构建了昆虫病原线虫共生菌Xenorhabdus nematophilus BP品系的粘粒文库并用生物测定的方法从中筛选到2个对棉铃虫初孵幼虫有口服抑杀作用的克隆cos83和cos76。为初步确定这两个克隆的毒素基因与已报道基因的差异程度，根据已发表的共生菌毒素基因序列资料设计了7对引物对这两个克隆进行PCR扩增并对扩增产物进行测序和分析。从毒力较强的cos83中7对引物均扩增出与目标片段大小一致的产物而从cos76中只有5对扩增到目标片段。测序结果显示cos76的5个PCR扩增产物与cos83的对应扩增产物DAN序列同源性为100%。以cos83的7个PCR扩增产物所编码氨基酸序列进行BLAST分析，它们与X. nematophilus PMF1296和Photorhabdus luminencenc W14毒素相应区间的平均同源性分别为94%和58%说明cos83毒素是共生菌口服毒素家族的一员但与同类的其它毒素有一定的差异，值得对其杀虫谱，作用机理等进行进一步的研究。

A new species of entomopathogenic nematode, Steinernema guangdongense sp. n. was recovered from a soil sample collected from Jijia town in the western part of Guangdong province, the Peoples Republic of China during a survey for entomopathogenic nematodes in 2001. The nematode can be separated from other described species of Steinernema, by morphological, morphometrical characteristics of different stages of the nematode, by crossbreeding tests and by characterizations and phylogeny of DNA sequences of either a partial 28S or the internal transcribed spacer regions of rDNA. This nematode is closest to S. longicaudum. It can be distinguished from that nematode by characteristics of different stages. For infective juveniles, although the body length is almost similar (1055 μm compared to 1063μm), body diameter of the new species is larger; values of EP (length from anterior end to excretory pore), NR (length from anterior end to nerve ring) and a body length/body width ratio are smaller, and tail with dorsal constriction. For male, the new species has longer spicule, not well curved, spicule head shorter, shaft not prominent or absent and spicule tip not suddenly tapered as shown in S. longicaudum. Also, the ratios SW (spicule length/anal body width) and GS (gubernaculums/spicule) are smaller. For female, the presence of a small double flapped epiptygma, a small projection on dorsal side of the tail tips and prominent post-anal swelling is typical for the new species.

Summary-Energy metabolism of the infective juveniles (IJ) of Steinernema carpocapsae under aerobic conditions and its relation to survival and infectivity of the IJ was studied by monitoring the changes in mean dry weight, levels of key energy reserve compounds, oxygen consumption rate, respiratory quotient, survival and infectivity of freshly harvested IJ incubated in tap water on a shaker at 28

Summary-A method using computer image analysis for the estimation of dry weight, lipid content, body size and length of individual infective juveniles (IJ) of entomopathogenic nematodes (EN) is described. Images of IJ, captured using a video camera, were digitalized and transferred to a computer for analysis. The longitudinal cross-sectional area of the nemat ode (Area), mean grey level (MGL) and perimeter of the nematode image (Perimeter) were measured automatically. Nematode samples having a range of sizes, mean dry weights and lipid contents, as well as a series of samples of different ages were examined. It was found that: i) the index Area £ MGL correlates well with the mean dry weight of the nematode, which is in turn a good indicator of the total energy reserve of the nematodes; ii) MGL correlates well with the lipid content (as % dry weight) which is a good indicator of the quality of the energy reserve; and iii) the perimeter of the nematode image can be used to estimate the length of an IJ. Compared to currently available methods, such as direct measurement of mean dry weight or lipid content, computerised image analysis gives more information concerning the samples tested and enables variations in dry weight, lipid content, body size and length between individual IJ to be estimated readily. A sample of 30 IJ can be examined in 30 to 60 min making this method ideal for quality control during production, processing, formulation and storage of EN-based biopesticide products. Résumé – Méthode rapide pour l’estimation du poids sec et du contenu lipidique moyens des juvéniles infestants de nematodes entomopathogènes basée sur l'analyse d'images-Il est décrit une méthode utilisant l'analyse numérisée d'images et permettant l'estimation du poids sec, du contenu lipidique, de la taille et de la longueur du corps des juvéniles infestants (IJ) de nematodes entomopathogènes. Les images des IJ, saisies par vidéo-camera, sont digitalisées et transférées pour analyse sur ordinateur. La surface de la coupe longitudinale du nématode (Area), le niveau moyen d'opacité (MGL) et le périmètre de l'image du nématode (Perimeter) sont mesurés automatiquement. Des échantillons de nématodes de taille, de poids sec et de contenu lipidique variés ainsi qu’une série représentant différents ages ont été examinés. Il a été constaté que: i) l'index Area£ MGL est bien corrélé au poidsmoyen du nématode, lequel représente un bon indicateur de la réserve énergétique totale du nématode; ii) MGL est bien corrélé au contenu lipidique (en % du poids sec), bon indicateur de la qualité de la réserve énergétique; iii) le périmètre de l’image peut être utilisé pour estimer la longueur des IJ. Comparée aux méthodes couramment utilisées-telle la mesure directe du poids sec et du contenu lipidique moyens-l'analyse numérisée des images donne une meilleure information sur les échantillons testés et permet une estimation rapide des variations individuelles du poids sec, du contenu lipidique, de la taille et de la longueur des IJ. Un échantillon de 30 IJ peut être examine en 30 à 60 min ce qui rend cette méthode idéale pour les contrôles de qualité pendant la production, les traitements, la formulation et le stockage des produits biopesticides à base de nématodes entomopathogènes.

Energy metabolism and its relation to survival of the infective juveniles (IJ) of S. carpocapsae under anaerobic and oxygen-deficient conditions were studied by monitoring changes in survival rate, levels of key energy reserve materials, oxygen consumption, and respiratory quotient (RQ). The effects of various factors on the survival of IJ under anaerobic conditions were also investigated. Under anaerobic conditions, the IJ were inactivated but could survive for several days in an immobile state, using the carbohydrate reserves glycogen and trehalose for energy supply. The survival time of IJ was mainly dependent on the availability of energy supply, which, in turn, was influenced by factors such as temperature and metabolic by-products. Surviving, anaerobically incubated IJ fully recovered upon return to aerobic conditions. Recovering IJ were characterized by regaining mobility and restoration of carbohydrate reserves consumed during the anaerobic period. Carbohydrate reserves were restored by conversion from lipid reserves and possibly from anaerobic metabolic by-products. The infectivity of IJ recovered from the anaerobic state was not affected. At 1% oxygen level, IJ were also immobile and mainly depended on carbohydrate reserves for energy supply and the RQ was greater than 1. However, some oxygen was consumed; the survival time of these IJ was shorter than those kept in natural air but longer than those under anaerobic conditions. When IJ were incubated at oxygen levels of 3% to 21%, the RQs were maintained at 0.7 to 0.8. Oxygen consumption rates and the reduction in both mean dry weight and lipid levels were proportional to oxygen levels while the survival time of IJ was inversely proportional to oxygen levels.

Two hypotheses on the synthesis of the protectants glycerol and trehalose of the infective juveniles (IJs) of Steinernema carpocapsae during osmotic dehydration were tested and utilised to evaluate the function and importance of glycerol on survival of the nematodes during osmotic dehydration. This was achieved by comparing the changes in survival, morphology, behaviour and levels of glycerol, trehalose and permeated compounds of the IJs dehydrated in seven hypertonic solutions at two temperature regimes: (1) 5 8C for 15 days; and (2) 23 8C for 1 day followed by 5 8C for another 14 days. The results substantiate both hypotheses tested: (1) the permeability of the IJs to various compounds, such as sucrose or ethylene glycol, when they are dehydrated in hypertonic solutions of these compounds; and (2) suppression of the synthesis of protectant glycerol but not trehalose when IJs are dehydrated at low temperature. The results also showed that: (1) although trehalose was the preferred dehydration protectant, glycerol played an important role in rapidly balancing the osmotic pressure when IJs were exposed in hypertonic solutions; (2) the presence of glycerol was essential for the IJs to survive and function properly even under moderate osmotic dehydration, especially when IJs were dehydrated in salt solutions; and (3) some exogenous compounds permeated into IJs during osmotic dehydration such as ethylene glycol, may function in the same way as glycerol and significantly improve the survival and function of the IJs. The results indicate that each of the protectants glycerol and trehalose has a specific function and neither is replaceable by the other.

Changes in survival, mean dry weight, levels of key energy reserve compounds and respiration of non-feeding infective juveniles (IJs) of Steinernema carpocapsae incubated in various ratios of D2O:H2O on a shaker at 28

Permeability of the sheath and cuticle of the infective juveniles (IJs) of Steinernema carpocapsae to glycerol and its effect on biochemical adaptation of the IJs to osmotic dehydration were examined by incubating both sheathed and exsheathed IJs in glycerol-d5 solution then monitoring the changes in levels of deuterium labelled and non-labelled glycerol and trehalose. Energy metabolism of the IJs during osmotic dehydration and subsequent rehydration and the effect of the permeated glycerol on this process were investigated by examining and comparing the changes in mean dry weight and key biochemical composition of the IJs dehydrated in glycerol and sodium chloride solutions. The results show: (1) similarly to evaporative dehydration, osmotic dehydration induces IJs to synthesise the protectants glycerol and trehalose; (2) glycerol permeates the sheath and the cuticle into the body of IJs during dehydration in glycerol solution. Part of the permeated glycerol plays a role as protectant like that synthesised by IJs from their energy reserve materials while part is incorporated into trehalose; (3) the sheath reduces the rate of permeation of glycerol and therefore affects the equilibrium glycerol and trehalose levels of the IJs and also the time needed to reach the equilibrium levels; (4) the reduction in mean dry weight and lipids of the IJs during dehydration in glycerol solution is substantially less than those dehydrated in sodium chloride solution. Both the total protectant level and the ratio of glycerol to trehalose of the IJs dehydrated in glycerol solution are higher than those dehydrated in sodium chloride solution; (5) glycogen reserves of the IJs play a role as a buffer reservoir of the protectants during both dehydration and rehydration but the principal sources of the protectants during dehydration are more likely to be lipids and proteins rather than glycogen.

对昆虫病原线虫共生菌杀虫毒素的种类，与口服毒性有关的杀虫毒素以及口服毒性与杀虫毒素基因的关系等研究进展进行了综述，并对未来的研究方向提出了作者的见解。