species are of ZW type, and the female is the heterozygous sex. This hybrid also showed significant heterosis with the heaviest pupal weight, and when it was backcrossed with H. armigera female, the sex ratio of the BC offspring was distorted as 1∶4. The potential utilization of this hybrid in genetic controlling H. armigera was finally discussed.     

Commercial production of transgenic Bt insect-resistant cotton varieties and the resistance management for bollworm (Helicoverpa armigera Hubner)

There are currently three kinds of transgenic Bt insect-resistant cotton germplasm lines, Shanxi 94-24, Zhongxin 94 and R19, in China. They showed high resistance to the neonate larvae of bollworm (Helicoverpa armigera). Transgenic Bt insect-resistant cotton varieties or hybrids have been bred using the three kinds of germplasm lines as parents. Our researches reveal that there exist different expressions in resistant level at different developmental stages in the three categories of germplasm lines. When neonate larvae are fed with leaves of cotton plant at the seeding stage with less than 10 leaves on the main stem, the mortality of the neonate larvae is 100%, but the resistance level will decline at later season. When Bt gene has been transferred to the cotton genome, it can be steadily transferred to the progeny, the level of resistance to bollworm keeps fundamentally uniform. Such insects as tobacco budworm (Heliothis virencens) in laboratory directive selection are very apt to produce resistance to the Bt insecticidal crystal protein. From the present crop system of cotton region in the Yangtze and Yellow River Valleys, and the expression characteristic of transgenic Bt resistant cotton, we suggest that the resistance to toxin protein in bollworm is not apt to be produced if the transgenic Bt insect-resistant cotton varieties are released and grown in the regions except in the Xinjiang cotton region. The managing strategies to delay or retard the resistance are discussed.     

苏云金芽孢杆菌营养期杀虫蛋白基因的克隆、表达及杀虫活性分析

选择本实验室分离的野生型苏云金芽孢杆菌菌株WY-197为出发菌株，用全长PCR方法从此菌株中克隆了2．3kb大小的vip3A基因,DNA序列比较发现所克隆的基因vip3A-197与已知的营养期杀虫蛋白基因存在很高的同源性．将基因vip3A-197亚克隆至原核表达载体pET33b构建了原核表达质粒pEVip，转化大肠杆菌BL21，转化子经IPTG诱导后可表达88kD大小的蛋白．该蛋白对甜菜夜蛾(Spodoptera exigua)棉铃虫(Helicoverpa armigera)的初孵幼虫进行生物测定，结果表明，营养期杀虫蛋白vip3A-197对夜蛾科害虫具有一定的杀虫活性。     

Induction of nicotine in tobacco by herbivory and its relation to glucose oxidase activity in the labial gland of three noctuid caterpillars

Tobacco Nicotiana tabacum L. is a host plant ofHelicoverpa armigera (Huibner), Helicoverpa assulta Guenee and Spodoptera litura (Fabricius) (Lepidoptera, Noctuidae). The difference in leaf nicotine response to the feeding by these three larvae and the mechanical simulation of their feeding was examined by HPLC. Results indicated that nicotine induction was suppressed by H. armigera and H. assulta larvae feeding or by simulated damage treated with their labial glands extracts. The production of nicotine was also suppressed by the glucose oxidase from Aspergillus niger when it was treated on mechanically wounded leaf area. On the contrary, the nicotine production was stimulated by S. litura larva feeding or by simulated damage treated with its labial gland extract. Heat denature can not counteract the stimulation effect of the S. litura labial gland extracts to tobacco nicotine production. The glucose oxidase activity was detected in labial gland extracts of both H. armigera and H. assulta, but the activity in H. armigera was significantly higher than that in H. assulta. No glucose oxidase activity was detected in labial gland extracts of S. litura. It is shown that the glucose oxidase activity in labial glands of caterpillars plays an important role in the nicotine response to herbivory. The glucose oxidase was mainly contained in the labial gland of H. armigera larva, and had the highest activity at pH 7.0. D-Glucose was the optimal substrate of the glucose oxidase. Labial gland glucose oxidase activities varied daily during larval development with high activities found when larvae were actively feeding.