褐色橘蚜全蛋白双向电泳体系条件优化

为优化有效分离褐色橘蚜Toxoptera citricida全蛋白的双向电泳体系,比较了饱和酚抽提法、三氯乙酸(trichloroacetic acid,TCA)/丙酮沉淀法、直接裂解液法和Tris-HCl法4种蛋白提取方法,以及蛋白上样量、聚焦条件和固相p H梯度(immobilized p H gradient,IPG)胶条对褐色橘蚜全蛋白双向电泳的影响。结果表明,TCA/丙酮沉淀法和直接裂解液法获得的全蛋白浓度分别为4.25、4.03μg/μL,显著高于饱和酚抽提法和Tris-HCl法,且TCA/丙酮沉淀法所提取的蛋白种类丰富度最高。采用TCA/丙酮沉淀法获得全蛋白,当上样量为100μg时,所得蛋白质点多、分辨率高、背景清晰,除盐3 h,等电聚焦量45 000 V·h,并选用p H 3~10非线性IPG胶条时,双向电泳图谱中横向拖尾现象少,蛋白点相对清晰,分布均匀。表明所建立的双向电泳体系,结合银染,可从褐色橘蚜全蛋白中识别出1 264个蛋白点,且重复性研究匹配率高,可用于褐色橘蚜蛋白质组学研究。     

甘草愈伤组织蛋白质组双向电泳技术的建立

以来源于野生乌拉尔甘草的愈伤组织为试材,对甘草愈伤组织蛋白质的提取、SDS-PAGE电泳、上样量、染色方法等关键步骤进行了优化,结果表明:采用改良丙酮法可提高提取液中蛋白质的含量及单向电泳图谱的分辨率;17 cm IPG胶条上样量为450μg时,采用银染法提高了双向电泳图谱的分辨率及分离效果,获得了1 132个蛋白点。建立了一套适用于乌拉尔甘草的愈伤组织蛋白质组分析的双向电泳方法。     

适用于双向电泳分析的链格孢菌体蛋白提取方法的筛选

为筛选出一套适用于双向电泳分析的链格孢菌蛋白的提取方法,以该菌为研究材料,首先采用TCA-丙酮法、磷酸-TCA-丙酮法、SDS提取法和TCA/丙酮-SDS/酚抽提法分别提取菌丝蛋白质,然后进行蛋白质浓度的测定和单向SDS-PAGE试验,并建立双向电泳体系。结果表明:TCA/丙酮-SDS/酚抽提法所获得的蛋白质浓度为15.9 mg/mL,分别是其他三种方法所得蛋白浓度的2.64、4.61和1.43倍;SDS-PAGE图谱中形成的条带数目最多、丰度最好;所获得的2-DE图谱背景清晰干净,无明显的横纵条纹,分辨率高,可辨别的蛋白质点数为1 138个,是TCA-丙酮法所分离蛋白点数的1.89倍。由此可知,TCA/丙酮-SDS/酚抽提法为链格孢菌蛋白提取的最优方法,该方法所提取的蛋白质适用于双向电泳分析及后续的蛋白质组学研究。     

南方小花蝽对为害蚕豆的三种蚜虫的捕食作用

豆蚜Aphis craccivora、豌豆蚜Acyrthosiphon pisum和豌豆修尾蚜Megoura crassicauda是为害豆科作物的主要害虫。南方小花蝽Orius strigicollis是重要捕食性食蚜天敌,广泛分布于我国南方。为明确南方小花蝽对豆科作物上3种蚜虫的捕食作用和生物防治潜能,本研究在实验室条件下,测定了南方小花蝽对3种蚜虫的捕食选择性和捕食功能反应。结果表明,南方小花蝽对3种蚜虫的捕食功能反应均符合HollingⅡ模型,不同龄期南方小花蝽对3种蚜虫捕食量由大到小依次为5龄若虫,雌成虫,低龄若虫。对豆蚜的捕食量均大于对豌豆修尾蚜的捕食量,对豌豆蚜的捕食量最小,雌成虫对以上3种蚜虫的日最大捕食量分别为豆蚜63.69头/d、豌豆蚜36.23头/d和豌豆修尾蚜49.26头/d。南方小花蝽对3种蚜虫的寻找效应随蚜虫密度的增加而降低,随小花蝽龄期的增加而升高,相同龄期的南方小花蝽对3种蚜虫的寻找效应由大到小依次为豌豆修尾蚜,豆蚜,豌豆蚜。南方小花蝽对猎物的捕食作用受自身密度的干扰,当猎物密度不变时,随着小花蝽自身密度的增加,单头小花蝽平均捕食量下降,其干扰反应符合H...     

麦长管蚜在不同温度下的年龄-龄期生命表

为了解温度对麦长管蚜Sitobion avenae (Fabricius) 种群动态的影响,采用生命表方法研究了16、19、22、25、28 ℃共5个不同温度下麦长管蚜的实验种群生命表,并分析了温度对麦长管蚜各生命表参数的影响。结果表明:麦长管蚜繁殖力(F)、净生殖率(R₀)、成蚜历期及寿命均随温度升高呈先增加后减小的趋势,在22 ℃时最大,分别为16.7±2.0头、14.11±1.90头、15.6±1.6 d、26.4±1.5 d。各龄若蚜发育历期和平均世代周期(T)随温度升高而缩短,在25 ℃时最短,其中T为12.83±0.30 d。内禀增长率(r)、周限增长率(λ)随温度升高而增大,在25 ℃时最大,分别为0.20±0.01/d和1.22±0.01/d。当温度达到28 ℃,内禀增长率出现负值为-0.14±0.02/d,周限增长率、净生殖率、繁殖力、寿命均最小,分别为0.87±0.02/d、0.16±0.04头、0.4±0.1头、15.5±0.3 d。除平均世代周期外,其它参数值在22 ℃均高于19 ℃,但无显著差异。表明麦长管蚜的生长繁殖在19 ℃和22 ℃较适,而在28 ℃受到抑制。     

福州地区柑桔蚜虫的种群动态

福州地区为害柑桔的蚜虫有7种。绣线菊蚜 Aphis citricola van der Goot为秋季重发型;棉蚜 A.gossypii Glover春、夏发生为主,桔蚜 Toxoptera citricidus(Kirkaldy)为全年轻发型。春季蚜虫的虫源以迁入为主。有翅蚜梢率和有翅胎生蚜的数量与受害程度之间有显著相关性。柑桔新梢的抽发时期、数量和成熟的快慢对蚜虫种群的变动产生重要影响。     

栗苞蚜的生活习性及其防治对策

为了对栗苞蚜Moritziella castaneivora Miyazaki进行有效监控,控制其蔓延及制定合理的防治策略,通过林间调查及室内观察研究栗苞蚜对日本栗的为害、主要生活习性和天敌对其捕食行为。结果表明,栗苞蚜对银寄和筑波为害炸苞率分别达(7.4±6.0)%和(8.6±5.5)%;4龄若蚜的耐饥力强于成蚜,二者离开寄主20 h后可分别存活50%和13%,且1龄若蚜迁移力明显强于成蚜;栗苞蚜卵在20:00的孵化率明显高于8:00和14:00。龟纹瓢虫、黑襟毛瓢虫、七星瓢虫和草蛉对栗苞蚜卵具有较强的捕食能力,龟纹瓢虫平均捕食栗苞蚜卵量约为43粒/h,平均取食1个卵为10.1 s,搜寻速率为8.5粒/min。此外,研究表明初春施用石硫合剂可有效减少虫口基数,防止当年栗苞蚜大发生。     

50g/L双丙环虫酯可分散液剂对草莓、马铃薯蚜虫的田间防效研究

探讨双丙环虫酯对草莓蚜虫、马铃薯蚜虫的田间防效.开展了50g/L双丙环虫酯可分散液剂防治草莓、马铃薯蚜虫的田间药效试验.药后7d,药剂处理区蚜量在清水对照区蚜量的<1/10,防效在90％～95％,可有效压低虫量,控制其危害.50g/L双丙环虫酯可分散液剂对草莓、马铃薯蚜虫具有良好的防效.     

蚜虫为害对五个燕麦品种苗期体内几种物质的影响

［目的］研究蚜虫为害后燕麦苗期植株体内的叶绿素、可溶性糖、可溶性蛋白、总酚含量的变化,评价这4种物质与燕麦抗蚜性的关系,探索燕麦抗蚜的生化机制,为选育抗蚜燕麦品种提供科学依据。［方法］ 选取田间具有不同抗蚜表现的5个燕麦品种,待幼苗长至3叶期接蚜,分别于接蚜时和接蚜后不同时间测定其体内的叶绿素、可溶性糖、可溶性蛋白、总酚的含量,与同期未接蚜植株做比较。［结果］ 随着感蚜时间增加,每个品种叶绿素含量均明显降低,作为抗蚜物质的总酚含量增加,可溶性糖和可溶性蛋白也有所增加。［结论］ 说明燕麦的抗蚜性与其体内的可溶性糖、可溶性蛋白、酚类物质含量有关。     

五种杀虫剂对苜蓿种子田中主要害虫和传粉昆虫的影响

为筛选高效、低毒的杀虫剂，测定1.5%除虫菊素、0.3%印楝素、4%阿维·啶虫脒、2.5%高效氯氟氰菊酯和70%吡虫啉5种杀虫剂对苜蓿种子田主要害虫牛角花齿蓟马Odontothrips loti和豌豆蚜Acyrthosiphon pisum的室内毒力和田间防效，并分析这5种药剂对主要传粉昆虫数量及多样性指数的影响。结果显示，0.3%印楝素对牛角花齿蓟马和豌豆蚜的毒力相对较高，LC₅₀分别为1.093 mg/L和2.864 mg/L。70%吡虫啉以及4%阿维·啶虫脒对牛角花齿蓟马和豌豆蚜均有较高的防效，最高防效均达到85.00%以上，但这2种药剂对传粉昆虫威胁较大，而0.3%印楝素则对传粉昆虫威胁较小。推荐花期苜蓿种子田使用0.3%印楝素1 000倍稀释液防控害虫，而非开花期可交替使用70%吡虫啉1 000倍稀释液和4%阿维·啶虫脒1 000倍稀释液防控害虫。     

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.     

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.     

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.     

Characterization of Sclerotinia minor populations in Texas peanut fields

In the summer of 2004 an epidemic of sclerotinia blight of peanut, a disease caused by Sclerotinia minor, occurred in Texas in fields where the disease was never previously detected. The disease was observed on many plants within one of the fields (>3000 disease foci), although most foci were <1 m. It is hypothesized that these observations were inconsistent with the recent introduction of a monocyclic pathogen, even if disease developed under conducive environmental conditions. The pattern of disease is most suggestive of the presence of foliar (ascospore) infections, although air temperature was above the known limits for apothecia development if the pathogen had arrived in the field in 2004 peanut seed. To further examine this epidemic, 232 isolates were collected, across a variety of spatial scales spanning this field and other Texas peanut fields, and evaluated for aggressiveness, fungicide sensitivity and genotypic diversity. There was wide variation among isolates for the phenotypic characteristics measured, but there was no evidence that a genotypically unique, highly aggressive, and fungicide resistant isolate had been introduced or evolved. The predominant genotype, TX1, which contained 154 isolates, was found in every county and field population.     

Tumours of Begonia and some other ornamentals,induced by Corynebacterium fascians

In 1975 many tumours were observed in plants ofBegonia Schwabenland grown in Aalsmeer. Submersion of the roots ofNicotiana megalosiphon seedlings in a homogenate of tumorous tissue, induced tumours after two weeks. Short periods of submergence yielded results similar to those obtained after longer periods. Tumour homogenates lost their infectivity after ten min at 50°C. Aphids transmitted the infectious agent.Treatment with propylene oxide did not inhibit infectivity completely. Filtration through a 450 nm filter removed the infectious agent.Tobacco tumor virus or a viroid could not be isolated. Cultures ofCorynebacterium fascians, isolated from tumours ofN. megalosiphon were highly infectious and induced tumours in healthyN. megalosiphon andBegonia. Tumorous tissue homogenates ofPelargonium zonale, Dahlia sp.,Gladiolus sp., andLilium sp. also caused tumours inN. megalosiphon, from whichC. fascians was isolated. It was not possible to produce tumours inN. megalosiphon with homogenates from roses with symptoms of bud proliferation.Samenvatting In 1975 werden vele tumoren waargenomen inBegonia Schwabenland op Aalsmeerse bedrijven (Fig. 1). De infectiositeit van tumorweefsel kon goed en snel worden vastgesteld door de wortels van zaailingen vanNicotiana megalosiphon in een homogenaat van tumorweefsel te dompelen. Tumoren ontstonden na twee weken, de eindbeoordeling geschiedde na een maand (Fig. 2). Ook verschillende andereNicotiana spp.,Melilotus officinalis (Fig. 3) enPisum odoratum (Fig. 4) werden aangetast.Bij de infectiositeitstoets gaven zeer korte dompeltijden even goede resultaten als langere (Tabel 1). Infectieus sap verloor zijn infectievermogen na 10 min verhitting bij 50°C. Bladluizen brachten de smetstof over. Propyleenoxide verminderde de infectiositeit wel, doch onderdrukte deze niet totaal. Bij filtratie door een 450 nm filter bleef het infectieuse agens op het filter achter. Het tumor-inducerende agens was ook aanwezig in die delen van planten met tumoren welke gezond leken en het ging voor een gering deel over met zaad (Tabel 2).Uit tumoren konden wij geen tabakstumorvirus of een viroïde isoleren. Culturen vanCorynebacterium fascians, geïsoleerd uit tumoren vanN. megalosiphon bleken zeer infectieus en veroorzaakten tumoren inN. megalosiphon enBegonia. Homogenaten van tumorweefsel vanPelargonium zonale, dahlia (Fig. 5), gladiool (Fig. 6) enLilium Mid Century Hybrid Enchantment (Fig. 7) veroorzaakten ook tumoren opN. megalosiphon, waaruitC. fascians werd geïsoleerd. Met sap van kroeskopzieke rozen konden wijN. megalosiphon niet besmetten.     

Differentiation of Bean pod mottle virus isolates based upon host symptoms

Accessions from Glycine, Phaseolus, and Vigna genera were screened for their reactions to different subgroups of isolates of Bean pod mottle virus (BPMV) in order to establish a differential host system. Screening results indicated that the BPMV isolates differed in pathogenic aggressiveness but not in virulence. No major resistance genes were found in soybean (Glycine max) or G. soja since all screened accessions showed mosaic or necrotic symptoms to BPMV inoculation. However, these accessions expressed differences in severity of symptoms when challenged by various BPMV isolates. The inoculation of G. tomentella accessions did not result in mosaic symptoms, and some accessions did not support systemic infection of some of the isolates. Resistance, presented as a hypersensitive reaction, was observed in some of Phaseolus and Vigna genotypes, and resistant response or susceptibility was stable to all the isolates used in the screening. In conclusion, the selected G. soja genotypes PI 407019, PI 464889A, and PI 464928, and ‘Amsoy 71’ soybean may help to separate severe (reassortant) from mild isolates of BPMV based upon their phenotypic reactions.