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．     

2-氨基苯并噻唑锌配合物对棉铃虫生长发育的影响

选取数百条正常三龄棉铃虫,分成5组,分别饲喂含不同剂量的2-氨基苯并噻唑锌配合物的人工配制饲料,期间测量每条虫的体重,直至生长到六龄使用血细胞计数板对血细胞进行计数,再测定血浆中的T-SOD活性.结果显示:与对照组相比,饲喂含低剂量2-氨基苯并噻唑锌配合物饲料的棉铃虫的血细胞数目较多,T-SOD活力较强;饲喂含高剂量配合物饲料的棉铃虫受到严重抑制,几乎处于不生长状态.以上结果表明低剂量2-氨基苯并噻唑锌配合物对棉铃虫生长发育有促进作用,而高剂量的则有抑制作用.     

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.     

Tissue-specific expression of glutathione S-transferases induced by 2-tridecanone or quercetin in cotton bollworms, Helicoverpa armigera (Hübner)

The tissue-specific expression of glutathione S-transferases (GSTs) in the cotton bollworm and the expression level induced by 2-tridecanone and quercetin were examined using the methods of biochemistry and the quantitative PCR. The relative expression level of GST mRNA was unanimous with the GSTs activity conjugaging with 1-chloro-2, 4-dimitro-benzene (CDNB) in fat bodies,midguts, heads and integuments of cotton bollworms. The GSTs activity in fat bodies was the highest, then midguts, heads and integuments in turn, which was in consistent with the relative expression level of GST mRNA. The specific activity of GSTs and the relative expression level of GST mRNA could be significantly induced by 2-tridecanone and quercetin, and after the induction the order of the GSTs activity and the relative expression level of GST mRNA in the above four tissues in cotton bollworms was not different from the control.The induction of GSTs by 2-tridecanone was stronger than by quercetin in all four tissues, which was in accordance with the relative expression level of GST mRNA. It suggested that the increase of GSTs activity induced by plant allelochemicals was associated with the elevated expression of GST mRNA in cotton bollworms.     

棉铃虫致病菌分离、鉴定及高活力苏云金杆菌的筛选

从山东省棉田自然死亡的棉铃虫上分离到常见致病菌至少有8种,经鉴定为:苏云金杆菌(Bacillusthuringiensis)、蜡状芽孢杆菌(Bacillus cereus)、铜绿假单胞菌(Pseudomonas aeruginosa)、球孢白僵菌(Beauveriabassiana)、卵孢白僵菌(B.tenella)、青霉菌属真菌(Penicilliumspp.)、拟青霉属真菌(Paecilimycesspp.)和蛾霉属真菌(Nomuraespp.);得到一株对棉铃虫高毒的苏云金杆菌菌株Hu,其毒力比普遍使用的Bt菌系7216高,二者的LC50分别为15.1μg/mL和26.28μg/mL。蛾霉属真菌对棉铃虫有明显的拒食作用和杀伤作用,但人工培养时生长十分缓慢。     

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。