Existence of different physiological forms within genetically diverse strains of <Emphasis Type="Italic">Ampelomyces quisqualis</Emphasis>

Powdery mildew fungi are parasitized by strains of the genetically distinct Ampelomyces quisqualis. To investigate whether differences in the phylogeny and other cultural, morphological and physiological characteristics of these different strains are related to differences in their geographic origins or the host species from which they were isolated, several A. quisqualis strains isolated from different species of Erysiphaceae collected in different countries and possessing different ITS rDNA sequences were selected and characterized. The results revealed some significant variation among the selected strains, which provides evidence for the existence of different physiological forms within the A. quisqualis species. Two groups that display differential growth on artificial media were identified. These groups also differ in the morphology of their mycelium, but not in the morphology of their pycnidia and conidia. Temperature greatly affected the in vitro growth of the A. quisqualis strains and growth rate was closely correlated to colony color. Differences in the conidial germination of distinct strains were observed during the recognition phase of the parasitic relationship. The germination of each of the investigated strains was greatly stimulated by all of the examined powdery mildew species and not only by the conidia of their original hosts. An Italian strain isolated from grapevine in the Trentino Alto-Adige region was identified as the strain that germinates the most quickly in the presence of powdery mildew conidia. Phylogenetic analysis revealed that these A. quisqualis strains can be classified into five different genetic groups, which generally correlate with the fungal host of origin and morphological and growth characteristics.     

Susceptibility of cocooned pupae and adults of the parasitoid <Emphasis Type="Italic">Microplitis mediator</Emphasis> to selected insecticides

The braconid parasitoid Microplitis mediator (Haliday) is a key natural enemy of the cabbage moth, Mamestra brassicae (L.), in Europe. In the context of an Integrated Pest Management approach, the use of selective insecticides is essential for the conservation of naturally occurring beneficial arthropods. The present laboratory study investigated the side effects of six insecticides applied at recommended field rates on adults and cocooned pupae of M. mediator. Male and female parasitoids were paired in drum cells contaminated with dry residues of insecticides. Besides lethal effects after 24 h, parasitization capacity and longevity of the surviving parasitoids was evaluated. Lethal effects on cocooned pupae were also investigated by assessing adult emergence from treated cocoons. Pirimicarb caused 100% adult mortality after 24 h, whereas the other tested insecticides caused no direct toxic effects. However, sub-lethal effects in terms of reduced parasitization activity, percentage of parasitism or female longevity were found for flonicamid, pymetrozine, spinosad and thiacloprid. Spirotetramat shortened only male longevity. Adult emergence from treated cocoons was reduced only by flonicamid and pymetrozine.     

Black nightshade (<Emphasis Type="Italic">Solanum nigrum</Emphasis>) emergence and development is affected by seed density and burial depth

Black nightshade (Solanum nigrum L.) is a troublesome weed worldwide, affecting a large number of crops. As weed development is affected by a variety of factors, advanced knowledge on the plant’s biological and ecological qualities can support favorable management strategies. Seed density is an ecological factor affecting emergence and early growth of many weed species. For that reason, the current study characterizes and quantifies the seed density impact on black nightshade seedling-emergence and early growth, and the interaction between seed density and burial depth on these growth parameters. Greenhouse studies conducted in 2005, 2006 and 2010 examined black nightshade seed densities of 1, 5, 10, 25 and 50 seeds per pot, buried at, 1, 2, 3, 4 and 5 cm. Impact of seed density on weed emergence and growth was characterized by number of emerged plants and leaves, followed by biomass measurements. Seed density had no impact on emergence at the tested burial depths. There was a strong logistic relationship (P < 0.001, and R ²  > 0.97) between time and emergence, and a strong linear relationship (P < 0.002 and R ²  > 0.86) between time and number of leaves produced in all depths and seed densities. A negative relationship was observed between seed density and plant development and growth rate. Results of this study extend the knowledge on black nightshade biology and support decision-making concerning rational weed management.     

Identification of <Emphasis Type="Italic">Pythium tracheiphilum</Emphasis> as the causal agent of vascular necrosis of endive (<Emphasis Type="Italic">Cichorium endivia</Emphasis>) in Spain

Pythium isolates were recovered from endive plants (Cichorium endivia) showing vascular necrosis collected from commercial fields located in Castellón province (eastern Spain). They were identified as Pythium tracheiphilum on the basis of their phenotypical and molecular profile. Pathogenicity tests conducted with two P. tracheiphilum isolates, obtained from endive and lettuce (Lactuca sativa), respectively, in this region, confirmed that both isolates were pathogenic to endive, with no significant differences in virulence between them. This is the first report of vascular necrosis caused by P. tracheiphilum on endive in Spain.     

Specific characters of 16S rRNA gene and 16S–23S rRNA internal transcribed spacer sequences of Xylella fastidiosa pear leaf scorch strains

Pear leaf scorch, the only Xylella fastidiosa-induced disease reported from Taiwan, was found in area where the variety Hengshan (Pyrus pyrifolia) was grown. Strains of pear leaf scorch Xyl. fastidiosa (XF-PLS) shared similarities to strains of other host origins in the requirement of complex medium and the exhibition of rippled cell walls, however, recent serological and molecular biology studies showed difference among them. Five strains of XF-PLS were compared with 20 other strains originally isolated from almond, oleander, pecan, plum, peach, mulberry, grapes, citrus, coffee, and sycamore by sequence analyses of the 16S rRNA gene and 16S–23S rRNA internal transcribed spacer region (ITS). When sequences of 16S rRNA gene based on fragment size of 1,537–1,540 bp were compared, the similarity index among 5 XF-PLS strains was 99.3–99.8%, whereas it was 97.8–98.6% between XF-PLS strains and strains from other hosts. When sequences of 16S–23S rRNA ITS based on fragment size of 510–540 bp were compared, the similarity index among 5 XF-PLS strains was 99.0–100%, whereas it was 80.7–82% between XF-PLS strains and strains from other hosts. Multiple sequence alignments led to the identification of 5 polymorphic nucleotides in the 16S rRNA gene among the 25 Xyl. fastidiosa strains, and there were considerable variations in the nucleotide sequences of 16S–23S rRNA ITS between XF-PLS and the other 20 Xyl. fastidiosa strains. The phylogenetic trees revealed that XF-PLS strains were separated from strains of other hosts. Strains of other hosts were divided into four subgroups: strains from (1) oleander, (2) grape, almond M23 and mulberry, (3) citrus and coffee, and (4) pecan, peach, plum, sycamore and almond M12. Results indicate that XF-PLS strains were not closely related to the above-mentioned strains from other hosts and could possibly belong to a new subspecies of Xyl. fastidiosa.     

Green-odour compounds have antifungal activity against the rice blast fungus <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

Four green-odour compounds—trans-2-hexenal, cis-3-hexenol, n-hexanal, and cis-3-hexenal—were applied (0.85 μg ml⁻¹ as vapour) to rice plants in laboratory conditions to observe their biological activity against the phytopathogenic fungus Maganporthe oryzae, which causes rice blast disease worldwide. Two compounds, trans-2-hexenal and cis-3-hexenal, showed remarkable disease suppression efficacy (99.7% and 100% suppression, respectively), while n-hexanal had moderate (86.5%) and cis-3-hexenol had weak (20.8%) disease-suppressing effects. Pre-application and post-application of trans-2-hexenal or cis-3-hexenal had slight effects on blast incidence, suggesting that these compounds had direct effects to suppress M. oryzae infection. In fact, trans-2-hexenal and cis-3-hexenal exhibited a growth suppression effect on M. oryzae. Interestingly, these two compounds inhibited appressorium formation at lower concentrations than the growth suppression. Studies on the hypersensitive response (HR)-like reaction and plant β-1,3-glucanase activity in rice plant confirmed that induced resistance was not the major factor involved in the disease suppression mechanism. Results of this study conclusively showed that trans-2-hexenal and cis-3-hexenal possess potent inhibitory activities against the growth and the appressorium formation of M. oryzae and could be used as antifungal agents to significantly reduce M. oryzae infections in rice.     

Airborne fungal succession in a rice field of Cuba

This study characterised the airborne mycobiota of a rice plantation in Bauta (Havana, Cuba), with a view to improving crop quality. Culturable airborne fungi were sampled between March 2007 and February 2008 using a volumetric method (SAS Super 100) with potato dextrose agar as culture medium. The total fungal count was 51,425 cfu/m³. The 39 filamentous genera identified included several major rice pathogens (Bipolaris, Curvularia, Alternaria, Pyricularia, Cercospora and Fusarium) as well as potentially mycotoxigenic species of Fusarium, Penicillium and Aspergillus. Analysis of the influence of weather-related factors on spore counts revealed that total counts and Penicillium counts correlated positively with daily relative humidity, mean temperature and dew point.     

Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Loose smut fungi of barley and wheat (Ustilago nuda and U. tritici, respectively) colonize the plant without causing obvious disease symptoms before heading. The availability of diagnostic methods to detect and follow the growth of these pathogens in the plant would therefore be highly advantageous for both resistance breeding and the development of effective seed treatments. Using seed lots of barley and wheat highly infected with loose smut, we studied the early establishment of the loose smut pathogens in the plant by fluorescence microscopy. In hand-cut sections stained with the fluorochrome Blankophor?, fungal hyphae were observed to invade the shoot apical meristem and leaf primordia during the first days after the onset of germination. At the first node stage the ear and leaf primordia were generally extensively colonized. Hyphae of U. nuda were also regularly observed in high density in the nodes. A protocol was developed for the specific amplification of U. nuda and U. tritici DNA extracted from infected plant tissue. PCR screening of U nuda in seedlings from infected and healthy seed lots was compared to ELISA, microscopy and ultimately head infection of mature plants derived from tillers of the tested seedlings. The results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.     

Screening new <Emphasis Type="Italic">Arachis</Emphasis> amphidiploids,and autotetraploids for resistance to late leaf spot by detached leaf technique

Cultivated groundnut is susceptible to late leaf spot (LLS) caused by Phaeoisariopsis personata [(Berk. & M.A. Curtis) Aex] and resistance is low to moderate in the primary gene pool of groundnut. Closely related wild species in the secondary gene pool are highly resistant to the disease. All the closely related Arachis species are diploid and cultivated groundnut is a tetraploid. Utilization of diploid Arachis species to transfer LLS resistance is time consuming and cumbersome. New sources of Arachis hypogaea (also called synthetic groundnut) were developed at ICRISAT. These are tetraploids and the present investigation has shown that they are resistant to LLS.     

Frequency of mutations associated with fungicide resistance and population structure of <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> in Tunisia

The occurrence of fungicide resistance in Mycosphaerella graminicola populations from Tunisia was investigated by examining mutations known to be associated with strobilurin and azole resistance. Few mutations associated with fungicide resistance were detected. No evidence for strobilurin resistance was found among 357 Tunisian isolates and only two among 80 sequenced isolates carried mutations associated with azole resistance. A network analysis suggested that these mutations emerged independently from mutations found in previously described European populations. The population genetic structure of M. graminicola in Tunisia was analyzed using variation at 11 microsatellite loci. Populations in Tunisia were characterized by high gene and genotype diversity. All populations were in gametic equilibrium and mating type proportions did not deviate from the 1:1 ratio expected under random mating, consistent with regular cycles of sexual reproduction. In combination with a high degree of gene flow among sampling sites, M. graminicola must be considered a pathogens with high evolutionary potential. Thus, control strategies against Septoria blotch in Tunisia should be optimized to reduce the emergence and spread of resistant isolates.