草莓内生细菌的分离及草莓灰霉病菌拮抗菌的筛选鉴定

本研究从健康的草莓植株内分离到54株内生细菌,其中根、茎、叶组织中分离内生细菌种群密度为2.8×101～3.95×104cfu/g鲜重不等。通过对峙试验,得到5株对草莓灰霉病菌有强拮抗作用的菌株,占所分离内生细菌总数的9.26%。来自草莓叶的SL6菌株抑菌效果最佳,抑菌半径达15mm,无菌发酵液对草莓灰霉病菌菌丝的抑制率为97.6%,具有作为生防菌的应用潜能。经形态、生理生化和16SrRNA等分析测定,并通过MEGA4方法构建其16SrDNA系统发育树,将SL6菌株鉴定为枯草芽胞杆菌(Bacillus subtilis)。     

枯草芽胞杆菌GB519在水稻植株中的定殖及对稻瘟病田间防效

枯草芽胞杆菌GB519是一株具有广谱抑菌活性的生防菌株。本研究利用绿色荧光蛋白标记的菌株GB519-GFP处理水稻种子、根和叶片,结合激光共聚焦显微镜观察和抗生素平板回收检测的方法,探究其在水稻根茎叶中的定殖动态。结果显示：经GB519-GFP发酵液处理水稻种子、根和叶片后,菌株均可内生定殖于植株的表皮、皮层和维管束中,表明其可在水稻植株内迁移和定殖。GB519-GFP在处理部位的定殖量通常呈现先减少后增多的趋势,非处理部位3～5 d后即可检测到标记菌株。浸种处理,3 d后在幼芽中可检测到标记菌株;20 d后在根中的菌量最多,达5.7×10⁵ cfu/g。灌根处理,1 d后根中菌量为5.4×10⁵ cfu/g; 20 d后根、茎和叶中菌量均达到最大值;处理80 d后,根中定殖数量仍达1.9×10⁵ cfu/g。叶面喷施处理,1 d后叶片菌量为4.2×10⁵ cfu/g; 20 d后叶片菌量达4.4×10⁵ cfu/g。不同处理方法在各部位的定殖量几乎均在处理20 d后达到峰值。...     

黄曲条跳甲幼虫致病菌的鉴定及其对黄曲条跳甲的杀虫活性研究

从自然死亡黄曲条跳甲幼虫体中分离出1株对黄曲条跳甲幼虫和成虫有较高致病性菌株PS-1。通过病原菌的形态观察、Biolog系统鉴定以及16S rDNA序列测定分析,确定菌株PS-1隶属于肠杆菌科、沙雷氏菌属的粘质沙雷氏菌Serratia marcescens族群。用菌株PS-1浓度为3×104、3×106和3×108 cfu/mL的菌悬液浸泡鲜萝卜块饲喂黄曲条跳甲幼虫24 h,4 d后的校正死亡率分别为36.1%、63.9%和75.0%;2、3和4 d的LC50分别是7.98×106、4.27×106和8.02×105 cfu/mL。用浓度1×108 cfu/mL的菌悬液处理菜心叶片饲喂黄曲条跳甲成虫,12 d的校正死亡率为23.25%。用浓度1×105、1×107和1×109 cfu/mL的菌悬液浸渍黄曲条跳甲卵,其孵化率与对照无显著差异。结果表明,粘质沙雷氏菌PS-1菌株对黄曲条跳甲成虫和幼虫有较强的毒力,在黄曲条跳甲的生物防治方面具有一定的应用前景。     

西藏砂生槐内生真菌的分离鉴定及其多样性分析

采用组织分离法分离纯化取自西藏砂生槐不同部位组织的内生真菌,依照其形态学特征将菌株初步归类;挑选代表性菌株进行核糖体内转录间隔区(r DNA-ITS)序列分子鉴定,采用MEGA5.10软件构建系统进化树,对菌株进行聚类分析;采用SPSS13.5软件进行砂生槐内生真菌的多样性分析。结果表明,从砂生槐不同部位共分离得到77株内生真菌,其荚果中的分离几率最高,达到45.68%,而叶片中内生真菌分离几率最低,仅为4.63%。其形态学特征和ITS-r DNA序列表明,供试的16株菌分别归属于11个不同的种,其中茎点霉属(Phoma sp.)为优势种群。系统进化树显示,4个菌株在96%相似水平上聚在一个分枝上,与茎点霉属关系最近;2个菌株在92%相似水平上聚在一个分枝上,与壳二孢属亲缘关系最近;2个菌株在97%相似水平上聚在一个分枝上,与盘菌属亲缘关系最近。荚果中的内生真菌多样性指数和均匀度指数均为最高,分别为1.078 1和0.601 7,刺中的内生真菌丰富度指数最高,达到0.836 8。     

商洛多花胡枝子根瘤菌生理生化特性和数值分类研究

选用分离自陕西商洛多花胡枝子的59株根瘤菌和15株参比菌,进行了125项表型性状测定和聚类分析。结果表明:未知菌能够利用的碳氮源比较广泛,生长的温度和pH范围较宽,对抗菌素、染料和重金属分别具有抗性的菌株以及产氧化酶菌株的数量比例较高,部分菌株具有耐高浓度盐或碱的性能,其中大约67%的菌株能耐受3.0%的NaCl,25%的菌株可在初始pH12的YMA培养基上生长。从数值分类树状图谱发现,在80.5%的相似性水平上,除CCNWSX0922外,供试菌株聚成8个表观群,群Ⅰ包含6株未知菌和3株已知中慢生根瘤菌属参比菌,群Ⅱ包含19株未知菌和6株中华根瘤菌属参比菌,群Ⅲ包含1株未知菌和3株根瘤菌属参比菌,群Ⅳ和群Ⅴ皆含根瘤菌属参比菌株。群Ⅵ、群Ⅶ和群Ⅷ三个类群未能与参比菌株聚在一起,为新类群,其分类地位需进一步研究确定。     

恶臭假单胞菌HB3S-20在棉花中的定殖及诱导抗性研究

植物内生菌是防治植物病害的重要生防资源。恶臭假单胞菌Pseudomonas putida HB3S-20是本实验室从棉花内生细菌中所筛选到的一株对棉花黄萎病具有显著防效的潜在生防菌株。本研究采用利福平抗性标记技术和绿色荧光蛋白(GFP)标记技术检测了菌株HB3S-20在棉花组织中的定殖动态和定殖位点,并检测了菌株HB3S-20接种棉花植株后植物防御相关酶SOD、POD和CAT酶活性的变化。利福平标记试验结果表明,菌株HB3S-20主要定殖于根部,棉株接种菌株HB3S-20后第3～30 d检测到棉花根部内的菌量范围为2.2×104～6.7×10⁴ cfu/g,检测到棉花茎部内的菌量为0.04×104～2.7×10⁴ cfu/g,棉花茎部内的菌量低于根部,但在叶片中未被分离到。GFP标记试验结果表明,菌株HB3S-20不仅能附着在棉花根系表面,还能定殖在根部维管组织中。此外,菌株HB3S-20能够提高棉花中植物防御相关酶SOD和POD的活性,说明菌株HB3S-20能够诱导棉花植株的系统抗性,增强棉花抗病性。本研究初步揭示了恶臭假单胞菌HB3S-2...     

西瓜枯萎病菌对西瓜种子发芽和幼苗生长发育的影响

分别采用滤纸法和浸胚根法测定了西瓜枯萎病菌对西瓜种子发芽和幼苗生长发育的影响。结果表明,枯萎病菌孢子悬浮液浓度为106～107个/ml时,种子的发芽势和发芽率均显著低于对照,并且对京欣1号品种和航兴1号品种种子发芽的影响大于黄小帅;枯萎病菌孢子悬浮液处理种子后,幼苗的茎长、子叶长度、子叶宽度、第3叶长度、第3叶宽度、百株鲜重等指标均有所降低,当浓度为106～107个/ml时,除百株鲜重和航兴1号品种幼苗的第3叶长度外,与对照相比,其他指标均显著降低。     

抱茎苦荬内生细菌的分离及其抑菌活性研究

为明确药用植物抱茎苦荬不同部位内生细菌分布及其在植物病害生物防治中的应用价值,采用组织分离法对抱茎苦荬不同部位内生细菌进行了分离.用平板对峙法筛选出具有抑菌活性的菌株,并用生长速率法测定了其代谢物的抑菌活性.结果表明,从抱茎苦荬不同部位共分离到24株内生细菌.其中,15株具有一定的抑菌活性,占分离总数的62.5％; 15株中有3株对梨黑斑病病菌、番茄早疫病病菌和番茄灰霉病病菌有较强的抑菌活性,其抑制率为84.2％～92.9％:3株内生细菌代谢物对6种植物病原真菌抑菌活性为8.1％～91.0％,经121℃高温处理后其抑菌活性在0～22.3％之间,表明其3株内生细菌具有较强的抑菌效果.     

我国部分地区土壤杆菌的数值分类及DNA同源性分析

 本研究对我国42株土壤杆菌待测菌株和28株参比菌进行了数值分类和DNA同源性分析,结果将41株土壤杆菌与2株根瘤菌分在3个土壤杆菌种内,其中23株来自内蒙等地葡萄的菌株分在葡萄土壤杆菌种内,10株来自山东等地樱桃、桃树的菌株为发根土壤杆菌,8株来自4个地区4种植物的菌株及分自宁夏刺果甘草的2株根瘤菌菌株参比菌分在根癌土壤杆菌种内。1株土壤杆菌和1株根瘤菌的分类地位有待进一步研究。     

辣椒内生细菌的分离及拮抗菌的筛选

对健康辣椒植株体内的内生细菌及拮抗菌进行了分离筛选。结果表明 ,不同品种、部位及种植地 ,内生细菌的数量有所不同 ,品种间内生细菌的数量变化为 2 83× 1 0 3 ～ 1 346×1 0 4cfu/g(FW) ;各品种以叶片中内生细菌的数量最多 ,其次为根 ,再次为茎 ,果最少 ;不同种植地中 ,前作为水稻田种植辣椒叶片中内生细菌的数量比旱地种植的多 ,而根、茎、果中内生细菌的数量正相反。拮抗、防病测定表明 ,1 0 8株辣椒内生细菌中 ,有 2 8 7%的菌株对香蕉枯萎菌和黄瓜枯萎菌有拮抗作用 ;来自辣椒叶片和茎杆内的BS 2和BS 1两菌株 ,对 1 3种植物病原真菌有较强和稳定的拮抗作用 ,其中对辣椒炭疽病有 57 34%～ 94 0 8%的防病效果 ,并可以在辣椒体内定殖。经鉴定该两菌株为枯草芽孢杆菌。     

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.     

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.     

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.     

Eyespot of Cereals Revisited: ITS phylogeny Reveals New Species Relationships

Four species so far classified in Pseudocercosporella or Ramulispora (hyphomycetes) are associated with eyespot disease symptoms of cereals. Two of these have been linked to teleomorphs that were described in Tapesia. Sequence data derived from the Internal Transcribed Spacer region (ITS1, 5.8S and ITS2) of the rDNA operon showed, however, that the eyespot fungi associated with Tapesia are not congeneric with Ramulispora sorghi, the type of Ramulispora. The genus name Tapesia is now rejected in favour of the conserved name Mollisia, which appears to comprise heterogeneous fungi. Tapesia yallundae is not closely related to the type of Mollisia, M. cinerea, but clusters separately, being more closely allied to species with Cadophora anamorphs. A new holomorph genus, Oculimacula, is therefore proposed for teleomorphs of the eyespot fungi, while the anamorphs are accommodated in Helgardia gen. nov.