Survey and characterization of potyviruses and their strains of Allium species

Nearly 5700 plants of 14 cultivated and 8 wildAllium species and varieties from the Netherlands and other parts of the world, were tested for infection with aphid-borne potyviruses by ELISA, electron microscope decoration tests and/or inoculation onto test plants. This resulted in the detection of two known viruses, viz. leek yellow stripe virus (LYSV) and onion yellow dwarf virus (OYDV), and the discovery and characterization of two new viruses, viz. shallot yellow stripe virus (SYSV) and Welsh onion yellow stripe virus (WoYSV), and of six strains of these viruses. ‘Garlic mosaic’, ‘barlic yellow streak’, ‘onion mosaic’, ‘shallot mosaic’, ‘shallot X’, and ‘shallot yellows’ viruses, incompletely described in the literature, are now reidentified as well-known viruses or as strains or mixtures of such viruses. ‘Garlic yellow stripe virus’ is also a complex containing a potyvirus possibly differing from the viruses found in this survey. The symptoms of the potyviruses studied varied widely and ranged from mild to severe chlorotic to yellow striping of leaves, and they are of little diagnostic importance.LYSV was found in vegetatively propagated pearl onion (A. ampeloprasum var.sectivum) from Europe and Asia. It has decreased in leek crops (A. ampeloprasum var.porrum) in the Netherlands since the 1970, apparently due to resistance in new cultivars. OYDV was common in onion (A. cepa var.cepa) from the former USSR and North Africa, and in European cultivars of shallot (A. cepa var.ascalonicum), with the exception of the highly resistant ‘Santé’, but was not detected during this survey in Asian shallot. European samples of ever-ready onion (A. cepa var.perutile), multiplier onion (A. cepa var.aggregatum) and tree onion (A. cepa var.viviparum) contained OYDV. It was also found in sand leek (A. scorodoprasum) from european gene collections. A strain of OYDV from onion and shallot in Morocco and Spain was virulent on onion and shallot cultivars resistant to common OYDV, as reported early for a similar isolate in the USA.Asian shallot appeared generally infected with the new SYSV, similar to OYDV in host range and symptoms but serologically distinct. It was not detected in onion and shallot from Europe or North Africa. A virulent strain of this virus caused striping in sap-inoculated garlic (A. sativum) and Formosan lily (Lilium formosanum). The new WoYSV, infecting Welsh onion in Indonesia and Japan, was earlier described in Japan as OYDV from rakkyo and Welsh onion. It appeared serologically closely related to SYSV and distantly to OYDV, but differed in its host range.Host-specific strains of LYSV and OYDV were detected in garlic, wild garlic (A. longicuspis), an unidentifiedAllium species (suffix-G), and great-headed garlic (A. ampeloprasum var.holmense) (suffix-GhG)., LYSV-G and OYDV-G infected on average 45% and 73%, respectively, of the garlic samples of worldwide origin. Symptoms of isolates of both strains varied in severity, implying the necessity of serological tests for disease diagnosis and health certification. LYSV-GhG was the cause of yellow striping in 93% of the great-headed garlic plants tested, mainly from the Mediterranean area. One sample was also infected with OYDV-GhG.Many samples from vegetatively propagated crops grown from non-certified planting stock contained a few plants free of potyviruses, implying the possibility to obtain healthy (and possibly resistant) selections of such cultivars avoiding meristem-tip culture. Cross-protection of garlic sets by a mild potyvirus isolate seems to be an alternative to the use of vulnerable virus-free sets.Generally, viruses and virus strains could not be transmitted to anyAllium species other than their natural host, except to the highly susceptible crow garlic (A. vineale). This species, and other predominantly vegetatively propagating wildAllium spp. (field garlic,A. oleraceum; ramsons,A. ursinum; sand leek), were found not to be reservoirs of viruses that might infectAllium crops in the netherlands. Streaking in vegetatively propagated wild leeks (A. ampeloprasum and closely related species) originating from the Mediterranean area and Asia was due to an undescribed miteborne virus. The survey confirmed that spread of potyviruses inAllium crops in the Netherlands is from planting sets, and from a neighbouring crop only if of the same species.     

Survey for viruses infecting onion, garlic and leek crops in Iran

Surveys to identify virus diseases affecting garlic ( Allium sativum ), onion ( Allium cepa ) and Persian leek ( Allium ampeloprasum var. persicum ) were conducted from 1999 to 2002. Surveys covered different regions of Iran (Tehran [different vegetable markets, farmer fields and cultivation areas], Noushahr, Chalous, Roudbar, Sari, Hamadan, Touyserkan, Ghazvin and Jiroft). A total of 2045 (1285 garlic, 525 onion and 230 leek) samples showing symptoms of virus infection were collected and tested by ELISA; and in some cases tests were also confirmed by immunoelectron microscopy (IEM) for the presence of Allium viruses. ELISA results showed that the following viruses were detected: Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV) (genus Potyvirus , family Potyviridae ), Garlic common latent virus (GarCLV), Shallot latent virus (SLV) (genus Carlavirus ), Garlic virus D (GarV-D), Garlic virus B (GarV-B) and Garlic virus C type (GarV-C) (genus Allexivirus ). None of the samples reacted with antibodies to Shallot yellow stripe virus (SYSV) genus Potyvirus , family Potyviridae ), Shallot virus X (ShVX) and Garlic virus A (GarV-A, genus Allexivirus ). GarCLV, SLV, GarV-D, GarV-B and GarV-C are reported for the first time from Allium crops in Iran.     

Virulence structure of the <Emphasis Type="Italic">Blumeria graminis</Emphasis> DC.f. sp. <Emphasis Type="Italic">avenae</Emphasis> populations occurring in Poland across 2010–2013

The aim of the present study was to determine the virulence structure of powdery mildew of oats (Blumeria graminis DC.f. sp. avena) in Poland in the years 2010–2013. For this purpose, powdery mildew isolates were collected from three experimental stations in Poland. To assess the virulence of the isolates, eight oat varieties with different responses to the pathogen were used. The results showed that a significant proportion of powdery mildew isolates found in Poland overcame the resistance genes of varieties Bruno (Pm6), Jumbo (Pm1) and Mostyn (Pm3). In contrast, lines Av1860 (Pm4), Am27 (Pm5) and Cc3678 (Pm2) were completely resistant to all pathogen isolates involved in the experiment. Changes constantly occurring in the powdery mildew population perfectly reflect diversity indexes, which were the smallest in the first year of observation, where in the following years these parameters were significantly higher. It is worth noting that the presence of powdery mildew is seasonal and local, which is reflected in the prevalence of the disease in a defined area of the country.     

Genetic variability within <Emphasis Type="Italic">Septoria carvi</Emphasis> Syd.- a pathogen of caraway <Emphasis Type="Italic">Carum carvi</Emphasis> L

Genetic variability within Septoria carvi isolates obtained from various organs of caraway cultivated in south-eastern and central Poland was studied using the RAPD-PCR technique. The tests were performed using randomly selected primers. The DNA profiles obtained using four primers proved useful in determining genetic variability among the genotypes of Septoria carvi isolates. The present study characterized the differences in the nucleotide sequence within the internal transcribed spacer region of rDNA (ITS1, 5.8S, ITS2) of selected S. carvi isolates and reference strains of Septoria spp. Moreover, eight isolates were sequenced for three loci: actin, calmodulin and translation elongation factor 1-alpha, and the obtained sequences were compared with the sequences of Septoria reference strains affecting other plants of the family Apiaceae. Phylogenetic analysis showed distinct differences of the tested isolates, which allowed to treat them Septoria carvi species affecting the above-ground organs of caraway Carum carvi L. This study is the first report on the genetic characteristics of the species S. carvi.     

Pathogenicity and Fungicide Sensitivity of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">R. cerealis</Emphasis> Isolates

The Rhizoctonia solani species consists of multinucleate isolates that belong to anastomosis groups AG1–AG3 and differ in virulence and host affinity. R. cerealis is a binucleate species of anastomosis group AG-D which causes sharp eyespot, a common plant disease in Poland. Rhizoctonia spp. is a ubiquitous soil pathogen that poses a significant threat for global crop production due to the absence of effective crop protection products. The aim of this study was to determine the virulence of R. solani and R. cerealis isolates towards Beta vulgaris, Zea mays, Triticum spelta and T. aestivum seedlings, to confirm the presence of endopolygalacturonase genes pg1 and pg5 in the genomes of the tested isolates and to evaluate the tested isolates’ sensitivity to triazole, strobilurin, imidazole and carboxamide fungicides. All tested isolates infected B. vulgaris seedlings. but none of them were virulent against Z. mays plants. R. solani isolates AG4 PL and AG2-2IIIB PL were characterized by the highest virulence (average infestation score of 2.37 and 2.53 points on a scale of 0–3 points) against sugar beet seedlings. The prevalence of infections caused by most of the analysed isolates (in particular R. solani AG4 J—11.8, and R. cerealis RC2—0.78) was higher in spelt than in bread wheat. The virulence of the analysed isolates was not correlated with the presence of pg1 and pg5 genes. The efficacy of the tested fungicides in controlling Rhizoctonia spp. infections was estimated at 100% (propiconazole + cyproconazole), 98.8% (penthiopyrad), 95.4% (tebuconazole) and 78.3% (azoxystrobin).     

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.     

In vitro and field efficacy of three fungicides against <Emphasis Type="Italic">Fusarium</Emphasis> bulb rot of garlic

Fusarium proliferatum has been identified as the main causal agent of bulb rot of garlic (Allium sativum L.). This disease occurs after the drying process and can rot almost 30 % of the bulbs. Few studies are available regarding the effectiveness of chemical treatments to reduce F. proliferatum incidence in garlic. The efficacy of three commercial fungicides of different chemical groups to reduce seven strains of F. proliferatum mycelial growth was tested in vitro. These three fungicides were also evaluated by foliar spreading of aqueous suspension in a field crop. Fluopyram 20 % + tebuconazole 20 % and tebuconazole 50 % + trifloxystrobin 50 % were highly effective at reducing mycelial growth in F. proliferatum with EC₅₀ values <2 ppm. In general, the effectiveness of the fungicides was enhanced with increasing dosage. Our results indicate that the fungicides evaluated in this study may lead to a risk of resistance appearing in F. proliferatum at low concentrations and this risk is maintained at higher doses for the fungicide dimethomorph 7.2 % + pyraclostrobin 4 %. Although several of the fungicides affected in vitro mycelial growth of F. proliferatum, as a part of an strategy to measure the efficacy of resistance management it is necessary to monitor the ongoing efficacy of fungicides under commercial conditions. All fungicidal treatments tested in field application failed to control garlic bulb rot during storage.     

Characterization of <Emphasis Type="Italic">Dickeya</Emphasis> and <Emphasis Type="Italic">Pectobacterium</Emphasis> strains obtained from diseased potato plants in different climatic conditions of Norway and Poland

Soft rot and blackleg of potato caused by pectinolytic bacteria lead to severe economic losses in potato production worldwide. To investigate the species composition of bacteria causing soft rot and black leg of potato in Norway and Poland, bacteria were isolated from potato tubers and stems. Forty-one Norwegian strains and 42 Polish strains that formed cavities on pectate medium were selected for potato tuber maceration assays and sequencing of three housekeeping genes (dnaX, icdA and mdh) for species identification and phylogenetic analysis. The distribution of the species causing soft rot and blackleg in Norway and Poland differed: we have demonstrated that mainly P. atrosepticum and P. c. subsp. carotovorum are the causal agents of soft rot and blackleg of potatoes in Norway, while P. wasabiae was identified as one of the most important soft rot pathogens in Poland. In contrast to the other European countries, D. solani seem not to be a major pathogen of potato in Norway and Poland. The Norwegian and Polish P. c. subsp. carotovorum and P. wasabiae strains did not cluster with type strains of the respective species in the phylogenetic analysis, which underlines the taxonomic complexity of the genus Pectobacterium. No correlation between the country of origin and clustering of the strains was observed. All strains tested in this study were able to macerate potato tissue. The ability to macerate potato tissue was significantly greater for the P. c. subsp. carotovorum and Dickeya spp., compared to P. atrosepticum and P. wasabiae.     

Identification and characterization of <Emphasis Type="Italic">Diaporthe vaccinii</Emphasis> Shear causing upright dieback and viscid rot of cranberry in Poland

During the summers of 2013–2014, symptoms similar to viscid rot and upright disease were observed on cranberries (Vaccinium macrocarpon) on one plantation in central Poland. The associated fungi were isolated from symptomatic plant tissue. On the basis of morphological and cultural characteristics and the ability of isolated fungi to elicit viscid rot symptoms on cranberry fruits, they were classified as the genus Diaporthe. Further analysis of ITS sequence data allowed for the classification of the newly obtained isolates as D. vaccinii. Additional analysis of genetic diversity using five RAPD and eight ISSR primers constituted additional confirmation of genetic distance existing between closely related D. vaccinii and D. eres species, enabling their differentiation.     

Resistance assessment and biochemical responses of sugar beet lines against <Emphasis Type="Italic"> Pythium aphanidermatum,</Emphasis> causing root rot

Wood-staining fungi, moulds and decay fungi colonize freshly cut wood. Of these, only ophiostomatoid fungi are considered major agents of sapstain on logs and freshly sawn lumber because they can cause permanent staining and discoloration. Sapstain in pine pallets significantly reduces their market value and use in the food industry. The aim of this study was to identify the sapstaining fungi that colonize Scots pine wood used in pallet production. In addition, we evaluated the growth and stain intensity of six isolates of ophiostomatoid fungi on freshly cut Scots pine billets. Fungi were isolated from samples of Scots pine and identified based on morphology and DNA sequence comparisons for three gene regions (ITS, β-tubulin, TEF-1α). A total of 1259 isolates representing 31 fungal species were obtained from the pine samples in Poland during July and September. The isolates represented different ecological and taxonomical groups and belonged to the categories of staining fungi, decay fungi and surface moulds. The most frequently isolated fungi were ophiostomatoid fungi (14 species including an unknown Leptographium sp. and Ophiostoma sp.) and moulds (mainly Trichoderma and Mucor spp.). Of the ophiostomatoid species, Ophiostoma floccosum, O. piceae, O. piliferum and Endoconidiophora pinicola were the predominant species. The results of pine billet inoculation showed that among the six fungal species tested, E. pinicola exhibited the fastest growth in all three directions on the billets. Ophiostoma minus and O. piliferum displayed moderate growth rates in pine billets, while O. floccosum, O. piceae, and Leptographium lundbergii grew very slowly, especially in the tangential and radial directions. The information provided in this paper will help develop more effective control strategies for sapstain prevention in Scots pine.     

Haplotypes of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter solanacearum’ identified in Umbeliferous crops in Spain

‘Candidatus Liberibacter solanacearum’ is a phloem-limited Gram-negative bacterium that causes serious damage to different crops of the botanical families Solanaceae and Apiaceae. Five haplotypes have been described: LsoA and LsoB are present in solanaceous crops in America and vectored by the tomato/potato psyllid Bactericera cockerelli; LsoC affects carrots from Northern and Central Europe, and is transmitted by the carrot psyllid Trioza apicalis; haplotypes LsoD and LsoE are present in Southern Europe and Morocco in carrot and celery, and are associated with the psyllid Bactericera trigonica. Thirty-four ‘Ca. L. solanacearum’ isolates were collected in six different regions of Spain from distinct Apiaceae hosts (carrot, celery, parsley and parsnip) in eight consecutive years and were analysed. Their haplotypes were determined by a sequence analysis of 16S ribosomal RNA, the 16S–26S ribosomal RNA intergenic spacer, and the 23S ribosomal RNA and rplJ and rplL genes. Both haplotypes LsoD and LsoE were found across Spain, and no host specificity appeared between these two haplotypes. This is the first report of ‘Ca. L. solanacearum’ associated with parsley and parsnip.     

Efficacy of a bacterial preparation of <Emphasis Type="Italic">Aneurinibacillus migulanus</Emphasis> against downy mildew of cucumber (<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis>)

The reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant roots and distributed in regions with Mediterranean, subtropical and tropical climates. In this study, we provide morphological and molecular characterisation of three out of 11 valid species of the genus Rotylenchulus: R. macrodoratus, R. macrosoma, and R. reniformis from Greece (Crete), Italy and Spain. The overall prevalence of reniform nematodes in wild and cultivated olives in Greece, Italy, and Spain was 11.5%, 19.0% and 0.6%, respectively. In Greece, R. macrodoratus and R. macrosoma were detected in cultivated olive with a prevalence of 8.2% and 6.2%, respectively, but none of them were found in wild olive. This is the first report of R. macrosoma in Greece. Only one reniform nematode species was detected in olive from Italy and Spain, viz. R. macrodoratus and R. macrosoma, respectively. The parasitism of R. macrosoma on hazelnut in northern Spain was also confirmed for the first time. This study demonstrates that R. macrodoratus and R. macrosoma have two distinct rRNA gene types in their genomes, specifically the two types of D2-D3 for R. macrosoma and R. macrodoratus, the two types of ITS for R. macrodoratus and the testing of the ITS variability in other R. macrosoma populations in different countries. Rotylenchulus macrosoma from Greece and Spain showed differences in nucleotide sequences in the ITS region and D2-D3 of 28S rRNA gene.     

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.     

First report of <Emphasis Type="Italic">Rhizoctonia</Emphasis> disease of lily caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-11 in Japan

Blight on leaves, stems and bulbs of lilies grown in a greenhouse were found in Hokkaido, Japan, in 2012. Two isolates obtained from the lesions were identified as Rhizoctonia solani anastomosis group (AG)-11 based on morphology and molecular analysis. Original symptoms were reproduced after artificial inoculation with the isolates. Except for R. solani AG-2-1 and AG-4 HG-I, none of the AGs have been reported as pathogens causing lily Rhizoctonia disease in Japan; therefore, we propose adding AG-11 as a pathogen of the disease. More importantly, we report the first appearance of crop disease caused by AG-11 in Japan.     

Molecular characterization and pathogenicity of <Emphasis Type="Italic">Alternaria</Emphasis> species on wheat and date palms in Oman

This study was conducted to investigate the Alternaria species associated with leaf spot of date palm and wheat in Oman. Out of 98 date palm leaf samples and 146 wheat leaf samples, Alternaria was isolated from 27 and 23% of the samples developing leaf spot symptoms, respectively. Identification of Alternaria isolates using sequences of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA), glyceraldehyde-3-phosphate dehydrogenase (GPDH), translation elongation factor (TEF) and RNA polymerase II subunit (RPB2) genes, showed that the isolates belong to seven Alternaria species or species complexes. A. burnsii - A. tomato and A. arborescens species complexes (58 and 4%, respectively) and A. alternata (38%) were the species recovered from the symptomatic date palm leaves. A. alternata (67%), A. burnsii - A. tomato species complex (15%), A. jacinthicola (3%), A. ventricosa (3%), A. slovaca (6%) and Alternaria caespitosa (6%) were isolated from wheat. Pathogenicity test showed that tested isolates of A. alternata (DPM19, WDK12), A. burnsii - A. tomato species complex (DPM31), A. jacinthicola (WBR4) and A. slovaca (WDK9, WDK7) were pathogenic on date palm, while A. alternata (DPM19, WDK12), A. burnsii - A. tomato species complex (DPM31, WDK11) and A. slovaca (WDK9, WDK7) were pathogenic on wheat. This is the first report of date palm and wheat as new hosts for A. burnsii - A. tomato species complex and the first reports of A. burnsii - A. tomato species complex, A. caespitosa A. slovaca, and A. ventricosa in Oman. The study shows that several species of Alternaria are associated with leaf spot in date palm and wheat in Oman, with some isolates having the ability to cause infection in both hosts.     

<Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">alliifistulosi</Emphasis> pv. nov., the causal agent of bacterial leaf spot of onions

In 1972, bacterial leaf spot of onion (BLSO) was first recorded in Japan by Goto. The pathogen was considered as a pathovar of Pseudomonas syringae specifically causing disease on onion and Welsh onion, but it has not been taxonomically investigated in detail. In 2012 and 2014, a disease suspected as BLSO re-emerged on onion in Shizuoka and Hyogo Prefectures, Japan, respectively. A pathogenic bacterium isolated from the infected onions was thought to be the BLSO agent after preliminary examinations. Strains isolated from BLSO in 1969, 1986, 1987, 2012 and 2014 were characterized and compared with the causal agent of bacterial blight of leek (P. syringae pv. porri), which causes similar symptoms on Allium plants. The result of rep-PCR distinguished the BLSO agent from P. syringae pv. porri. Multilocus sequence analysis on housekeeping genes and hrp genes encoding the type-III secretion system revealed that the strains of the BLSO agent clustered independently of P. syringae pv. porri. The BLSO agent and P. syringae pv. porri also differed in utilization of erythritol, dl-homoserine, glutaric acid and other bacteriological characteristics and caused different reactions on onion, Welsh onions, chives, shallot, rakkyo, leek, garlic and Chinese chive. Thus, the BLSO agent clearly differs from P. syringae pv. porri and is considered to be a new pathovar of P. syringae. The name P. syringae pv. alliifistulosi is proposed with pathotype strain ICMP3414.     

Characterization of <Emphasis Type="Italic">Trichoderma</Emphasis> species isolated in Ecuador and their antagonistic activities against phytopathogenic fungi from Ecuador and Japan

Native Trichoderma spp. were isolated from agricultural fields in several regions of Ecuador. These isolates were characterized via morphological observation as well as molecular phylogenetic analysis based on DNA sequences of the rDNA internal transcribed spacer region, elongation factor-1α gene and RNA polymerase subunit II gene. Fifteen native Trichoderma spp. were identified as T. harzianum, T. asperellum, T. virens and T. reesei. Some of these strains showed strong antagonistic activities against several important pathogens in Ecuador, such as Fusarium oxysporum f. sp. cubense (Panama disease) and Mycosphaerella fijiensis (black Sigatoka) on banana, as well as Moniliophthora roreri (frosty pod rot) and Moniliophthora perniciosa (witches’ broom disease) on cacao. The isolates also showed inhibitory effects on in vitro colony growth tests against Japanese isolates of Fusarium oxysporum f. sp. lycopersici, Alternaria alternata and Rosellinia necatrix. The native Trichoderma strains characterized here are potential biocontrol agents against important pathogens of banana and cacao in Ecuador.     

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.