HOST SPECIFICITY OF LAGENIDIUM GIGANTEUM

Lagenidium giganteum 的宿主特异性

基本信息

批准号：
2633528
负责人：
JAMES L KERWIN
金额：
$ 20.08万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-01-01 至 1999-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2633528
关键词：
Culicidae arthropod nonpollutant control chemical binding fluorescence microscopy gas chromatography gas chromatography mass spectrometry genetic manipulation genetic strain guinea pigs host organism interaction invertebrate cuticle larva membrane proteins nuclear magnetic resonance spectroscopy parasitism quail receptor binding virulence

项目摘要

Lagenidium giganteum (Oomycetes: Lagenidiales), a facultative parasite of mosquito larva, is the only biological control agent (this category does not include the microbial insecticides produced by various Bacillus spp.) that is approaching operational use for mosquito control. This is significant because the use of chemicals commonly applied for mosquito control is being increasingly restricted due to regulatory action or development of resistance. The infective stage of the fungus is a motile zoospore, produced by either sexual or asexual reproduction, which specifically attaches to, encysts, and penetrates the cuticle of mosquito larvae. Use of L. giganteum is limited, therefore, to mosquito habitats in which sporulation, host recognition by the zoospore, and subsequent events in the infection process can occur. The long-term objective of the proposed research is to identify the molecular bases for host recognition by the fungus, so that selection for more virulent isolates can proceed systematically. This work will provide basic information (protoplast generation, proteins involved in host infection) for genetic engineering should this be necessary to develop more virulent strains or to modify host range. The project encompasses two main areas; 1) Elucidation of the chemical components on the cuticular surface of larvae and pupae of Culex tarsalis which trigger L. giganteum zoospore attachment, encystment and penetration; and 2) Identification of the putative proteins on the zoospore anterior flagellum and cyst cell wall which are involved in the recognition process. Investigations of cuticular chemistry will entail isolation and structural identification of lipids, (poly)phenols, chitin and its derivatives, and other components using high pressure liquid chromatography, gas chromatography - mass spectrometry, nuclear magnetic resonance and other standard spectroscopic techniques. Selective modification or removal of cuticular components will be followed by assaying the effects of these treatments on zoospore behavior. Zoospore and cyst proteins, and peroxidase isozymes will be purified and characterized. Monoclonal antibodies will be made to these proteins, and fluorescence microscopy used to confirm their spatial distribution on the zoospore/cyst surface. Documentation of zoospore/cyst behavior following incubation with the various monoclonals will confirm a role for the complementary proteins in host recognition.

lagenidium giganteum（卵肉：lagenidiales），这是一种兼性寄生虫蚊子幼虫是唯一的生物控制剂（此类别确实不包括各种芽孢杆菌产生的微生物杀虫剂。）那就是接近蚊子控制的操作用途。这是意义重大，因为使用通常用于蚊子的化学物质由于监管行动或阻力的发展。真菌的感染性阶段是一种运动杂种，由性行为或无性繁殖，专门附加于并穿透蚊子幼虫的角质层。使用L。因此，Giganteum被限制在蚊子栖息地中，其中孢子形成动物孢子的宿主识别以及感染中的随后事件可以发生过程。拟议研究的长期目标是识别真菌识别宿主识别的分子碱基，以便选择更多有毒的分离株可以系统地进行。这项工作将提供基本信息（原生质体生成，涉及的蛋白质宿主感染）基因工程的宿主感染是否需要发展更具毒的菌株或修改宿主范围。该项目包括两个主要领域。 1）阐明化学物质 Culex tarsalis的幼虫和p的表面上的成分这会触发L. giganteum zoo孢子的附件，encystment和渗透； 2）鉴定在前孢子上的假定蛋白参与识别过程的鞭毛和囊细胞壁。表皮化学的研究将需要隔离和结构鉴定脂质，（多）酚，几丁质及其衍生物，以及使用高压液相色谱法，气体的其他组件色谱 - 质谱，核磁共振和其他标准光谱技术。选择性修改或去除表皮组件将随后分析这些作用关于动物孢子行为的治疗。 zoo孢子和囊肿蛋白，以及过氧化物酶同工酶将被纯化和表征。单克隆将对这些蛋白质制作抗体，并使用荧光显微镜确认其在Zoo孢子/囊肿表面上的空间分布。与孵化后的动物孢子/囊肿行为的文档各种单克隆人将确认互补蛋白在主机识别。