4种葫芦科植物病毒复合胶体金免疫层析试纸条的研制

使用柠檬酸三钠还原法制备胶体金颗粒,在pH值7.6,蛋白用量10～15μg/mL条件下,制备形成4种葫芦科植物病毒多克隆抗体的稳定胶体金蛋白复合物;设计了双向复合试纸条,使用微定量喷头在试纸条两侧的硝酸纤维素膜上分别喷2条病毒检测线和1条质控线,组装制成免疫层析检测试纸条。结果表明经过条件优化后制备的双向复合试纸条特异性好,可在10min内同时检测4种葫芦科植物病毒,且对不同病毒阳性材料的检测灵敏度可达到稀释103倍以上。     

番茄褐色皱果病毒胶体金免疫试纸条的研制

 番茄褐色皱果病毒(tomato brown rugose fruit virus, ToBRFV)是一种新发病毒,严重威胁番茄的安全生产。为了快速、简便地检测该病毒,我们制备了ToBRFV胶体金免疫试纸条。本研究以ToBRFV粒子为免疫原,通过杂交瘤技术制备了17个抗ToBRFV的单克隆抗体。将不同单抗两两组合分别作为胶体金标记抗体和硝酸纤维素膜检测线上的捕获抗体,共获得272个配对组合的胶体金试纸条。通过特异性测定筛选到一组配对抗体制备的试纸条能够在5 min内特异识别ToBRFV,而与番茄斑驳花叶病毒、番茄花叶病毒、烟草花叶病毒、黄瓜花叶病毒、辣椒轻斑驳病毒、马铃薯X病毒、马铃薯Y病毒、番茄褪绿病毒、番茄斑萎病毒、番茄黄化曲叶病毒等无交叉反应。灵敏度分析表明,该试纸条可从稀释12 800倍的番茄叶片病汁液中检测到ToBRFV,也可检测到50 ng ToBRFV粒子。本研究制备的胶体金试纸条使用方便,灵敏度高,特异性强,适合田间大批量样品检测,可用于ToBRFV的精准监测及早期预警。     

建兰花叶病毒和齿兰环斑病毒联合免疫胶体金试纸条的研制及应用

为了实现兰花种植苗圃和口岸一线直接检测样品中建兰花叶病毒Cymbidium mosaic virus(CymMV)和齿兰环斑病毒Odontoglossum ringspot virus(ORSV)的目的,我们研制了上述两种病毒联合免疫胶体金检测试纸条。采用柠檬酸三钠还原法制备胶体金颗粒,标记单抗C10并固定于胶金垫上;将抗体5B7和4F3分别固定于硝酸纤维素膜上作为检测线,将羊抗鼠抗体包被固定于硝酸纤维素膜上作为质控线,经优化后组装胶体金试纸条。结果表明研制的联合试纸条特异性强,能够在10min之内同时检出两种病毒,操作简便。对两个种植苗圃的93份兰花样品进行实际检测所得结果与ELISA的检测结果符合性好(Kappa值分别为84.6%和86.7%),能满足兰花样品中建兰花叶病毒和齿兰环斑病毒的初筛要求。     

黄瓜细菌性角斑病免疫胶体金检测试纸条的研制

 应用免疫胶体金技术进行黄瓜细菌性角斑病(Pseudomonas syringae pv.lachrymans)的快速检测研究。为了研制黄瓜细菌性角斑病的免疫胶体金检测试纸条,通过柠檬酸三钠还原法制备胶体金,选择25 nm胶体金标记黄瓜细菌性角斑病菌多克隆抗体(CMb)。采用双抗夹心法,将CMb-胶体金复合物包被在胶体金结合垫上,将羊抗兔二抗和CMb包被在硝酸纤维素膜上,分别作为质控线(C)和检测线(T),组装成试纸条。该试纸条检测灵敏度为106 cfu/mL,与唐菖蒲疮痂病菌(Pseudomonas fluorescens biovarⅡ)等26个菌株无交叉反应,特异性强,检测时间为15 min。稳定性试验表明试纸条在37℃条件下放置15 d可保持检测结果的可靠性。用试纸条对田间采集的病叶进行检测,C线和T线清晰可见,缓冲液对照呈阴性反应。本试纸条可应用于生产上黄瓜细菌性角斑病早期快速检测,进而指导病害防治。     

马铃薯X病毒和马铃薯Y病毒胶体金免疫层析试纸条的研制

采用柠檬酸三钠还原法制备胶体金颗粒,标记马铃薯X病毒和马铃薯Y病毒的兔多克隆抗体。用微定量喷头在硝酸纤维素膜上喷好病毒检测线和羊抗兔抗体质控线,制成免疫层析检测试纸条。马铃薯X病毒试纸条检测烟草病汁液可稀释10000倍(重量/体积)马铃薯Y病毒试纸条检测烟草病汁液可稀释1000倍。10min内即可出检测结果。两种试纸条相互测试,未出现交叉反应。用健康和缓冲液对照测试,两种试纸条结果都为阴性。田间采集的马铃薯叶片用试纸条检测的结果和酶联结果相吻合。在密封、干燥、低温的条件下保存,有利于维持试纸条的稳定性。     

番茄环斑病毒和烟草环斑病毒复合型胶体金免疫层析试纸条的研制

采用柠檬酸三钠还原法制备胶体金颗粒,标记番茄环斑病毒和烟草环斑病毒的兔多克隆抗体,用微定量喷头在硝酸纤维素膜上喷好2条病毒检测线（T线）和1条羊抗兔抗体质控线（C线）,制成复合型免疫层析检测试纸条。一张试纸条在10min内,可同时检测出这两种病毒。试纸条检测番茄环斑病毒和烟草环斑病毒粗提纯液,检测灵敏度在1μg/mL,试纸条检测番茄环斑病毒和烟草环斑病毒的混合病汁液可稀释1000倍（重量/体积）。用健康叶片和缓冲液对照测试,试纸条结果都为阴性。     

黄瓜细菌性白枯病病菌检测试纸条的研制

应用胶体金免疫层析技术研制了黄瓜细菌性白枯病病菌[Pseudomonas viridiflava (Burkholder 1930) Dowson1939]检测试纸条.采用柠檬酸三钠法还原氯金酸制备胶体金,标记黄瓜细菌性白枯病病菌多克隆抗体,将金标抗体喷涂在结合垫上,将黄瓜细菌性白枯病病菌抗体和羊抗兔二抗包被在硝酸纤维素膜上作检测线和质控线,组装制成黄瓜细菌性白枯病病菌检测试纸条.用试纸条检测黄瓜细菌性白枯病病菌的结果表明,制备的试纸条特异性好,与其他常见植物病原细菌等无交叉反应,对黄瓜叶片中黄瓜细菌性白枯病病菌的最低检测限为106 cfu/mL,能在5～15 min内快速检测出黄瓜细菌性白枯病病菌,适合田间现场快速检测黄瓜细菌性白枯病病菌.     

马铃薯Y病毒CP基因的原核表达及免疫金标试纸条的研制

选取马铃薯Y病毒（potato virus Y,PVY）代表株（NC＿001616.1）外壳蛋白基因37～281区间，采用化学合成方法合成目标序列，将其连接到原核表达载体pET-32a（+），并转化至大肠杆菌BL21(DE3)，经IPTG诱导表达出45 kDa融合蛋白，回收后免疫日本大耳白兔制备抗血清。Western Blot分析多克隆抗体特异性良好，后制备25 nm胶体金，标记PVY CP37-281多克隆抗体，制备胶体金免疫层析试纸条。结果表明，试纸条能在15 min内检出PVY,PVY病株汁液检测稀释限度可达106倍（W/V），使用常见复合侵染烟草检测，未出现交叉反应。由此可知，该试纸条操作简便，反应灵敏特异，适用于田间一线及种薯生产中马铃薯Y病毒的检测。     

黄瓜绿斑驳花叶病毒磁免疫层析试纸条的研制

用磁性纳米颗粒标记黄瓜绿斑驳花叶病毒的兔多克隆抗体建立了简便快速的磁免疫层析试纸条检测方法。该方法检测提纯病毒的灵敏度为0.1 μg/mL,检测葫芦病汁液可稀释10 000倍(重量／体积)。试纸条和黄瓜绿斑驳花叶病毒具有特异性反应,和同属的烟草花叶病毒、番茄花叶病毒、辣椒轻斑驳病毒和齿兰环斑病毒,未出现交叉反应。该试纸条可用于田间病害检测和诊断。     

检测南方水稻黑条矮缩病毒胶体金免疫试纸条的建立

由白背飞虱Sogatella furcifera(Horváth)传播的南方水稻黑条矮缩病毒(Southern rice blackstreaked dwarf virus,SRBSDV)是目前我国南方水稻上危害最严重的病毒,为开发简便、快速、准确的SRBSDV病毒检测技术和检测试剂,以感染SRBSDV的植物粗提液为免疫原,利用杂交瘤技术制备了2株抗SRBSDV的单抗(14A8和15G6),并利用制备的单抗建立了可快速、特异、灵敏地检测SRBSDV的胶体金免疫试纸条。结果表明,2株制备单抗的抗体类型及亚类均为Ig G1、kappa链,单抗腹水的间接ELISA效价均达到10~(-7);Western blot分析表明,2株单抗均与SRBSDV的外壳蛋白亚基有特异反应,而不与水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV)反应。以制备14A8和15G6单抗分别为捕获抗体和胶体金标记抗体,开发成能在5 min内准确、特异地检测水稻植物和白背飞虱传毒介体体内SRBSDV的胶体金免疫试纸条;灵敏度分析表明,该检测试纸条的检测水稻病叶的灵敏度达到1∶6 400倍(g/m L),检测单头携毒白背飞虱的灵敏度达到1∶51 200倍(单头/μL)。田间样品检测结果表明,该试纸条的检测结果与RT-PCR的符合率达到100%。建立的SRBSDV胶体金免疫试纸条可对南方水稻黑条矮缩病毒进行快速、特异、灵敏的诊断和检测。     

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.     

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     

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.     

The autumn leafroller: Phenology,damage and parasitoids in a Dutch apple orchard

The phenology of the autumn leafroller,Syndemis musculana, a local pest of apple, was studied in order to forecast larval emergence. From 1983–1986, peak flight as determined with sexpheromone traps was always between 13–18 May. The duration of embryonic development was determined at various constant temperatures and used to estimate the periods of egg hatch in these four years. Each year, most eggs should have hatched in the second decade of June.Differences in attack rates between apple cultivars seem to be explained largely by the variation in picking time. Larvae are only half grown at the beginning of harvest (cv. James Grieve), and have gone into hibernation when the latest variety (cv. Golden Delicious) is picked. Moreover, the varieties Cox's Orange Pippin and Belle de Boskoop, picked about half time, are liable to receive additional damage by caterpillars brought with the picked fruits into storage.Various hymenopterous parasites were reared from caterpillars. As the only leafroller in the orchard which hibernates as mature larva,S. musculana may promote winter survival of some parasitoids, like the eulophidColpyclypeus florus.Samenvatting De fenologie van de herfstbladroller (Syndemis musculana Hübner), een incidentele plaag op appel, werd nader bepaald met het doel het uitkomen van de eieren te kunnen voorspellen. In 1983–1986 viel de piekvlucht, bepaald met behulp van feromoonvallen, steeds tussen 13 en 18 mei.De ontwikkelingsduur van de eieren bij verschillende constante temperaturen werd gebruikt om de periode van uitkomen te schatten. De meeste eieren zullen ieder jaar in de eerste helft van juni uitkomen.Geconstateerde verschillen in schade tussen appelrassen blijken goeddeels terug te voeren op verschillen in pluktijdstip. De rupsen van de herfstbladroller zijn pas half-was als de eerste appels eind augustus geplukt worden, terwijl tegen het einde van de oogst begin oktober de meeste al in winterslaap zijn. Met name tussentijdse rassen, als Cox's Orange Pippin and Schone van Boskoop, lopen extra schade op doordat grotere rupsen met de geplukte vruchten in de kist terecht komen.Uit de rupsen werden negen, al van andere boomgaardbladrollers bekende, sluipwespen gekweekt, Omdat deze bladrollersoort, als enige in de boomgaard, als volgroeide rups overwintert, lijkt zij bij uitstek geschikt als winterwaard.This study was carried out at the Experimental Orchard De Schuilenburg, Schuilenburg 3, 4041 BK Kesteren, the Netherlands, to which address correspondence should be addressed.     

The parasitoid complex ofLiriomyza cicerina on chickpea (Cicer arietinum)

Liriomyza cicerina (Diptera: Agromyzidae) is an important pest on chickpea in Turkey. The objective of this study was to determine the parasitoids and rates of parasitism ofL. cicerina on chickpea (Cicer arietinum L.) during the 2005 and 2006 seasons in ?anl?urfa province, Turkey. Leaves with mines were sampled weekly and kept in the laboratory to observe and count emerging leafminer and parasitoid adults. Eight parasitoid species were collected: the braconidsOpius monilicornis Fischer andOpius tersus Foerster and the eulophidsDiaulinopsis arenaria (Erdös) andNeochrysocharis formosa (Westwood), which occurred in both the winter and summer seasons;Diglyphus crassinervis Erdös,Neochrysocharis ambitiosa Hansson,Neochrysocharis sericea (Erdös) andPediobius metallicus (Nees), which occurred only in the summer growing areas.Diaulinopsis arenaria was the predominant parasitoid with 4–7.7% parasitism rate whileN. ambitiosa andO. monilicornis were the second and third most predominant species. The results of these trials show that sinceDia. arenaria occurred throughout every season, it could potentially be used for control of the leafminerL. cicerina.