大豆对大豆胞囊线虫侵染的应答机制研究

大豆胞囊线虫病(Heterodera glycines,soybean cyst nematode,SCN)是大豆生产上的重要病害,其特点为危害重、分布广、难防治,每年对大豆生产造成极大的损失。种植大豆抗性新品种是防治SCN目前最为有效的措施,研究大豆对SCN侵染的应答机制,是培育大豆持久抗病品种的前提,对加快抗线虫品种选育及SCN的防控具有重要的意义。本文综述了大豆对SCN侵染的组织细胞学应答机制;介绍了大豆在SCN侵染后酶系变化及酚类代谢的生理生化应答机制;从分子水平阐明了SCN侵染后大豆的基因转录变化,差异蛋白及DNA甲基化的应答机制,以期为大豆胞囊线虫病害的进一步研究与防治提供参考。     

捕食线虫性真菌的分离培养及分布规律

对自然界中的捕食线虫性真菌进行了分离和种属区分，并对分布规律进行了研究。从土壤、粪便等材料中获得了2类活性较强的捕食线虫性菌株：即套捕型(hooping)捕食线虫性真菌和粘捕型(sticking)捕食线虫性真菌。主要的代表种类有3种。这些捕食线虫性真菌在捕食性结构分生孢子形态及某些生物学特性上有一定差异。套捕型捕食线虫性真菌代表种为少孢节丛孢菌(Arthrobotrys oligospara)和梨形指环菌(Dactylaria pyriformis)，它们以菌环、菌网作为捕食性结构(器官)，以套捕方式杀灭线虫幼虫。梨形指环菌可产生多量厚垣孢子(chlamyalospore)。粘捕型捕食线虫性真菌代表种为纺锤隔指孢菌(Dactylella ellipsospora)。可形成菌结捕食线虫性结构，以粘捕的方式杀灭线虫幼虫。结果表明：捕食线虫性真菌在土壤和家畜粪便中的分离率是40．41％；此类菌适合于生存在阴暗、潮湿、富含腐植质的环境中。在温暖季节时，采用0．4g／L玉米粉琼脂培养基(CMA)易于对其进行分离培养。     

捕食线虫性真菌菌种保存技术的研究

本试验对捕食线虫性真菌菌种保存技术进行了研究,测定了在不同介质保存条件下,捕食线虫真菌分生孢子和菌丝的存活情况及其不是寄生性线虫的能力.结果表明:在设定的保存条件下,分别经过1周冷冻融化或2周反复冷冻融化,被保存的分生孢子接种于玉米粉琼脂培养基上能生长发育:在加入线虫第3期幼虫后,能生产捕食器,对虫体发挥捕食作用.     

胃肠道线虫的黏膜免疫

从机体抗胃肠道线虫的黏膜免疫、胃肠道黏膜的抗原加工与递呈、胃肠道黏膜抵御寄生虫的效应机制等几方面论述了胃肠道线虫的免疫机理和免疫学研究的进展，旨在进一步阐明用免疫介入方法防治胃肠道线虫的理论基础，以解决胃肠道线虫治疗过程中的药物残留以及寄生虫的抗药性问题。     

Spring invasion of Heterodera schachtii in sugarbeet; a simulation study

A simulation model was developed for the spring invasion of the beet cyst nematode,Heterodera schachtii Schmidt, into sugarbeet roots, according to the state variable approach. This model describes the processes of egghatch, emergence of second stage larvae from cysts, migration to roots and penetration into roots quantitatively, using published data.In 1983 a field experiment was conducted to test this model.H. schachtii cysts were introduced at depths 6–29 cm in PVC-cylinders, buried in the soil. The rooting depth of sugarbeet seedlings, growing in these cylinders, was limited to 5 cm by 50 m mesh nylon gauze. Every 10 days the second stage larvae, which had penetrated into the roots of these seedlings were counted. After 50 days, about 40% of the eggs had hatched. More than 20% of the emerged larvae penetrated if the cysts had been buried undeeply, and only 4% if the cysts had been buried at 29 cm depth.The model predicted the course of penetration into the root during the first 40 days with reasonable accuracy (r²=0.79), but in the 5th period of 10 days the model made an overestimation of more than 100%. Egghatch after 50 days was correctly simulated. The differences in penetration into the root between the model and the experiment might result from an oversimplified simulation of the penetration success or the neglection of mortality of second stage larvae. Detailed experiments should be done to provide better parameters for these factors.Samenvatting Volgens de toestandsvariabele-benadering werd een simulatiemodel ontwikkeld van de voorjaarspenetratie van het bietecystenaaltje. Het model beschrijft aan de hand van literatuurgegevens het uikomen van de eieren, het verlaten van de cyst door de larven, de migratie naar en de penetratie in de wortel.In 1983 werd een veldproef uitgevoerd om het model te toetsen. Cysten vanH. schachtii werden op 5 dieptes tussen 6 en 29 cm ingegraven in PVC-cylinders, welke waren verzonken in de bodem. De bewortelingsdiepte van de suikerbiete-zaailingen die hierin groeiden werd beperkt tot 5 cm door nylon gaas van 50 m maaswijdte. Elke 10 dagen werden de larven geteld die in de wortels van deze plantjes waren gepenetreerd. Na 50 dagen was 40% van de eieren uitgekomen. Meer dan 20% van de gelokte larven penetreerden als de cysten ondiep waren ingegraven, en slechts 4% als de cysten op 29 cm diepte waren ingegraven.Gedurende de eerste 40 dagen werd het verloop van de penetratie in de wortel met redelijke nauwkeurigheid door het model voorspeld (r²=0.79). In de 5e periode van 10 dagen maakte het model echter een overschatting van meer dan 100%. Het uitkomen van de eieren werd correct gesimuleerd. De verschillen in penetratie tussen het model en de proef zouden het gevolg kunnen zijn van een oververeenvoudigde simulatie van het penetratiesucces of van het verwaarlozen van de mortaliteit van de migrerende larven. Betere gegevens hierover zullen moeten komen uit detailproeven.     

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。     

Identification of a major gene (Mex-1) from Coffea canephora conferring resistance to Meloidogyne exigua in Coffea arabica

Among the most damaging root-knot nematode species, Meloidogyne exigua is especially common in Latin America and constitutes a major agronomic constraint in all major coffee-growing ( Coffea arabica ) areas. Growing nematode-resistant coffee represents the most promising option for control of the pest. The present study aimed to determine the mode of inheritance of the M. exigua resistance transferred into C. arabica from a related species, Coffea canephora , and to identify associated molecular markers. Segregation data analysis of F ₂ progeny derived from a cross between the resistant introgression line T5296 and the susceptible accession Et6 showed that the resistance to M. exigua is controlled by a simply inherited major gene (designated the Mex -1 locus). The gall index distribution exhibited by the F ₂ individuals suggested incomplete dominant expression. Fourteen AFLP markers were found associated with the resistance to M. exigua and a localized genetic map of the chromosome segment carrying Mex -1 was constructed. Furthermore, the association of the identified AFLP markers with Mex -1 was confirmed by analysis of a set of genotypes involving 28 introgression Arabica lines either resistant or susceptible to M. exigua in field conditions. These results represent an important starting point to enhance backcross breeding programmes and to perform an early selection of resistant seedlings.     

昆虫病原线虫和共生细菌培养系统中噬菌体的检测

本文利用紫外照射、温度、pH、盐度诱导五株溶源性共生菌株Xenorhabdus和Photorhabdus同时测定斯氏和异小杆属线虫Steinernema carpocapsae A24,S.carpacapsae ALL,S.feltiae English,S.feltiae SN,Heterorhabditis bacterophora H06在体外培养过程中是否存在感染共生细菌的噬菌体。结果未发现噬菌斑，说明实验菌株在实验诱导条件下以及昆虫病原线虫固体培养系统中不可能诱发噬菌体的危害。     

淡紫拟青霉发酵滤液对大豆胞囊线虫趋化性的影响

 实验室条件下,研究了大豆胞囊线虫幼虫对大豆根和根浸出液的趋化性以及淡紫拟青霉代谢产物对其趋化性的影响。结果表明,大豆胞囊线虫二龄幼虫(J₂)对大豆根存在着一定的趋化性,而淡紫拟青霉发酵滤液对J₂存在明显的驱避性。在根浸出液、真菌滤液及其混合液剂处理下,大豆胞囊线虫J₂在WA平板上靠近处理液的0~1cm区间中分布率存在极显著差异(P <0.0 1),而加样时间(液剂与线虫同时放置或提前2 4h处理)对其分布率没有显著影响。大豆幼根蘸取滤液后对线虫J₂在该区间的分布与不经处理的幼根差异显著(P <0.0 5),证明淡紫拟青霉发酵滤液可以明显地降低线虫与大豆根的亲和力,并强烈抑制线虫对大豆根的侵染。