scholarly journals КУЛЬТУРАЛЬНО-МОРФОЛОГІЧНІ ХАРАКТЕРИСТИКИ КСИЛОТРОФНИХ БАЗИДІОМІЦЕТІВ У ГЛИБИННІЙ КУЛЬТУРІ

Author(s):  
A. V. Chaika ◽  
O. V. Fedotov

<p>The aim of the present study was to evaluate the growth specificities of 79 strains of 18 species xylotrophic basidiomycetes under submerged fermentation on glucose-peptone medium. Most of the strains (85%) were isolated from the fruiting bodies collected in different localities of Donetsk and its region. The macro- and microscopic structure of pellets was studied, pH of the culture filtrate was determined by potentiometric method, ovendry biomass was determined by weighing method and biomass increase and specific growth rate were calculated. The results obtained allowed to ascertain the strain and species character of culture-morphological characteristics of the studied basidiomycetes. On the basis of the research the higher basidiomycetes submerged cultivation method was optimized making the period of fermentation significantly reduced. The growth peculiarities revealed can be used for further studies of biosynthetic activity of the xylotrophic basidiomycetes submerged cultures.</p> <p><em>Key words: xylotrophic basidiomycetes, submerged fermentation, growth and morphological characteristics</em></p>

Author(s):  
A. V. Chaika ◽  
O. V. Fedotov

<p>The effect of different carbon and nitrogen nutrition sources on the efficiency of xenobiotic biodegradation was investigated by means of submerged xylotrophic basidiomycetes cultures. We cultivated the prospective strains of xylotrophic basidiomycetes like <em>Flammulina velutipes</em> F-1105, <em>Pleurotus eryngii</em> P-er, <em>Trametes hirsuta</em> Th-11, and <em>Daedalea quercina</em> Dq-08 on the lab shaker on modified glucose-peptone medium. Some 24 of the carbon and 15 of the nitrogen nutrition sources were investigated. Absolutely dry biomass of the mycelium was determined by weighting method. The pollutants biodegradation efficiency of submerged cultures was determined by a modified method on a model compound – <em>Methyl Orange</em>. The individual carbon- and nitrogen-containing components of the modified glucose-peptone medium, the cultivation on which provides induction of model compound <em>Methyl Orange</em> biodegradation efficiency of the strains was also determined. They mostly were the carbon sources like glucose, fructose, xylose, starch, glycerol, PEG -1500, and the nitrogen sources like urea and peptone. It is suggested that the biodegradation efficiency of induction could caused either by inductive action of the nutrient component on metabolism of culture or by the manifestation of an adaptive response to adverse conditions.</p> <p><em>Keywords: xylotrophic basidiomycetes, biodegradation, Methyl Orange.</em></p>


Author(s):  
A. V. Chaika ◽  
O. V. Fedotov

<p>The aim of the study was to determine the lipid peroxidation (LP) intensity of xylotrophic basidiomycetes cultures under submerged fermentation on glucose-peptone medium. The materials were submerged mycelium and culture filtrate of 79 strains of 18 species xylotrophic basidiomycetes. Among the studied strains, 60 strains belong to the order <em>Agaricales</em> and 19 – to order <em>Polyporales</em>. Most of the strains (85%) were isolated from the fruiting bodies collected in different localities of Donetsk city and its region. The following methods were used. Oven-dry biomass was determined by gravimetric method and biomass increase and specific growth rate were calculated. The pH of the culture filtrate was determined by potentiometric method. The intensity of lipid peroxidation processes was estimated with the modified thiobarbituric acid test. The study concluded that among the studied strains high level of LP in the mycelium is typical for the <em>F. velutipes</em> cultures, particularly the F-vv, F-03 and F-1. The low intensity of LP in the mycelium was established for the <em>F. fomentarius</em> Ff-09, Ff-1201 strains and the <em>L. edodes</em> Le-2, Le-4, Le-6, Le-7 strains. The high content of LP products in the culture filtrate is typical in most <em>Polyporales</em> cultures, for example, in the strains of <em>I. lacteus</em> IL-1201, <em>T. hirsuta</em> Th-11, <em>D. quercina</em> Dq-08 and <em>F. fomentarius</em> T-10. Paucity of the LP products in the culture filtrate established for some strains of <em>F. velutipes</em>, for example, F-204, F-11, <em>S. commune</em> Sc-1102 strain and <em>G. lucidum</em> cultures. Direct dependence between the content of LP products in the mycelium and culture filtrate was not established. Calculated correlation coefficients for the investigated strains showed the dependence of the growth rates and the intensity of LP in the mycelium and culture filtrate. Selected strains with high growth rate and a significant LP level in the culture filtrate are promising in technologies of lignocellulosic wastes and xenobiotics biodegradation and environment bioremediation.</p> <p><em>Key words: xylotrophic basidiomycetes, submerged fermentation, lipid peroxidation</em></p>


Author(s):  
O. V. Fedotov

<p>The article deals with the efficiency of pollutants biodegradation by xylotrophic basidiomycetes submerged cultures grown on standard glucose-peptone medium (GPM). The efficiency of pollutants biodegradation was determined by the model compound – dye <em>Methyl Orange</em>. The purpose of the work is screening of 19 species 81 strains xylotrophic basidiomycetes cultures on the indicator of the dye oxidative degradation efficiency and exploring the possibility of induction of this indicator by modifying the culture medium. The biodegradation efficiency was determined by following method. Assigned amount of culture filtrate (experiment) or medium (control) was added to the 0.001% solution of <em>Methyl Orange</em> in sodium acetate buffer. pH of the reaction mixture was 4.4 units. Samples were incubated at +40°C for 48 hours. Then pH of the reaction mixture was set up at 3.1 units using sodium acetate buffer and the optical density of solutions at a wavelength of 506 nm was measured. The efficiency of biodegradation was calculated by the difference of the optical density of control and experiment as a percentage. The most promising strains – <em>F. velutipes</em> F-1105, <em>P. eryngii</em> P-er, <em>T. hirsuta</em> Th-11 and <em>D. quercina</em> Dq-08 were selected. The composition of the glucose-peptone medium was modified for these strains by the introduction in the medium lignosulfonate, Tween 80, Kirk’s minerals solution and selecting the concentration of these components. According to the study for the purpose of pollutants degradation it is advisable to cultivate  <em>F. velutipes</em> F-1105 strain on modified GPM, which further comprises at 1 l: lignosulfonate – 3.5 g; Tween 80 – 1.0 g, Kirk’s minerals solution – 70 ml; <em>P. eryngii</em> P-er strain – 5.0 g, 1.0 g, 70 ml; <em>T. hirsuta</em> Th-11 strain – 5.0 g, 1.0 g, 105 ml; and <em>D. quercina</em> Dq-08 strain – 6.5 g, 1.0 g, 105 ml, respectively. This allowed to increase the model compound degradation efficiency by the culture filtrate of strain <em>F. velutipes</em> F-1105 in 9,3; <em>D. quercina</em> Dq-08 – in 9,6; <em>P. eryngii </em>P-er – in 13,3 and T. hirsuta Th-11 – 19,2 times. Thus, GPS modifications were designed that enhance the model compound oxidative degradation efficiency and are the basis for further optimization of the selected xylotrophic basidiomycetes strains submerged cultivation conditions to increase<strong> </strong>biodegradation of xenobiotics.</p> <p><em>Keywords:</em> xylotrophic basidiomycetes, submerged cultivation, biodegradation, <em>Methyl Orange</em>.</p>


Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1662-1662 ◽  
Author(s):  
Q. R. Bai ◽  
S. Han ◽  
Y. Y. Xie ◽  
J. Gao ◽  
Y. Li

Veronica sibirica (Veronicastrum sibiricum) is an erect perennial herb, an ornamental, and a traditional Chinese medicine plant distributed mostly in northeastern, northern, and northwestern China. It has dehumidifying and detoxifying properties, and is mainly used for the treatment of cold, sore throat, mumps, rheumatism, and insect bites (4). In June 2008 through 2012, leaf spots of V. sibirica were observed in the Medicinal Herb Garden of Jilin Agricultural University (43°48′N, 125°23′E) and the medicinal plantations of Antu County (43°6′N, 128°53′E), Jilin Province. Leaf spots were amphigenous, subcircular, angular-irregular, brown, and 1 to 10 mm in diameter; they occasionally merged into a larger spot with an indefinite margin or with a pale center and dark border. Pale conidiomata were hypophyllous and scattered on the spots. The conidiophores were 100 to 400 μm high and clustered together to form synnemata 20 to 50 μm in diameter, which splayed out apically and formed loose to dense capitula. Conidiophores occasionally emerged through the stomata individually and produced conidia on the surface of the infected leaves. The conidiogenous cell terminal was geniculate-sinuous with somewhat thickened and darkened conidial scars. Conidia were solitary or catenulate, ellipsoid-ovoid or subcylindric-fusiform, hyaline and spinulose, 4.01 to 7.18 × 11.16 to 20.62 μm with obtuse to somewhat attenuated ends, and slightly thickened, darkened hila. Six isolates were obtained from necrotic tissue of leaf spots and cultured on potato dextrose agar at 25°C. After incubation for 14 days, colony surfaces were white to pinkish. The colony diameter increased by 12 mm after 21 days' incubation. Hyphae were hyaline, septate, and branched. Conidiophores grew individually or fascicularly. The symptoms and morphological characteristics were consistent with previous descriptions (1,2), and the fungus was identified as Phacellium veronicae (Pass.) (U. Braun 1990). The internal transcribed spacer (ITS) region of the nuclear rDNA was amplified using primers ITS4/ITS5 (3). The ITS was identical among all six isolates (HE995799) and 98% identical to that of P. veronicae (JQ920427, HQ690097). Pathogenicity was confirmed by spraying five 1-year-old V. sibirica seedlings with a conidial suspension (106 conidia/ml) of each isolate and five seedlings with sterile water as a control treatment. Plants were grown in the greenhouse at 20 to 25°C and were covered with plastic bags to maintain humidity on the foliage for 72 h. After 15 days, the same symptoms appeared on the leaves as described earlier for the field-grown plants; the control plants remained healthy. The same fungus was reisolated from the leaf spots of inoculated plants. Currently, the economic importance of this disease is limited, but it may become a more significant problem, as the cultivated area of V. sibirica is increasing. To our knowledge, although P. veronicae was recorded on the other species of Veronica (V. austriaca, V. chamaedrys, V. grandis, V. longifolia, V. paniculata, and V. spicata ssp. incana) in Europe (Germany, Denmark, Ireland, Romania) and V. wormskjoldii in North America (Canada) (1), this is the first report of V. sibirica leaf spots caused by P. veronicae in the world, and it is a new disease in China. References: (1) U. Braun. A monograph of Cercosporella, Ramularia and allied genera (phytopathogenic Hyphomycetes) 2, IHW-Verlag, Germany, 1998. (2) U. Braun. Nova Hedwigia 50:499, 1990. (3) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (4) Jiangsu New Medical College. Dictionary of Chinese Materia Medica. Shanghai: Shanghai Scientific and Technical Publishers, China, 1977.


Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1654-1654 ◽  
Author(s):  
A. L. Vu ◽  
M. M. Dee ◽  
J. Zale ◽  
K. D. Gwinn ◽  
B. H. Ownley

Knowledge of pathogens in switchgrass, a potential biofuels crop, is limited. In December 2007, dark brown to black irregularly shaped foliar spots were observed on ‘Alamo’ switchgrass (Panicum virgatum L.) on the campus of the University of Tennessee. Symptomatic leaf samples were surface-sterilized (95% ethanol, 1 min; 20% commercial bleach, 3 min; 95% ethanol, 1 min), rinsed in sterile water, air-dried, and plated on 2% water agar amended with 3.45 mg fenpropathrin/liter (Danitol 2.4 EC, Valent Chemical, Walnut Creek, CA) and 10 mg/liter rifampicin (Sigma-Aldrich, St. Louis, MO). A sparsely sporulating, dematiaceous mitosporic fungus was observed. Fungal plugs were transferred to surface-sterilized detached ‘Alamo’ leaves on sterile filter paper in a moist chamber to increase spore production. Conidia were ovate, oblong, mostly straight to slightly curved, and light to olive-brown with 3 to 10 septa. Conidial dimensions were 12.5 to 17 × 27.5 to 95 (average 14.5 × 72) μm. Conidiophores were light brown, single, multiseptate, and geniculate. Conidial production was polytretic. Morphological characteristics and disease symptoms were similar to those described for Bipolaris oryzae (Breda de Haan) Shoemaker (2). Disease assays were done with 6-week-old ‘Alamo’ switchgrass grown from seed scarified with 60% sulfuric acid and surface-sterilized in 50% bleach. Nine 9 × 9-cm square pots with approximately 20 plants per pot were inoculated with a mycelial slurry (due to low spore production) prepared from cultures grown on potato dextrose agar for 7 days. Cultures were flooded with sterile water and rubbed gently to loosen mycelium. Two additional pots were inoculated with sterile water and subjected to the same conditions to serve as controls. Plants were exposed to high humidity by enclosure in a plastic bag for 72 h. Bags were removed, and plants were incubated at 25/20°C with 50 to 60% relative humidity. During the disease assay, plants were kept in a growth chamber with a 12-h photoperiod of fluorescent and incandescent lighting. Foliar leaf spot symptoms appeared 5 to 14 days post-inoculation for eight of nine replicates. Control plants had no symptoms. Symptomatic leaf tissue was processed and plated as described above. The original fungal isolate and the pathogen recovered in the disease assay were identified using internal transcribed spacer (ITS) region sequences. The ITS region of rDNA was amplified with PCR and primer pairs ITS4 and ITS5 (4). PCR amplicons of 553 bp were sequenced, and sequences from the original isolate and the reisolated pathogen were identical (GenBank Accession No. JQ237248). The sequence had 100% nucleotide identity to B. oryzae from switchgrass in Mississippi (GU222690, GU222691, GU222692, and GU222693) and New York (JF693908). Leaf spot caused by B. oryzae on switchgrass has also been described in North Dakota (1) and was seedborne in Mississippi (3). To our knowledge, this is the first report of B. oryzae from switchgrass in Tennessee. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/, 28 June 2012. (2) J. M. Krupinsky et al. Can. J. Plant Pathol. 26:371, 2004. (3) M. Tomaso-Peterson and C. J. Balbalian. Plant Dis. 94:643, 2010. (4) T. J. White et al. Pages 315-322 in: PCR Protocols: a Guide to Methods and Applications. M. A. Innis et al. (eds), Acad. Press, San Diego, 1990.


Botany ◽  
2012 ◽  
Vol 90 (9) ◽  
pp. 866-875 ◽  
Author(s):  
Deana L. Baucom ◽  
Marie Romero ◽  
Robert Belfon ◽  
Rebecca Creamer

New species of Undifilum , from locoweeds Astragalus lentiginosus Vitman and Astragalus mollissimus Torr., are described using morphological characteristics and molecular phylogenetic analyses as Undifilum fulvum Baucom & Creamer sp. nov. and Undifilum cinereum Baucom & Creamer sp. nov. Fungi were isolated from dried plants of A. lentiginosus var. araneosus , diphysus , lentiginosus , and wahweapensis collected from Arizona, Oregon, and Utah, USA, and A. mollissimus var. biglovii , earleii , and mollissimus collected from New Mexico, Oklahoma, and Texas, USA. Endophytic fungi from Astragalus locoweeds were compared to Undifilum oxytropis isolates obtained from dried plant material of Oxytropis lamberteii from New Mexico and Oxytropis sericea from Arizona, Colorado, New Mexico, Utah, and Wyoming. Extremely slow growth in vitro was observed for all, and conidia, if present, were ellipsoid with transverse septa. However, in vitro color, growth on four different media, and conidium size differed between fungi from Astragalus spp. and U. oxytropis. Neighbor-joining analyses of internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase (GPD) gene sequences revealed that U. fulvum and U. cinereum formed a clade distinct from U. oxytropis. This was supported by neighbor-joining analyses of results generated from random amplified polymorphic DNA (RAPD) fragments using two different primers.


Phytotaxa ◽  
2016 ◽  
Vol 270 (2) ◽  
pp. 71 ◽  
Author(s):  
SLAVOMÍR ADAMČÍK ◽  
MIROSLAV CABOŇ ◽  
URSULA EBERHARDT ◽  
MALKA SABA ◽  
FELIX HAMPE ◽  
...  

The current generally accepted concept of Russula maculata defines the species by yellow-brownish spots on the basidiomata, an acrid taste, a yellow spore print and a red pileus. This concept was tested using collections originating from various geographical areas mainly in Europe. Analyses of the ITS region suggested that there were three species within this broad concept. One of them, R. maculata, was identified based on the sequence from the epitype. Two other species, R. nympharum and R. sp., are described here as newly identified species. The European species R. maculata and R. nympharum grow in deciduous forests, are similar in their field aspect and are distinctly different in micro-morphological characteristics of spores, pleurocystidia and pileipellis. An Asian species, R. sp., is associated with pine and has smaller basidiomata and spores. These three species form the R. maculata complex and represent the sister clade to the R. globispora complex. This clade consists of species also characterized by a yellow-brownish context discolouration but with a different type of spore ornamentation. All of the other tested species had an acrid taste and yellow spore print but did not have a conspicuous yellow-brownish context discolouration and were placed in various unrelated clades.


Foods ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2774
Author(s):  
Neda Rousta ◽  
Coralie Hellwig ◽  
Steven Wainaina ◽  
Lukitawesa Lukitawesa ◽  
Swarnima Agnihotri ◽  
...  

New food sources are explored to provide food security in sustainable ways. The submerged fermentation of edible filamentous fungi is a promising strategy to provide nutritious and affordable food that is expected to have a low environmental impact. The aim of the current study was to assess the novel use of Aspergillus oryzae cultivated in submerged fermentation on oat flour as a source for food products that do not undergo secondary fermentation or significant downstream processing. The fungus was cultivated in a pilot-scale airlift bioreactor, and the biomass concentration and protein content of the biomass were assessed. A tasting with an untrained panel assessed consumer preferences regarding the taste and texture of minimally processed vegetarian and vegan burger patties made from the biomass, and how the patties fared against established meat-alternative-based patties. The cultivation of Aspergillus oryzae resulted in a yield of 6 g/L dry biomass with a protein content of 37% on a dry weight basis. The taste and texture of the minimally processed fungal burger patties were to the liking of some participants. This was also reflected in diverse feedback provided by the participants. The cultivation of the fungus on oat flour and its utilization in developing burger patties shows its promising potential for the production of nutritious food. The applications of the fungus can be further developed by exploring other favorable ways to texture and season this relatively new functional food source to the preferences of consumers.


Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 843-843 ◽  
Author(s):  
N.-H. Lu ◽  
Q.-Z. Huang ◽  
H. He ◽  
K.-W. Li ◽  
Y.-B. Zhang

Avicennia marina is a pioneer species of mangroves, a woody plant community that periodically emerges in the intertidal zone of estuarine regions in tropical and subtropical regions. In February 2013, a new disease that caused the stems of A. marina to blacken and die was found in Techeng Island of Zhanjiang, Guangdong Province, China. Initial symptoms of the disease were water-soaked brown spots on the biennial stems that coalesced so whole stems browned, twigs and branches withered, leaves defoliated, and finally trees died. This disease has the potential to threaten the ecology of the local A. marina community. From February to May 2013, 11 symptomatic trees were collected in three locations on the island and the pathogen was isolated as followed: tissues were surface disinfected with 75% ethanol solution (v/v) for 20 s, soaked in 0.1% mercuric chloride solution for 45 s, rinsed with sterilized water three times, dried, placed on potato dextrose agar (PDA), and incubated for 3 to 5 days at 28°C without light. Five isolates (KW1 to KW5) with different morphological characteristics were obtained, and pathogenic tests were done according Koch's postulates. Fresh wounds were made with a sterile needle on healthy biennial stems of A. marina, and mycelial plugs of each isolate were applied and covered with a piece of wet cotton to maintain moisture. All treated plants were incubated at room temperature. Similar symptoms of black stem were observed only on the stems inoculated the isolate KW5 after 35 days, while the control and all stems inoculated with the other isolates remained symptomless. An isolate similar to KW5 was re-isolated from the affected materials. The pathogenic test was repeated three times with the same conditions and it was confirmed that KW5 was the pathogen causing the black stem of A. marina. Hyphal tips of KW5 were transferred to PDA medium in petri dishes for morphological observation. After 48 to 72 h, white, orange, or brown flocculence patches of KW5 mycelium, 5.0 to 6.0 cm in diameter, grew. Tapering and spindle falciform macroconidia (11 to 17.3 μm long × 1.5 to 2.5 μm wide) with an obviously swelled central cell and narrow strips of apical cells and distinctive foot cells were visible under the optical microscope. The conidiogenous cells were intertwined with mycelia and the chlamydospores were globose and formed in clusters. These morphological characteristics of the isolate KW5 are characteristic of Fusarium equiseti (1). For molecular identification, the ITS of ribosomal DNA, β-tubulin, and EF-1α genes were amplified using the ITS4/ITS5 (5), T1/T2 (2), and EF1/EF2 (3) primer pairs. These sequences were deposited in GenBank (KF515650 for the ITS region; KF747330 for β-tubulin region, and KF747331 for EF-1α region) and showed 98 to 99% identity to F. equiseti strains (HQ332532 for ITS region, JX241676 for β-tubulin gene, and GQ505666 for EF-1α region). According to both morphological and sequences analysis, the pathogen of the black stem of A. marina was identified as F. equiseti. Similar symptoms on absorbing rootlets and trunks of A. marina had been reported in central coastal Queensland, but the pathogen was identified as Phytophthora sp. (4). Therefore, the disease reported in this paper differs from that reported in central coastal Queensland. To our knowledge, this is the first report of black stems of A. marina caused by F. equiseti in China. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual, 1st ed. Wiley-Blackwell, Hoboken, NJ, 2006. (2) K. O'Donnell and E. Cigelnik. Mol. Phylogenet. Evol. 7:103, 1997. (3) K. O'Donnell et al. Proc. Natl. Acad. Sci. USA. 95:2044, 1998. (4) K. G. Pegg. Aust et al. Plant Pathol. 3:6, 1980. (5) A. W. Zhang et al. Plant Dis. 81:1143, 1997.


Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 429-429
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

Lupinus polyphyllus (Leguminosae), Washington lupine, is a perennial herbaceous plant. In March 2008, in a campus greenhouse at the University of Torino, Grugliasco (northern Italy), a leaf blight was observed on 20% of potted 30-day-old plants. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Severely infected plants died. Plants were grown in a sphagnum peat/perlite/clay (70:20:10) substrate at temperatures between 18 and 25°C and relative humidity of 60 to 80%. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani (4) was consistently and readily recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 20 ± 1°C appeared light brown, rather compact, and exhibited radial growth. The isolates of R. solani successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (3). Pairings were also made with tester isolates AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 660-bp fragment showed 100% homology with the sequence of R. solani. The nucleotide sequence has been assigned GenBank Accession No. FJ486272. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old L. polyphyllus were grown in 10-liter containers (10 plants per container) on a steam disinfested sphagnum peat/perlite/clay (70:20:10) medium. Inoculum, consisting of an aqueous suspension of mycelium disks prepared from PDA cultures (5 g of mycelium per plant), was placed at the collar of plants. Plants inoculated with water and PDA fragments alone served as control treatments. Three replicates were used. Plants were maintained in a greenhouse at temperatures between 18 and 23°C. First symptoms, similar to those observed in the nursery, developed 10 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was repeated twice. The susceptibility of L. polyphyllus to R. solani was reported in Poland (2). This is, to our knowledge, the first report of leaf blight of L. polyphyllus caused by R. solani in Italy. The importance of the disease is at the moment limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) W. Blaszczak. Rocz. Nauk. Roln. Ser A 85:705, 1962. (3) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (4) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.


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