scholarly journals New Anastomosis Groups, AG-T and AG-U, of Binucleate Rhizoctonia spp. Causing Root and Stem Rot of Cut-Flower and Miniature Roses

2005 ◽  
Vol 95 (7) ◽  
pp. 784-792 ◽  
Author(s):  
Mitsuro Hyakumachi ◽  
Achmadi Priyatmojo ◽  
Mayumi Kubota ◽  
Hirokazu Fukui
Keyword(s):  
Sequence Data ◽  
Its Region ◽  
Restriction Enzymes ◽  
Anastomosis Group ◽  
Stem Rot ◽  
Cut Flower ◽  
Binucleate Rhizoctonia ◽  
Rdna Its ◽  
Rdna Its Region ◽  
Hyphal Anastomosis

Root and stem rot of cut-flower roses (Rosa spp.) was observed in commercial glasshouse-grown roses in 10 prefectures of Japan from 1998 through 2001. Binucleate-like Rhizoctonia spp. were isolated mainly from the disease plants. In all, 670 isolates were divided into two types based on cultural appearance; 168 isolates of light brown to brown type and 502 isolates of whitish type. A hyphal anastomosis reaction using representative isolates from each type revealed that the light brown to brown type belonged to anastomosis group G (AG-G), whereas the whitish type (AG-CUT) failed to anastomose with tester strains of binucleate Rhizoctonia AG-A through AG-S. Neither isolates of AG-G nor AG-CUT anastomosed with tester strains of a previously reported unknown AG (AG-MIN) of binucleate Rhizoctonia spp. collected from miniature roses. In pathogenicity tests, randomly selected isolates of the three groups caused root and stem rot on cut-flower and miniature roses. To differentiate AG-CUT and AG-MIN from known AGs of binucleate Rhizoctonia spp., restriction fragment length polymorphism (RFLP) and sequence analyses of a ribosomal (r)DNA internal transcribed spacer (ITS) region were conducted. Among the eight restriction enzymes used, HaeIII produced DNA banding patterns for AG-CUT that differed from those of tester strains and AG-MIN. Additionally, restriction profiles of AG-MIN differed from those of all tester strains. AG-G isolates from cut-flower roses had the same RFLP pattern as the tester strains of AG-G. Based on the results of hyphal anastomosis and RFLP and sequence analysis of an rDNA-ITS region, we propose that AG-CUT be designated AG-T and AG-MIN be designated AG-U, two new AGs of binucleate Rhizoctonia spp. The phylogenetic tree based on the sequence data of the rDNA-ITS region showed that isolates of AG-MIN were in a distinct clade from other AGs, whereas isolates of AG-CUT were in the same clade as those of AG-A. More detailed phylogenetic analysis besides rDNA-ITS region might be necessary for AG classification of binucleate Rhizoctonia spp.

scholarly journals A New Subgroup of Rhizoctonia AG-D, AG-D III, Obtained from Japanese Zoysia Grass Exhibiting Symptoms of a New Disease

Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1389-1394 ◽  
Author(s):  
Toshihiro Hayakawa ◽  
Takeshi Toda ◽  
Qu Ping ◽  
Joseph M. Mghalu ◽  
Shigeharu Yaguchi ◽  
...  
Keyword(s):  
Growth Rate ◽  
Phylogenetic Trees ◽  
Its Region ◽  
Hyphal Growth ◽  
Anastomosis Group ◽  
Hyphal Fusion ◽  
Rdna Its ◽  
Light Yellow ◽  
Different Temperatures ◽  
Rdna Its Region

Isolates of an unidentified Rhizoctonia sp. (UN isolates) were obtained from Japanese zoysia grass (Zoysia japonica Steud) that exhibited symptoms of a new sheath rot disease. UN isolates were binucleate and showed hyphal fusion with tester isolates of Rhizoctonia anastomosis group (AG)-D. Those isolates were compared with isolates of subgroups I and II of Rhizoctonia AG-D based on cultural morphology, hyphal growth rate at different temperatures, anastomosis frequency, pathogenicity, and sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA genes (rDNA-ITS region). The mycelial color of UN isolates was light yellow which differs from AG-D I but is similar to AG-D II. Sclerotia of UN isolates were dark brown in color and larger in size (1 to 3 mm in diameter) than those of AG-D subgroup I (1 mm in diameter), whereas isolates of AG-D II produced white mycelial clamps 4 to 5 mm in size. Hyphal growth rate of UN isolates was slower than that of two AG-D subgroups at several temperatures, especially 25°C. In pathogenicity tests on Japanese zoysia grass, UN isolates showed moderate disease severity and lower pathogenicity than isolates of AG-D subgroups I and II. Sequences of the rDNA-ITS region within UN isolates were almost homologous, but had lower homology with subgroups AG-D I or II. Phylogenetic trees constructed using ITS sequences showed that UN isolates formed an individual cluster that differed from the clusters of the two subgroups. We propose that UN isolates are a new subgroup of Rhizoctonia AG-D, subgroup III, and the name of the disease is “spring-rot” on Japanese zoysia grass.

scholarly journals First Report of Binucleate Rhizoctonia AG-L Causing Root and Stem Rot of Wishbone Flower (Torenia fournieri) in Taiwan

Plant Disease ◽  
2021 ◽  
Author(s):  
Chang Xian Chen ◽  
Yea-Fang Wu ◽  
Hsi Huan Gong ◽  
Yi Jyun Lin ◽  
Chi Yu Chen
Keyword(s):  
Anastomosis Group ◽  
Stem Rot ◽  
Its Sequence ◽  
Pathogenicity Test ◽  
Torenia Fournieri ◽  
Agar Block ◽  
Potato Dextrose Broth ◽  
First Report ◽  
Binucleate Rhizoctonia ◽  
Rdna Its

Wishbone flower (Torenia fournieri L.) is a common ornamental plant for flower bed in Taiwan. In August 2018, root and stem rot of wishbone flower occurred on the flower bed in the campus of National Chung Hsing University, Taichung city, with 100% incidence. Symptoms were dark brown discoloration of basal stems and brown necrosis of roots. Lesions from base of stems were excised into 5 mm long fragments, which were then surface sterilized in 1% sodium hydrochloride for 1 min, rinsed in sterile distilled water, dried on filter paper and thereafter placed onto 2% water agar. After 24 h, hyphae characteristic of Rhizoctonia (Sneh et al. 1991) appeared and dominated in every isolation. Hyphal tips were transferred to potato dextrose agar (PDA). After 5 days of incubation at 28°C, characteristic brown colonies of Rhizoctonia (Sneh et al. 1991) were observed. Hyphal width was 4.29±0.52 μm. No sclerotia were visibly present after 21 days of growth on PDA at 28°C. Hyphae were stained by 0.3% safranin-O and 1% KOH. There were two nuclei in each hyphal compartment, suggesting a binucleate Rhizoctonia fungus. ITS sequence has been used as the best tool to identify specific anastomosis group (AG) of Rhizoctonia as shown by Sharon et al. (2006, 2008). ITS sequence was amplified using the primers Bd1a and ITS4 (Goka et al. 2009; White et al. 1990). Blast search analysis of this acquired sequence (acc. no. LC498494) revealed the highest similarity (98.75 to 99.83%) with the reference sequences (acc. nos. AB286934, AB286933, and AB196653) of binucleate Rhizoctonia AG-L, namely Ceratobasidium sp. AG-L. Pathogenicity test was carried out using seedlings of 4-week-old wishbone flower each grown in a pot of 6.35 cm diameter. To prepare the inoculum, a PDA agar block (6 mm in diameter) excised from the growing front of 5-day-old colony was transferred into a flask with 200 ml of potato dextrose broth (PDB) incubated in a shaker at 26°C and 120 rpm for 6 days. The PDB broth was then blended into slurry. Ten pots each with a seedling were inoculated by pouring 50 ml of slurry onto the potting medium. Five pots were served as the controls by pouring PDB only. Pots were maintained in a greenhouse (26 to 33°C). Three days after inoculation, all inoculated plants exhibited symptom of root and stem rot. The same fungus was reisolated and confirmed by sequencing rDNA-ITS. This is the first report of root and stem rot of wishbone flower caused by binucleate Rhizoctonia AG-L in Taiwan and in the world. Although this is the second cases, since Wang and Hsieh (1993), for binucleate Rhizoctonia AG-L to be pathogenic, this study shows that this fungus has the potential to cause damages and is worth of further investigations.

scholarly journals First report of Rhizoctonia solani Kühn AG 2-2 LP in Zoyzia japonica Steud in Brazil

2020 ◽  
Vol 46 (4) ◽  
pp. 289-298
Author(s):  
Maria Aurea Saboya Chiaradia Picarelli ◽  
Flavia Rodrigues Alves Patricio ◽  
Ricardo Harakava ◽  
Eliana Borges Rivas ◽  
Addolorata Colariccio
Keyword(s):  
Rhizoctonia Solani ◽  
Mycelial Growth ◽  
Its Region ◽  
Anastomosis Group ◽  
Large Patch ◽  
First Report ◽  
Rdna Its ◽  
Rdna Its Region ◽  
Important Disease

ABSTRACT The use of cultivated grasses in Brazil has grown by 40% between 2010 and 2015, and the species Zoysia japonica Steud, especially the cultivar ‘Esmeralda’, corresponds to 81% of the grass market in the country. The most important disease affecting zoysia grass, known as large patch, is caused by Rhizoctonia solani and occurs in the Brazilian lawns particularly during winter months. The aim of this study was to contribute to the identification and characterization of the anastomosis group of R. solani isolates from lesions typical of large patch collected from ‘Esmeralda’ grass at gardens and golf courses in the states of São Paulo and Bahia, Brazil. All 12 obtained isolates presented dark-brown colonies with aerial mycelial growth, multinucleated hyphae and absence of concentric zonation or sclerotia, and showed their greatest mycelial growth rate at 25°C. In pathogenicity experiments, except three out of R. solani isolates, reduced the growth of zoysia grass. Based on the analysis of sequences of the rDNA-ITS region, the isolates clustered with reference isolates of the anastomosis group AG 2-2 LP. Phylogenetic inference showed that the Brazilian isolates are grouped into two clades that shared the same common ancestral with 96% bootstrap. One of the clades includes only Brazilian isolates while the other one also includes American and Japanese R. solani isolates AG 2-2 LP. This is the first report and characterization of R. solani AG 2-2 LP in zoysiagrass in Brazil.

Morphological, pathogenic and molecular characterization of Phoma spp. isolated from onion field soils in Puerto Rico.

1969 ◽  
Vol 92 (1-2) ◽  
pp. 73-86 ◽  
Author(s):  
Ismael E. Badillo-Vargas ◽  
Lydia I. Rivera-Vargas ◽  
Juan Calle-Bellido
Keyword(s):  
Puerto Rico ◽  
Disease Incidence ◽  
Its Region ◽  
Restriction Enzymes ◽  
Molecular Characteristics ◽  
Base Pairs ◽  
Rdna Its ◽  
Field Soils ◽  
Rdna Its Region

Thirteen Phoma spp. isolates collected during a survey conducted in onion field soils in Santa Isabel, Puerto Rico, were examined on the basis of morphology, pathogenicity and molecular characteristics. Twelve isolates were identified as Phoma putaminum Speg. and one isolate as an atypical Phoma macrostoma var. incolorata (section Phyllostictoides).This is the first report of P. putaminum and P. macrostoma var. incolorata for Puerto Rico and the Caribbean. In vitro, Phoma putaminum isolates were pathogenic to onion cvs. Mercedes and Excalibur, resulting in necrosis of young bulbs and roots seven days after inoculation. Disease incidence caused by P. putaminum was higher in cultivar Mercedes, ranging from 75 to 100%, than in P. macrostoma var. incolorata (0 to 25%). No symptoms were observed on cultivar Excalibur inoculated with P. macrostoma var. incolorata or on control plants. In vitro inoculations of commercial onion bulbs and field inoculations of roots failed to reproduce symptoms, thus showing that P. putaminum and P. macrostoma var. incolorata are weak pathogens. Sequence size of the nuclear internal transcribed spacer (ITS) of ribosomal DNA (rDNA) gene ranged from 458 to 610 base pairs (bp) for P. putaminum and was 456 bp for P. macrostoma var. incolorata isolate. Analysis of rDNA ITS region by PCR-RFLP showed that restriction enzyme, Hindlll, among other restriction enzymes evaluated (Alul, EcoRI, Clal and Seal), differentiate between P. putaminum and P. macrostoma var. incolorata isolates.

scholarly journals PCR-RFLP ITS-based analysis of wine yeast autochthonous strains isolated from different grape cultivars in Taman subregion

2021 ◽  
Vol 285 ◽  
pp. 05020
Author(s):  
Elena Lobodina ◽  
Ivan Suprun ◽  
Natalya Ageeva ◽  
Ekaterina Al-Nakib
Keyword(s):  
Genetic Analysis ◽  
Restriction Analysis ◽  
Its Region ◽  
Wine Yeast ◽  
Product Size ◽  
Rdna Its ◽  
Hanseniaspora Uvarum ◽  
Pcr Rflp ◽  
Autochthonous Yeast ◽  
Rdna Its Region

The studies present the results of morphological, cultural and genetic analysis of the ITS1-ITS4 region of the autochthonous yeast strains genome by using the HaeIII restriction enzyme. On the red and white grapes varieties, based on the morphology of the cells, autochthonous strains belonging to the genus Saccharomyces prevail – 83.3%, what is confirmed by genetic analysis of rDNA ITS region. Restriction analysis showed that all strains of the genus Saccharomyces belong to the species Saccharomyces cerevisiae / S. paradoxus. The percentage of Saccharomyces isolated on the Pervenets Magaracha variety is 86.7%, Krasnostop Anapsky - 80%. The non-Saccharomyces yeast had a product size of 750 bp, presumably of the species Hanseniaspora uvarum.

scholarly journals Phylogenetic Analysis and Molecular Diversity of Capsicum Based on rDNA-ITS Region

Horticulturae ◽  
2020 ◽  
Vol 6 (4) ◽  
pp. 87
Author(s):  
Kumpei Shiragaki ◽  
Shuji Yokoi ◽  
Takahiro Tezuka
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
Wild Species ◽  
Morphological Characteristics ◽  
Its Region ◽  
Internal Transcribed Spacers ◽  
Nuclear Ribosomal Dna ◽  
Rdna Its ◽  
Rdna Its Region ◽  
Genus Capsicum

The genus Capsicum is comprised of 5 domesticated and more than 30 wild species. The region of nuclear ribosomal DNA internal transcribed spacers (rDNA-ITS) has widely been used for species identification, but has rarely been used in Capsicum. In this study, the evaluation of genetic diversity and a phylogenetic analysis were conducted using rDNA-ITS of 28 Capsicum accessions, including five domesticated and two wild species. We surveyed six conventional keys of domesticated species and another five traits in Capsicum accessions. Specific morphological characteristics were found in C. annuum, C. baccatum, and C.pubescens. Three subclones of each accession were sequenced, and rDNA-ITS polymorphisms were detected in all accessions excluding C. annuum, suggesting that incomplete concerted evolution occurred in rDNA-ITS of Capsicum. The genetic diversity was evaluated using nucleotide polymorphism and diversity. C. annuum had the lowest genetic diversity of all species in this study. The phylogenetic tree formed a species-specific clade for C. annuum, C. baccatum, and C. pubescens. The C. chinense clade existed in the C. frutescens clade, implying that it was a cultivated variant of C. frutescens. C. chacoense likely belonged to the C. baccatum complex according to its morphologic and genetic features. This study indicated that the rDNA-ITS region can be used for simple identification of domesticated Capsicum species.

scholarly journals Molecular identification and artificial cultivation of a wild isolate of oyster mushroom in Albania

2016 ◽  
Vol 10 (1s) ◽  
pp. 35
Author(s):  
Jordan Merkuri ◽  
Stefania Mirela Mang ◽  
Ippolito Camele ◽  
Magdalena Cara ◽  
Gian Luigi Rana
Keyword(s):  
Its Region ◽  
Edible Mushroom ◽  
Biological Efficiency ◽  
Potato Dextrose Agar Medium ◽  
Wild Isolate ◽  
Rdna Its ◽  
Pure Cultures ◽  
Rdna Its Region ◽  
And Diffusion ◽  
Artificial Cultivation

Basidiomata of a wild mushroom macroscopically recognised as <em>Pleurotus ostreatus</em> were observed on an oak trunk in a mixed wood of northern Albania. Pure cultures of the fungus were then obtained on potato-dextrose-agar medium. Molecular analyses of genomic DNA of the fungus confirmed its identification. The rDNA ITS region nucleotide sequence of the studied <em>Pleurotacea</em> matched at 99% those of two <em>P. ostreatus</em> strains already present in NCBI GenBank database. The rDNA ITS nucelotide sequences of two pure cultures of the Albanian <em>P. ostreatus</em> were deposited in EMBL database under the accession numbers LN849458 and LN849459. One of the fungus isolates was subsequently cultivated under protected and semi-natural conditions. Productivity and biological efficiency of the Albanian <em>P. ostreatus</em> ranged from about 10% to 16% and from 33 to 53.33%, respectively. This seems to be the first report on the artificial cultivation of <em>P. ostreatus</em> in Albania and could have, in the next future, a high economic impact on development and diffusion of this important edible mushroom over the country.

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