稻瘟病菌一假定几丁质酶多克隆抗体的制备及其检测

 为检测几丁质酶的表达,制备稻瘟病菌的一假定几丁质酶多克隆抗体,探索其可能运用。将稻瘟病菌(Magnaporthe oryzae) 几丁质酶基因克隆入融合表达载体pET\|32a,并在大肠杆菌Escherichia coli BL21(DE3)中进行诱导表达。表达菌株经0.5 mmol/L IPTG诱导4～6 h后,用Ni²⁺\|NTA亲和柱纯化蛋白,得到可溶的几丁质酶\|His融合蛋白。以该融合蛋白免疫新西兰雄兔,制备多克隆抗体。ELISA分析表明该抗体效价达1∶204 800,特异性良好。用该抗体ELISA检测了稻瘟病菌4个不同田间菌株中,不同的菌株表达量不同。对稻瘟病菌侵染水稻3、5和7d后的病叶进行抗原Western验证,结果表明,稻瘟病菌不同的侵染时间其几丁质酶表达量有明显差异。 几丁质酶表达的差异是否与菌株间致病型等生物学和生理学差异有关,目前正在进一步研究。     

溴氰菊酯酶联免疫吸附分析方法研究

建立了定量测定溴氰菊酯间接竞争酶联免疫吸附分析方法(ic-ELISA)。合成了半抗原1-羧基-(3'-苯氧基苯基)甲基-3-(2',2'-二溴乙烯基)-2,2-二甲基环丙基羧酸酯(Med)和N-2-(羧基丙基)氨基甲酰基-(3'-苯氧基苯基)甲基-3-(2',2'-二溴乙烯基)-2,2-二甲基环丙基羧酸酯(Di)。分别采用碳二亚胺法和混合酸酐法将半抗原与牛血清蛋白(BSA)和卵清蛋白(OVA)偶联制备了免疫原Di-BSA和包被原Di-OVA、Med-OVA。将制得的溴氰菊酯免疫原免疫动物获得多克隆抗体,经间接非竞争ELISA法测得其效价为2.5×105。通过对甲醇含量、离子强度、pH值等影响因素进行异源分析条件的优化,确立了溴氰菊酯间接竞争酶联免疫分析方法的最佳检测条件(30%甲醇、氯化钠0.4 mol/L、pH 7.5),并建立了标准竞争曲线。该方法的IC50值为0.55±0.05 mg/L,检测限(IC10)为3.76±0.35 μg/L,对大多数拟除虫菊酯无交叉反应。分别在自来水、河水和土壤样品中添加0.05 ~5.0 mg/L(或mg/kg)的溴氰菊酯,回收率分别为89.7% ~106.8%、82.4% ~101.7%、75.6% ~97.8%。     

高效氯氰菊酯对小菜蛾阿维菌素抗性、敏感种群的亚致死效应

在实验室条件下研究了高效氯氰菊酯亚致死剂量LC10(43.44mg/L)(小菜蛾阿维菌素抗性种群)和LC10(12.61mg/L)(敏感种群)对小菜蛾阿维菌素抗性(AV-R)、敏感(AV-S)种群的亚致死效应。结果表明:高效氯氰菊酯亚致死剂量12.61mg/L处理小菜蛾阿维菌素敏感、抗性种群3龄幼虫后,处理组AV-S(SLC10)和AV-R(SLC10)的化蛹率、羽化率显著低于AV-S(ck)对照组,与AV-R(ck)对照组无显著性差异;用高效氯氰菊酯亚致死剂量43.44mg/L处理抗性种群3龄幼虫后,其处理组AV-R(RLC10)的化蛹率、蛹重、羽化率都显著低于对照组。AV-S(SLC10)组的单雌产卵量为176.45±36.11粒,显著高于对照组和处理的抗性种群,AV-R(SLC10)和AV-R(RLC10)组的单雌产卵量与对照组间无显著性差异,AV-S(SLC10)和AV-R(RLC10)组的产卵期明显缩短。AV-S(SLC10)组的内禀增长率(rm)、周限增长率(λ)和净增殖率(R0),显著高于对照和处理的抗性种群。结果表明高效氯氰菊酯亚致死剂量对小菜蛾阿维菌素敏感种群具有一定的刺激增殖作用。     

虫酰肼对苜蓿银纹夜蛾核型多角体病毒的增效作用

研究了以甜菜夜蛾为靶标害虫时,昆虫生长调节剂虫酰肼对苜蓿银纹夜蛾核型多角体病毒(AcNPV)的增效作用.结果表明:虫酰肼在作用于甜菜夜蛾时对AcNPV有显著增效作用,不仅可提高杀虫毒力,而且也能提高杀虫速度.作用于3龄和4龄幼虫时,6.67 mg/L虫酰肼对AcNPV的增效比分别是1.31和3.62,使害虫因表现感染病毒症状而死亡的LT50(致死中时)分别缩短10.8%和6.5%,使害虫因病毒和虫酰肼作用而死亡的LT50分别缩短25%和20.4%.虫酰肼和病毒协同作用还显著抑制了幼虫的生长发育,在协同作用1天后,幼虫体重即显著低于单用病毒或对照,并且随着作用时间的延长,协同作用组幼虫体重与病毒单剂组及对照组的差异越来越大.但6.67mg/L虫酰肼和2.7×105PIB/mL AcNPV协同作用于幼虫期时,其对存活幼虫发育为成虫后的繁殖力无影响,成虫产卵量、交配率、卵孵化率以及卵孵化幼虫数等均与病毒单剂及虫酰肼单剂无显著差异,也与对照无显著差异.     

Dermestid beetles in ‘evolution Canyon’, lower Nahal Oren, Mt. Carmel, including new records for Israel

Fifteen species of dermestid beetles were recorded at ‘Evolution Canyon’ (EC), Lower Nahal Oren, Mt. Carmel, Israel. They represent ~35% of known Israeli dermestid species. The following three species were recorded for the first time in Israel:Trogoderma svriaca Dalla Torre, 1911;Ctesias svriaca Ganglbauer, 1904; andAnthrenus (s.str.) jordaniens Pic, 1934. Adults of 13 species were collected on the more solar radiated, warmer and climatically more fluctuating south-facing slope (SFS); ten species were collected on the opposite, north-facing slope (NFS), which was cooler and climatically more stable. The abundance of adult dermestid beetles was 1.9 times higher on the SFS than on the NFS (86 and 47, respectively). Species richness and abundance distribution at EC (three collecting stations on each slope and one at the valley bottom) were significantly negatively correlated with the plant cover that consisted of trees and bushes (Spearmanr _s ,P=0.007 and 0.039, respectively) and perennials (Spearmanr _s ,P=0.039 and 0.077, respectively), indicating that non-woody plants were preferred by adult dermestid beetles.     

Epidemiology of Toxigenic Fungi and their Associated Mycotoxins for Some Mediterranean Crops

Recent data on the epidemiology of the common mycotoxigenic species of Fusarium, Alternaria, Aspergillus and Penicillium in infected or colonized plants, and in stored or processed plant products from the Mediterranean area are reviewed. Emphasis is placed on the toxigenicity of the causal fungal species and the natural occurrence of well known mycotoxins (aflatoxins, ochratoxins, fumonisins, trichothecenes, zearalenone, patulin, Alternaria-toxins and moniliformin), as well as some more recently described compounds (fusaproliferin, beauvericin) whose toxigenic potential is not yet well understood. Several Fusarium species reported from throughout the Mediterranean area are responsible of the formation of mycotoxins in infected plants and in plant products, including: Fusarium graminearum, F. culmorum, F. cerealis, F. avenaceum, F. sporotrichioides and F. poae, which produce deoxynivalenol, nivalenol, fusarenone, zearalenone, moniliformin, and T-2 toxin derivatives in wheat and other small grains affected by head blight or scab, and in maize affected by red ear rot. Moreover, strains of F. verticillioides, F. proliferatum, and F. subglutinans, that form fumonisins, beauvericin, fusaproliferin, and moniliformin, are commonly associated with maize affected by ear rot. Fumonisins, were also associated with Fusarium crown and root rot of asparagus and Fusarium endosepsis of figs, caused primarily by F. proliferatum. Toxigenic A. alternata strains and associated tenuazonic acid and alternariols were commonly found in black mould of tomato, black rot of olive and citrus, black point of small cereals, and black mould of several vegetables. Toxigenic strains of A. carbonarius and ochratoxin A were often found associated with black rot of grapes, whereas toxigenic strains of A. flavus and/or P. verrucosum, forming aflatoxins and ochratoxin A, respectively, were found in moulded plant products from small cereals, peanuts, figs, pea, oilseed rape, sunflower seeds, sesame seeds, pistachios, and almonds. Finally, toxigenic strains of P. expansum and patulin were frequently found in apple, pear and other fresh fruits affected by blue mould rot, as well as in derived juices and jams.     

Diagnostics,genetic diversity and pathogenic variation of ascochyta blight of cool season food and feed legumes

Molecular diagnostic techniques have been developed to differentiate the Ascochyta pathogens that infect cool season food and feed legumes, as well as to improve the sensitivity of detecting latent infection in plant tissues. A seed sampling technique was developed to detect a 1% level of infection by Ascochyta rabiei in commercial chickpea seed. The Ascochyta pathogens were shown to be genetically diverse in countries where the pathogen and host have coexisted for a long time. However, where the pathogen was recently introduced, such as A. rabiei to Australia, the level of diversity remained relatively low, even as the pathogen spread to all chickpea-growing areas. Pathogenic variability of A. rabiei and Ascochyta pinodes pathogens in chickpea and field pea respectively, appears to be quantitative, where measures of disease severity were based on aggressiveness (quantitative level of infection) rather than on true qualitative virulence. In contrast, qualitative differences in pathogenicity in lentil and faba bean genotypes indicated the existence of pathotypes of Ascochyta lentis and Ascochyta fabae. Therefore, reports of pathotype discrimination based on quantitative differences in pathogenicity in a set of specific genotypes is questionable for several of the ascochyta-legume pathosystems such as A. rabiei and A. pinodes. This is not surprising since host resistance to these pathogens has been reported to be mainly quantitative, making it difficult for the pathogen to overcome specific resistance genes and form pathotypes. For robust pathogenicity assessment, there needs to be consistency in selection of differential host genotypes, screening conditions and disease evaluation techniques for each of the Ascochyta sp. in legume-growing countries throughout the world. Nevertheless, knowledge of pathotype diversity and aggressiveness within populations is important in the selection of resistant genotypes.     

Hymenopteran Parasitoids on Fruit-infesting Tephritidae (Diptera) in Latin America and the Southern United States: Diversity, Distribution, Taxonomic Status and their use in Fruit Fly Biological Control

We first discuss the diversity of fruit fly (Diptera: Tephritidae) parasitoids (Hymenoptera) of the Neotropics. Even though the emphasis is on Anastrepha parasitoids, we also review all the information available on parasitoids attacking flies in the genera Ceratitis, Rhagoletis, Rhagoletotrypeta, Toxotrypana and Zonosemata. We center our analysis in parasitoid guilds, parasitoid assemblage size and fly host profiles. We also discuss distribution patterns and the taxonomic status of all known Anastrepha parasitoids. We follow by providing a historical overview of biological control of pestiferous tephritids in Latin American and Florida (U.S.A.) and by analyzing the success or failure of classical and augmentative biological control programs implemented to date in these regions. We also discuss the lack of success of introductions of exotic fruit fly parasitoids in various Latin American countries. We finish by discussing the most pressing needs related to fruit fly biological control (classical, augmentative, and conservation modalities) in areas of the Neotropics where fruit fly populations severely restrict the development of commercial fruit growing. We also address the need for much more intensive research on the bioecology of native fruit fly parasitoids.     

A survey of the genera of<Emphasis Type="Italic">Microgaster</Emphasis> latreille and<Emphasis Type="Italic">Hygroplitis</Emphasis> thomson (Hymenoptera: Braconidae: Microgastrinae) from China

The genera ofMicrogaster Latreille 1804 andHygroplitis Thomson 1895 from China are presented systematically in this paper. Thirty-two species ofMicrogaster and three species ofHygroplitis are known in China. Diagnosis, character variation, distribution and host of each species among the two genera are presented, including its host and distribution. Keys to the species ofMicrogaster andHygroplitis are given. http://www.phytoparasitica.org posting Dec. 19, 2006.     

Plant Viruses Transmitted by Whiteflies

One-hundred and fourteen virus species are transmitted by whiteflies (family Aleyrodidae). Bemisia tabaci transmits 111 of these species while Trialeurodes vaporariorum and T. abutilonia transmit three species each. B. tabaci and T. vaporariorum are present in the European–Mediterranean region, though the former is restricted in its distribution. Of the whitefly-transmitted virus species, 90% belong to the Begomovirus genus, 6% to the Crinivirus genus and the remaining 4% are in the Closterovirus, Ipomovirus or Carlavirus genera. Other named, whitefly-transmitted viruses that have not yet been ranked as species are also documented. The names, abbreviations and synonyms of the whitefly-transmitted viruses are presented in tabulated form together with details of their whitefly vectors, natural hosts and distribution. Entries are also annotated with references. Whitefly-transmitted viruses affecting plants in the European–Mediterranean region have been highlighted in the text.     

Brief Guide for Authors

正Journal of Arid Land(JAL)is an international journal(ISSN 1674-6767;CN 65-1278/K)for the natural sciences,sponsored by the Xinjiang Institute of Ecology and Geography,Chinese Academy of Sciences and Science Press.It is published by Science Press and Springer-Verlag Berlin Heidelberg bimonthly.JAL publishes original,innovative,and integrative research from arid and semiarid regions,ad     

The use of Solacol® (validamycin) as a growth retardant in the isolation of soil fungi

Solacol^®, a formulation of the antibiotic validamycin, at 0.33% in 2% malt extract agar, reduced the spread of fungi on dilution plates drastically and allowed twice as much incubation time before subculturing; this resulted in an elevated number of species isolated. Using pure cultures of 62 common soil fungi, it was shown that all fast-growing species (exceptPythium ultimum) were efficiently inhibited but not completely suppressed. Inhibition was comparable to that by 0.5% oxgall, though, while this substance completely suppressed several species, Solacol very strongly inhibited onlyGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum andRhizoctonia solani. In a further experiment each separate constituent of Solacol was tested against 22 fungi at equivalent concentrations. Validamycin strongly inhibitedChaetomium globosum and two Basidiomycetes, though hardly more than the non-ionic detergent which mainly inhibited the other fungi. A few species were, however, more inhibited by Solacol than by the detergent alone. Solacol at 0.33% is a suitable aid in dilution plating of soil fungi, by increasing the number of colonies and species observed.Samenvatting Solacol^®, een formulering van het antibioticum validamycine, remde de groei van schimmels in verdunningsplaten met een concentratie van 0.33% in 2% moutagar en maakte het mogelijk de periode tot afenten met een factor 2 te verlengen; daardoor was het aantal geïsoleerde soorten duidelijk toegenomen. Met reincultures van 62 algemene grondschimmelsoorten werd aangetoond, dat alle snelgroeiende soorten (met uitzondering vanPythium ultimum) voldoende geremd, maar niet volkomen onderdrukt werden. Het remmingspercentage was vergelijkbaar met dat van 0.5% ossegal, hoewel dit laatste sommige soorten volkomen onderdrukte; Solacol remde alleenGaeumannomyces graminis, Gerlachia nivalis, Harzia acremonioides, Verticillium biguttatum enRhizoctonia solani zeer sterk. In een volgend experiment werden de componenten van Solacol t.o.v. 22 fungi apart getoetst in concentraties equivalent aan 0.33% Solacol. Validamycine remde alleenChaetomium globosum en twee basidiomyceten behoorlijk, maar nauwelijks meer dan de niet-ionische uitvloeier, die in hoofdzaak de overige remeffecten veroorzaakte. Enkele soorten werden echter door het complete Solacol veel sterker geremd dan door de uitvloeier alleen. Solacol in een verdunning van 0,33% wordt aanbevolen bij verdunningsplaten voor het isoleren van grondschimmels ten einde het aantal kolonies en soorten te verhogen.     

Broad bean mottle virus in Morocco; curculionid vectors,and natural occurrence in food legumes other than faba bean (Vicia faba)

Broad bean mottle virus (BBMV) was transmitted from infected to healthy faba-bean plants by the curculionid weevilsApion radiolus Kirby,Hypera variabilis Herbst,Pachytychius strumarius Gyll,Smicronyx cyaneus Gyll, andSitona lineatus L. The latter appeared to be an efficient vector: acquisition and inoculation occurred at the first bite, the rate of transmission was c. 41%, and virus retention lasted for at least seven days.S. lineatus transmitted the virus from faba bean to lentil and pea, but not to the three genotypes of chickpea tested. This is the first report on the generaHypera, Pachytychius, andSmicronyx as virus vectors, and onA. radiolus, H. variabilis, P. strumarius, andS. cyaneus as vectors of BBMV.Out of 351 samples of food legumes with symptoms suggestive of virus infection, 16, 11, 19, and 17% of the samples of chickpea, lentil, pea, and common bean, respectively, were found infected when tested for BBMV in DAS-ELISA. This is the first report on the natural occurrence of BBMV in chickpea, lentil, pea, and common bean. The virus should be regarded as a food-legume virus rather than a faba-bean virus solely, and is considered an actual threat to food legume improvement programmes.     

Characterization of potato potyvirus Y (PVY) isolates from seed potato batches. Situation of the NTN, Wilga and Z isolates

A collection of 38 PVY isolates from seed potato batches, originating from several Western European countries, was characterized by using current biological, serological and molecular tools differentiating PVY strains and groups. The correlation between the three kinds of tests was good but not absolute. No single serological or PCR method was able to discriminate among the five isolate groups found. Twenty-nine isolates belonged to the PVY^N strain and six to the PVY^O strain. No PVY^C was found. Two other isolates reacted serologically like PVY^O, but were unable to elicit a hypersensitive response from the Ny_tbr gene and probably represent the PVY^Z group. At the molecular level, these two isolates showed a combination of both PVY^O and PVY^N and could be recombinants of these strains. Another isolate reacted serologically like PVY^O, but induced vein necrosis in tobacco, like PVY^N-Wilga. Some PVY^N isolates caused tuber ring necrosis in glasshouse conditions. These might belong to the PVY^NTN group. The PVY^NTN, PVY^N-Wilga and PVY^Z groups probably represent pathotypes within strains PVY^N and PVY^O, respectively. The present study also confirms previous reports showing a high genetic variation at the 5 end within the PVYN strain.