Pathogenicity and toxigenicity of <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> isolates collected from maize roots,stems and ears in South Africa

Sunflower (Helianthus annuus L.) is an important oilseed crop in South Africa, and is grown in rotation with maize in some parts of North West, Limpopo, Free State, Mpumalanga and Gauteng provinces. Alternaria leaf blight is currently one of the major potential disease threats of sunflower and is capable of causing yield losses in all production regions. Alternaria helianthi was reported as the main cause of Alternaria leaf blight of sunflower in South Africa; however small-spored Alternaria species have been consistently isolated from leaf blight symptoms during recent surveys. The aim of this study was to use morphological and molecular techniques to identify the causal agent(s) of Alternaria blight isolated from South African sunflower production areas. Alternaria helianthi was not recovered from any of the sunflower lesions or seeds, with only Alternaria alternata retrieved from the symptomatic tissue. Molecular identification based on a combined phylogenetic dataset using the partial internal transcribed spacer regions, RNA polymerase second largest subunit, glyceraldehyde-3-phosphate dehydrogenase, translation elongation factor and Alternaria allergen gene regions was done to support the morphological identification based on the three-dimensional sporulation patterns of Alternaria. Furthermore, this study aimed at evaluating the pathogenicity of the recovered Alternaria isolates and their potential as causal agents of Alternaria leaf blight of sunflower. Pathogenicity tests showed that all the Alternaria alternata isolates tested were capable of causing Alternaria leaf blight of sunflower as seen in the field. This is the first report of A. alternata causing leaf blight of sunflower in South Africa.     

<Emphasis Type="Italic">Alternaria alternata</Emphasis>: A new seed-transmitted disease of coriander in South Africa

This is the first report of Alternaria leaf spot disease on coriander (Coriandrum sativum L.) in South Africa. Using the agar plate method, Alternaria alternata was isolated from coriander seed lots together with four other fungal genera, which included Aspergillus, Fusarium, Penicillium and Rhizopus. Standard seed germination tests of coriander seed lots infected with seed-borne mycoflora showed a positive correlation with the number of diseased seedlings (r?=?0.239, p?<?0.01). Pathogenicity tests demonstrated that this seed-borne A. alternata was pathogenic on coriander and symptoms on leaves first appeared as small, dark brown to black, circular lesions (<5 mm diam.) that enlarged and coalesced to form dark brown blotches as time progressed. Leaf spot disease was most severe (64%) on wounded leaves inoculated with A. alternata. Re-isolation of A. alternata from diseased coriander plants satisfied the Koch’s postulates, thus confirming it as the causal agent of Alternaria leaf spot disease. Parsimony analysis based on rpb2 (GenBank Accession No. KT895947), gapdh (KT895949) and tef-1α (KT895945) sequences confirmed identity of the Alternaria isolate, which grouped within the A. alternata clade. Alternaria alternata was shown to be transmitted from infected coriander seed to the developing plants.     

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.     

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.     

Identification and characterization of <Emphasis Type="Italic">Alternaria</Emphasis> species causing leaf spot on cabbage,cauliflower, wild and cultivated rocket by using molecular and morphological features and mycotoxin production

Alternaria species are common pathogens of fruit and vegetables able to produce secondary metabolites potentially affecting human health. Twenty-nine isolates obtained from cabbage, cauliflower, wild and cultivated rocket were characterized and identified based on sporulation pattern and virulence; the phylogenetic analysis was based on the β-tubulin gene. Isolates were identified as A. alternata, A. tenuissima, A. arborescens, A. brassicicola and A. japonica. Pathogenicity was evaluated on plants under greenhouse conditions. Two isolates showed low level of virulence on cultivated rocket while the other isolates showed medium or high level of virulence. Isolates were also characterized for their mycotoxin production on a modified Czapek-Dox medium. Production of the five Alternaria toxins, tenuazonic acid, alternariol, alternariol monomethyl ether, altenuene and tentoxin were evaluated. Under these conditions, about 80% of the isolates showed the ability to produce at least one mycotoxin.     

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.     

Multi-resistance to thiophanate-methyl,diethofencarb, and procymidone among <Emphasis Type="Italic">Alternaria alternata</Emphasis> populations from tobacco plants,and the management of tobacco brown spot with azoxystrobin

In 2014 and 2015, a total of 151 tobacco brown spot (Alternaria alternata) isolates were collected from Guizhou Province in China to evaluate their resistance to the benzimidazole thiophanate-methyl, the carbamate diethofencarb, and the dicarboximide procymidone. Resistance to thiophanate-methyl and diethofencarb was observed in all isolates. Resistance to all the three fungicides, thiophanate-methyl, diethofencarb, and procymidone was detected at a frequency of 6.0%. The F167Y single mutation in the β-tubulin gene was found to be associated with resistance to thiophanate-methyl,but no mutation was found in the coiled-coil region of the histidine kinase-encoding gene OS1, a fungal gene for dicarboximide resistance. Procymidone applied at the rate of 20 mg l^?1 inhibited spot lesion formation on tobacco leaves with an efficacy of 51.7% for the low resistance (LR) isolates and 74.2% for the procymidone-sensitive isolates. Thiophanate-methyl applied at 100 mg l^?1, however, slightly promoted the expansion of disease lesions with an efficacy of ?7.7%. Azoxystrobin applied at 10 and 20 mg l^?1 provided efficacies of 91.1 and 100%, respectively, regardless of whether the isolates were thiophanate-methyl resistant or procymidone-LR. Further studies suggested that azoxystrobin exhibited excellent protective activity and good curative activity against A. alternata in plants. The baseline sensitivity to azoxystrobin was then determined. In the presence SHAM, the mean EC₅₀ values for conidial germination inhibition were 0.49?±?0.22 (Mean?±?SD) mg l^?1. Interestingly, no resistance was recovered through UV irradiation or Agrobacterium tumefaciens-mediated mutagenesis. This research indicated widespread resistance to thiophanate-methyl and diethofencarb, low frequency of (6.0%) resistance to procymidone in A. alternata populations from tobacco, and suggested that azoxystrobin could potentially constitute a good alternative for the management of tobacco brown spot disease.     

First report of leaf spot caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> on <Emphasis Type="Italic">Drimia maritima</Emphasis>

Drimia maritima (squill) is a historically important medicinal plant. During the spring of 2016, small, yellow leaf spots, which became brown and finally necrotic, were observed on squill plants in Kohgiluyeh and Boyer-Ahmad Provinces in Iran. A fungus was consistently isolated from infected leaves and identified as Alternaria alternata based on morphological and phylogenetic analyses. Pathogenicity tests confirmed A. alternata to be the causal agent of the newly observed leaf spot disease. This is the first report of leaf spot on D. maritima caused by A. alternata in the world.     

Biological and molecular characterization of field isolates of <Emphasis Type="Italic">Alternaria alternata</Emphasis> with single or double resistance to respiratory complex II and III inhibitors

Field isolates of Alternaria alternata collected from tomato processors were characterized for sensitivity to respiration inhibitors using in vitro mycelial growth assays. Pyraclostrobin (QoI), boscalid, fluopyram and isopyrazam (SDHIs) mean EC₅₀ values were 0.32, 1.43, 2.21, and 3.53 μg/ml respectively. Of the 42 isolates, 36 were sensitive to all respiration inhibiting fungicides tested whereas three isolates were less sensitive to boscalid, one to pyraclostrobin and two were simultaneously resistant to both inhibitors and isopyrazam. Correlation analysis between fungicide sensitivities revealed a positive cross-resistance between pyraclostrobin and tebuconazole, and between cyprodinil and mancozeb. There was no cross-resistance between QoIs, SHDIs or any other mode of action. Sequencing of the QoI and SDHI targets revealed the G143A cytochrome b resistance mutation in all pyraclostrobin-resistant isolates while analysis of the succinate dehydrogenase coding gene revealed point mutations in two of three of the gene subunits analyzed in boscalid-resistant isolates. Specifically, two isolates carried the H277Y and three the H133Q resistance mutations located in the sdhB and sdhD subunits of the respiration complex II, respectively. Isolates bearing the H277Y mutation also carried the G143A cytochrome b resistance mutation. Boscalid and pyraclostrobin-resistant isolates exhibited greater pathogenicity and sporulation compared to sensitive isolates, respectively. Isolates with cross-resistance exhibited greater pathogenicity and sporulation but slower mycelial growth compared to sensitive isolates. This is the first report of field isolates of A. alternata with single or double resistance to QoIs and SDHIs in Greece and should be considered in planning and implementing effective anti-resistance strategies.     

<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.     

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).     

Ethylene production by <Emphasis Type="Italic">Alternaria alternata</Emphasis> and its association with virulence on inoculated grape berries

Ethylene has been shown to promote spore germination and hyphal growth in the phytopathogenic fungus Alternaria alternata. However, little is known about the ethylene biosynthetic pathway in this fungus. In the present study, the ethylene biosynthetic pathway in A. alternata was investigated to explore ethylene-associated virulence of this fungus. The strain A0 of A. alternata did not produce ethylene on basal medium with different possible precursors or intermediates for ethylene biosynthesis (glutamate, aspartate, 2-oxoglutarate and 1-aminocyclopropano ?1-carboxylic acid). However, ethylene production was observed when methionine was added as a precursor to the medium and was further promoted by continuous light illumination. Furthermore, addition of 2-keto-4-methylthiobutyric acid (KMBA) promoted ethylene production in the absence of methionine, indicating that the KMBA pathway was mainly responsible for ethylene biosynthesis in this fungus. The strain A0 was inoculated into grape berries to examine the effect of ethylene production on its virulence (as assessed by lesion formation at the inoculation site). The results indicated that higher ethylene production caused larger lesion formation. Similar results were also obtained when isolates of A. alternata, obtained from infected grapes, were inoculated. Thus, the present study thus demonstrated that A. alternata produces ethylene via the KMBA pathway and utilizes it for enhanced virulence expression during infection.     

Characterization of <Emphasis Type="Italic">Phaeoacremonium</Emphasis> isolates associated with Petri disease of table grape in Northeastern Brazil,with description of <Emphasis Type="Italic">Phaeoacremonium nordesticola</Emphasis> sp. nov.

Severe outbreaks of Alternaria leaf blotch and fruit spot were recently observed in cv. Pink Lady apples in northern Israel, especially on fruit. Such severe outbreaks have not been reported from other countries. Symptoms involved cracks and rot around the calyx and external rot of the fruit body. Up to 80 % of the fruit in some orchards were affected by the disease. Microscopic examinations, fulfillment of Koch’s postulates and molecular (genetic) analyses confirmed the causal agent as Alternaria alternata f. sp. mali. The incidence of Alternaria increased as the degree of calyx cracking increased, or if fruit were both cracked and rotted. Injecting spore suspensions into the fruit produced typical rot symptoms. Injection assays of detached fruit of eight apple cultivars showed that cvs. Pink Lady and Golden Delicious were susceptible whereas cv. Jonathan was resistant. Pink Lady and Golden Delicious produced more fruit rot as the inoculum concentration increased. Rot in all three cultivars was moderate close to the skin but more severe close to the seed locule. Aqueous extracts taken from Jonathan fruit peel inhibited germ tube elongation of A. alternata f. sp. mali in vitro. This is the first report on heavy infection of Pink Lady fruit in Israel caused by A. alternata f. sp. mali.     

Morphological and molecular variability among Indian isolates of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> causing banded leaf and sheath blight in maize

Rhizoctonia solani, a devastating soil borne fungus inciting banded leaf and sheath blight (BLSB) disease is a constraint in maize production and improvement program. Rhizoctonia isolates collected from seven diverse maize cropping zones of India were examined for morphological and molecular variability. All the tested isolates caused symptoms of BLSB on maize and were also cross infective on rice and sugarcane hosts, but showed significant variability in hyphal diameter, mean hyphal cell size, weight, size and distribution of scleorotia, culture pigmentation, incubation period, pathogenicity and expression of symptoms. Neighbour joining cluster analysis placed the 62 isolates of R. solani into four major groups, A, B, C and D. Group A was more diverse and included isolates of diverse agro-ecological zones. The cluster analysis corresponded well with principle component analysis. Pathogenicity testing of R. solani isolates on maize genotype (CM 501) revealed highly variable virulence pattern of the pathogen population suggesting its high evolutionary potential, and hence adaptability to diverse geographical regions. The study reveals a strong evidence of inherent potential of the R. solani isolates to survive in diverse ecological zones and its probable spread to other maize cultivars across India. Sequence comparisons of the internal transcribed sequence-ribosomal DNA region of 62 isolates did not reveal much diversity among the isolates. Majority of the isolates (n?=?61) clustered together with anastomosis group (AG) AG1-IA used as reference strain in the phylogram, distinct from AG1-IB, AG2–2IIIB and Waitea circinata used as reference strains. BLSB isolates representing distinct geographical locations shared identical sequences indicating long-distance dispersal of the pathogen. The study confirms that the genetic flexibility of the pathogen allows for its adaptation to variable ecological niches and long-distance introduction of new genotypes into the region. The study emphasizes that epidemiological studies may complement the molecular studies.     

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.     

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.     

Activity of <Emphasis Type="Italic">ß</Emphasis>-1,3-glucanase and <Emphasis Type="Italic">ß</Emphasis>-1,4-glucanase in two potato cultivars following challenge by the fungal pathogen <Emphasis Type="Italic">Alternaria solani</Emphasis>

Early blight of potato, caused by Alternaria solani, is a ubiquitous disease in many countries around the world. Our previous screening of several Iranian potato cultivars found that variation in resistance exists between two cultivars: ‘Diamond’ and ‘Granula’. Cultivar Diamond is more resistant to multiple isolates of A. solani when compared to cv. Granula. Furthermore, we have found that different pathogen isolates have varying degrees of infection. We monitored the activities of two pathogen-related (PR) glucanase proteins in Diamond and Granula in response to two isolates of A. solani with different degrees of virulence. ß-1,3-glucanase and ß-1,4-glucanase activities were recorded in healthy and diseased leaves of potatoes up to 10 days after inoculation. Their activities were found to be higher in diseased leaves when compared to those of uninfected leaves. Our data suggest that significantly reduced activities of theses enzymes in potato could be related to a lower degree of resistance or an increased ability of a more aggressive isolate to suppress PR protein expression.     

Screening for Antifungal Potential of Plant Extracts of Fifteen Plant Species Against Four Pathogenic Fungi Species

The antifungal activity of fifteen wild plant species grown in the Al-Qassim region, Saudi Arabia, was investigated against the fungi species Fusarium solani, Botrytis cinerea, Alternaria alternata and Stemphylium botryosum at concentrations of 10, 20, 40 and 80?g/L. The inhibitory effect of test extracts varied among examined fungi, and Fusarium solani exhibited the least sensitivity compared to other fungi species. Antifungal activity of the tested extracts was proportional to the applied dose. Extracts of Lactuca virosa, Neurada procumbens, Ochradenus baccatus and Cyperus conglomerates showed relatively low inhibition effects on the fungi species. The most effective plants were Pulicaria undulata, Artemisia monosperma, Prosopis juliflora, Withania somnifera and Rumex vesicarius. At 80?g/L, extracts of these plant species reduced mycelial growth of Botrytis cinerea by 16.5–32.2%, of Fusarium solani by 11.1–27.9%, of Alternaria alternaae by 26.9–63.5% and of Stemphylium botryosum by 22–40%. The methanolic extract of the most effective plant species was further fractioned with hexane, methylene chloride and ethyl acetate. The obtained fractions varied in their effects on mycelial growth of the four tested fungi. Using the same fraction resulted in different inhibition effects on mycelial growth of all tested fungi. The antifungal activity of each crude extract tended to be distributed among its three fractions, probably because the bioactive components were also distributed among the fractions.     

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.     

Phylogenetic relationship and fungicide sensitivity of members of the <Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis> species complex from apple

The members of the Colletotrichum gloeosporioides species complex (CGSC), the dominant pathogens of apple bitter rot in Nagano prefecture, Japan, were reidentified and the relationship between the species and fungicide sensitivity was revealed. Based on phylogenetic analysis of the ApMat locus with the neighbor-joining (NJ) method, isolates from apple contained three species of the CGSC; C. fructicola, C. aenigma, C. siamense, and three clades of the CGSC: Clade V, S and K. Colletotrichum fructicola and Clade S dominated in Nagano Prefecture. Isolates of C. siamense, C. aenigma and Clade V, S and K remained sensitive to benomyl and quinone outside inhibitor (QoI) fungicides, while C. fructicola often developed resistance to benomyl and QoI fungicides. These results suggest that the development of fungicide resistance differs among members of the CGSC.