土壤样品中长喙壳属真菌诱导分离方法的研究

 长喙壳属真菌(Ceratocystis spp.)是一类重要的土传植物病原真菌,可危害多种作物。由于技术方法的限制,对于土壤样品中该病原菌的分离比较困难。本研究通过比较以胡萝卜、马铃薯、甘薯作为诱导富集载体对土壤中长喙壳属真菌的富集分离效果,确定了胡萝卜为最优诱导富集载体。进一步通过对胡萝卜片厚度、土壤相对湿度、培养温度、光照条件、保湿方式等培养条件的优化,建立了从土壤中分离长喙壳属真菌的有效方法,具体为:以厚度为3~5 mm的胡萝卜片作为诱导富集载体;土壤样品相对含水量为10%~30%;12 h光暗交替或24 h黑暗;在22℃~26℃条件下不保湿进行培养,8 d后可从携带最低数量为10个孢子/g的土壤样品中分离到靶标菌。采用该方法,对采自新疆、河南、湖南、云南等地共计108份自然或耕作土壤样品进行了分离,结果表明从采自云南蒙自市、临沧市的45份土壤样品中均分离获得了长喙壳属真菌。该方法具有操作简便、所需仪器设备简单、灵敏度较高、成本低和高通量等特点,适用于田间调查,可对样本量较大的土壤样品进行快速及时分离,以获得不同土壤生境中的长喙壳属真菌分离物。     

Storability at room temperature of Steinernema yirgalemense (Rhabditida: Steinernematidae) in diatomaceous earth and the effect of antifungal agents

Journal of Plant Diseases and Protection - The commercial use of entomopathogenic nematodes after mass production requires the development of formulation techniques that extend nematode survival...     

A survey of entomopathogenic nematodes of the families Steinernematidae and Heterorhabditidae (Nematoda: Rhabditida) in Damascus countryside of Syria

A survey of entomopathogenic nematodes was conducted in Damascus countryside from January 2011 to December 2013. Soil samples were tested for the presence of steinernematid and heterorhabditid nematodes by baiting with Galleria mellonella larvae. Of the 189 soil samples studied 17 were positive for entomopathogenic nematodes (9%), with 11 of these positive samples (65%) containing Heterorhabditis and 6 (35%) Steinernema isolates. Morphological studies were carried out to characterize eight isolates. The Heterorhabditis isolates collected in Syria were identified as Heterorhabditis zealandica (Poinar, 1990), Heterorhabditis indica (Poinar et al., 1992) and Heterorhabditis bacteriophora (Poinar, 1990). Heterorhabditis zealandica was isolated from 4 sites. Heterorhabditis indica and H. bacteriophora were isolated from two sites each. Entomopathogenic nematodes were mainly found in stone fruit orchards and apple orchards but also in citrus groves, vineyards, and walnut orchards.     

对比分析不同方法提取土壤中的二氯喹啉酸

本文采用4种常用方法分别对福建田土中添加的二氯喹啉酸进行了提取,并采用高效液相色谱法对其添加回收率进行了测定,对比分析表明,当二氯喹啉酸添加浓度为1.25μg/g时,方法二即氢氧化钠溶液提取、乙酸乙酯净化法回收率最高,约为70%,且杂质峰较小,基线平稳,峰形较对称;重复性高,两次生物学试验的平均回收率无显著差异,均略高于70%。方法一即乙腈磷酸水溶液提取、氯化钠净化法,回收率较高,但杂质峰较高,基线不平稳。方法三即硼砂甲醇溶液提取、二氯甲烷净化法,虽然杂质峰小、基线平稳、峰形较好、但回收率低,约为50%。方法四即甲醇磷酸水溶液直接提取法,因提取液难以在旋转蒸发仪中浓缩,因此未继续检测。综合结果表明,方法二最适用于福建省水稻田二氯喹啉酸的提取。     

Phylogenetic relationships of some spirurine nematodes (Nematoda: Chromadorea: Rhabditida: Spirurina) parasitic in fishes inferred from SSU rRNA gene sequences

Abstract: Small subunit rRNA sequences were obtained from 38 representatives mainly of the nematode orders Spirurida (Camallanidae, Cystidicolidae, Daniconematidae, Philometridae, Physalopteridae, Rhabdochonidae, Skrjabillanidae) and, in part, Ascaridida (Anisakidae, Cucullanidae, Quimperiidae). The examined nematodes are predominantly parasites of fishes. Their analyses provided well-supported trees allowing the study ofphylogenetic relationships among some spirurine nematodes. The present results support the placement of Cucullanidae at the base of the suborder Spirurina and, based on the position of the genus Philonema (subfamily Philoneminae) forming a sister group to Skrjabillanidae (thus Philoneminae should be elevated to Philonemidae), the paraphyly of the Philometridae. Comparison of a large number of sequences of representatives of the latter family supports the paraphyly of the genera Philometra, Philometroides and Dentiphilometra. The validity of the newly included genera Afrophilometra and Caranginema is not supported. These results indicate geographical isolation has not been the cause of speciation in this parasite group and no coevolution with fish hosts is apparent. On the contrary, the group of South-American species ofAlinema, Nilonema and Rumai is placed in an independent branch, thus markedly separated from other family members. Molecular data indicate that the skrjabillanid subfamily Esocineminae (represented by Esocinema bohemicum) should be either elevated to the rank of an independent family or Daniconematidae (Mexiconema africanum) should be decreased to Daniconematinae and transferred to the family Skrjabillanidae. Camallanid genera Camallanus and Procamallanus, as well as the subgenera Procamallanus and Spirocamallanus are confirmed to be paraphyletic. Paraphyly has also been found within Filarioidea, Habronematoidea and Thelazioidea and in Cystidicolidae, Physalopteridae and Thelaziidae. The results of the analyses also show that Neoascarophis, Spinitectus and Rhabdochona are monophyletic, in contrast to the paraphyletic genus Ascarophis. They further confirm the independence of two subgenera, Rhabdochona and Globochona, in the genus Rhabdochona. The necessity of further studies of fish-parasitizing representatives of additional nematode families not yet studied by molecular methods, such as Guyanemidae, Lucionematidae or Tetanonematidae, is underscored.     

Comparison of a baiting method and PCR for the detection of Phytophthora fragariae var. rubi in certified raspberry stocks*

In winter 2002/2003, a total of 136 root samples from 57 different raspberry stocks in Scotland were examined for the presence of raspberry root rot caused by the fungus‐like pathogen Phytophthora fragariae var. rubi. All stocks had been planted as propagation material entered at different grades in the Scottish certification scheme or had applied for plant passports. For detection, a modified ‘Duncan bait test’ was compared to a nested PCR method. The two tests identified the same infected stocks: PCR detected 10 positive samples from four different stocks, while the bait test picked up two additional positive samples coming from the same four stocks. The two tests had a similar level of reliability in this examination and a recommendation for one or the other depends mainly on the technical equipment and skills available in the laboratory.     

Characterization of Longidorus helveticus (Nematoda: Longidoridae) from the Czech Republic

Longidorus helveticus was found at two out of 285 sampling sites for the first time in the Czech Republic. Females, males and juvenile stages were analyzed morphologically and morphometrically. The morphological identification of samples was verified by polymerase chain reaction using a species specific primer. Four markers of ribosomal DNA (18S, ITS1, ITS2, D2-D3 expansion segments of 28S rRNA) and two markers of mitochondrial DNA (cox1 and nad4) were sequenced and analyzed and compared with published gene sequences of other populations of L. helveticus. The partial mitochondrial cytochrome c-oxidase subunit 1 gene and partial nicotinamide dehydrogenase subunits 4 gene showed relatively high genetic variation within the species compared with ribosomal DNA markers.     

Capillariids (Nematoda: Capillariidae) from passeriform birds of Nepal).

A total of 83 hosts of the order Passeriformes from Nepal were examined for nematodes. Nine of them were found to be infected with 4 nematode species, Capillaria exilis, C. ovopunctata, Capillaria sp. and Thominx rigidula. The last species is redescribed in detail. Three of the definitive hosts are new for the above-mentioned nematode species and this is the first recorded from Nepal.     

Comparison of the Fenwick can and Schuiling centrifuge methods for the extraction of potato cyst nematodes from soil1

An evaluation of the efficacy of the Schuiling centrifuge and traditional Fenwick can methods of extracting Globodera cysts from infested field soils confirmed the accuracy of the Schuiling centrifuge extraction method. Experience has shown that samples with a high organic matter content may reduce extraction efficiency. Improvements in the mechanical and electrical construction of the present Schuiling apparatus are needed, but once they have been made the choice of extraction method in large-scale statutory and advisory programmes need only depend on considerations such as convenience of use and cost-effectiveness.     

Oulema melanopus (L.) (Coleoptera: Chrysomelidae) adults are susceptible to entomopathogenic nematodes (Rhabditida) attack: results from a laboratory study

Journal of Plant Diseases and Protection - Three species of entomopathogenic nematodes from Slovenia (Steinernema feltiae B30, S. carpocapsae C101, and Heterorhabditis bacteriophora D54) and...     

格氏线虫表皮蛋白和分泌蛋白抑制昆虫免疫活性的比较

格氏线虫表皮蛋白和分泌蛋白都具有抑制昆虫免疫反应的能力,能够帮助侵染期线虫侵染寄主,但两者所含蛋白组分并不相同,从而导致蛋白活性也有所差异。本研究采用乙醇萃取和昆虫匀浆诱导,分别从侵染期的格氏线虫体表及分泌物中得到了表皮蛋白和分泌蛋白,并比较了这两者之间的异同。结果表明,无论是在昆虫体内还是体外,侵染期线虫表皮蛋白和分泌蛋白都具有抑制昆虫免疫反应的生物活性,并且分泌蛋白的活性更强。本研究为进一步研究线虫侵染与昆虫免疫间的相互关系和线虫抑制昆虫免疫反应的作用机理提供了科学佐证。     

The enzymes of glycogen and trehalose catabolism from Hysterothylacium aduncum (Nematoda: Anisakidae)

The activity of alpha-amylase, glucoamylase, maltase, trehalase, glycogen phosphorylase and trehalose phosphorylase was measured in extracts from larval and adult Hysterothylacium aduncum (Rudolphi, 1802), parasitic nematode of marine fish. The content of glycogen and trehalose in the worm's body was also determined. Both the hydrolytic and phosphorolytic paths of sugar decomposition are present in H. aduncum. In the larvae glycogen was utilised mainly via the hydrolytic path. In the adults the activities of phosphorolytic enzymes were higher than in the larvae. In both stages the activity of trehalose phosphorylase is present. In adult nematodes it is uncommonly high. The dominating sugars in the adults were glucose and glycogen, while in the larvae it was trehalose.     

The influence of herbicides on the viability of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae)

Entomopathogenic nematodes (EPNs) are agents that can be used for the biological control of pests associated with pesticides in a tank mix. Compatibility studies need to be conducted to analyse which products are compatible with EPNs. The survival of infective juveniles (IJs) of four species of EPNs (Steinernema, Heterorhabditis) was determined after exposure to eight chemical herbicides. The effect of direct IJs exposure to herbicides for 1, 4 and 24 h was tested in a Petri dish at 15, 20 and 25 °C. The study showed that Steinernema kraussei was the most tolerant among the tested EPN species, while S. carpocapsae was the most sensitive to all tested herbicides. The lowest mortality of IJs was at 15 °C (19%). Our investigation showed, overall, the herbicides negatively affected EPN survival. The results confirmed that the compatibility is a species-specific characteristic, influenced by the temperature and time of exposure. Application of two different control ingredients (insecticide and herbicide) at the same time would reduce cost and time consumption in pest/weed control.     

Two new species of Rhabdochona (Nematoda: Rhabdochonidae) from freshwater fishes in Thailand

Two new species ofrhabdochonid nematodes are described from the intestine of freshwater fishes in Chiang Mai Province, northern Thailand: Rhabdochona (Rhabdochona) pseudomysti sp. n. from the catfish Pseudomystus siamensis (Regan) (Bagridae, Siluriformes) in the Fang Brook, a tributary of the Kok River (the Mekong River basin), Fang District and Rhabdochona (Globochona) thaiensis sp. n. from the cyprinid Mystacoleucus marginatus (Valenciennes) (Cyprinidae, Cypriniformes) in the Ping River (the Chao Phraya River basin), Muang District. Rhabdochona pseudomysti is mainly characterized by simple, leaf-like oval deirids (a unique feature among Rhabdochona spp.), a prostom with 14 anterior teeth, the presence of basal prostomal teeth, the length ratio of the muscular and glandular portions of oesophagus (1:2.1-2.6), an unusually long left spicule (1.10-1.22 mm), length ratio of spicules (1:11.5-14.7), arrangement of genital papillae, and conspicuously elevated vulval lips. Rhabdochona thaiensis differs from other representatives of the subgenus Globochona Moravec, 1972 possessing caudal projections on the tail tip in that it has only 2 claw-shaped projections located ventrally on the tail tip of both males and females; the species is mainly characterized by the presence of distinct pseudolabia, 8 anterior prostomal teeth, absence of basal teeth, bifurcated deirids, length ratio of the muscular and glandular portions of oesophagus (1:11.3-11.9), conspicuously short (135-141 microm) left spicule, arrangement of genital papillae, and somewhat elevated vulval lips. Fully developed eggs of R. pseudomysti and R. thaiensis remain unknown. These are the first nominal species of Rhabdochona reported from Thailand.     

半轮线虫属新种——拟中华半轮线虫Hemicriconemoides parasinensis sp. nov.的描述

 拟中华半轮线虫新种(Hemicriconemoides parasinensis sp. nov.)模式标本采集于辽宁省沈阳市沈阳农业大学校园的丁香(Syringa oblata)根际土中,在河南省安阳市安阳公园的小叶女贞(Ligustrum quihoui)根际土中也有分布。主要鉴别特征是虫体头区圆形,头环2个,第1头环略小于或等于第2头环,排泄孔位于肠的前端(Rex=29~33),体长中等,口针中等(76~86 μ m),体环数R值中等(96~110),卵巢单生前伸,受精囊长圆形,尾部宽圆锥形、尾端半球形圆;幼虫具指状的棘片,约12~16排,后翻;雄虫未见。该新种虫体形态特征与中国半轮线虫(H. sinensis Vovlas,1988),芒果半轮线虫(H. mangiferae Siddiqi,1961)和微小半轮线虫(H. minutus Esser,1960)相近似。     

A new species of the genus Xiphinema Cobb 1913 (Nematoda: Longidoridae) from north and northwest Iran

European Journal of Plant Pathology - Five populations of a new dagger nematode species were recovered from natural grasslands and forests of north and northwest Iran, and described based upon...     

In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method

Macrophomina phaseolina (Tassi) Goid. is the most important pathogen of jute and primarily causes seedling blight, leaf spot and stem rot. The pathogen was detected from field samples by a simple method of direct PCR (dPCR) which obviates the steps of DNA extraction. The leaf bits were treated with a lysis buffer at 65°C for 25 min, whereas the stem pieces were initially incubated at 65°C for 5 min followed by incubation at 60°C for 25 min and the lysate was used as PCR template. Based on the type of tissue, the composition and concentration of lysis buffer systems were optimized. For leaf samples the optimized buffer system composed of 20 mmol l ^-1 tris (hydroxymethyl aminomethane (Tris)-Cl (pH 8.0), 1.5 mmol l ^-1 ethylene diamine tetra acetate (EDTA) (pH 8.0), 1.4 mol l ^-1 sodium acetate and 200 μg/ml proteinase K. Further, 3% PVP (w/v) and β-mercaptoethanol (1% w/v) were added into the buffer. In case of stem samples, PVP was not applied and higher concentrations were used for other components. M. phaseolina could be detected from both leaf and stem samples generating amplicon of 350 bp. This is the first report of detecting M. phaseolina by a direct PCR method without DNA extraction.