Automated and Unbiased Image Analyses as Tools in Phenotypic Classification of Small-Spored Alternaria spp

ABSTRACT For more than 25 years, controversy has surrounded the characterization and differentiation of small-spored Alternaria spp. And, therefore, the application of names of several species that are involved in the pathology of diseases related to host-specific toxin production. The name A. alternata often has been broadly applied to various morphologically and chemically distinct groups of isolates from different hosts. The purpose of this study was to develop and evaluate automated and unbiased image analysis systems that will analyze different phenotypic characters and facilitate testing and application of the morphological species concept in Alternaria taxonomy. Host-specific toxin-producing Alternaria isolates assigned to five morpho-species were compared with representative isolates of morphologically distinct A. alternata. Combined results of growth rates at different temperatures, colony morphology, and metabolite profiles were found to be useful in characterization and differentiation of small-spored Alternaria spp. when standardized conditions are applied and representative isolates employed for comparison.     

Neotropical Monogenoidea 37. Redescription of Gyrodactylus superbus (Szidat, 1973) comb. n. and description of two new species of Gyrodactylus (Gyrodactylidea: Gyrodactylidae) from Corydoras paleatus and C. ehrhardti (Teleostei: Siluriformes: Callichthyidae) of Southern Brazil

Two new species of Gyrodactylus (Gyrodactylidae) are described from Corydoras paleatus (Jenyns) and Corydoras ehrhardti Steindachner (Callichthyidae) of Rio Piraquara, Municipality of Piraquara, near Curitiba, Paraná, Brazil: Gyrodactylus anisopharynx sp. n. and Gyrodactylus samirae sp. n. These species are unique among the known Neotropical species of Gyrodactylus by the combination of several characters. Gyrodactylus anisopharynx is characterised by having hooks with point evenly curved, toe moderately pointed, round convex heel, straight sloping shelf, superficial bar with anterolateral projections and male copulatory organ armed with two rows of spinelets. Two variants of G. anisopharynx, of distinct pharynx size, are recognised: forma "large-pharynx" and forma "small-pharynx". Gyrodactylus samirae is characterised by having hook with straight shaft, short slightly recurved point, truncate toe, trapezoidal heel, shelf slightly concave, orthogonal to shaft; anchors with relatively short superficial root; superficial bar with extremities lightly expanded; and male copulatory organ armed with two rows of spinelets. Paragyrodactyloides superbus (Szidat, 1973) is redescribed and transferred to Gyrodactylus based on examination of type and new specimens. Features previously considered as diagnostic for the genus are misinterpretations or primitive characters for Gyrodactylus spp.     

Biological control of pigweeds (Amaranthus retroflexus, L. A. Powellii, S. Watson and A. bouchonii Thell.) with phytophagous insects, fungal pathogens and crop management

Pigweeds (Amaranthus spp.) are of economic importance worldwide. In Europe, Amaranthus retroflexus is one of the ten weed species of greatest economic importance. It is a serious problem weed in several field crops (e.g. maize), as well as in vegetables, orchards and grape vines. It is an annual spreading by seeds which have a long viabilityand are dispersed principally by wind and water, but also by machinery. There is great variability in seed germination which renders control with post-emergence herbicides difficult. In addition, triazine herbicide-resistant populations occur in ten European countries. The aim of this subproject of COST action 816 is to investigate the possibilities of classical and inundative biological control of Amaranthus spp., to characterize potentialbiological control agents and to develop methods for their integration with current phytosanitary measures in the target crops. The project was initiated with an extended literaturesurvey followed by field surveys for insects and pathogens associated with Amaranthus spp. in several European countries. Promising isolates of fungal pathogens have been tested ondetached leaves and whole plants, and initial studies on the application of pathogens causing damping off in seedlings have been made. Further, the variability of different provenances ofAmaranthus spp. in response to fungal attack has been investigated     

茶尺蠖和灰茶尺蠖幼虫两种共有寄生蜂的鉴定

茶尺蠖Ectropis obliqua (Prout)及其近缘种灰茶尺蠖E. grisescens Warren是我国茶树上最为重要的两种食叶性害虫,茶尺蠖绒茧蜂Apanteles sp.和单白绵绒茧蜂Apanteles sp.是茶尺蠖Ectropis spp.幼虫期最重要的两种寄生性天敌昆虫。为了明确这两种绒茧蜂的物种名称及形态差别,同时探明这两种绒茧蜂是否是茶尺蠖和灰茶尺蠖的共有寄生蜂,本研究采集了我国8省22个地方的茶尺蠖或灰茶尺蠖幼虫,通过室内饲养获得两种绒茧蜂的标本,对标本进行形态鉴定;同时将两种绒茧蜂的COI基因序列与Bold数据库中已知物种进行比对。结果表明,从野生茶尺蠖和灰茶尺蠖幼虫中均饲养出之前报道的茶尺蠖绒茧蜂Apanteles sp.和单白绵绒茧蜂Apanteles sp.两种寄生蜂;形态鉴定和分子比对结果表明,这两种广义绒茧蜂Apantelesspp.实为尺蠖原绒茧蜂Protapantelesimmunis(Haliday,1834)和单白绵副绒茧蜂Parapanteles hyposidrae (Wilkinson, 1928)。本研究明确了40多年前国内报道...     

关于植物病原生物名称汉译的几点意见

 Many plant pathogens have their scientific names after being identified. Most of them have their Chinese names which being given or translated by Chinese scientists correctly. But, some of them were not correct or standard, or have the same Chinese names. Such as Apiosporium and Pyricularia;Colletogloeum and Myxosporium;Aureobasidium and Napicladium, etc. A few genus names have changed several times since new taxonomy system being published, for example, the pathogen of tomato bacterial wilt pathogen has 4 genus names, Bacterium, Pseudomonas, Burkholderia, and Ralstonia, the Ralstonia is valuable and updated. The term of MLO has been used not longer, and the "Candidatus Phytoplasma" is adopted now."The Latin-Chinese and Chinese-Latin Names of Plant Pathogens" is publishing, which is edited and contributed by seven professors engaged in education and plant protection in China. Main target of the book is(trying) to make unification when Chinese name of pathogen is used.     

Distinct Species Exist Within the Cercospora apii Morphotype

ABSTRACT The genus Cercospora is one of the largest genera of hyphomycetes. Cercospora apii sensu lato is the oldest name for a large complex of morphologically indistinguishable Cercospora spp. occurring on a wide host range. There are currently 659 recognized Cercospora spp., and names of another 281 morphologically identical species are included in the synonymy of C. apii sensu lato. Two of the species that belong to the C. apii complex, C. apii and C. beticola, cause Cercospora leaf spot on Apium graveolens (celery) and Beta vulgaris (sugar beet), respectively. In the present study, multilocus sequence data, amplified fragment length polymorphism analysis, and cultural characteristics were used as additional features to characterize morphologically similar Cercospora strains occurring on celery and sugar beet. From the data obtained, it is shown that C. apii and C. beticola, although morphologically similar and able to cross-infect each others' hosts, are distinct functional species that should be retained as separate entities. Furthermore, a third, as yet undescribed species of Cercospora was detected in celery fields in Korea and Venezuela, suggesting that additional undescribed species also may be found to cause Cercospora leaf spot on celery. A polymerase chain reactionbased diagnostic protocol distinguishes all three Cercospora spp.     

Fungicide resistance: definitions and use of terms

On the basis of an analysis of current use of terms concerning resistance to fungicides, a series of definitions is proposed for discussion. Consistent use of such terms in scientific publications will facilitate understanding and avoid possible misinterpretations.     

西瓜种子带菌检测及杀菌剂消毒处理效果

采用离体平皿法对来自新疆等4个省区的14个西瓜品种进行种子带菌检测、分离纯化和鉴定,并测定了7种杀菌剂和1种种衣剂对种子带菌的消毒处理效果。结果表明,种子表面携带的优势菌群主要为青霉属Penicillium spp.、根霉属Rhizopus spp.、曲霉属Aspergillus spp.、交链孢属Alternaria spp.和镰孢属Fusarium spp.;种子内部寄藏真菌主要为青霉属、根霉属和曲霉属;不同品种之间种子表面携带真菌种类差异较大,种子内部寄藏真菌种类差异不明显;种壳带菌率一般高于种仁带菌率。15% FDDF ·霜·福悬浮种衣剂、福美双和代森锰锌对种子带菌消毒效果优于多菌灵、敌磺钠、 FDDF 霉灵、拌种灵和甲霜灵。     

To what extent are soil amendments useful to control Verticillium wilt?

The genus Verticillium includes several species that attack economically important crops throughout the world. The control of Verticillium spp. becomes especially difficult when they form microsclerotia that can survive in the field soil for several years. It has been common practice to fumigate soil with chemicals such as methyl bromide and/or chloropicrin to control soil‐borne fungal pathogens. Other chemicals that are used against Verticillium spp. are the antifungal antibiotic aureofungin, the fungicides benomyl, captan, carbendazim, thiram, azoxystrobin and trifloxystrobin and the plant defence activator acibenzolar‐S‐methyl. However, the potential risks involved in applying phytochemicals to crop plants for both the environment and human health, together with their limited efficacy for controlling Verticillium‐induced diseases, support the need to find alternatives to replace their use or improve their efficacy. Soil amendment with animal or plant organic debris is a cultural practice that has long been used to control Verticillium spp. However, today the organic farming industry is becoming a significant player in the global agricultural production scene. In this review, some of the main results concerning the efficacy of several soil amendments as plant protectors against Verticillium spp. are covered, and the limitations and future perspectives of such products are discussed in terms of the control of plant diseases. Copyright © 2009 Society of Chemical Industry     

Farmers' knowledge and perceptions of termites as pests of yam (Dioscorea spp.) in Central Benin

Destruction of yam (Dioscorea spp.) tubers by termites is a major constraint to yam production in Central Benin. To obtain the basic information for the development of appropriate strategies of termites management in yam, a structured questionnaire was used to interview 142 farmers about their knowledge of termite species of yam, susceptibility of yam landraces, and management practices in 14 villages through the study area. A total of 45 vernaculars names of termites were recorded corresponding to 10 species. All the names given to termite species had a signification, mainly related to morphological aspects, type of damage, and termite caste. Eleven features were used by farmers to identify yam termites. The infestation of yam fields mostly occurred from October to April with Amitermes evuncifer and Trinevitermes oeconomus being the most damaging termite species. Overall, eight factors were identified by farmers as favouring the proliferation of termites in yam fields with drought being the most frequently reported factor. Farmers identified 11 yam landraces that were highly susceptible to termites attack, while 12 yam landraces were listed as resistant. Application of chemicals was the most commonly reported control method, followed by destruction of termite nest, and application of botanical extracts. According to farmers, the high multiplication rate of termites is the most important constraint in managing the issue of termites in yam.     

Molecular Detection of Phytophthora ramorum, the Causal Agent of Sudden Oak Death in California, and Two Additional Species Commonly Recovered from Diseased Plant Material

ABSTRACT Sudden oak death is a disease currently devastating forest ecosystems in several coastal areas of California. The pathogen causing this is Phy-tophthora ramorum, although species such as P. nemorosa and P. pseudo-syringae often are recovered from symptomatic plants as well. A molecular marker system was developed based on mitochondrial sequences of the cox I and II genes for detection of Phytophthora spp. in general, and P. ramorum, P. nemorosa, and P. pseudosyringae in particular. The first-round multiplex amplification contained two primer pairs, one for amplification of plant sequences to serve as an internal control to ensure that extracted DNA was of sufficient quality to allow for polymerase chain reaction (PCR) amplification and the other specific for amplification of sequences from Phytophthora spp. The plant primers amplified the desired amplicon size in the 29 plant species tested and did not interfere with amplification by the Phytophthora genus-specific primer pair. Using DNA from purified cultures, the Phytophthora genus-specific primer pair amplified a fragment diagnostic for the genus from all 45 Phytophthora spp. evaluated, although the efficiency of amplification was lower for P. lateralis and P. sojae than for the other species. The genus-specific primer pair did not amplify sequences from the 30 Pythium spp. tested or from 29 plant species, although occasional faint bands were observed for several additional plant species. With the exception of one plant species, the resulting amplicons were smaller than the Phytophthora genus-specific amplicon. The products of the first-round amplification were diluted and amplified with primer pairs nested within the genus-specific amplicon that were specific for either P. ramorum, P. nemorosa, or P. pseudo-syringae. These species-specific primers amplified the target sequence from all isolates of the pathogens under evaluation; for P. ramorum, this included 24 isolates from California, Germany, and the Netherlands. Using purified pathogen DNA, the limit of detection for P. ramorum using this marker system was approximately 2.0 fg of total DNA. However, when this DNA was spiked with DNA from healthy plant tissue extracted with a commercial miniprep procedure, the sensitivity of detection was reduced by 100- to 1,000-fold, depending on the plant species. This marker system was validated with DNA extracted from naturally infected plant samples collected from the field by comparing the sequence of the Phytophthora genus-specific amplicon, morphological identification of cultures recovered from the same lesions and, for P. ramorum, amplification with a previously published rDNA internal transcribed spacer species-specific primer pair. Results were compared and validated with three different brands of thermal cyclers in two different laboratories to provide information about how the described PCR assay performs under different laboratory conditions. The specificity of the Phytophthora genus-specific primers suggests that they will have utility for pathogen detection in other Phytophthora pathosystems.     

First interception of the asiatic Bamboo longhorn, <Emphasis Type="Italic">Chlorophorus annularis</Emphasis> (F., 1787) (Coleoptera,Cerambycidae) in Italy

This study provide information on the first occurrence in Italy of the Coleoptera Cerambycidae Chlorophorus annularis (Fabricius, 1787). It is a species naturally and widely distributed in Asia and in oriental tropical and subtropical areas. This cerambyx is a post-harvest pest of bamboo belonging to several genera as Bambusa, Dendrocalamus, Phyllostachys, Sinocalamus, but it can also infest cultivated plants as Citrus spp., Gossypium, Liquidambar spp., Saccharum officinarum L., Vitis spp., and Zea mays L.     

A universal microarray detection method for identification of multiple Phytophthora spp. using padlock probes

The genus Phytophthora consists of many species that cause important diseases in ornamental, agronomic, and forest ecosystems worldwide. Molecular methods have been developed for detection and identification of one or several species of Phytophthora in single or multiplex reactions. In this article, we describe a padlock probe (PLP)-based multiplex method of detection and identification for many Phytophthora spp. simultaneously. A generic TaqMan polymerase chain reaction assay, which detects all known Phytophthora spp., is conducted first, followed by a species-specific PLP ligation. A 96-well-based microarray platform with colorimetric readout is used to detect and identify the different Phytophthora spp. PLPs are long oligonucleotides containing target complementary sequence regions at both their 5' and 3' ends which can be ligated on the target into a circular molecule. The ligation is point mutation specific; therefore, closely related sequences can be differentiated. This circular molecule can then be detected on a microarray. We developed 23 PLPs to economically important Phytophthora spp. based upon internal transcribed spacer-1 sequence differences between individual Phytophthora spp. Tests on genomic DNA of many Phytophthora isolates and DNA from environmental samples showed the specificity and utility of PLPs for Phytophthora diagnostics.     

First report of the papaya mealybug,Paracoccus marginatus (Coccomorpha: Pseudococcidae), in Tahiti,French Polynesia

The papaya mealybug, Paracoccus marginatus Williams and Granara de Willing (Coccomorpha: Pseudococcidae), is a polyphagous pest that damages many tropical crops. It is native to Central America and spread to the Caribbean region and South America in the 1900s. Since then, it has accidentally been introduced to several islands in the Pacific region and some countries in Africa and Asia. The first record of this mealybug in Polynesia was in Hawaii, in 2004. This paper presents the first report of this pest in Tahiti (French Polynesia), where it was found in March 2017 on papaya (Carica papaya) and frangipani trees (Plumeria spp.). Its presence, which is mainly in the inhabited part of the island of Tahiti, suggests the recent introduction of the pest in French Polynesia. Several coccinellid predators have been found feeding on the mealybug in Tahiti: Cryptolaemus montrouzieri, Hyperaspis pantherina and Scymnus spp. None of its parasitoids have been found yet.     

Effect of 2,4-diacetylphloroglucinol on pythium: cellular responses and variation in sensitivity among propagules and species

ABSTRACT The antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) plays an important role in the suppression of plant pathogens by several strains of Pseudomonas spp. Based on the results of this study, there is variation within and among Pythium spp. to 2,4-DAPG. Also, various propagules of Pythium ultimum var. sporangiiferum, that are part of the asexual stage of the life cycle, differ considerably in their sensitivity to 2,4-DAPG. Mycelium was the most resistant structure, followed by zoosporangia, zoospore cysts, and zoospores. Additionally, we report for the first time that pH has a significant effect on the activity of 2,4-DAPG, with a higher activity at low pH. Furthermore, the level of acetylation of phloroglucinols is also a major determinant of their activity. Transmission electron microscopy studies revealed that 2,4-DAPG causes different stages of disorganization in hyphal tips of Pythium ultimum var. sporangiiferum, including alteration (proliferation, retraction, and disruption) of the plasma membrane, vacuolization, and cell content disintegration. The implications of these results for the efficacy and consistency of biological control of plant-pathogenic Pythium spp. by 2,4-DAPG-producing Pseudomonas spp. are discussed.     

Wild Lactuca species,their genetic diversity,resistance to diseases and pests,and exploitation in lettuce breeding

Current knowledge of wild Lactuca L. species, their taxonomy, biogeography, gene-pools, germplasm collection quality and quantity, and accession availability is reviewed in this paper. Genetic diversity of Lactuca spp. is characterized at the level of phenotypic and phenological variation, variation in karyology and DNA content, biochemical traits, and protein and molecular polymorphism. The reported variation in reaction to pathogens and pests of wild Lactuca spp. is summarized, including the viral pathogens (Lettuce mosaic virus-LMV, Mirafiori lettuce virus/Lettuce big vein virus-LBV, Beet western yellows virus-BWYV, Tomato spotted wilt virus-TSWV, Cucumber mosaic virus-CMV, Lettuce necrotic stunt virus-LNSV), bacterial pathogens (corky root-Rhizomonas suberifaciens, bacterial leaf spot-Xanthomonas campestris pv. vitians), fungal pathogens (downy mildew-Bremia lactucae, powdery mildew-Golovinomyces cichoracearum, anthracnose-Microdochium panattoniana, stemphylium leaf spot-Stemphylium spp., sclerotinia drop-Sclerotinia spp., verticillium wilt-Verticillium dahliae, fusarium wilt-Fusarium spp., pythium wilt-Pythium tracheiphylum, P. uncinulatum), nematodes (potato cyst nematode-Globodera rostochiensis, root-knot nematode-Meloidogyne spp., incognita, hapla, javanica, enterolobii), insects and mites (the green lettuce aphid-Nasonovia ribisnigri, the green peach aphid-Myzus persicae, the potato aphid-Macrosiphum euphorbiae, leafminer-Liriomyza spp., L. langei). The approaches used to exploit wild Lactuca spp. in lettuce breeding (interspecific hybridization, cell and tissue culture, transformation) are dicussed, and known examples of lettuce cultivars with traits derived from wild Lactuca spp. are described.     

Induced Systemic Resistance and Promotion of Plant Growth by Bacillus spp

ABSTRACT Elicitation of induced systemic resistance (ISR) by plant-associated bacteria was initially demonstrated using Pseudomonas spp. and other gram-negative bacteria. Several reviews have summarized various aspects of the large volume of literature on Pseudomonas spp. as elicitors of ISR. Fewer published accounts of ISR by Bacillus spp. are available, and we review this literature for the first time. Published results are summarized showing that specific strains of the species B. amyloliquefaciens, B. subtilis, B. pasteurii, B. cereus, B. pumilus, B. mycoides, and B. sphaericus elicit significant reductions in the incidence or severity of various diseases on a diversity of hosts. Elicitation of ISR by these strains has been demonstrated in greenhouse or field trials on tomato, bell pepper, muskmelon, watermelon, sugar beet, tobacco, Arabidopsis sp., cucumber, loblolly pine, and two tropical crops (long cayenne pepper and green kuang futsoi). Protection resulting from ISR elicited by Bacillus spp. has been reported against leaf-spotting fungal and bacterial pathogens, systemic viruses, a crown-rotting fungal pathogen, root-knot nematodes, and a stem-blight fungal pathogen as well as damping-off, blue mold, and late blight diseases. Reductions in populations of three insect vectors have also been noted in the field: striped and spotted cucumber beetles that transmit cucurbit wilt disease and the silver leaf whitefly that transmits Tomato mottle virus. In most cases, Bacillus spp. that elicit ISR also elicit plant growth promotion. Studies on mechanisms indicate that elicitation of ISR by Bacillus spp. is associated with ultrastructural changes in plants during pathogen attack and with cytochemical alterations. Investigations into the signal transduction pathways of elicited plants suggest that Bacillus spp. activate some of the same pathways as Pseudomonas spp. and some additional pathways. For example, ISR elicited by several strains of Bacillus spp. is independent of salicylic acid but dependent on jasmonic acid, ethylene, and the regulatory gene NPR1-results that are in agreement with the model for ISR elicited by Pseudomonas spp. However, in other cases, ISR elicited by Bacillus spp. is dependent on salicylic acid and independent of jasmonic acid and NPR1. In addition, while ISR by Pseudomonas spp. does not lead to accumulation of the defense gene PR1 in plants, in some cases, ISR by Bacillus spp. does. Based on the strains and results summarized in this review, two products for commercial agriculture have been developed, one aimed mainly at plant growth promotion for transplanted vegetables and one, which has received registration from the U.S. Environmental Protection Agency, for disease protection on soybean.     

入侵杂草虉草在云南的发生、危害和防除状况调查

采用倒置"W"九点取样法和5级目测法,调查了云南冬季作物田杂草虉草的发生和危害状况。结果表明,截至2017年在云南有7个地州33个区县发现有虉草危害,其中保山地区危害最为严重,占全省受害面积的92%,其次是大理白族自治州。虉草在冬季作物田大麦、小麦、油菜和蚕豆田均有发生。危害的虉草有两个种,分别是小籽虉草Phalaris minor和奇异虉草P.paradoxa。其中小籽虉草的发生较为普遍,奇异虉草为零星发生。虉草造成的危害指数在6%~50%之间,对农作物造成的产量损失在40~110kg/667m2之间。虉草的防除以化学防除为主,人工防除为辅,但目前尚缺乏经济有效的防除农田虉草的除草剂。     

Prevalence and diversity of plant parasitic nematodes in Northern Ireland grassland and cereals,and the influence of soils and rainfall

The prevalence and diversity of plant parasitic nematodes in Northern Ireland cereal and grassland was determined from 191 agricultural fields. A total of 18 nematode genera were detected, including economically important pests, Meloidogyne spp., Heterodera spp. and Pratylenchus spp., each of which were above economic damage thresholds in a significant proportion of the sites (92.4%, 70% and 28.6%, respectively). The detection of the root knot nematode, Meloidogyne minor (6% prevalence), was significant given its recent emergence across the turf grass sector and the prospect of M. minor becoming a common agricultural pest. Analyses of nematode prevalence and abundance highlighted significant associations with grass and cereals, soil types, soil grade (proxy for soil quality) and rainfall levels. Specifically, nematode populations varied between the two major soils (brown earths and gleys), while significant trends for increased nematode diversity and greater prevalence of both Meloidogyne and Pratylenchus with increasing rainfall were also observed. Multivariate analyses were performed to determine interactive effects and the relative importance of the factors affecting nematode populations. Notably, rainfall, in combination with either crop type or soil grade, had a significant effect on nematode abundance and diversity. The findings suggest significant changes in nematode populations have occurred over the last several decades and the possibility that these are linked to changing climate and cropping practices are discussed, as well as future concerns for plant parasitic nematode management.     

New Anastomosis Groups, AG-T and AG-U, of Binucleate Rhizoctonia spp. Causing Root and Stem Rot of Cut-Flower and Miniature Roses

ABSTRACT Root and stem rot of cut-flower roses (Rosa spp.) was observed in commercial glasshouse-grown roses in 10 prefectures of Japan from 1998 through 2001. Binucleate-like Rhizoctonia spp. were isolated mainly from the disease plants. In all, 670 isolates were divided into two types based on cultural appearance; 168 isolates of light brown to brown type and 502 isolates of whitish type. A hyphal anastomosis reaction using representative isolates from each type revealed that the light brown to brown type belonged to anastomosis group G (AG-G), whereas the whitish type (AG-CUT) failed to anastomose with tester strains of binucleate Rhizoctonia AG-A through AG-S. Neither isolates of AG-G nor AG-CUT anastomosed with tester strains of a previously reported unknown AG (AG-MIN) of binucleate Rhizoctonia spp. collected from miniature roses. In pathogenicity tests, randomly selected isolates of the three groups caused root and stem rot on cut-flower and miniature roses. To differentiate AG-CUT and AG-MIN from known AGs of binucleate Rhizoctonia spp., restriction fragment length polymorphism (RFLP) and sequence analyses of a ribosomal (r)DNA internal transcribed spacer (ITS) region were conducted. Among the eight restriction enzymes used, HaeIII produced DNA banding patterns for AG-CUT that differed from those of tester strains and AG-MIN. Additionally, restriction profiles of AG-MIN differed from those of all tester strains. AG-G isolates from cut-flower roses had the same RFLP pattern as the tester strains of AG-G. Based on the results of hyphal anastomosis and RFLP and sequence analysis of an rDNA-ITS region, we propose that AG-CUT be designated AG-T and AG-MIN be designated AG-U, two new AGs of binucleate Rhizoctonia spp. The phylogenetic tree based on the sequence data of the rDNA-ITS region showed that isolates of AG-MIN were in a distinct clade from other AGs, whereas isolates of AG-CUT were in the same clade as those of AG-A. More detailed phylogenetic analysis besides rDNA-ITS region might be necessary for AG classification of binucleate Rhizoctonia spp.