Assessment of recent outbreaks of <Emphasis Type="Italic">Dickeya</Emphasis> sp. (syn. <Emphasis Type="Italic">Erwinia chrysanthemi</Emphasis>) slow wilt in potato crops in Israel

Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.     

Evaluation of biocontrol preparations and plant extracts for the control of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on potato leaves

The potential of biocontrol products and plant extracts for control of late blight on potato plants, caused by Phytophthora infestans was evaluated in detached leaf assays and on potted plants. Based on an initial screening of 22 preparations and plant extracts, the 10 most active treatments were selected for further investigation. In the detached leaf assays the commercial preparations Elot-Vis, Serenade and Trichodex, and plant extracts of Rheum rhabarbarum and Solidago canadensis showed a significant effect on the level of infestation by P. infestans. However, none of the treatments was as effective as copper. In the case of Serenade, the metabolites produced by its active micro-organism, Bacillus subtilis, were demonstrated to be the effective component of the formulation, and not the micro-organism itself. In order to take curative and protective modes of action into account, the test substances were applied 24 h before, or 90 min after inoculation with P. infestans. Generally, better effects were obtained when the applications were made 24 h before inoculation. For defining the optimum time of application, potted plants were treated 72 or 24 h before, and 1 and 24 h after inoculation with P. infestans. In these tests, Trichodex showed no activity, while Elot-Vis gave best results when applied 1 day before inoculation. Serenade and the extracts of R. rhabarbarum and S. canadensis (all at 5% concentration) however, were effective when applied up to 3 days before and just after inoculation with P. infestans. The results of the experiments on potted plants indicated direct effects on the pathogen for all agents except the extract of S. canadensis, but other mode of actions, e.g. induced resistance, could not be ruled out. None of the treatments had a curative effect.     

Note: Tachinid parasitoids of<Emphasis Type="Italic">Ancyrosoma leucogrammes</Emphasis> and notes on parasitization rates of<Emphasis Type="Italic">Clytiomya dupuisi</Emphasis>

The study deals with the tachinid parasitoids ofAncyrosoma leucogrammes (Gmelin) (Heteroptera: Pentatomidae):Clytiomya dupuisi Kugler,Clytiomya sola (Rondani) andGymnosoma clavatum (Rohdendorf) (Diptera: Tachinidae). All species are recorded for the first time as parasitoids ofA. leucogrammes, andC. dupuisi was reared for the first time from a host. The parasitization rates ofC. dupuisi on adults ofA. leucogrammes varied from 7% to 9% between 1994 and 1999.C. sola andG. clavatum were reared in only small numbers. http://www.phytoparasitica.org posting July 10, 2003.     

Structural conservation of the <Emphasis Type="Italic">hrp</Emphasis> gene cluster in <Emphasis Type="Italic">Xanthomons oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis>

The clustered hrp genes encoding the type III secretion system in the Japanese strains MAFF301237 and MAFF311018 of Xanthomonas oryzae pv. oryzae were sequenced and compared. The strains differ in their pathogenicity, location, and year of isolation. A 30-kbp sequence comprising 29 open reading frames (ORFs) was identical in its structural arrangement in both strains but differed from X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines in certain genes located between the hpaB-hrpF interspace region. The DNA sequence and the putative amino acid sequence in each ORF was also identical in both X. oryzae pv. oryzae strains as were the PIP boxes and the relative sequences. These facts clearly showed that the structure of the hrp gene cluster in X. oryzae pv. oryzae is unique.     

Use of a diagnostic medium for<Emphasis Type="Italic">in situ</Emphasis> determination of the response of<Emphasis Type="Italic">Erwinia amylovora</Emphasis> strains to bactericides

Erwinia amylovora, the causal agent of fire blight, is managed by application of bactericides to protect fruit tree blossoms from infection. Monitoring the response ofE. amylovora strains to bactericides is crucial for adequate disease management. The coliform agar medium produced by Merck was recently reported as an effective tool for rapid diagnosis ofE. amylovora (RD-medium). The objective of the present study was to examine the possibility of using the RD-medium forin situ determination of the response ofE. amylovora strains to oxolinic acid and streptomycin. The phenotypic response of 48E. amylovora strains isolated in 2002 to both bactericides was determined with the RD-medium and, for comparison, by a routine laboratory test. The results of 45 samples (93.7%) were in agreement with the findings of the routine laboratory test. Aχ ² test rejected the null hypothesis that the phenotypic characteristics as determined by the two respective methods differed significantly (P=0.389). Thein situ test was implemented on a national scale in 2003 and the results were in agreement with those obtained in laboratory tests, which suggests that this medium can be usedin situ for monitoring the appearance of resistance inE. amylovora populations. http://www.phytoparasitica.org posting Feb. 11, 2004.     

Inhibition of germination of<Emphasis Type="Italic">Orobanche ramosa</Emphasis> seeds by<Emphasis Type="Italic">Fusarium</Emphasis> toxins

Eighteen toxins produced byFusarium species were tested at different concentrations onOrobanche ramosa seeds to evaluate their effectiveness in inhibiting germination. Many of the toxins were active at the highest concentration used. Seven of them,viz. fusarenon X, nivalenol, deoxynivalenol, T-2 toxin, HT-2 toxin, diacetoxyscirpenol and neosolaniol, were highly active at 10μM, causing 100% inhibition of germination. Many of them were still active when assayed at a concentration ten times lower, with T-2, HT-2, nivalenol, neosolaniol and diacetoxyscirpenol still able to cause total inhibition; the last mentioned was very active also at 0.1μM, causing more than 90% inhibition. The results show that the use of toxic secondary metabolites could represent a useful alternative strategy in the management of parasitic weeds, by interfering with the induced germination process, and that fungal culture extracts could be an interesting source of new compounds acting as natural and original herbicides. http://www.phytoparasitica.org posting Sept. 18, 2002.     

Note: Evaluation of antibacterial activity of essential oil of<Emphasis Type="Italic">Rosa damascena</Emphasis> on<Emphasis Type="Italic">Erwinia amylovora</Emphasis>

Significant antibacterial activity was observed in the essential oil (E.O.) ofRosa damascena Mill. and the Minimum Bactericidal Concentration (MBC) of the E.O. was determined as 1386.5 μg ml⁻¹ forErwinia amylovora, the causal agent of fire blight disease. Ooze formation on immature pears and lesion formation in artificially inoculated shoots were completely (100%) prevented by the essential oil ofR. damascena (1500 μg ml⁻¹), essential oil ofThymbra spicata var.spicata (500 μg ml⁻¹) and streptomycin (100 μg ml⁻¹). Copper oxychloride plus maneb significantly reduced ooze formation and lesion formation, but the control was less than that obtained with the essential oils or streptomycin. The essential oil ofR. damascena may be a useful natural bactericide for the control of the fire blight pathogen,E. amylovora. http://www.phytoparasitica.org posting July 14, 2004.     

Insecticidal activity of crude seed extracts of<Emphasis Type="Italic">Annona</Emphasis> spp.,<Emphasis Type="Italic">Lansium domesticum</Emphasis> and<Emphasis Type="Italic">Sandoricum koetjape</Emphasis> against lepidopteran larvae

Crude ethanolic seed extracts ofAnnona muricata, A. squamosa (Annonaceae),Lansium domesticum andSandoricum koetjape (Meliaceae) collected from different locations and years in Maluku, Indonesia, were screened for inhibition of larval growth against the polyphagous lepidopteranSpodoptera litura (Noctuidae). Extracts ofA. squamosa were significantly more active (20-fold) than those ofA. muricata. A. squamosa collected from Namlea yielded the extracts with the greatest inhibitory activity. There were significant differences among locations for bothA. squamosa andA. muricata but not forL. domesticum andS. koetjape. Extracts ofA. squamosa, collected from Namlea, inhibited larval growth in a dose-dependent manner, with a dietary EC₅₀ (effective concentration to inhibit growth by 50% relative to controls) of 191.7 ppm fresh weight. Extracts ofA. squamosa collected from individual trees in Namlea also varied in growth inhibitory effect againstS. litura andTrichoplusia ni larvae. This species is a candidate for development of a botanical insecticide for local use in Indonesia. http://www.phytoparasitica.org posting Dec. 1, 2003.     

Oospores associated with tomato seed may lead to seedborne transmission of<Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Tomato fruits at the green mature stage were inoculated with a mixed sporangial suspension of A₁ and A₂ isolates ofPhytophthora infestans. Other fruits were inoculated with either A₁ or A₂ sporangia. Seeds were extracted from the blighted fruits and sown in soil or on agar media to test for the transmission of late blight to the emerging seedlings. Only 23 (0.09%) of approximately 25,000 seedlings developed symptoms. All blighted seedlings originated from fruits inoculated with mixed A₁ + A₂ sporangia. Isolates ofP. infestans recovered from the emerging blighted seedlings were seemingly of oosporic origin, as they differed phenotypically (mating type, virulence, sensitivity to metalaxyl) from the parent isolates used to inoculate the fruits. The results suggest that transmission ofP. infestans might occur by seeds extracted from fruits carrying oospores and less probably by seeds extracted from fruits having no oospores. http://www.phytoparasitica.org posting April 30, 2004.     

<Emphasis Type="Italic">Trichoderma harzianum</Emphasis> elicits defence response genes in roots of potato plantlets challenged by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Biological control of Rhizoctonia solani with Trichoderma harzianum has been demonstrated in several studies. However, none have reported the dynamics of expression of defence response genes. Here we investigated the expression of these genes in potato roots challenged by R. solani in the presence/absence of T. harzianum Rifai MUCL 29707. Analysis of gene expression revealed an induction of PR1 at 168 h post-inoculation (hpi) and PAL at 96 hpi in the plants inoculated with T. harzianum Rifai MUCL 29707, an induction of PR1, PR2 and PAL at 48 hpi in the plants inoculated with R. solani and an induction of Lox at 24 hpi and PR1, PR2, PAL and GST1 at 72 hpi in the plants inoculated with both organisms. These results suggest that in the presence of T. harzianum Rifai MUCL 29707, the expression of Lox and GST1 genes are primed in potato plantlets infected with R. solani at an early stage of infection. Mycothèque de l’Université catholique de Louvain of S. Cranenbrouck's affiliation is part of the Belgian Coordinated Collections of Micro-organisms (BCCM).     

Identification and pathogenicity of<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and binucleate<Emphasis Type="Italic">Rhizoctonia</Emphasis> anastomosis groups isolated from forage legumes in Erzurum,Turkey

Abstrast  Three-hundred-twenty-five isolates ofRhizoctonia (215R. solani and 110 binucleateRhizoctonia) were obtained from roots and crowns of alfalfa, sainfoin and common vetch grown in Erzurum, Turkey. The isolates were assigned to five anastomosis groups (AG) ofR. solani (AG-2-1, AG-3, AG-4, AG-5, and AG-10) and two anastomosis groups of binucleateRhizoctonia (AG-I and AG-K). In pathogenicity tests on alfalfa, sainfoin and common vetch, the highest disease severities were caused by isolates of AG-4 and AG-5. Isolates of AG-10 and AG-I were not pathogenic on the three tested forage legumes, whereas isolates of AG-K on alfalfa and sainfoin, and of AG-2-1 on sainfoin, were moderately virulent. Alfalfa isolate AG-3 was moderately virulent on sainfoin. This is the first report ofR. solani AG-3, AG-5, AG-10 and binucleateRhizoctonia AG-I on alfalfa. In addition, all theR. solani and binucleateRhizoctonia groups isolated from sainfoin and common vetch were recovered from these crops for the first time in Turkey. http://www.phytoparasitica.org posting Dec. 16, 2002.     

A new leaf blight disease of <Emphasis Type="Italic">Trifolium dasyurum</Emphasis> caused by <Emphasis Type="Italic">Botrytis fabae</Emphasis>

A new disease was observed on Trifolium dasyurum, with symptoms beginning as a halo spot and developing into a leaf blight. The causal organism was identified by microscopy and DNA sequence studies as Botrytis fabae. This strain of B. fabae was also demonstrated to cause disease on foliage of a range of pulse crops, including Vicia faba, Pisum sativum, and Lens culinaris. This study demonstrates the potential of this strain of B. fabae to not only pose a significant threat to T. dasyurum but also to pulses grown in rotation with T. dasyurum that are susceptible to this strain of B. fabae.     

Reaction of <Emphasis Type="Italic">Leersia</Emphasis> grasses to <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> collected from Japan and Asian countries

The present study was conducted to determine if there is specificity in the host-pathogen relationship between the isolates of Xanthomonas oryzae pv. oryzae, the causal bacterium for rice blight and Leersia grasses, the alternative weed hosts of the disease. Plants of three species of Leersia, namely, L. sayanuka, L. oryzoides and L. japonica, were collected from various parts of Japan and were inoculated with the X. oryzae pv. oryzae isolates obtained from various locations in Japan and from 11 Asian countries. Four L. sayanuka plants were found susceptible to all Race II isolates and some Race I isolates, but were resistant to all Race III isolates. Race III is known to have a wider range pathogenicity to rice cultivar groups compared with Race I and II. Although the reactions of two L. oryzoides plants to Race I and II isolates were similar to that of L. sayanuka, the L. oryzoides plant collected from Niigata Prefecture showed a susceptible reaction to some Race III isolates. On the other hand, L. japonica plants gave reactions different those of L. sayanuka and L. oryzoides, with two plants of L. japonica found to be resistant to all test isolates collected from Japan. The Asian isolates exhibited a wide host range against the international differential rice cultivars, but almost all of them were avirulent to Leersia plants. These results indicate that the relationship between the pathogenicity of the causal bacterium and the resistance of host plants is very complex, and suggest that pathogenic diversity of X. oryzae pv. oryzae might be related to the resistance of Leersia spp.     

Tachinidae parasitoids of<Emphasis Type="Italic">Traumatocampa ispartaensis</Emphasis> from Turkey

Tachinid parasitoids ofTraumatocampa ispartaensis Doğanlar & Avcı (Lepidoptera, Thaumetopoeidae), which was found to be a new species, were collected from the cedar forests around Isparta-Kapıdağ. The species found wereBlondelia nigripes (Fall.),Compsilura concinnata (Meig.),Pales processioneae (Ratz.),Phryxe caudata (Rond.),Exorista segregata (Rond.) andCarcelia iliaca (Ratz.). Within the six species of Tachinidae,B. nigripes was the most common one, parasitizing up to 4.6% ofT. ispartaensis pupae. http://www.phytoparasitica.org     

Infection of<Emphasis Type="Italic">Ceratitis capitata</Emphasis> by two species of the<Emphasis Type="Italic">Entomophthora muscae</Emphasis> species complex (Zygomycetes: Entomophthorales) in the field

Adult Mediterranean fruit flies (Ceratitis capitata), collected in the field, were infected with entomophthoralean fungi. The fungi sporulated poorly on the cadavers, and resting spores, rhizoids and cystidia were not observed. Measurements of conidia and nuclei and counts of nuclei per conidium from different specimens suggest that the causative agents wereEntomophthora muscae sensu stricto andEntomophthora schizophorae, species recently separated from theEntomophthora muscae species complex. This is the first report ofC. capitata as a host for entomopathogenic fungi. http://www.phytoparasitica.org posting Jan. 29, 2003.     

Cytological events in tomato leaves inoculated with conidia of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> and <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01

Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.     

Synergistic interaction between BABA and mancozeb in controlling<Emphasis Type="Italic">Phytophthora infestans</Emphasis> in potato and tomato and<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> in cucumber

Spray mixtures consisting of the plant activator BABA (DL-3-aminobutyric acid) and the protectant fungicide mancozeb were significantly more effective than BABA or mancozeb alone in controlling late blight (Phytophthora infestans) in potato and tomato and downy mildew (Pseudoperonospora cubensis) in cucumber. A mixture composed of 5 parts BABA and 1 part mancozeb (w/w, a.i.) exhibited a higher synergy factor than the 1+1 or the 1+5 (BABA + mancozeb) mixtures. No synergistic interaction was detected between BABA plus mancozeb in controlling sporangial or cystospore germination, nor mycelial growth ofP. infestans in vitro. The results showed enhanced effect of mancozeb in BABA-induced plants, suggesting, therefore, that lower dosages of this fungicide may be sufficient to control late blight or downy mildew under field conditions. http://www.phytoparasitica.org posting July 15, 2003.     

Fumigant toxicity of<Emphasis Type="Italic">Eucalyptus blakelyi</Emphasis> and<Emphasis Type="Italic">Melaleuca fulgens</Emphasis> essential oils and 1,8-cineole against different development stages of the rice weevil<Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Essential oils extracted fromEucalyptus blakelyi (1,8-cineole, 77.5%),Melaleuca fulgens (1,8-cineole, 56.9%) and 1,8-cineole were shown to have fumigant toxicity against different development stages ofSitophilus oryzae. The eggs ofS. oryzae were the most tolerant, followed by pupae, larvae and adults in that order.M. fulgens oil,E. blakelyi oil and 1,8-cineole at 100 μl per liter of air gave, respectively, LT₅₀ values of 16.2, 17.4 and 9.1 h for adults, 31.1, 19.3 and 17.5 h for larvae, 55.6, 75.2 and 39.7 h for pupae, and required >7 days for eggs. Only 1,8-cineole (200 μl ⁻¹ air) gave a significant egg kill by 7 days and the LT₉₅ was 134.5 h. 1,8-Cineole could be a useful new fumigant. http://www.phytoparasitica.org posting Oct. 3, 2004.     

Toxigenicity and pathogenicity of <Emphasis Type="Italic">Fusarium poae</Emphasis> and <Emphasis Type="Italic">Fusarium avenaceum</Emphasis> on wheat

In a field experiment between 2004 and 2006, 14 winter wheat varieties were inoculated with either a mixture of three isolates of F. poae or a mixture of three isolates of F. avenaceum. In a subsequent climate chamber experiment, the wheat variety Apogee was inoculated with individual single conidium isolates derived from the original poly conidium isolates used in the field. Disease symptoms on wheat heads were visually assessed, and the yield as well as the fungal incidence on harvested grains (field only) was determined. Furthermore, grains were analysed using LC-MS/MS to determine the content of Fusarium mycotoxins. In samples from field and climate chamber experiments, 60 to 4,860 μg kg⁻¹ nivalenol and 2,400 to 17,000 μg kg⁻¹ moniliformin were detected in grains infected with F. poae and F. avenaceum, respectively. Overall, isolate mixtures and individual isolates of F. avenaceum proved to be more pathogenic than those of F. poae, leading to a higher disease level, yield reductions up to 25%, and greater toxin contamination. For F. poae, all variables except for yield were strongly influenced by variety (field) and by isolate (climate chamber). For F. avenaceum, variety had a strong effect on all variables, but isolate effects on visual disease were not reflected in toxin production. Correlations between visual symptoms, fungal incidence, and toxin accumulation in grains are discussed.     

Dry mycelium of<Emphasis Type="Italic">Penicillium chrysogenum</Emphasis> protects cucumber and tomato plants against the root-knot nematode<Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Incorporation into soil of dry mycelium ofPenicillium chrysogenum, a waste product of the pharmacological industry, enhanced plant growth and reduced root galling caused by the root-knot nematodeMeloidogyne javanica in cucumber and tomato plants. Incorporation into sandy loam soil in pots of dry mycelium at a concentration of 0.25% (w/w) resulted in complete protection of cucumber plants from the nematode. The number of juveniles recovered from soils containing dry mycelium was greatly reduced even at a concentration of 0.1% (w/w). In microplot studies conducted at two sites in two seasons, with three or four doses, dry mycelium caused a dose-dependent reduction in root galling index (GI) and promotion of plant growth of cucumber and tomato plants. Inin vitro studies, the water extract of dry mycelium immobilized nematode juveniles and reduced the egg hatching rate, but these effects were partly reversible after a rinse in water. Soil-drenching of cucumber and tomato seedlings with water extract of dry mycelium did not reduce GI or number of root-invading juveniles. The results show that dry mycelium promotes plant growth and protects plants against nematode infection. Protection, however, does not operatevia induced resistance. http://www.phytoparasitica.org posting April 6, 2003.