Effects of cover crops, weeds and plant debris on development of Mycocentrospora acerina in caraway

This paper reports on the search for inoculum sources of Mycocentrospora acerina on caraway (Carum carvi L.). Obvious suspects are cover crops of biennial caraway and preceding crops of annual caraway. Other suspects are weeds in or alongside the field. Finally, survival structures of the fungus, chlamydospore chains, packed in plant debris or naked, are suspected. M. acerina is able to infect many plant species, including cover crops of caraway such as spinach for seed production and peas. However, the agronomical suitability of a crop to serve as a cover crop of biennial caraway proved to be a more important factor in determining caraway yield than the susceptibility of the cover crop to M. acerina. This finding was corroborated by the fact that spinach and peas as preceding crops had no significant effects on M. acerina development in spring caraway sown the next year. Dill, barley and four weed species were found as new hosts of M. acerina. The role of weed hosts, susceptible crops and plant debris in the survival of the fungus in years without caraway is discussed. Caraway sown on soil containing infested caraway straw, infested debris of other plant species or chlamydospores grown in pure culture, became infected by M. acerina. Only high inoculum densities of chlamydospores in the soil caused severe damping-off of caraway seedlings. The opportunity for disease management by agronomical means is quite limited.     

Doubled haploid production via anther culture in annual,winter type of caraway (<Emphasis Type="Italic">Carum carvi</Emphasis> L.)

Caraway (Carum carvi L.) is a traditional medicinal and spice cross-pollinated plant species. Although in vitro techniques are recently extensively applied in plant breeding programmes, these are not commonly utilized in caraway. Therefore, based on the protocol for anther culture in carrot (Daucus carota L., a closely related species of caraway in Daucaceae family), in vitro androgenesis in caraway has been studied with the aim to produce completely homozygous inbred lines. Various induction conditions, such as temperature pretreatments, carbon sources and combination of growth regulators in a culture medium as well as the effect of genotype on in vitro androgenesis were examined. Ten breeding lines of winter caraway representing third generation of forced (artificial) self-pollination were used as donor plant material. Cultured anthers produced embryogenic calli, and subsequently two types of regenerated plants were obtained, namely haploids with evident microspore origin, and diploids which may represent somatic (anther wall) regenerants or spontaneous doubled haploids. The ploidy status of regenerated plants was determined by flow cytometry. This is the first report on androgenic doubled haploid production in caraway.     

Characteristics of <Emphasis Type="Italic">Phomopsis sclareae</Emphasis> obtained from sage (<Emphasis Type="Italic">Salvia officinalis</Emphasis>)

In the years 2004–2006 the species P. sclareae was isolated from sage stems showing the symptoms in the form of bark peeling off and breaking. On the basis of 5 isolates randomly chosen from the fungus population, morphology and the conditions of temperatures favourable for the most intensive growth and creation of the fungus infectious material were studied. The temperature of −6°C was viewed as unfavourable for the fungus growth, and that of 32°C was considered to prevent the formation of picnidia and conidia. The dynamic growth of the colonies and the formation of numerous picnidia and conidia were observed at the temperatures ranging from 20°C to 28°C.     

Biocides application as a natural and safe alternative for treating and protecting citrus fruits

Liquid formulations (emulsifiable concentrates) were prepared from the essential oils of fennel, peppermint and caraway. Different concentrations of the formulated oils were prepared. The prepared concentrations were tested in vivo for their inhibitory activity against the growth of Penicillium digitatum (the casual agent of green rot on citrus fruits). The results demonstrated that different concentrations of the formulated fennel, peppermint and caraway oils exerted highly toxicity against the inoculated fungus. Moreover, all formulated oils with different forms showed high activity for controlling the decay when applied as protective or therapeutic agents.     

A molecular approach to explore the extent of the threatened fungus Hypocreopsis rhododendri within wood

Hypocreopsis rhododendri is a rare fungus that grows on woody stems in hyperoceanic climax scrub on the west coasts of Britain, Ireland, and France. Knowledge of the distribution and abundance of the fungus is based entirely on sporocarp records; it does not account for any occurrence as vegetative mycelia. To address this issue, a H. rhododendri-specific polymerase chain reaction (PCR) was developed and used to assay Corylus avellana (hazel) stems for the presence of H. rhododendri mycelia. The primers ITSHrF and ITSHrR were designed within the internal transcribed spacer 2 region, and their specificity to H. rhododendri was established by their failure to amplify DNA extracted from 14 other Hypocreaceae species. The sensitivity of the assay was demonstrated by amplifying DNA extracted from 4 mg C. avellana wood spiked with 0.0013 % H. rhododendri mycelium. Samples of wood and bark were then taken from around and directly underneath 11 H. rhododendri sporocarps and assayed for the presence of H. rhododendri. PCR products were obtained from a third of the surface bark samples, but only one faint product was obtained from 70 samples taken from beneath the outer bark. The results support the view that H. rhododendri does not form mycelia within stems. We suggest that H. rhododendri is not a saprotrophic fungus, but instead appears to be a parasitic on the wood decay fungus Hymenochaete corrugata, with which it always occurs. Evidence that tissue of H. corrugata is present within the sporocarps of H. rhododendri is discussed.     

Kohninia linnaeicola, a new genus and species of the Sclerotiniaceae pathogenic to Linnaea borealis

A new genus and species is described to accommodate a newly discovered fungus pathogenic to Linnaea borealis. The fungus forms true sclerotia on stems and leaves of its host and apothecia arise singly or gregariously on the surface of ripe sclerotia. The new fungus was collected together with a stromatic conidiomal fungus that occurred on the same host. A putative teleomorph-anamorph connection of the observed taxa was ruled out by sequence comparison of the nuclear ribosomal internal transcribed spacer DNA sequences (ITS rDNA). Based on morphology and pathogenicity, the new fungus belongs in the family Sclerotiniaceae, Helotiales, Ascomycota. A phylogenetic analysis of ITS rDNA sequences from 26 taxa of the family Sclerotiniaceae was performed to conclude on the systematic position of the new fungus. The small tuberoid sclerotia, brownish subsessile to substipitate apothecia, four-spored asci, ellipsoid to isthmoid ascospores, inability to grow on PDA culture media and a number of ITS rDNA sequence autapomorphies characterize and distinguish the fungus from other taxa of the Sclerotiniaceae.     

罕见角膜炎致病真菌的体外药敏试验

目的探索角膜炎罕见致病真菌对于临床常用抗真菌药物的敏感性。方法应用美国临床实验室标准化委员会制订的标准M38-A方案进行体外药敏试验。结果束状刺盘孢对4种抗真菌药的MIC值最低,≤4.0μg/ml;茄病镰刀菌次之;淡紫拟青霉的MIC值最高;茄病镰刀菌和淡紫拟青霉对氟康唑耐药。结论束状刺盘孢对氟康唑、酮康唑、伊曲康唑和两性霉素B体外试验敏感;淡紫拟青霉对酮康唑、伊曲康唑和两性霉素B的MIC值较高;茄病镰刀菌和淡紫拟青霉对氟康唑耐药。     

A Glomerella species phylogenetically related to Colletotrichum acutatum on Norway maple in Massachusetts

A fungus isolated from Norway maple (Acer platanoides) in the Boston, Massachusetts, area was determined to be a species of Glomerella, the teleomorph of Colletotrin chum acutatum. Pure cultures of the fungus were obtained from discharged ascospores from perithecia in leaf tissue. This fungus was determined to be homothallic based on the observation of perithecial development in cultures of single-spore isolates grown on minimal salts media and with sterile toothpicks. A morphological and molecular analysis was conducted to determine the taxonomic position of this fungus. Parsimony analyses of a combined nucleotide dataset of the ITS and LSU rDNA region, and of the D1-D2 LSU rDNA region, indicated that this species has phylogenetic affinities with Colletotrichum acutatum, C. acutatum f. sp. pineum, C. lupini, C. phormii and G. miyabeana. These results are significant because C. acutatum has not been reported on Acer platanoides. In addition the consistent presence of perithecia on leaf tissue and in culture is unusual for Colletotrichum, suggesting that the teleomorphic state is important in the life cycle of this fungus.     

Utilisation of biocides for controlling pest attacks on potato tubers

Essential oils of fennel, peppermint and caraway were formulated in the form of emulsifiable concentrates. Different concentrations of the formulated oils were prepared. The prepared concentrations were tested in vivo for their inhibitory activity against the growth of Fusarium oxysporum (the casual agent of dry rot on potato). The results showed that different concentrations of the formulated fennel, peppermint and caraway oils exerted highly toxicity against the inoculated fungus. In addition, all formulated oils with different forms showed high activity for controlling the decay when applied as protective and therapeutic agents.     

Effect of fungicidal essential oils against Botrytis cinerea and Rhizopus stolonifer rot fungus in vitro conditions

The development of natural crop protection products as alternatives to the use of synthetic fungicides is currently popular. The aim of this study is to evaluate the antifungal effects of several essential oils against the fungal pathogens, Botrytis cinerea and Rhizopus stolonifer, under in vitro condition. Four essential oils (fennel, black caraway, peppermint and thyme) were each tested at five concentrations (0, 200, 400, 600 or 800 μl l^?1). In vitro results showed that the essential oil of black caraway and fennel had the highest fungicidal effect against B. cinerea and R. stolonifer, respectively. The growth of B. cinerea was completely inhibited by the essential oil of black caraway at 400 μl l^?1. Fennel oil perfectly inhibited growth of R. stolonifer fungus colonies at concentration higher than 600 μl L^?1 in potato dextrose agar medium. Percentage of spores germination was the lowest in medium of Fennel and black caraway essential oils, and was the highest in Thyme ones. These results show that plant essential oils can have a strong effect on reducing post-harvest decay. These plant essential oils could provide an alternative to synthetic chemicals to control post-harvest phytopathogenic fungi on fruit.     

Interaction between the black yeast Aureobasidium pullulans and the gall midge Lasioptera ephedricola in gall formation on the desert shrub Ephedra trifurca

Galls produced by the cecidomynd Lastoptera ephedncola on Ephedia trifurca always have a black ring associated with them while galls produced by the congener L ephedrae never do Black ring material after microscopic examination and culture proved to be Aureobasidium pullulans In addition to lacking black ring material neither L ephedrae galls nor healthy stems consistently yielded Aureobasidium on culture Gall and larva size measurements indicated that continued larval presence is not necessary for gall development, suggesting fungus initiated gall formation However inoculation of healthy stems with Aureobasidium caused lesions hut not galls The mycelium m galls did not appear grazed and neither larvae nor pupae contained Aureobasidium propagules suggesting that larvae do not feed directly on fungi These data also suggest that there is no trans-pupal passage of fungus from larvae or pupae to adults Newly emerged females do not carry fungal propagules suggesting that thcy are not inoculated upon exiting the gall Gall position leaf culture and stem culture data suggest that the fungus is picked up from leaves prior to oviposition     

Scytalidium dimidiatum an opportunistic fungus for both man and Mangifera indica trees in Venezuela

The purpose of this study was to confirm the presence of Scytalidium dimidiatum on Mangifera indica (mango) trees, in a plantation managed by a diabetic patient with a white grain mycetoma of the foot caused by the same fungus. Samples from necrotic apices, roots, burned leaves and rotten stems from eight trees were processed by the Smith and Furcolow's mineral oil technique (modified). Several isolates from the apex material and clinical samples from the diabetic patient isolated in pure culture a fungus with the macroscopic and microscopic characteristics of that in S. dimidiatum. This fungus should be considered as an opportunistic microorganism for both humans and M. indica.     

A new species of <Emphasis Type="Italic">Discostroma</Emphasis> and its anamorph <Emphasis Type="Italic">Seimatosporium</Emphasis> with two morphological types of conidia,isolated from the stems of <Emphasis Type="Italic">Paeonia suffruticosa</Emphasis>

Conidia of two morphologically different types, one with a basal appendage only and the other with appendage at both ends, were isolated from the stems of Paeonia suffruticosa. Single conidial isolates of both types of conidia yield identical colonies, which then produced both types of conidia on agar media depending on temperature, thus showing that both types of conidia belong to the same fungus. Seimatosporium botan is described based on its morphological characteristics. The teleomorph of the fungus was first found on sterilized P. suffruticosa stems placed on water agar, when grown at 5°C for 2 months in 12-h photoperiod. Discostroma botan is described for this fungus. The teleomorph is also found on the same host in the field.     

番茄灰霉病拮抗内生细菌的筛选、鉴定及其活性

对安徽省淮北市番茄植株根、茎、叶中内生细菌及其数量进行了调查和筛选,并测定了其抑菌活性。结果表明,番茄根、茎和叶中的内生细菌的数量分别为5.69×105、5.16×105和2.83×105CFU.g-1鲜重。根据分离部位和表型特征,从健康番茄植株体内分离到267株内生细菌,通过对峙实验,筛选到11株对番茄灰霉病菌有拮抗作用的菌株,占所分离内生细菌总数的4.12%。来自茎组织中的菌株XF136的抑菌效果最佳,抑菌带宽度达32.2mm。根据形态特征、生理生化特性、(G+C)mol%和16SrDNA序列分析,将菌株XF136鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。室内测定菌株XF136发酵滤液对灰霉病菌菌丝生长及分生孢子萌发的抑制作用,结果表明,菌株XF136发酵滤液可以抑制灰霉病菌菌丝生长和分生孢子萌发,且浓度越高,抑制能力越强;当发酵滤液浓度为20%时则完全抑制灰霉病菌菌丝生长和分生孢子萌发。盆栽防效试验结果表明,10%菌株XF136发酵滤液对番茄灰霉病防治效果与50%多菌灵600倍液相当,20%发酵滤液对番茄灰霉病的防治效果高于50%多菌灵600倍液。本研究表明,菌株XF136是防治番茄灰霉病潜在的优良生防菌株,具有良好的开发应用价值。     

Oxalic Acid Production and Mycelial Biomass Yield of Sclerotinia minor for the Formulation Enhancement of a Granular Turf Bioherbicide

The fungus Sclerotinia minor is presently under development in this laboratory as a granular bioherbicide for broadleaf weed species. With a view to enhancing the virulence of the fungus, the effect of increasing endogenous oxalic acid concentration through modification of the growth conditions was investigated. S. minor was grown in 125 ml of eight different liquid culture media in shake flasks incubated at 20°C for 7 days. The final pH, mycelial dry weight, and oxalic acid content of the spent growth media were determined and the virulence of S. minor grown on each solid culture medium was screened on detached dandelion leaves. A 330% increase in oxalic acid was obtained plus 56 mM of sodium succinate to Modified Richard's solution (MRS) as compared to MRS alone. A concomitant increase in virulence of 218% was expressed as increased lesion diameter.     

Effect of culture conditions on mycelial growth, antibacterial activity, and metabolite profiles of the marine-derived fungus Arthrinium c.f. saccharicola

The effects of culture conditions and competitive cultivation with bacteria on mycelial growth, metabolite profile, and antibacterial activity of the marine-derived fungus Arthrinium c.f. saccharicola were investigated. The fungus grew faster at 30°C, at pH 6.5 and in freshwater medium, while exhibited higher antibacterial activity at 25°C, at pH 4.5, 5.5, and 7.5, and in 34 ppt seawater medium. The fungus grew faster in a high-nitrogen medium that contained 0.5% peptone and/or 0.5% yeast extract, while exhibiting higher bioactivity in a high-carbon medium that contained 2% glucose. The fungal growth was inhibited when it was co-cultured with six bacterial species, particularly the bacterium Pseudoalteromonas piscida. The addition of a cell free culture broth of this bacterium significantly increased the bioactivity of the fungus. Metabolite profiles of the fungus revealed by gas chromatography (GC)-mass spectrometry showed clear difference among different treatments, and the change of relative area of three peaks in GC profile followed a similar trend with the bioactivity variation of fungal extracts. Our results showed clear differences in the optimal conditions for achieving maximal mycelial growth and bioactivity of the fungus, which is important for the further study on the mass cultivation and bioactive compounds isolation from this fungus.     

产喜树碱喜树内生真菌的筛选及喜树内生真菌的SRAP分析

生活在宿主植物里的内生真菌是很重要的药用资源。喜树是中国的传统药用植物。从喜树植物中分离得到了大约50种菌株,其中一株产喜树碱的菌株通过形态学鉴定为青霉属,这是首次在喜树植物中发现产喜树碱的青霉属菌株。为研究简单序列重复相关序列扩增多态性(SRAP)方法在喜树内生真菌中应用的可行性,选择了十株喜树内生真菌进行SRAP多态性分析。SRAP引物共扩增出1 295条带,而这些菌株也被分为三大类。这些结果表明,SRAP研究喜树内生真菌具有高效性,是讨论喜树内生真菌的遗传多样性的有效方法。     

A new species, Embellisia astragali sp. nov., causing standing milk-vetch disease in China

A new species, Embellisia astragali sp. nov., is described from necrotic leaves, petioles and stems of Astragalus adsurgens in China. The morphology of E. astragali is compared and contrasted to that of four similar species, E. abundans, E. oxytropis, E. phragmospora and E. telluster. The fungus grew intercellularly in stems and leaf blades and intracellularly in leaves. It was isolated from most sources of seeds at frequencies of 0.1-44.6%. Growth rates of colonies on potato-carrot agar, potato-dextrose agar, wheat hay decoction (WHDA) and V8 at 25 C were 0.48, 0.32, 0.68 and 0.27 mm d(-1), respectively. The optimal temperature for colony growth on WHDA was 20-25 C, and no growth was measured above 30 C. Five week old standing milk-vetch seedlings were inoculated with E. astragali by dipping whole roots and pruned roots in a conidial suspension and pouring the suspension onto the soil surface in which two seedlings had been planted. After 20 wk 66.5%, 62.1% and 85.0% plants were diseased and 24.1%, 20.7% and 17.5% plants were dead, respectively. Symptoms included the development of more side shoots with small, curved, necrotic and yellowed young leaves, plant stunting, reddish brown lesions, stem browning, dieback, shoot blight, crown rot, root black rot and plant death. This is first report of a pathogenic Embellisia on legumes.     

Mulberry anthracnose antagonists (iturins) produced by Bacillus amyloliquefaciens RC-2

Bacillus amyloliquefaciens strain RC-2 produced seven antifungal compounds (1-7) secreted into the culture filtrate. These compounds inhibited the development of mulberry anthracnose caused by the fungus, Colletotrichum dematium. Chemical structural analyses by NMR and FAB-MS revealed that all these compounds were iturins (cyclic peptides with the following sequence: L-Asn --> D-Tyr --> D-Asn --> L-Gln --> L-Pro --> D-Asn --> L-Ser --> D-beta-amino acid -->) and compounds 1-6 are identical to iturins A-2-A-7, respectively. Compound 7 (iturin A-8) is a new iturin, which has a -(CH(2))(10)CH(CH(3))CH(2)CH(3) group as a side chain in the beta-amino acid in the molecule.     

Effects of temperature and fungicides on Chrysosporium pannorum (Link) Hughes

C. pannorum was isolated in low frequencies and only in the colder months from untreated soil and leaves and from those treated with Captan, Dicloran and Thiram. The fungus was isolated throughout the year as the major species from Verdasan-treated substrates and it occurred with increased frequencies immediately after application of the fungicide. The abundance of C. pannorum on Verdasan-treated substrates was attributed to the ability of the fungus to utilise the fungicide rather than to the absence of faster growing competitor species. C. pannorum was shown to be a slow growing and moderately cellulolytic fungus with maximum rates of growth, germination and cellulose clearing at between 15° and 20°C. Increasing concentrations of the fungicides retarded or prevented growth and activity of the fungus. C. pannorum could grow, germinate and clear cellulose in higher concentrations of Verdasan than could other species studied. C. pannorum was more tolerant of HgCl₂ than of Verdasan. The fungus could detoxify up to 3 g/ml active ingredient of Verdasan (=120 g/ml of the formulated fungicide) in liquid culture.