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.     

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.     

Assessing the Belgian potato <Emphasis Type="Italic">Alternaria</Emphasis> population for sensitivity to fungicides with diverse modes of action

Early blight and brown spot, caused by respectively Alternaria solani and Alternaria alternata, can lead to severe yield losses in potato-growing areas. To date, fungicide application is the most effective measure to control the disease. However, in recent years, a reduced sensitivity towards several active ingredients has been reported. To shed light on this issue, Alternaria isolates were collected from different potato fields in Belgium during two growing seasons. Subsequently, the sensitivity of these isolates was assessed using four widely used fungicides with different modes of action. Demethylation inhibitors, quinone outside inhibitors, a dithiocarbamate and a carboxylic acid amide were included in this study. Although all fungicides reduced spore germination and vegetative growth of Alternaria species to some extent, the interspecies sensitivity was very variable. In general, A. solani was more suppressed by the fungicides compared to A. alternata. The effectiveness of the dithiocarbamate mancozeb was high, whereas the quinone outside inhibitor azoxystrobin showed a limited activity, especially towards A. alternata. Therefore, a subset of the A. alternata and A. solani isolates was tested for the presence of, respectively, the G143A substitution and the F129L substitution in the cytochrome b. The frequency of A. alternata isolates bearing the resistant G143A allele (approximately 65%) was comparable in both sampling years, although sensitivity of isolates decreased during the growing season. This finding points to a shift of the population towards resistant isolates. Both the European genotype I and American genotype II were present in the A. solani population, with genotype I being the most prevalent. None of the genotype I isolates carried the F129L substitution, whereas in 83% of the genotype II isolates this substitution was present. Our results demonstrate for the first time that the Belgian Alternaria population on potato comprises a considerable broad spectrum of isolates with different sensitivity to fungicides.     

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">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.     

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.     

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.     

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.     

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.     

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.     

<Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> subsp. <Emphasis Type="Italic">michiganens</Emphasis>is strains virulence and genetic diversity. a first study in Argentina

Tomato leaves showing severe leaf spot symptoms have been observed and sampled in the central west and southwest Taiwan during 2015 and 2016. The symptoms were similar to those of bacterial leaf spot/late blight diseases, but only Stemphylium-like fungi were consistently isolated from the diseased tomato. Upon spray inoculation of tomato, Stemphylium-like isolates caused leaf spot symptoms identical to those of naturally infected plants, and the pathogenic isolates were successfully re-isolated from inoculated leaves. The tomato-pathogenic isolates were identified as S. lycopersici based on morphological characterization and molecular identification. S. lycopersici has been previously reported to cause gray leaf spot of tomato in the temperate regions, but the majority of S. lycopersici-caused lesions were black/dark brown rather than gray in our surveillance. Accordingly, it is suggested that S. lycopersici-caused disease of tomato is named Stemphylium leaf spot of tomato more appropriately than tomato gray leaf spot. Moreover, S. lycopersici-caused leaf spot disease on tomato has been distributed in major tomato production regions in Taiwan. The information provided by our study will be important for future breeding of tomato cultivars, especially for tomato producers in Taiwan.     

Foliar diseases of willows (<Emphasis Type="Italic">Salix</Emphasis> spp.) in selected locations of the Karkonosze Mts. (the Giant Mts)

The species of Salix herbacea L. and Salix lapponum L. are the rare relicts within the genus of Salix. With respect to their phylogeography, they are classified into the taxa of the Arctic-Alpine distribution. They can be found in the Arctic zone (Greenland, Scandinavia, Spitsbergen) as well as in such geographically separated areas as the mountains of lower latitudes like the Alps, the Sudetes or the Carpathians. Another species of willow occurring in the mountains of Europe, and more specifically on montane positions of the Balkan-Carpathian distribution is S. silesiaca. The aim of the study was to determine the severity of the diseases of the willows growing in selected locations in the Karkonosze Mountains. The research was conducted in the years 2014 and 2015 in the Kocio? ?omniczki [the ?omniczka Cirque] and the Wielki and Ma?y ?nie?ny Kocio?, [the Great and Small Snowy Cirques] at intervals of 4 weeks, from April to October. Field observations included the species of the genera Salix: S. herbacea, S. silesiaca and S. lapponum. Disease symptoms were identified with the percentage of infected plants at selected locations, as well as the percentage of infected leaves, recorded. In the course of the research, symptoms of rust caused by Melampsora epitea and M. alpina (S. herbacea and S. silesiaca), black spot blight caused by Rhytisma salicinum (S. silesiaca) and leaf spot, probably caused by a complex of fungal taxa (S. herbacea), were detected on the leaves of Salix spp. growing in post glacial cirques. During the study period, there were no disease symptoms on S. lapponum. A total of 13 species of fungi were isolated from the infected leaves of S. herbacea. In the first year, the highest share in the fungal assembly was taken by Penicillium notatum, followed by Alternaria alternata and an unidentified species of the genus Ceuthospora. In the second year of the research, Ceuthospora spp. and A. alternata predominated in the species assembly of fungi in the infected leaves of the herb willow.     

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">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.     

Fungicidal activity of Thai medicinal plant extracts against <Emphasis Type="Italic">Alternaria brassicicola</Emphasis> causing black spot of Chinese kale

Crude ethanol extracts and six organic solvent fractions of 10 Thai medicinal plants were evaluated for their antifungal activity against Alternaria brassicicola in laboratory and under greenhouse conditions. The results showed that the ethanol extracts of Coscinium fenestratum, Piper betle, Syzygium aromaticus and Zingiber cassumunar displayed complete mycelial growth inhibition of A. brassicicola at a concentration of 0.1%. Meanwhile, the crude ethanol extract and methanol fraction obtained from the stems of C. fenestratum revealed the greatest inhibition against A. brassicicola at 10%, forming inhibition zones 2.55–2.58 cm in diameter. In the greenhouse experiments, crude ethanol extracts of C. fenestratum and P. betle at 1% significantly (P?<?0.05) reduced the disease incidence at up to 67%, indicating promising preventive and curative activities against A. brassicicola. This activity is similar to that of iprodione, a widely used commercial fungicide. Interestingly, Illicium verum extract showed a greater curative effect (58% disease reduction) than protective effect (47% disease reduction). Because the C. fenestratum extract showed the highest activity against the black spot pathogen both in vitro and under greenhouse conditions, its methanol fraction was further analyzed by spectroscopic techniques. We found that berberine is a key active substance inhibiting mycelial growth of A. brassicicola. The results of this study showed the potential of Thai medicinal plants as alternatives to the use of synthetic fungicides for controlling black spot in Chinese kale caused by A. brassicicola.     

Effect of fumonisin B<Subscript>1</Subscript> on the emergence,growth and ceramide synthase gene expression of cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp)

Rice production is currently expanding from the south-eastern regions of Australia into northern Australia where indigenous species of wild rice occur widely. A survey of fungal diseases on wild (Oryza australiensis, Oryza spp.) and cultivated rice (Oryza sativa) in North Queensland, Australia, in May 2014 revealed a diverse range of fungal genera species, including important pathogens of cultivated rice. Whilst a single isolate of Magnaporthe oryzae (causal agent of rice blast) was obtained from wild rice, Bipolaris oryzae (causal agent of brown spot) was the predominant pathogen detected under North Queensland conditions. For the first time for Australia, we report Rhizoctonia oryzae-sativae (causal agent of aggregate sheath spot) occurring on wild rice. Other pathogens detected on wild rice included Curvularia lunata, Cochliobolus intermedius, Cochliobolus geniculatus, and Fusarium equiseti present in the majority of wild rice samples. Nearby cultivated rice fields harboured additional pathogens not found in wild rice including Fusarium graminearum, Leptosphaeria spegazzinii and Cochliobolus lunatus, causing scab disease, glume blight and leaf blight, respectively. We also confirmed that Bipolaris oryzae from wild rice can infect cultivated rice. This study highlights the importance of wild rice species as alternative hosts harbouring pathogens of cultivated rice and the likely disease threats to expansion of cultivated rice into the same region(s) where wild rice is endemic.     

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.     

Diversity and potential impact of Botryosphaeriaceae species associated with <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> plantations in Portugal

Eucalyptus globulus, a non-native species, is currently the most abundant forest species in Portugal. This economically important forest tree is exploited mainly for the production of pulp for the paper industry. The community of Botryosphaeriaceae species occurring on diseased and healthy E. globulus trees was studied on plantations throughout the country. Nine species from three different genera were identified, namely Botryosphaeria (B. dothidea), Diplodia (D. corticola and D. seriata) and Neofusicoccum (N. australe, N. algeriense, N. eucalyptorum, N. kwambonambiense, N. parvum and Neofusicoccum sp.). Of these, N. algeriense, D. corticola and D. seriata are reported for the first time on E. globulus, while N. algeriense, N. eucalyptorum and N. kwambonambiense correspond to first reports in Portugal. The genus Neofusicoccum was clearly dominant with N. australe and N. eucalyptorum being the most abundant species on both diseased and healthy trees. In artificial inoculation trials representative isolates from all nine species were shown to be pathogenic to E. globulus but there were marked differences in aggressiveness between them. Thus, D. corticola and N. kwambonambiense were the most aggressive while B. dothidea and D. seriata were the least aggressive of the species studied.     

Evaluation of oilseed rape seed yield losses caused by <Emphasis Type="Italic">Leptosphaeria biglobosa</Emphasis> in central China

Phoma stem canker of oilseed rape (Brassica napus), caused by Leptosphaeria maculans/L. biglobosa is a globally important disease. Severe phoma stem canker symptoms have been observed on winter oilseed rape in China but the seed yield loss caused by this disease remains unknown. In May 2012 and May 2013, 17 and 13 crops were surveyed, respectively, in seven counties of Hubei Province, central China. Stems with phoma stem canker disease symptoms were sampled for pathogen isolation and identification. Only L. biglobosa was identified by culture morphology and species-specific PCR; no L. maculans was found. To evaluate the yield losses, yield components (number of branches per plant, number of pods per plant, 1000-seed weight, number of seeds per pod) were assessed on healthy and diseased plants sampled from crops in four counties and on plants from inoculated pot experiments (plants of three cultivars were inoculated at the green bud stage by injecting L. biglobosa conidia into the stem between the first and second leaf scars). Results of the field surveys showed that diseased plants had 14–61% less branches and 32–83% less pods than healthy plants, respectively. The estimated seed yield loss varied from 10% to 21% and from 13% to 37% in 2012 and 2013, respectively. In the pot experiments, there were no differences in numbers of branches or pods but there were differences in number of seeds per pod between inoculated and control plants. For the three cultivars tested, the inoculated plants had yield losses of 29–56% compared with the control. This study indicates that L. biglobosa could cause substantial seed yield loss in China.     

Rutin promoted resistance of tomato against <Emphasis Type="Italic">Xanthomonas perforans</Emphasis>

Xanthomonas perforans is the causal agent of bacterial spot, one of the most devastating diseases of tomato that results in considerable yield losses worldwide. Rutin, as a polyphenolic substance, was used to induce resistance in tomato against X. perforans. Rutin at concentration of 2 mM had ability to reduce the disease severity of bacterial spot. On the other hand, 2 mM rutin had no antibacterial activity in vitro. Expression profiling of pathogenesis-related gene 5 (PR-5), Phenylalanine ammonia-lyase (PAL) and lipoxygenase (LOX) was probed during the enhanced resistance by rutin. Pretreatment with rutin (rutin/ X. perforans) led to induction of PR-5, PAL and LOX compared to controls (water/ X. perforans). Our results suggest that rutin-induced resistance against X. perforans in tomato might be mediated through stimulation of some defense genes such as PR-5, PAL and LOX.