Identification and characterization of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. associated with chili anthracnose in peninsular Malaysia

Anthracnose fruit rot caused by Colletotrichum spp. is a serious post-harvest disease of chili fruits (Capsicum spp.). One hundred-thirty isolates of Colletotrichum spp. were isolated from anthracnose of green and red cayenne pepper (Capsicum annuum) and bird’s eye chili (Capsicum frutescens). The isolates were morphologically identified as Colletotrichum acutatum sensu lato (62 isolates), Colletotrichum truncatum (54 isolates), and Colletotrichum gloeosporioides sensu lato (14 isolates). Molecular identification and phylogenetic analyses were based on internal transcribed spacer regions, β-tubulin, actin, and glyceraldehyde-3-phosphate dehydrogenase genes, and the isolates were re-identified as C. scovillei (58 isolates), C. truncatum (54 isolates), C. siamense (11 isolates), C. fioriniae (four isolates), and C. fructicola (3 isolates). Maximum likelihood trees using combined sequences showed that isolates of the same species grouped in the same main clade with the isolates used for comparison. Pathogenicity testing showed that the tested isolates from each species were pathogenic towards green and red Capsicum annuum and Capsicum frutescens upon treatment of wounded fruit, using both the mycelial plug and conidial suspension methods. Only five isolates of C. truncatum and seven isolates of C. scovillei were found to be pathogenic upon treatment of unwounded fruit. The occurrence of five Colletotrichum spp. (C. siamense, C. fructicola, C. scovillei, C. fioriniae, and C. truncatum) associated with chili anthracnose in Peninsular Malaysia indicates that correct species identification is important to formulate not only effective disease management, but also effective quarantine policy.     

Markers to differentiate species of anthracnose fungi identify <Emphasis Type="Italic">Colletotrichum fructicola</Emphasis> as the predominant virulent species in strawberry plants in Chiba Prefecture of Japan

Colletotrichum fungi belonging to the Colletotrichum gloeosporioides species complex include a number of economically important postharvest pathogens that often cause anthracnose. Until now, different species within this group could only be distinguished from one another reliably using multigenic phylogenetic analyses. Using a comparative genomics approach, we developed a marker that can differentiate Colletotrichum fructicola, Colletotrichum aenigma and Colletotrichum siamense within the C. gloeosporioides species complex based on PCR amplicon size differences. When we used this marker to classify 115 isolates collected over 20 years from strawberries in Chiba Prefecture, Japan, the isolates were predominantly C. fructicola. To our knowledge, this is the first report characterizing different species of Colletotrichum infecting strawberries in Japan and contributes to our understanding on the diversity of anthracnose pathogens in Japan.     

Molecular and phenotypic characterization revealed six <Emphasis Type="Italic">Colletotrichum</Emphasis> species responsible for anthracnose disease of small cardamom in South India

Small cardamom (Elettaria cardamomum Maton) is extensively cultivated in the Western Ghats of South India either as a monocrop under the forest trees or as an intercrop along with arecanut and coffee plantations. Colletotrichum species responsible for severe outbreaks of anthracnose on small cardamom in South India are reported. Small cardamom anthracnose, popularly known as “Chenthal”, manifests itself on the foliage as yellowish lesions, which later coalesce to form large blighted areas. In advanced stages, the affected leaves dry up giving a burnt appearance to the plant. Twenty-five isolates of Colletotrichum were isolated from leaves of small cardamom in Karnataka, Kerala and Tamil Nadu states of India. The isolates were characterized through morphological studies and multilocus phylogenetic analysis (ITS, ACT, CHS-1, GAPDH, TUB2, CYLH3, GS and ApMat gene regions) to test whether different species are present and identified: C. karstii (2 isolates), C. gloeosporioides (1), C. siamense (7), C. syzygicola (6), Colletotrichum sp (5), and C. guajavae (4), as the cause of anthracnose on small cardamom for the first time. Pathogenicity of the six species was confirmed. To our knowledge, this is the first detailed study of Colletotrichum species which cause anthracnose diseases on small cardamom.     

Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> species in coffee (<Emphasis Type="Italic">Coffea arabica</Emphasis>) plantations in Mexico

The aim of this study was to identify the Colletotrichum species associated with anthracnose symptoms in coffee (Coffea arabica L.) plantations in northern Puebla, Mexico. In 2013, five surveys were conducted in different production areas and at different altitudes. Symptomatic leaves, shoots, and ripe and unripe fruits of the coffee variety Red Caturra were collected. Isolates were obtained and the Colletotrichum species were identified morphologically and characterized by multilocus sequence analyses of the ACT, CAL, GAPDH, and TUB2 genes and the rDNA region. Additionally, pathogenicity tests were conducted using six isolates. We identified C. gigasporum, C. gloeosporioides, C. karstii (two isolates), C. siamense, and C. theobromicola. This is the first report of these five species infecting leaves of coffee. The symptoms caused by these species were characterized, but the species causing Coffee Berry Disease was not found. This is the first report of a complex of species affecting coffee plants in the same geographical area in Mexico, and suggests that other complexes of species may be important pathogens in coffee-producing areas elsewhere.     

Role of secondary metabolites in the biocontrol activity of <Emphasis Type="Italic">Pseudomonas corrugata</Emphasis> and <Emphasis Type="Italic">Pseudomonas mediterranea</Emphasis>

Colletotrichum gloeosporioides is the causal agent of Camellia oleifera anthracnose, mainly infecting fruits and leaves. The fungus secretes degrading enzymes to destroy the cuticle of aerial plant parts and help infect the host successfully. To validate whether a cutinase gene (CglCUT1) was required for cutinase activity and pathogenicity of C. gloeosporioides, the CglCUT1 gene was cloned and analyzed. The characterization of CglCUT1 predicted protein suggests that the cloned DNA encoded a cutinase in C. gloeosporioides affecting C. oleifera. The CglCUT1 showed a high homology to those from C. gloeosporioides causing papaya anthracnose and C. capsici causing pepper anthracnose, as well as those of other ascomycetes. The whole CglCUT1 gene was knocked-out and the knockout mutant (?CglCUT39) was subsequently complemented using Agrobacterium tumefaciens mediated transformation. The knockout transformants exhibited significant decreases in cutinase activity and virulence compared with the wild-type strain. The complemented transformants of the disrupted transformant ?CglCUT39 showed a significant increase in cutinase activity and virulence compared with the disrupted transformant ?CglCUT39. This study suggests that the CglCUT1 gene has a positive effect on fungal virulence of the hemibiotrophic C. gloeosporioides on C. oleifera.     

Construction of a recombinant strain of <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> producing both phenazine-1-carboxylic acid and cyclic lipopeptide for the biocontrol of take-all disease of wheat

Four Neofabraea species are responsible for bull’s eye rot, which is an important postharvest disease of apples and pears. The species diversity of its causal agents in Europe has not been thoroughly explored using molecular genetic methods. Eighty-one Neofabraea isolates were obtained mostly from apples with bull’s eye rot symptoms in the Czech Republic over a two year period. The isolates were identified using PCR fingerprinting and DNA sequencing of the ITS rDNA region, the mitochondrial SSU rDNA and the β-tubulin and EF1α genes. The most common species was N. alba (89 %), followed by N. perennans (5 %) and N. kienholzii (5 %). This is the third published record of N. kienholzii in Europe. The species identity of the isolate CPPF507, which was placed close to N. kienholzii, remains unclear. EF1α was shown to be a suitable marker for the identification of species of the genus Neofabraea and was comparable to the previously used β-tubulin gene. Furthermore, the aggressiveness of individual species was compared and species distribution across Europe was summarized. N. perennans and isolate CPPF507 proved to be the most aggressive, whereas the least aggressive was N. kienholzii. Two N. alba isolates isolated from symptomless apple fruits and leaves were pathogenic to apples in the infection tests.     

Pathogenicity and characterization of <Emphasis Type="Italic">Colletotrichum lentis</Emphasis>: A causal agent of anthracnose in common vetch (<Emphasis Type="Italic">Vicia sativa</Emphasis>)

A series of studies were carried out on Colletotrichum lentis which had been been identified in 2015 based largely on the distinctive shape of conidia and ITS sequences, and which has been causing severe anthracnose disease symptoms on common vetch plants (Vicia sativa) in Gansu Province in the northwest region of China. A key focus of the present studies was to determine how vetch crops become infected. The addition of residues from harvested common vetch crops to land being prepared as a seedbed was shown to result in the highest levels of disease severity indicating that this management practice was the most likely way for crops to become severely infected. Seed transmission was unlikely to be the cause of severe outbreaks as less than 5% of seeds harvested from severely infected plants carried C. lentis. To verify that the species causing the severe outbreaks of anthracnose disease of vetch crops was C. lentis, sequence analysis of the ITS, TUB2, ACT, HIS3 and GAPDH genes was conducted. C. lentis isolates from common vetch and lentil (Lens culinaris) formed a distinctive group among Colletotrichum species, including those species that infect other forage and field crops. The unique shape of conidia of C. lentis, straight with only one end curved, was confirmed as being reliable for rapid identification of disease outbreaks caused by this damaging fungal pathogen.     

Pathogenicity,Morpho-Species and Mating Types of <Emphasis Type="Italic">Alternaria</Emphasis> spp. causing Alternaria blight in <Emphasis Type="Italic">Pistacia</Emphasis> spp. in Turkey

Alternaria genus includes many plant pathogens on numerous hosts, causing leaf spots, rots and blights. Alternaria blight has been observed as one of the important fungal diseases of pistachio (Pistacia vera L.) as well as its wild relatives (P. terebinthus, P. lentiscus, P. khinjuk, P. atlantica, P. mutica) in Turkey. Alternaria species were sampled from Pistacia spp. hosts from different geographic regions in Turkey during field trips in late spring to early fall of 2013. Alternaria blight symptoms were observed mainly on fruits and rarely on leaves. Four hundred and twenty two of the isolates were morphologically defined as A. alternata, A. tenuissima, A. arborescens and also intermediate morpho-species between A. alternata/A. arborescens. Pathogenicity of the isolates was confirmed with host inoculations on detached fruits. Mating types of 270 isolates of Alternaria spp. from the collection were identified using a PCR-based mating type assay that amplifies either a MAT1-1 or a MAT1-2 fragment from the mating locus. Although a strongly clonal population structure was expected due to the putative asexual reproduction of these fungi, both idiomorphs were detected at equal frequencies at several different spatial scales. The distribution of mating types within each geographic region, within host species as well as in overall collection was not significantly different from 1:1. Amplified fragments of partial idiomorph sequences were obtained for representative isolates. Parsimony trees were depicted based on sequence data of mating type genes for these representative isolates as well as some other Alternaria species obtained by Genebank. Several point mutations presented a few clusters which are supported by high bootsrapped values. The Alternaria blight disease agents both from cultivated and wild hosts were pathogenic on pistachio which may cause difficulties to control the disease because of extensity of pathogen sources. Besides, equal mating type distribution of the pathogen at both geographic and host species levels suggests a potential for sexual reproduction of Alternaria spp. in Turkey.     

<Emphasis Type="Italic">Fibulorhizoctonia psychrophila</Emphasis> is the causal agent of lenticel spot on apple and pear fruit in the Netherlands

In a survey for postharvest diseases of apples and pears in the Netherlands, an unknown postharvest fruit rot was observed. The disease appeared to originate from infected lenticels. A fungus was consistently isolated from the decayed fruits. The fungal pathogen was isolated on potato dextrose agar, and at low temperatures development of a fast-growing whitish mycelium was observed. Growth of this fungus was observed between 1 and 20 °C with an optimum at 15 °C, while incubation of mycelium at 25 °C resulted in no growth. The isolates did not produce asexual or sexual spores. The isolates were characterized and identified by morphology and molecular phylogenetic analysis. Genomic DNA was isolated and amplified using ITS1-ITS4, EF1 and RPB2 primers, and BLAST searches in GenBank placed the fungus taxonomically in the genus Fibulorhizoctonia, with the highest matches to F. psychrophila. Pathogenicity of representative isolates from apple and pear fruit was confirmed under laboratory conditions. To the best of our knowledge this is the first report of F. psychrophila causing lenticel spot on apple and pear, and also the cause of a whitish mould on storage bins.     

Fungicide sensitivity,growth rate,aggressiveness and frost hardiness of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> and <Emphasis Type="Italic">Monilinia laxa</Emphasis> isolates

Monilinia fructicola, the most destructive pathogen of the genus Monilinia, has recently been introduced into Serbia and many other European countries. Since then, many studies have been conducted to evaluate the characteristics of Monilinia species that have a role in the establishment and survival of the pathogen in new areas. The present study assessed the capacity of M. fructicola to repress and replace Monilinia laxa in Serbia based on: fungicide sensitivity, growth rate and aggressiveness at different temperatures, as well as frost hardiness of the isolates of both species. The results showed that the isolates of M. fructicola, compared to M. laxa, were significantly less sensitive to the following fungicides: iprodione, tebucanozole, chlorothalonil, azoxystrobin, fluopyram, and boscalid. In addition, M. laxa isolates exhibited little variation in sensitivity to all of the tested fungicides, whereas M. fructicola isolates displayed a wide range of sensitivity. The temperature of 5°C favored M. laxa growth and aggressiveness, while at 30°C M. fructicola grew faster and had higher lesion expansion rate. These results support an assumption that M. fructicola will continue to spread in Serbian orchards in coming years, particularly on stone fruits harvested during hot summer weather.     

First report of flyspeck of mango caused by <Emphasis Type="Italic">Stomiopeltis</Emphasis> sp. in Japan

Flyspeck symptoms were found on mango (Mangifera indica L.) in Okinawa Prefecture, Japan in June 2014. Just before harvest, surfaces of fruits and green branches developed groups of tiny black dots within dark smudges. A fungus was isolated from the dots on both fruits and branches. The isolates were identified as Stomiopeltis sp. based on pathogenicity, morphology and molecular phylogenetic analyses with rDNA-ITS and LSU sequences. This is the first report of flyspeck (susuten-byo in Japanese) on mango caused by Stomiopeltis sp. in Japan.     

Histological observation of cucumber infected with <Emphasis Type="Italic">Corynespora cassiicola</Emphasis>

Erwinia amylovora is the causative agent of fire blight, which is a destructive bacterial disease of rosaceous plants. In Hungary Erwinia amylovora (Burrill) Winslow et al. was first detected in 1996. Since the appearance of fire blight, E. amylovora samples have been collected from different host plats from various geographic locations. A motif of eight nucleotides (ATTACAGA) is repeated 3–15 times in the PstI fragment of the pEa29 plasmid in Erwinia amylovora strains, and represents a valuable tool for strain classification. The number of short-sequence DNA repeats in plasmid pEa29 of 30 Hungarian isolates were determined by PCR assays and they ranged from five to ten. The SSR test is suitable for distinguishing the individual strains between the E. amylovora isolates. The examined isolates showed high pathogenicity on immature pear fruits. Several biochemical techniques, such as miniaturized API 20E, were applied on the samples. Differences were also revealed in microbiological assays like levan formation and colony morphology on semi-selective media. Examining the Hungarian Erwinia amylovora population by molecular analysis we can draw the conclusion that the population consists of different strains, which shows great diversity. E. amylovora is a widespread pathogen in Hungary, which is supported by the 30 strains isolated from various host plants from many parts of the country. The phenotypic diversity-evaluation of the E. amylovora strains showed, that they differ metabolically, like other plant pathogenic bacteria as reported by several authors. This is the first report on the diversity of E. amylovora strains isolated from Hungary.     

Population features of <Emphasis Type="Italic">Xanthomonas arboricola</Emphasis> pv. <Emphasis Type="Italic">pruni</Emphasis> from <Emphasis Type="Italic">Prunus spp.</Emphasis> orchards in northern Italy

Bacterial leaf/fruit spot and canker of stone fruits, caused by Xanthomonas arboricola pv. pruni, is a recurrent disease in Italy. A set of 23 strains has been isolated in peach and plum orchards in an intensively stone fruit cultivated area located in north-eastern Italy. They were all identified as X. arboricola pv. pruni by means of phytopathological and serological features: hypersensitive reaction on bean pods, pathogenicity test on immature peach or plum fruitlets, identification by immunofluorescence assay and conventional PCR. Phylogenetic analysis based on sequencing of the gyrB housekeeping gene of the isolates showed that they formed a unique clade, well characterised and separated from other xanthomonads. An insight into the genetic population features was attempted by rep-PCR analysis, using the ERIC, REP and BOX primers. The combined rep-PCR fingerprints showed a slight intra-pathovar variation within our isolates, which grouped in five close clusters. Copper resistance has been assessed in vitro for our whole X. arboricola pv. pruni collection, highlighting that two isolates show a level of resistance in vitro up to 200 ppm of copper. Nonetheless, the copLAB gene cluster, present in many other species of Xanthomonads, was not detected in any isolate, confirming the presence of a still unknown mechanism of copper detoxification in our Xanthomonads arboricola pv. pruni tolerant/resistant strains.     

Postharvest evaluation of natural coatings and antifungal agents to control <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in <Emphasis Type="Italic">Rosa</Emphasis> sp.

Botrytis cinerea is a fungal pathogen that limits rose production and commercialization worldwide. Therefore, we evaluated a novel postharvest treatment against Botrytis cinerea in roses (Rosa sp. cv Vendela) using coating bases and antifungal agents of natural origin. Aloe vera pulp, cassava starch and gelatin were used as coating bases. Oregano essential oil (Origanum vulgare), thyme essential oil (Thymus vulgaris) and chitosan were used as natural antifungal agents. The coating bases were evaluated in different concentrations to observe effects of toxicity and opening diameter in rose buds. Gelatin and cassava starch coatings inhibited rose opening and showed petal damage in all concentrations tested. However, Aloe vera pulp at 25% allowed normal buds’ opening and no damage was observed, indicating that Aloe vera could be an ideal coating base for rose postharvest treatments. During in vitro assays, natural antifungal agents efficiently inhibited Botrytis cinerea growth in the concentrations tested. Further, mixture treatments of Aloe vera pulp (25%) with oregano essential oil (1%), thyme essential oil (0.1%) and chitosan (0.1%) showed independently neither damage nor opening inhibition in rose buds. Selected combinations of Aloe vera pulp and natural antifungal agents were applied in roses infected with Botrytis cinerea to evaluate their control of this pathogen. Unfortunately, the selected combinations did not reduce pathogen growth during postharvest treatments since they were similar to untreated controls. Further research has to be performed to find ideal combinations with Aloe vera that could inhibit B. cinerea during postharvest treatments in roses.     

Comparison among Japanese isolates of <Emphasis Type="Italic">Pseudomonas savastanoi</Emphasis> pv. <Emphasis Type="Italic">savastanoi</Emphasis>, causal agent of olive knot disease

Olive knot disease in Japan was first reported in Shizuoka Prefecture in 2014, and the causal agent was identified as Pseudomonas savastanoi pv. savastanoi. Subsequently, olive trees having knots were also found in Aichi and Kanagawa Prefectures in 2015, and the isolates from knots were also suspected to be P. savastanoi pv. savastanoi through preliminary examinations. Therefore, the Aichi and Kanagawa isolates were identified through comparison of isolates from three prefectures. Phylogenic analysis based on 16S rDNA and housekeeping genes (gyrB, rpoD, gltA and gap1) revealed that the isolates belonged to the same cluster as the pathotype strain, ICMP4352^PT. The iaaM, H and L genes, which are involved in promotion of symptoms, and the ina gene coding the ice nucleation protein, were detected by PCR from all the isolates. In rep-PCR (ERIC and REP) analyses, the isolates yielded DNA fragment-banding patterns that were nearly identical to that of ICMP4352^PT, but slight variations in banding patterns were observed among them. In a pathogenicity test, the isolates formed distinct knots on olive and pink jasmine. Phenotypic properties of the isolates were almost identical to those of ICMP4352^PT, with the exception of d-sorbitol utilization. Consequently, Aichi and Kanagawa isolates from olive were identified as P. savastanoi pv. savastanoi, and several genetic diversities in terms of rep-PCR were found in the Japanese population of P. savastanoi pv. savastanoi, indicating their heterogeneity.     

Control of infection of tomato fruits by <Emphasis Type="Italic">Alternaria</Emphasis> and mycotoxin production using plant extracts

Tomato fruits are susceptible to infection by Alternaria species. In addition, Alternaria species may contaminate the fruits with mycotoxins. There is thus interest in control systems to minimise pathogenicity and control toxin production. The objectives of this study were to examine the effect of plant extracts of Eucalyptus globulus and Calendula officinalis on the growth of strains of Alternaria alternata and A. arborescens, on pathogenicity of tomato fruits and mycotoxin production. The growth bioassays showed that the ethanolic and chloroformic fractions of E. globulus were the most effective in reducing growth of A. alternata (66–74 %) and A. arborescens (86–88 %), respectively at 2500 μg/g. The effects of plant extracts on mycotoxin biosynthesis were variable and strain dependent. The most effective fractions in decreasing mycotoxin accumulation were the ethanolic and chloroformic extracts of E. globulus, which reduced tenuazonic acid by 89 %, alternariol by 75–94 % and almost complete inhibition of alternariol monomethyl ether. All the tested fractions reduced percentage of infected tomato fruits when compared to the controls. The ethanolic and chloroformic fractions of E. globulus completely inhibited growth of A. alternata and A. arborescens on unwounded fruits and reduced the aggressiveness on wounded fruits of strains of both species significantly.     

The occurrence of postharvest gray-mold rot of sweet persimmon caused by <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in Korea

In this study, we identified the causative agent of postharvest gray-mold rot in sweet persimmon fruit that were collected from Gangneung, Gangwon Province, Korea in October 2016. Symptoms included extensive growth of mycelia on post harvested fruit. The fungus was isolated from infected fruit and cultured on potato dextrose agar (PDA). For identification of the fungus, we examined morphology characteristics and rDNA sequencing analysis of the fungus and confirmed its pathogenicity according to Koch’s postulates. The results of morphological examinations, pathogenicity tests, 5.8S rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the five nuclear protein-coding genes G3PDH, HSP60, RPB2, MS547 and TUB revealed that the causal agent of postharvest gray-mold rot on sweet persimmon fruit in Korea was Botrytis cinerea.     

Occurrence and characterisation of <Emphasis Type="Italic">Rhizoctonia</Emphasis> species causing diseases of ornamental plants in Italy

During surveys conducted in 2010–2012 Rhizoctonia symptoms were observed on 30 ornamental species in different nurseries located in eastern Sicily (Southern Italy). Eighty-eight isolates of Rhizoctonia spp. were obtained from symptomatic leaves, roots and stems. Fifty-six of the isolates were binucleate and 32 were multinucleate Rhizoctonia. Characterisation of anastomosis groups (AGs) was performed using morphological characteristics and sequence analysis of the internal transcribed spacer of ribosomal DNA ( rDNA-ITS) region. Most isolates collected were Rhizoctonia solani AG-4 HG-I (35.2% of all isolates) and one isolate was AG-2-2 IIIB. The binucleate isolates belonged to AG-R (27.3%), AG-A (21.6%), AG-G (12.5%), AG-V (1.1%) and AG-Fb (1.1%). The pathogenicity of 38 representative isolates collected from each host was tested on seedlings or cuttings grown in a growth chamber. All R. solani AG-4 HG-I isolates, most of the binucleate AG-R, AG-A and AG-G and AG-V were pathogenic and reproduced symptoms identical to that observed in nurseries, while binucleate AG-Fb and R. solani AG-2-2 IIIB isolates were nonpathogenic. This is the first report of the occurrence of Rhizoctonia species on some ornamental plants and the first report of binucleate Rhizoctonia AG-R and AG-V in Europe.     

Biochemical,physiological, and molecular characterization of <Emphasis Type="Italic">Dickeya dianthicola</Emphasis> (formerly named <Emphasis Type="Italic"> Erwinia chrysanthemi</Emphasis>) causing potato blackleg disease in Japan

The taxonomic assignment of Japanese potato blackleg isolates of Dickeya spp. has not been confirmed after the changes in their former name, Erwinia chrysanthemi. Therefore, we investigated and identified 23 representative isolates of Dickeya spp. from symptomatic stems of potatoes in Japan, with biochemical tests and phylogenetic sequence analysis using recA, dnaX, rpoD, gyrB, and 16S rDNA sequences. Results of our biochemical tests showed that all isolates can be assigned to phenon 5 and biovar 1, which are associated with D. dianthicola. Based on the recA, dnaX, rpoD, gyrB, and 16S rDNA sequences, all isolates are in the same clade with D. dianthicola and were clearly distinguished from D. chrysanthemi, D. dadantii, D. dadantii subsp. dieffenbachiae, D. solani, D. zeae, and D. paradisiaca. Therefore, we conclude that Dickeya spp. isolated from potatoes with blackleg symptoms in Japan are D. dianthicola.