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．     

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.     

抗棉铃虫组织蛋白酶B单克隆抗体的制备及鉴定

分别以棉铃虫组织表达的组织蛋白酶B和基因重组的大肠杆菌表达的组织蛋白酶B为抗原免疫BALB／c小鼠，选择血清抗体滴度高的免疫小鼠，取其脾细胞和Sp2／0骨髓瘤细胞进行细胞融合．经ELISA法筛选阳性克隆和有限稀释法克隆细胞，共获得分泌抗组织蛋白酶B的单克隆抗体杂交瘤细胞3株，分别命名为9D5，100E2和100H6．以ELISA法和Western blotting法对3株杂交瘤细胞株分泌的单克隆抗体进行了特异性鉴定，其中9D5和100E2能够同时识别两种不同来源的组织蛋白酶B，而100H6只能识别基因重组的大肠杆菌表达的组织蛋白酶B．     

抗性棉铃虫神经细胞的离体培养和超微结构分析

对棉铃虫幼虫神经细胞进行了离体培养,并建立了其体外培养的实用方法。神经细胞在增补棉铃虫幼虫血淋巴的TC-100培养基中可贴壁生长,存活5-7d,培养基中不加血淋巴,而增补谷氨酰胺时,细胞也可贴壁生长,但接种24h后出现老化现象,增补适量葡萄糖,细胞可存活22-28d。细胞存活的最适pH值为6。4.90%的细胞具多条突起,与在体内生长有明显区别。棉铃虫抗性种群、对照种群、同种群不同龄期幼虫以及同一个体胸、腹神经节的神经细胞体外培养时形态无明显差异,但抗生品系幼虫运动神经元内线粒体明显增多,且神经围鞘与神经世膜厚度之比明显增加。     

An intracellular toxic protein (Xin) isolated from Xenorhabdus nematophilus strain BJ

An intracellular toxic protein, designated Xin and with remarkable inhibitory effect on growth of Helicoverpa armigera, was isolated from X. Nematophilus strain BJ by ion exchange chromatography and hydrophobic interaction chromatography. After placing neonates on the artificial diet containing Xin at concentration of 0 or 3.48μg/g for 7 days, the average weight of insects was 23.50mg and 0.55mg, respectively. The activity of Xin was lost by heating, freeze-drying, digestion with proteases or urea. The Xin molecular weight, over kD, was determined by gel filtration on Superose 12, and there were six component proteins analyzed by SDS-PAGE. The results show that Xin is a high molecular protein complex inhibiting growth of insect pest.     

高浓度稀土 Nd(NO3)3等毒性研究

文章研究了稀土Nd(NO3)3对棉铃虫生长及家兔小肠运动和呼吸的影响．研究表明：低浓度稀土溶液对棉铃虫的生长影响很小，而中浓度、高浓度稀土溶液对棉铃虫的生长影响较大，它可以使其死亡率增大，生长速度变慢、化蛹率降低．轻稀土Nd(NO3)3能使兔呼吸频率加快．小肠收缩运动，随着浓度的增加，幅度减慢得越快。     

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.     

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.     

不同寄主植物对棉铃虫生长发育影响因素的分析

为了探索棉花叶片、加工番茄叶片、加工番茄青果和加工番茄红果对棉铃虫幼虫生长发育的影响,测定棉花叶片和加工番茄的果实等组织器官的主要物质含量,分析影响棉铃虫幼虫生长发育的因素。结果表明:以加工番茄叶片饲喂的棉铃虫上幼虫发育历期最短16.04 d、幼虫存活率最高54.44%,蛹羽化率最高83.58%,以加工番茄红果发育历期最长26.17 d,幼虫成活率最低3.67%,蛹羽化率最低0.00%。因此,4种食料中棉铃虫的适合度为:加工番茄叶片棉花叶片加工番茄青果加工番茄红果。棉花叶片、加工番茄叶片和果实中的营养成分差异较大,其中加工番茄叶片的可溶性蛋白含量最高为15.14%、C/N为0.91;加工番茄红果可溶性蛋白含量最低为3.04%、C/N为9.65。因此,植物组织内的可溶性糖含量低、C/N小、含水量和可溶性蛋白含量高有利于棉铃虫生长发育。     

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.