Evaluation of Greek grapevine cultivars for resistance to <Emphasis Type="Italic">Phaeomoniella chlamydospora</Emphasis>

In the present study, four Greek (Agiorgitiko, Asyrtiko, Roditis and Xinomavro) and one international (Soultanina) grapevine cultivars (Vitis vinifera L.) were screened for their resistance to Phaeomoniella chlamydospora. Artificial inoculation was carried out by drilling a hole into the trunk and injecting a concentrated conidial suspension into the vessels. Disease reactions were evaluated in an 87-day assessment period, on the basis of external symptoms (disease incidence, disease severity and mortality) and by calculating the relative areas under disease progress curves (relative AUDPC). The extension of vascular browning as well as the isolation ratio along the inoculated vine trunks were also taken into account as additional parameters for evaluating resistance. The results indicated that the resistance of grapevine cultivars to P. chlamydospora varied significantly. ‘Agiorgitiko’ and ‘Soultanina’ were susceptible, whereas ‘Asyrtiko’ and ‘Xinomavro’ were resistant; ‘Roditis’ showed an intermediate level of resistance. Cultivars’ resistance was mostly distinguished in terms of the extension of vascular browning and pathogen isolation ratio. On the contrary, the disease incidence, final disease severity, mortality and relative AUDPC provided less distinctive efficiency in resistance evaluation. The robust methodology presented here could be useful in rapid evaluation experiments for future screening programs to search and recognize natural resistant sources within grapevine genotypes against P. chlamydospora.     

The chemotaxis regulator <Emphasis Type="Italic">pilG</Emphasis> of <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> is required for virulence in <Emphasis Type="Italic">Vitis vinifera</Emphasis> grapevines

Type IV pili of X. fastidiosa are regulated by pilG, a response regulator protein putatively involved in chemotaxis-like operon sensing stimuli through signal transduction pathways. To elucidate the roles of pilG in pathogenicity of X. fastidiosa, the pilG-deletion mutant XfΔpilG and complemented strain XfΔpilG-C were generated. While all strains had similar growth curves in vitro, XfΔpliG showed significant reduction in cell-matrix adherence and biofilm production compared with wild-type X. fastidiosa and XfΔpilG-C. The genes pilE, pilU, pilT, and pilS were down-regulated in XfΔpliG when compared with its complemented strain and wild-type X. fastidiosa. Finally, no Pierce’s disease symptoms were observed in grapevines inoculated with XfΔpilG, whereas grapevines inoculated with the wild-type X. fastidiosa and complemented strain of XfΔpilG-C developed typical Pierce’s Disease (PD) symptoms. The results indicate that pilG has a role in X. fastidiosa virulence in grapevines.     

Chitin elicitor receptor kinase 1 (CERK1) is required for the non-host defense response of <Emphasis Type="Italic">Arabidopsis</Emphasis> to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. Sp. <Emphasis Type="Italic">cubense</Emphasis>

Banana wilt disease is a typical vascular disease caused by the fungal pathogen Fusarium oxysporum f. sp. cubense 4 (Foc 4). Pattern recognition receptors in the plant cell membrane can recognize pathogen-associated molecular patterns (PAMPs) to activate multi-layer defense responses, including defense gene expression, stomatal closure, reactive oxygen species (ROS) burst and callose deposition, to limit pathogen growth. In the present study, we found that chitin elicitor receptor kinase 1 (CERK1) was required for the non-host resistance of Arabidopsis thaliana to Foc B2 (a strain of Foc 4). The cerk1 mutant had weaker defense responses after Foc B2 treatment, including lower expression of PAMP- and salicylic acid-responsive genes, no stomatal closure, lower ROS level and less callose deposition, than that of the wild-type plant. Consistent with this, the cerk1 mutant plants exhibited higher susceptibility to non-host pathogen Foc B2. These results suggest the crucial importance of CERK1 in Foc B2-triggered non-host resistance.     

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.     

Conidial sporulation of <Emphasis Type="Italic">Stemphylium solani</Emphasis> under laboratory conditions and infectivity of the inoculum produced <Emphasis Type="Italic">in vitro</Emphasis>

Potato virus Y (PVY) is the type-species of the genus Potyvirus, family Potyviridae, being reported as a major tomato (Solanum lycopersicum L.) pathogen in several regions of the world. Pepper yellow mosaic virus (PepYMV) was originally described as a resistance-breaking Potato virus Y (PVY) isolate on Capsicum annuum L. cultivars, and afterwards it was also reported infecting tomatoes in Brazil. In the present work, a search for sources of resistance to both PepYMV and PVY was conducted in a collection of 119 accessions belonging to seven Solanum (section Lycopersicon) species. This germplasm was initially evaluated to PepYMV reaction by mechanical inoculation followed by symptom observations and ELISA. Potential PepYMV resistance sources were identified for the first time in S. habrochaites, S. peruvianum, S. corneliomuelleri, S. chilense, S. pimpinellifolium, and one accession derived from an interspecific cross (S. lycopersicum x S. peruvianum). A sub-group of 24 accessions with negative serology for PepYMV was also challenged with a PVY isolate, followed by serological and molecular detection with universal primers. Solanum habrochaites ‘L.03683’ and ‘L.03684’ were the only accessions found with stable resistance to both viruses. These results confirm S. habrochaites as the most important source of multiple resistance factor(s) to distinct Potyvirus species.     

Antagonistic effects of biofilm-forming bacterial strains co-inoculated with blood disease pathogenic strain, <Emphasis Type="Italic">Ralstonia syzygii</Emphasis> subspecies <Emphasis Type="Italic">celebesensis</Emphasis> in banana plants

A blood disease pathogenic strain, Ralstonia syzygii subspecies celebesensis was used to study the possible association of biofilm-forming bacteria with the development and severity of blood disease in banana plants. Therefore, the objective of this study was to determine the effects of mono-culture and co-culture inoculation of isolated biofilm-forming bacteria with the blood disease pathogen in banana pseudostems in glasshouse conditions. Putative biofilm-forming bacteria were isolated from an infected banana plant and were further identified using 16SrRNA sequencing. Four isolates, identified as Enterobacter hormaechei, Enterobacter cloacae, Kosakonia radicincitans and Klebsiella pneumoniae, were inoculated as a mono- and co-culture with R. syzygii subsp. celebesensis into 2 months old banana plants. The observation after the 8 weeks of post inoculation showed that plants which were co-inoculated with the pathogen and K. radicincitans, a biofilm-forming bacterium, were the most susceptible towards the infection. In contrast, plants under two treatments (which were co-inoculated with the pathogen and E. cloacae and the pathogen with E. hormaechei) were less susceptible towards the infection. This study revealed the antagonistic effects of two biofilm-forming strains which reduced the severity of infection caused by the pathogenic agent. Scanning electron micrographs of the cross section of plant rhizomes indicated the dissimilarity of adhesion and host colonization conditions of the pathogen in each infected plant from different treatments.     

AlgU contributes to the virulence of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">tomato</Emphasis> DC3000 by regulating production of the phytotoxin coronatine

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen to investigate the molecular basis of plant–pathogen interactions. The function of many potential virulence factors encoded in the Pst DC3000 genome and their modes of action are not fully understood. P. syringae is known to produce the exopolysaccharide alginate. Although AlgU, a sigma factor, is known to regulate the expression of genes such as algD related to alginate biosynthesis, the molecular mechanisms of AlgU in the virulence of Pst DC3000 is still unclear. To investigate the function of AlgU and alginate in plant–bacterial pathogen interactions, we generated ΔalgU and ΔalgD mutants. After inoculation with ΔalgU but not ΔalgD, host plants of Pst DC3000 including tomato and Arabidopsis had milder disease symptoms and reduced bacterial populations. Expression profiles of Pst DC3000 genes revealed that AlgU can regulate not only the expression of genes encoding alginate biosynthesis, but also the expression of genes related to type III effectors and the phytotoxin coronatine (COR). We also demonstrated that the ΔalgU mutant showed full virulence in the Arabidopsis fls2 efr1 double mutant, which is compromised in the recognition of PAMPs. Further, the application of COR was able to restore the phenotype of the ΔalgU mutant in the stomatal response. These results suggest that AlgU has an important role in the virulence of Pst DC3000 by regulating COR production.     

Genetic structure of <Emphasis Type="Italic">Pyrenophora teres</Emphasis> f. <Emphasis Type="Italic">teres</Emphasis> and <Emphasis Type="Italic">P. teres</Emphasis> f. <Emphasis Type="Italic">maculata</Emphasis> populations from western Canada

Infection by Pyrenophora teres f. teres (Ptt) or P. teres f. maculata (Ptm), the causal agents of the net and spot forms of net blotch of barley, respectively, can result in significant yield losses. The genetic structure of a collection of 128 Ptt and 92 Ptm isolates from the western Canadian provinces of Alberta (55 Ptt, 27 Ptm), Saskatchewan (58 Ptt, 46 Ptm) and Manitoba (15 Ptt, 19 Ptm) were analyzed by simple sequence repeat (SSR) marker analysis. Thirteen SSR loci were examined and found to be polymorphic within both Ptt and Ptm populations. In total, 110 distinct alleles were identified, with 19 of these shared between Ptt and Ptm, 75 specific to Ptt, and 16 specific to Ptm. Genotypic diversity was relatively high, with a clonal fraction of approximately 10 % within Ptt and Ptm populations. Significant genetic differentiation (PhiPT = 0.230, P = 0.001) was found among all populations; 77 % of genetic variation occurred within populations and 23 % between populations. Lower, but still significant genetic differentiation (PhiPT = 0.038, P = 0.001) was detected in Ptt, with 96 % of genetic variation occurring within populations. No significant genetic differentiation (PhiPT = 0.010, P = 0.177) was observed among Ptm populations. Isolates clustered in two distinct groups conforming to Ptt or Ptm, with no intermediate cluster. The high number of haplotypes observed, combined with an equal mating type ratio for both forms of the fungus, suggests that P. teres goes through regular cycles of sexual recombination in western Canada.     

Gene-based analysis of <Emphasis Type="Italic">Puccinia</Emphasis> species and development of PCR-based marker to detect <Emphasis Type="Italic">Puccinia striiformis</Emphasis> f. sp. <Emphasis Type="Italic">tritici</Emphasis> causing yellow rust of wheat

Stripe rust is considered as the current major rust disease affecting winter cereal production across the world. A quick, reliable PCR-based marker was developed here to detect, identify and rapidly monitor Puccinia striiformis f. sp. tritici (Pst) in wheat-growing areas. Three respective sets of primers, designed from β-tubulin, squalene monooxygenase and ketopantoate reductase genes selected from the full genome of Puccinia striiformis f. sp. tritici, amplified sequences of 239, 358 and 1518 bp, respectively, in Pst pathotypes. A fragment of 1518 bp unique to Pst pathotypes was amplified using primer set PstKeto F1_30/Pst KetoR1_1547 and distinguished the pathogen clearly from different Puccinia spp. and other fungal pathogens. The detection limit of the marker (KetoPstRA₁₅₀₀, accession no. KU240073) by conventional PCR assay was 10 pg. This marker could detect the pathogen in the host before symptom expression. The sensitivity and utility of the marker were further enhanced in a qPCR-based assay that was developed with a newly designed primer set PstKeto F1_1246/Pst KetoR1_1547, which amplified a product of 302 bp and detected as little as 10 fg of DNA. This PCR/qPCR based marker is suitable for studying cultivar resistance, which requires accurate quantification of the pathogen in diseased host tissue.     

Important genetic diversity of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma solani’ related strains associated with bois noir grapevine yellows and planthoppers in Azerbaijan

Bois noir (BN) is an important grapevine yellows endemic to the Euro-Mediterranean basin caused by ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’), a non culturable plant pathogenic Mollicute. Bois noir symptoms could be associated with ‘Ca. P. solani’ in two Azerbaijanian vineyards where disease incidence and severity were recorded for five local Vitis vinifera cultivars. In order to gain insight into the epidemiology of Bois noir in Azerbaijan, ‘Ca. P. solani’ isolates infecting plants were characterized by multi-locus sequence analysis and their secY and stamp gene sequences compared to that of the strains detected in other plants and in local Cixiidae planthoppers. Genotypes were determined for two non-ribosomal house-keeping genes, namely tuf and secY, as well as two variable markers namely Stamp and mleP1 genes, that respectively encode the antigenic membrane protein AMP and a 2-Hydroxycarboxylate transporter. The Azerbaijanian BN phytoplasma isolates corresponded to three tufB and secY genotypes. A finer differentiation of Azerbaijanian ‘Ca. P. solani’ isolates was obtained with mleP1 as five different mleP1 genetic variants were found. Finally, Stamp gene allowed differentiating four new genotypes in grapevine among the 10 new Stamp genotypes detected in various plants in Azerbaijan. The preliminary survey for infected insects conducted in northern Azerbaijan, led to the identification of Hyalesthes obsoletus and Reptalus noahi as potential vectors for two ‘Ca. P. solani’ new genotypes phylogenetically distant from the known genetic clusters. Altogether these results indicate an important genetic diversity of BN phytoplasmas in Azerbaijan that certainly result from spread through local insect vectors.     

Rice <Emphasis Type="Italic">OsPBL1</Emphasis> (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) enhanced defense of <Emphasis Type="Italic">Arabidopsis</Emphasis> against <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> DC3000

The receptor-like cytoplasmic kinases (RLCK family VII) are required for plant defense against various pathogens. Previously, OsPBL1 (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) was isolated from rice as a potential RSV (rice stripe virus) resistant factor, but its physiological roles in plant defense are yet to be investigated. In this study, we demonstrated that OsPBL1increased defense against P. syringae in transgenic Arabidopsis. To ascertain the role of OsPBL1 gene in plant defense, OsPBL1 tagged with HA (i.e. Hemagglutinin) was overexpressed in Arabidopsis and examined for the resistance against Pseudomonas syringae pv. tomato DC3000 (i.e. Pst DC3000). At 3 dpi of Pst DC3000, transgenic Arabidopsis lines exhibited the reduced chlorotic lesion and propagation of P. syringae, compared to wild type. Elevated pathogen resistance of transgenic lines was correlated with increased H₂O₂ accumulation and callose deposition on the infected leaves. It was also revealed that expression levels of salicylic acid dependent genes such as PR1, PR2, and PR5, were induced higher in transgenic lines than wild type. Taken together, our data suggested that OsPBL1 exerted the role in defense against pathogen attacks in plant via mainly facilitating salicylic acid dependent pathway.     

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.     

<Emphasis Type="Italic">Fusarium ershadii</Emphasis> sp. nov., a Pathogen on <Emphasis Type="Italic">Asparagus officinalis</Emphasis> and <Emphasis Type="Italic">Musa acuminata</Emphasis>

Two Fusarium strains, isolated from Asparagus in Italy and Musa in Vietnam respectively, proved to be members of an undescribed clade within the Fusarium solani species complex based on phylogenetic species recognition on ITS, partial RPB2 and EF-1α gene fragments. Macro- and micro-morphological investigations followed with physiological studies done on this new species: Fusarium ershadii sp. nov can be distinguished by its conidial morphology. Both isolates of Fusarium ershadii were shown to be pathogenic to the monocot Asparagus officinalis when inoculated on roots and induced hollow root symptoms within two weeks in Asparagus officinalis seedlings. In comparison mild disease symptoms were observed by the same strains on Musa acuminata seedlings.     

<Emphasis Type="Italic">In vivo</Emphasis> evaluation of essential oils and biocontrol agents combined with heat treatments on basil cv Genovese Gigante seeds against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">basilici</Emphasis>

Seed treatments with essential oils (from savory and thyme) and biocontrol agents (Pseudomonas spp. and Fusarium oxysporum) have been evaluated in vivo after dry hot air treatments against Fusarium oxysporum f. sp. basilici on basil seeds. The savory and thyme essential oils showed a significant pathogen control activity because of their innate antifungal activity and because of the seed application method, but the dry hot pre-treatment did not show any obvious effect on the performance of the essential oil treatments. The dry heat treatment improved the Pseudomonas seed dressing effect against F.oxysporum f. sp. basilici, and showed important reductions in plant infection and the disease index on the treated seed plants, without any negative effect on seed germination. However, the pathogen control provided by the heat treatments combined with the application of the biocontrol agents never reached the same performance as the chemical treatments considered as the reference. Thus, short dry heat treatments on basil seeds have been shown to be a valid but complementary seed disinfection method against Fusarium wilt.     

Overexpression of <Emphasis Type="Italic">SlARG2</Emphasis> or <Emphasis Type="Italic">SlTD2</Emphasis> in Arabidopsis enhances resistance against <Emphasis Type="Italic">Plutella xylostella</Emphasis> L.

Tomato (Solanum lycopersicum L.) ARGINASE2 (ARG2) and THREONINE DEAMINASE2 (TD2) are involved in plant defense. These enzymes act in the midgut of herbivores fed on tomato plants to degrade the essential amino acids Arg and Thr, respectively. Although it has been demonstrated that overexpression of the SlARG2 gene in tomato enhanced its resistance against M. sexta larvae, knock-down the expression of SlTD2 reduced the resistance of tomato to lepidopteran herbivores; it remains unclear whether overexpression of SlTD2 could enhance the resistance of the host plants to herbivores, or whether combined overexpression of SlARG2 and SlTD2 could lead to synergistically enhanced resistance to insects. Here, we generated transgenic Arabidopsis plants overexpressing SlARG2 (SlARG2 OE) and SlTD2 (SlTD2 OE) individually as well as in combination (SlARG2-SlTD2 OE). Overexpression of these genes did not affect Arabidopsis development, seed yield, or Arg and Thr content. Insect-feeding bioassay was performed by feeding diamondback moth (Plutella xylostella L.) larvae on detached leaves of wild-type, SlARG2 OE, SlTD2 OE, and SlARG2-SlTD2 OE plants. Larvae fed on SlARG2 OE leaves showed approximately 31% to 35% reduction in weight and 6% to 10% reduction in survival rate compared to those fed on wild-type leaves. Although larvae fed on SlTD2 OE leaves showed no reduction in survival rate, they gained less weight. Whereas larvae fed on SlARG2-SlTD2 OE leaves showed neither reduction in weight nor reduction in survival rate. We further investigated the arginase enzymatic activity of the SlARG2 OE and SlARG2-SlTD2 OE transgenic plants. The SlARG2 OE line most resistant to diamondback moth larvae displayed the highest arginase activity. Our data indicate that overexpression of SlARG2 or SlTD2 in Arabidopsis can enhance its resistance against diamondback moth, whereas combined overexpression of SlARG2 and SlTD2 did not generate synergistically increased resistance to diamondback moth.     

Morphological and molecular characterization of <Emphasis Type="Italic">Diaporthe (</Emphasis>anamorph <Emphasis Type="Italic">Phomopsis)</Emphasis> complex and pathogenicity of <Emphasis Type="Italic">Diaporthe aspalathi</Emphasis> isolates causing stem canker in soybean

The complex of Diaporthe (asexual morph) species occurring on soybean constitutes an important pathogenic group associated with diseases such as pod and stem blight, seed decay and stem canker. Stem canker, caused by Diaporthe aspalathi, has been reported as the most aggressive form of canker and its occurrence has limited soybean crop productivity in the southern United States. The main form of pathogen control is the use of stem canker resistant soybean varieties. In this study, strains of Diaporthe and Phomopsis were isolated from stem and seeds of soybean in different locations in South America during the years 1989–2014. Genomic DNA from 26 isolates were analyzed by PCR-restriction fragment length polymorphism (RFLP) and Amplified Fragment Length Polymorphism (AFLP) techniques, and sequencing of internal transcribed spacer (ITS) regions of ribosomal DNA. The molecular analysis of ITS sequences by alignment with those of ex-type strains deposited in GenBank and morphological characteristics allowed the identification of Phomopsis longicolla, D. phaseolorum var. sojae, D. caulivora and D. aspalathi. An analysis of the pathogenicity of 13 isolates of D. aspalathi inoculated in soybean genotypes carrying different resistance genes to stem canker (Rdm1, Rdm2, Rdm3, Rdm4, Rdm5 and Rdm?) enabled us to identify the occurrence of at least three races of D. aspalathi occurring in Brazil. Among the isolates identified as D. aspalathi, both molecular and phenotypic analyses showed clustering depending on the date of collection and pathogenicity, which revealed the existence of variability of the pathogen.     

Comparative colonisation by virulent versus avirulent <Emphasis Type="Italic">Pyricularia oryzae</Emphasis> on wild <Emphasis Type="Italic">Oryza australiensis</Emphasis>

Pyricularia oryzae (rice blast) conidial development at pre-penetration stage determines success or otherwise of infection inside the rice host plants. Studies on conidial germination and growth on the leaf surface in commercial rice (Oryza sativa) report differently, dependent upon host type and level of blast resistance. Although wild rice (O. australiensis) is known to be an alternative host of blast, the interaction between P. oryzae conidia and wild O. australiensis on its leaf surface has not been previously studied. We found significant (P?<?0.001) differences in conidial development between two blast isolates with different virulence in terms of conidial germination, germ tube growth and appressoria formation on both wild and cultivated rice. Conidial germination at 6 h post-inoculation (hpi) for the virulent isolate was significantly (P?<?0.001) delayed. Germ tubes of the avirulent isolate conidia grew significantly (P?<?0.001) faster and with significantly (P?<?0.001) longer germ tubes than from virulent conidia. Appressoria development for the virulent isolate was significantly (P?<?0.001) faster at its later growth stages of 12 and 18 hpi when approximately 100% of germ tubes formed appressoria. In contrast, formation rate of appressoria for the avirulent isolate was significantly (P?<?0.001) slower and only reached 76% of germ tubes forming appressoria. Appressoria formation on O. australiensis was significantly (P?<?0.001) greater than the formation on O. sativa for both virulent and avirulent P. oryzae at 12 hpi, a clear indication that host type influences the extent of appressoria formation.     

<Emphasis Type="Italic">Alternaria alternata</Emphasis> apple pathotype (<Emphasis Type="Italic">A. mali</Emphasis>) causes black spot of European pear

A disease caused by Alternaria alternata occurred on the leaves of European pear cultivar Le Lectier in Niigata Prefecture, Japan, and was named black spot of European pear. In conidial inoculation tests, the causal pathogen induced not only small black lesions on the leaves of European pear cultivar Le Lectier, but severe lesions on the leaves of apple cultivar Red Gold, which is susceptible to the A. alternata apple pathotype (previously called A. mali) causing Alternaria blotch of apple. Interestingly, the apple pathotype isolate showed the same pathogenicity as the European pear pathogen. HPLC analysis of the culture filtrates revealed that A. alternata causing black spot of European pear produced AM-toxin I, known as a host-specific toxin of the A. alternata apple pathotype. AM-toxin I induced veinal necrosis on leaves of Le Lectier and General Leclerc cultivars, both susceptible to the European pear pathogen, at 5?×?10^?7 M and 10^?6 M respectively, but did not affect leaves of resistant cultivars at 10^?4 M. PCR analysis with primers that specifically amplify the AM-toxin synthetase gene detected the product of expected size in the pathogen. These results indicate that A. alternata causing black spot of European pear is identical to that causing Alternaria blotch of apple. This is the first report of European pear disease caused by the A. alternata apple pathotype. This study provides a multiplex PCR protocol, which could serve as a useful tool, for the epidemiological survey of these two diseases in European pear and apple orchards.     

Detection of <Emphasis Type="Italic">Xanthomonas axonopodis</Emphasis> pv. <Emphasis Type="Italic">manihotis</Emphasis>, the causal agent of cassava bacterial blight diseases in cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis>) in Ghana by polymerase chain reaction

Xanthomonas axonopodis pv. manihotis (Xam) is the causal agent of cassava bacterial blight (CBB) disease. CBB is a major constraint to cassava cultivation in Ghana. In this study, a survey was conducted in eight regions of Ghana to assess the presence of CBB disease. Out of the eight regions visited, CBB, though at different prevalence, was observed in five regions. Cassava plants samples showing suspected bacterial blight symptoms were collected for analysis by Polymerase Chain Reaction (PCR). The results of the analysis showed that Ashanti region had the highest prevalence in percentage of CBB, which recorded (70%), followed by Volta region (60%); Brong Ahafo region (40%); Eastern region (40%) and Greater Accra region (20%). Morphological examination of the putative pathogen was carried out on Cefazolin trehalose agar (CTA) and Nutrient agar (NA) media. The isolates were subjected to conventional PCR using Xanthomonas genus specific primer, RST2/RST3, Xam specific Variable Number Tandem Repeat (VNTRs) loci, XaG1_67F/R and X-gumD primers, which produced 840, 446 and 402 bp, respectively. The isolates also tested positive with SYBR Green fluorescent dye, using Real-time PCR. The resulting PCR products were sequenced and analyzed using a BLASTn program, which revealed homology between 93 and 100% with several other Xam strains retrieved from GenBank nucleotide database. The pathogenicity test of the isolates on the susceptible Esam cassava variety produced symptoms typical of Xam and the pathogen was consistently re-isolated from the inoculated cassava plants and thereby satisfying the Koch’s postulates.     

Five plant families support natural sporulation of <Emphasis Type="Italic">Cronartium ribicola</Emphasis> and <Emphasis Type="Italic">C. flaccidum</Emphasis> in Finland

Fusarium is one of the most destructive fungal genera whose members cause many diseases on plants, animals, and humans. Moreover, many Fusarium species secrete mycotoxins (e.g. trichothecenes and fumonisins) that are toxic to humans and animals. Fusarium isolates from date palm trees showing disease symptoms, e.g. chlorosis, necrosis and whitening, were collected from seven regions across Saudi Arabia. After single-sporing, the fungal strains were morphologically characterized. To confirm the identity of morphologically characterized Fusarium strains, three nuclear loci, two partial genes of translation elongation factor 1 α (tef1α) and β-tubulin (tub2), and the rDNA-ITS region, were amplified and sequenced. Of the 70 Fusarium strains, 70 % were identified as F. proliferatum that were recovered from six regions across Saudi Arabia. Fusarium solani (13 %), as well as one strain each of the following species: F. brachygibbosum, F. oxysporum, and F. verticillioides were also recovered. In addition, five Fusarium-like strains were recognized as Sarocladium kiliense by DNA-based data. The preliminary in vitro pathogenicity results showed that F. proliferatum had the highest colonization abilities on date palm leaflets, followed by F. solani. Although F. oxysporum f. sp. albedinis is the most serious date palm pathogen, F. proliferatum and F. solani are becoming serious pathogens and efforts should be made to restrict and control them. In addition, the potential toxin risks of strains belonging to F. proliferatum should be evaluated.