野蚕DH—PBAN基因启动子的克隆及序列分析

为研究昆虫滞育激素－性信息素合成激活肽基因在进化过程中的变化以及基因表达调控的异同，用ＰＣＲ方法克隆了野蚕的ＤＨ－ＰＢＡＮ基因的启动子并测序，     

野蚕DHPBAN基因启动子的克隆及序列分析

为研究昆虫滞育激素性信息素合成激活肽基因在进化过程中的变化以及基因表达调控的异同,用ＰＣＲ方法克隆了野蚕的ＤＨＰＢＡＮ基因的启动子并测序,这是第二个被克隆的昆虫ＤＨＰＢＡＮ基因启动子．与家蚕相比,在核苷酸水平上同源性达９４％．野蚕ＤＨＰＢＡＮ基因启动子的ＴＡＴＡ框的保守序列为ＴＡＴＡＴＡＡＡ,位于－４７—－４０处;ＣＡＡＴ框是ＧＧＣＣＣＡＴＣＴ,位于－８３—－７５处;野蚕与家蚕一样具有ＴＡＡＴ保守序列,这段序列是一类调控蛋白的结合核心位点,家蚕的位于－１８５—－１７６,野蚕的位于－１７６—－１６７．在－６４—－５６部位,核苷酸序列表现出较大的不同．     

昆虫滞育相关激素调节的研究进展 (动物科学)

昆虫的生长发育离不开激素的调节。滞育作为昆虫特殊的发育状态,其特征是代谢活动显著降低,发育停滞。在滞育发育过程中,各类激素通常相互协调形成网络,发挥着重要的调节作用。在神经内分泌系统中,由神经细胞分泌的神经肽类激素通常作为上游调控激素,通过调节下游激素的合成和分泌从而影响昆虫的滞育发育;蜕皮激素和保幼激素作为下游激素参与到神经肽类激素的调控网络中。目前已克隆得到滞育激素-性信息素合成激活肽和促前胸腺激素的基因,并对二者在不同滞育型昆虫中的调节作用进行了深入研究。此外,激素受体的研究将为解释激素之间的相互作用提供重要依据;而通过滞育昆虫体内激素水平的调节将为有害昆虫生物防治提供新的方法。     

昆虫性信息素合成方法的进展

本文介绍了近几年来昆虫性信息素合成方法的进展.阐述了聚合物Wittig试剂法、有机硼试剂法、炔键移位法等的优缺点.同时评介了这些合成方法对昆虫性信息素合成在工业化生产发展中的实际意义.     

用PCR法检测人巨细胞病毒DNA

用聚合酶链式反应（ＰＣＲ）法检测患者尿液中人巨细胞病毒（ＨＣＭＶ）ＤＮＡ．结果表明，自行设计合成的引物位于ＨＣＭＶ基因组早期蛋白基因区，经ＰＣＲ仪扩增一段长４３０ｂｐ的特异序列片段，对正常人基因组ＤＮＡ或其它疱疹病毒ＤＮＡ无交叉反应．此法可检测出少至１０－１６ｇ（０．１ｆｇ）的病毒ＤＮＡ．通过对３５份尿液标本的检测，比较ＰＣＲ法和组织培养法的检测结果完全一致     

昆虫性信息素的固相合成

昆虫性信息素是昆虫分泌的一种化学物质,它传递昆虫性的信息.各种不同昆虫的性行为分别受其相应的性信息素的控制.对各种昆虫性信息素的研究给人们提供了一个控制害虫的美好前景.到目前为止,对鳞翅目昆虫(蛾和蝴蝶)性信息素作了较详细的研究,它们绝大部分为直链的单烯或多烯醇、其乙酸酯或相应的醛.其有关的化学和合成方法近年来已有很多评述.鳞翅目昆虫性信息素的结构虽然看起来很简单,但由于高度立体化纯     

新型抗菌肽基因设计、合成及其在酵母中的表达(Ⅰ)——杂合抗菌肽基因的设计与合成

以化学合成并证实抗菌活性和抗菌谱都高于天然抗菌肽的杂合肽ＣｅｃｒｏｐｉｎＡ１－１１Ｄ１２－３７的氨基酸序列为基础,选用酵母高频使用密码子设计了一种新型抗菌肽基因,基因合成采用二次ＰＣＲ（聚合酶链式扩增）方法．设计和合成的基因全长１４０个碱基对,包括氨基酸编码序列、起始密码子、终止密码子和两端限制性内切酶ＢａｍＨＩ、ＥｃｏＲＩ、ＳａｌＩ识别顺序,合成的基因克隆于ＰＣＲＴＭ２．１载体上．经ＤＮＡ序列分析证实,合成基因碱基序列与设计序列完全一致．     

天然产物农药昆虫性信息素的提取与结构鉴定——以亚洲玉米螟信息素的鉴定为例

昆虫性信息素昆虫求偶过程中释放的微量信号化合物,模拟这种信号系统可以制成专一高效的新型天然产物农药。本文以亚洲玉米螟性信息素的鉴定为例,阐述了鳞翅目昆虫性信息素的提取、结构鉴定的一般过程。     

榆跳象性信息素的提取及组分分析

榆跳象(Orchestes alni Linnaeus)是一种危害榆树叶片的毁灭性害虫,成虫和幼虫均危害榆树叶片.采用有机溶剂浸提法在4—6月分3次获得榆跳象雌性成虫性信息素的粗提取物,并分别通过气相色谱-质谱联用仪进行分析.结果表明,4—6月份的性信息素疑似物分别有5,6,6个.通过与鳞翅目及鞘翅目象甲科等昆虫性信息素比较,并基于多数昆虫性信息素的共同特征,最终确定2种性信息素疑似组分,即11,13-二甲基-12-乙酸酯、顺-9-十四碳烯酸异丁酯.该研究初步明确榆跳象雌性成虫性信息素的主要成分,可为下一步性信息素的合成和林间诱捕实验提供参考.     

PCR扩增纳豆激酶基因的程序优化

研究了利用ＰＣＲ从纳豆菌基因组ＤＮＡ中扩增纳豆激酶基因的最优化条件，得到ＰＣＲ反应在本研究中的最重要的三个参数值为：模板量１０ｎｇ；Ｍｇ＾２＋浓度１．５ｍｍｏｌ／Ｌ；退火温度６０℃。在这种优化程序下进行ＰＣＲ反应，获得了特异、高效和忠实的纳豆激酶基因产物。     

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.     

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。     

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．     

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.     

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.     

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.     

Reconstruction of HaSNPV with helicoverpa hormone receptor 3

In order to develop a more efficient virus for controlling the cotton bollworm Helicoverpa armigera, Helicoverpa hormone receptor 3 （HHR3）, which is involved in the ecdysteroid regulatory pathway, was used to genetically modify wild HaSNPV. HaSNPV-HHR3 budded virus and occlusion body virus were constructed in three steps： preparation of pFastBacHaPhpP10-HHR3 donor plasmid, transposition of HHR3 into the HaBacHZ8 bacmid, and transfection of HzAM1 cells to get HaSNPV-HHR3 virus.HHR3 was proved to be expressed in the HaSNPV-HHR3 virus infected HzAM1 cells by immunoblotting. Results of bioassay indicated that the body weight of the HaSNPV-HHR3 infected larvae was lower than the larvae infected with wild virus and uninfected normal larvae, which suggests that HaSNPV-HHR3 delayed larval growth.     

新疆昌吉市棉花害虫种类及防治现状调查

对昌吉市棉花害虫种类进行了调查.结果表明:棉花害虫(螨)有17种,其中害螨4种,为害严重的害虫(螨)有棉铃虫、棉蚜、棉叶螨等.文章阐述了昌吉市棉花主要害虫的危害及防治现状,提出了今后对棉花害虫(螨)综合治理的意见和建议.     

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.