粘虫颗粒体病毒及其增效因子对粘虫核型多角体病毒的增效作用

生物测定的结果表明,粘虫颗粒体病毒ＰｕＧＶ－Ｐｓ及其增效因子对粘虫核型多角体病毒ＰｓＮＰＶ均有明显的增效作用．粘虫颗粒体病毒不仅能提高粘虫感染ＰｓＮＰＶ的死亡率,而且ＮＰＶ、ＧＶ两种病毒混合感染使粘虫幼虫代谢发育受到抑制,表现为生长缓慢、体重减少等．增效因子的增效作用表现在能显著提高粘虫幼虫的死亡率,降低核型多角体病毒的半致死浓度ＬＣ５０,并缩短半致死时间ＬＴ５０．     

棉铃虫核多角体病毒基因组限制酶酶切分析及其多角体基因的定位

应用１１种限制性内酶ＢａｍＨⅠ、ＢｓｔＥⅡ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ＨｉｎｄⅢ、ＫｐｎⅠ、ＰｓｔⅠ、ＳａｌⅠ、ＳｓｔⅠ、ＸｂａⅠａⅠ和ＸⅠⅠ和ＸｈｏⅠ分别将中国棉铃虫核多角体病毒（单粒包埋ＨａＳＮＰＶ）湖北株基因组Ｄ组ＤＮＡ酶切为１０、１２、２２、２１、１３、６、６、４０、６、２１、６个片段，并求得基因组大小平均为１什什Ｍｒ≈７９．１×１０６．以家委核多角体病毒ＢｍＮＰＶ多角体基因为探针，利用Ｓｏｕｔｈｅｒｎ杂交技术将病毒多角体基因定位在ＳａｌⅠ４．２×１０３ｂ左右的片段上．与棉铃虫核多角体病毒其它株系酶切图谱比较结果表明，本株病毒与上海等株系及美洲棉铃虫核多角体病毒ＨａＳＮＰＶＥｌｋａｒ株系酶切图谱相似，它们之间的条带数和大小差异较小，而与已发现的所有多粒包埋型病毒ＨａＳＮＰＶ酶切图主谱差异较大．据此认为ＨａＳＮＰＶ和ＨａＭＮＰＶ属于基因型不同的两类病毒，而ＨａＳＮＰＶ不同分离株的同源性很高     

昆虫病毒增效蛋白的研究进展

杆状病毒是专化性很强的一类双链ＤＮＡ病毒,主要感染无脊椎动物,特别是鳞翅目昆虫．杆状病毒科又分为３个亚组：核型多角体病毒ＮＰＶ（Ｎｕｃｌｅａｒｐｏｌｙｈｅｄｒｏｓｉｓｖｉｒｕｓ）、颗粒体病毒ＧＶ（Ｇｒａｎｕｌｏｓｉｓｖｉｒｕｓ）和非包涵体型杆状病毒（...     

温度和pH值对重组杆状病毒(AcMNPV-BmKIT-Chi)的毒力影响

研究了温度、酸碱度对重组杆状病毒(AcMNPV-BmKIT-Chi)毒力的影响.结果显示,多角体病毒在温度4~50℃,pH6.0~8.0范围内可以保持较高的毒力稳定性;不同虫龄棉铃虫幼虫对重组病毒的毒力敏感性差异显著,低虫龄对重组核型多角体病毒更为敏感;病毒对幼虫存在亚致死作用,饲毒组与对照组幼虫体重差异显著.图3表3参13     

EoNPV的限制性酶切图谱及polyhedrin基因和egt基因的定位

用８种限制性内切酶酶解茶尺蠖Ｅｃｔｒｏｐｉｓｏｂｌｉｑｕａ核型多角体病毒（ＥｏＮＰＶ）基因组ＤＮＡ,获得大于０．８８ｋｂ的片段数分别为７、３８、１９、１７、１６、９、３２和１４．由此统计,基因组平均大小大于１１８．５ｋｂ,即Ｍｒ约大于７８．６×１０６．用ＢｍＮＰＶ的多角体蛋白（ｐｈ）基因和ＡｃＭＮＰＶ的ｅｇｔ基因为探针,应用Ｓｏｕｔｈｅｒｎ杂交方法,将ＥｏＮＰＶ的ｐｈ基因定位于ＢａｍＨＩＡ、ＣｌａＩＤ、ＥｃｏＲＩＭ、ＥｃｏＲＶＬ、ＨｉｎｄⅢＦ、ＰｓｔＩＡ、ＳａｌＩＨ和ＸｈｏＩＢ片段上,将ｅｇｔ基因定位于ＢａｍＨＩＡ、ＣｌａＩｌ、ＥｃｏＲＩＫ、ＥｃｏＲＶＡ、ＨｉｎｄⅢＨ、ＰｓｔＩＡ、ＳａｌＩＢ、ＸｈｏＩＩ片段上．通过克隆和酶切鉴定ＥｃｏＲＩＭ和ＥｃｏＲＶＬ片段,测定ｐｈ基因部分序列,并以ＥｃｏＲＩＭ为探针对基因组酶切印迹进行杂交,制作了ＥｏＮＰＶｐｈ基因和ｅｇｔ基因区的酶切图谱,推定ｅｇｔ基因保守区位于ｐｈ基因上游约４．５５ｋｂ处．     

HaNPV多角体蛋白在宿主不同组织中的时相性表达

应用亲和凝胶过滤层析对多角体蛋白多克隆抗血清进行纯化,辣根过氧化物酶进行标记,用于ＥＬＩＳＡ法检测中肠、血淋巴、脂肪体中的多角体蛋白的含量变化．结果：中肠在感染２４ｈ含量最高,４８ｈ最低,７２ｈ较４８ｈ略有升高,９６ｈ和１２０ｈ与７２ｈ基本保持一致;血淋巴在感染４８ｈ含量最高,然后下降至９６ｈ,１２０ｈ又稍有升高;脂肪体在感染７２ｈ开始增加,直至１２０ｈ．三种组织中多角体蛋白含量变化并不同步．从表达的时相性看,与ＮＰＶ的感染过程基本一致,即从中肠至血腔再到脂肪体．免疫电镜观察表明,感染７２ｈ的中肠,在胞浆中合成的多角体蛋白此时已转运到细胞核中     

杆状病毒包涵体蛋白的共同抗原性

在比对已公布的37种杆状病毒包涵体蛋白氨基酸序列的基础上,选定棉铃虫核型多角体病毒(HaNPV)多角体蛋白的两个高度保守多肽(54-113aa和206-245aa)制备多克隆抗体,用Western blot法检测这两个高度保守多肽与多种杆状病毒包涵体蛋白之间的血清学关系.结果表明,HaNPV多角体蛋白的两个多肽的抗体与14种核型多角体病毒的多角体蛋白和5种颗粒体病毒的颗粒体蛋白均有明显的杂交信号,表明杆状病毒的包涵体蛋白之间具有共同的抗原决定簇.根据杆状病毒包涵体蛋白之间的这种血清学关系,进一步讨论了利用免疫金试纸技术检测病毒杀虫剂中包涵体含量的可行性.     

荧光增白剂对重组苜蓿银纹夜蛾核型多角体病毒的增效作用

以β-半乳糖苷酶基因为标志基因的重组苜蓿银纹夜蛾核型多角体(AcMNPV-hsp70／lacz)研究光增白剂对该病毒的增效作用及其作用方式．通过病毒毒力的生物测定表明：光增白剂FB-28能提高甜菜夜蛾幼虫的死亡率，降低半致死浓度，缩短半致死时间；光增白剂的增效作用与感染虫龄有关．此外，应用组织切片技术对中肠组织病理以及血淋巴细胞感染观察表明：在幼虫的病毒接种物中添加合适浓度的荧光增白剂不会改变病毒在幼虫中肠组织中的感染部位，却可以提高幼虫对病毒的敏感性，产生更多的病灶．图版1图4表3参19     

3种典型微生物农药对家蚕的毒性研究

苏云金芽孢杆菌、球孢白僵菌以及棉铃虫核型多角体病毒是细菌类、真菌类和病毒类微生物农药的典型代表。为了明确微生物农药对家蚕的毒性影响,选取以上3种典型微生物农药,分别探究其对家蚕的毒性。结果显示:苏云金杆菌原药对家蚕LC50值为1.33×105CFU·m L-1;球孢白僵菌原药对家蚕LC50值为1.14×106CFU·m L-1;棉铃虫核型多角体病毒母药对家蚕LC50值大于1.00×108PIB·m L-1。与此同时,通过进一步试验,明确了以上3种微生物农药对家蚕结茧率、死笼率、全茧量、茧层量等关键指标的影响。结果显示:除了棉铃虫核型多角体病毒母药,高浓度苏云金杆菌原药、球孢白僵菌原药对家蚕的结茧率和死笼率均具有显著影响;3种微生物农药对家蚕产茧能力指标全茧量、茧层量以及蛹重均无显著性影响。     

团花锦蛇、绿锦蛇和三索锦蛇的核型及Ag-NORs

报道了锦蛇属团花锦蛇（Ｅ．ｄａｖｉｄｉ）、绿锦蛇（Ｅ．ｐｒｖｓｉｎａ）和三索锦蛇（Ｅ．ｒａｄｉａｔａ）３个种的核型和Ａｇ－ＮＯＲｓ,团花锦蛇的型２ｎ＝３６（８Ｖ＋６ｓＶ＋２Ｉ＋２０Ｍ）,绿锦蛇的核型２ｎ＝３６（７Ｖ＋７ｓＶ＋２Ｉ＋２０ｍ）,三索锦蛇的核型２ｎ＝４０（６Ｖ＋３ｓＶ＋２ｓＩ＋５Ｉ＋２４ｍ）;①团花锦蛇和绿锦蛇的染色体数目和结构与此属大多数咱的相似,;②此种为雄性,未见染色体异型。③Ａｇ     

Antifeedant activity of aqueous extract of Gnidia glauca Gilg. and Toddalia asiatica Lam. on the gram pod borer, Helicoverpa armigera (Hbn)

Aqueous leaf extracts of two plants namely Gnidia glauca Gilg. and Toddalia asiatica Lam., have been screened for their antifeedant activity against the sixth instar larvae of Helicoverpa armigera (Hbn) by applying the aqueous leaf extracts at various concentrations viz., 0.2, 0.4, 0.6, 0.8 and 1.0 percent on young tomato leaves. The larval mortality of more than 50 percent at higher concentrations (0.8 and 1.0 percent) was observed in the aqueous extracts. Among the two aqueous leaf extracts tested, T. asiatica was found to show higher rate of mortality (86.1%) at 1.0 percent concentration. A reduction in the rate of food consumption and growth was observed in the larvae of H. armigera after 48 hours of treatments in both the aqueous extracts. Since this insect pest species have developed resistance and resurgence to synthetic insecticides, the only alternate is the usage of bio-pesticides for they are eco-friendly, pollution free and easily degradable.     

菊酯类杀虫剂抗性棉铃虫para型钠离子通道基因的部分序列测定与突变

昆虫神经系统para型钠离子通道是菊酯类杀虫剂的主要靶标，对10多种昆虫的研究表明，钠离子通道基因发生点突变与昆虫对菊酯类杀虫剂的抗性密切相关．通过RT-PCR扩增的方法，我们获得了编码溴氰菊酯抗性和敏感品系棉铃虫钠离子通道域Ⅱ-Ⅳ的cDNA片段．核苷酸序列及推导的氨基酸序列分析结果表明，溴氰菊酯抗性品系棉铃虫钠离子通道基因不存在其它昆虫中报道的kdr和super-kdr抗性突变，但是我们发现了一个甲硫氨酸(M)到亮氨酸(L)的新突变，该突变位于域Ⅱ和Ⅲ之间．由于昆虫钠离子通道高度保守，因此，该突变很可能与棉铃虫对溴氰菊酯的抗性有关．另外，通过PCR扩增的方法，克隆了棉铃虫钠离子通道域Ⅱ-Ⅳ的基因片段，该片段全长7334bp，包含有13个外显子和12个内含子，与果蝇和烟蚜夜蛾相比，内含子在基因中的位置基本一致，进一步表明昆虫钠离子通道基因在进化过程中高度保守．图5表1参22     

Rotational application of bioinsecticide with deltamethrin-An antilarval measure for the control of filarial vector, Culex quinquefasciatus (Culicidae: Diptera)

A laboratory investigation was undertaken to study the cyclic usage of field recommended doses of Bacillus thuringiensis israelensis (Bti), Bacillus sphaericus (Bsp) and combination of Bti and Bsp (half the recommended dose of each) with deltamethrin 2.8 EC to attain better control of mosquito larvae. To understand the susceptibility status of the Culex quinquefasciatus larvae in the field, early fourth instar field collected larvae were exposed to the above treatments of biopesticides and were monitored for their persistency till 50% mortality was obtained. The larvae those survived in each treatment were transferred to the concentration of deltamethrin that gave 50% mortality. Observations on larval mortality after the treatment with deltamethrin were recorded after 24 h. The results revealed that Bti excels Bsp, as it recorded 54% mortality only on 17th day after application. The other salient finding of this study is LC50 of deltamethrin is sufficient to follow the biopesticides application for an effective control of Culex larvae.     

味精工业高浓度有机废水培养苏云金芽孢杆菌的研究

利用味精工业发酵废水进行了培养苏云金芽孢杆菌（Bacillus thuringiensis,Bt）以生产生物农药的实验研究.经过对废水成分分析和预处理、Bt菌种驯化、摇瓶机发酵配方正交优化实验和生物毒性测定,探讨了一条经济可行的工艺路线.该工艺在利用发酵废水生产生物农药时不会产生二次污染问题,具有较高的工业应用价值,对于其他发酵工业废水的资源化处理也有一定参考价值.     

Green leaf volatiles enhance sex attractant pheromone of the tobacco budworm,Heliothis virescens (Lep.: Noctuidae)

Summary Components of the green leaf volatile complex (Z-3-hexenyl acetate andE-2-hexenyl acetate) were shown to enhance responses of tobacco budworm,Heliothis virescens, males to the sex attractant pheromone of conspecific females in the field. The results are discussed with regard to green leaf volatiles which enhance the attractant pheromone of a cohabiting species, and serve as attractants of a parasitoid of conspecific larvae.     

Patch study of mortality,growth and feeding of the larvae of the southern gadoid Macruronus novaezelandiae

A discrete dense patch of eggs and larvae of hoki (Macruronus novaezelandiae) within the hoki spawning grounds off Westland, New Zealand, was sampled to examine prey selectivity by larvae and to obtain estimates of larval mortality and growth. The patch was tracked using a free-drifting drogue, and surveys of the horizontal distribution of larvae before and after the patch study indicated that the drogue had successfully followed the patch. Modal analysis of the size-frequency distributions of hoki larvae revealed up to six cohorts within the patch at any one sampling time, and a growth rate of 0.21 mm standard length per day. The daily mortality coefficient for larvae within the patch was 0.19, although this is considered to be an overestimate. Differences in the mean length between cohorts suggest that hoki have a synchronised, diel spawning periodicity, and results of a simple cellular design model revealed that ten continuous days of spawning were required to yield the observed size structure of the hoki larvae population within the patch. Diet analysis of larvae in the patch showed that copepods of the genus Calocalanus are actively selected, and are especially important in the diet of early-stage larvae. Based on aspects of larval diet, morphology, and rates of mortality and growth, it is hypothesised that hoki larvae are adapted to a low-food environment, and that predation is likely to be more important as a source of mortality than starvation.