• Title/Summary/Keyword: rDNA ITS region

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Phylogenetic Relationships of the Mutualistic Fungi Associated with Macrotermes subhyalinus in Oman

  • Hilal S. AlShamakhi;Abdullah M. Al-Sadi;Lyn G. Cook
    • Mycobiology
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    • v.51 no.5
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    • pp.281-287
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    • 2023
  • The symbiotic association between fungus-gardening termites Macrotermes and its fungal symbiont has a moderate degree of specificity-although the symbiotic fungi (Termitomyces) form a monophyletic clade, there is not a one-to-one association between termite species and their fungus-garden associates. Here, we aim to determine the origin and phylogenetic relationships of Termitomyces in Oman. We used sequences of the internal transcribed spacer region (ITS) and the nuclear large subunit ribosomal RNA (LSU rRNA, 25S) gene and analyzed these with sequences of Termitomyces from other geographic areas. We find no evidence for more than a single colonization of Oman by Termitomyces. Unexpectedly, we find Termitomyces in Oman is most closely related to the symbiont of M. subhyalinus in West Africa rather than to those of geographically closer populations in East Africa.

The Incidence of Alternaria Species Associated with Infected Sesamum indicum L. Seeds from Fields of the Punjab, Pakistan

  • Nayyar, Brian Gagosh;Woodward, Steve;Mur, Luis A.J.;Akram, Abida;Arshad, Muhammad;Naqvi, S.M. Saqlan;Akhund, Shaista
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.543-553
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    • 2017
  • Sesame (Sesamum indicum) is an important oil seed crop of Asia. Yields can be negatively impacted by various factors, including disease, particularly those caused by fungi which create problems in both production and storage. Foliar diseases of sesame such as Alternaria leaf blight may cause significant yield losses, with reductions in plant health and seed quality. The work reported here determined the incidence of Alternaria species infecting sesame seeds grown in the Punjab, Pakistan. A total of 428 Alternaria isolates were obtained from 105 seed samples and grouped into 36 distinct taxonomic groups based on growth pattern and morphological characters. Isolation frequency and relative density of surface sterilized and non-surface sterilized seeds showed that three isolates (A13, A47 and A215) were the most common morphological groups present. These isolates were further identified using sequencing of the Internal Transcribed Spacer (ITS) region of ribosomal DNA (rDNA) and the Alternaria major allergen gene (Alt a 1). Whilst ITS of rDNA did not resolve the isolates into Alternaria species, the Alt a 1 sequences exhibited > 99% homology with Alternaria alternata (KP123850.1) in GenBank accessions. The pathogenicity and virulence of these isolates of Alternaria alternata was confirmed in inoculations of sesame plants resulting in typical symptoms of leaf blight disease. This work confirms the identity of a major source of sesame leaf blight in Pakistan which will aid in formulating effective disease management strategies.

Phylogenetic Analysis on Wild Cordyceps Collected from Miryang Region of South Korea (밀양근교에서 채집한 야생 동충하초 계통의 PCR 산물에 근거한 계통 유전학적 연구)

  • Park, Hyeancheal;Lee, Sangmong;Park, Namsook
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.1-16
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    • 2021
  • The phylogenetic relationships among thirty-two strains (P1~P32; including Cordyceps sp., Paecilomyces sp., Beauveria sp., Aranthomyces sp., Isaria sp. and Himenostilbe sp.) in Miryang region located in the southern part of Korea, were investigated based on internal transcribed spacer (ITS) sequences of ribosomal DNA. A fragment of ITS region was amplified by polymerase chain reaction (PCR) using the specific primer pairs ITS1 and ITS4. After obtained same size of PCR products from various strains, we cloned them into a pGEM-T easy vector to determine their sequences. BLAST analyses of the nucleotide sequence ITS1, 5.8S and ITS2 gene fragments revealed the identity and their phylogenetic relationship. Among 32 strains isolated from Miryang region, Cordyceps militaris was shared 100% sequences with Genbank (AY49191, EU825999, AY491992), while some species are not shared perfectly with reported sequences. For example, strain P17 (P. tenuipes in Ulju-gun Gaji Mountain) has some differences among the other strains of P. tenuipes (Miryang-si Jocheon-eup, Miryang-si Gaji Mountain) and those of gene bank. We conclude that ITS analyses with strains in the suburbs of Miryang in this study can be effectively used as a tool for classification, evaluation and collection of the natural eco-type genetic resources.

Occurrence of Gray Mold Caused by Botrytis cinerea on Common Fig in Korea (Botrytis cinerea에 의한 무화과 잿빛곰팡이병)

  • Cheong, Seong-Soo;Choi, In-Young;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.38-41
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    • 2013
  • In 2010 and 2011, gray mold was found on common fig (Ficus carica) fruit grown at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. Gray mold symptoms on common fig fruit mainly occurred after harvest season until December. The typical symptom included brown water-soaked rot and fruit decay. The diseased fruit was covered by gray to brown colored conidiophore and conidia. The conidiophores were tree shape and measured $15-33{\times}2{\mu}m$. Conidia on conidiophore were ellipsoidal or lemon shape, colorless, single cell, and measured $7.3-14.6{\times}6.8-11.1{\mu}m$. The nucleotide sequences of the rDNA ITS region obtained from the pure culture of the gray mold on common fig were 100% similar to the sequences of the GenBank accession number HQ171052, EU519210, HQ171053, FN812726, HM849615, and EU563120 of B. cinerea isolates. In phylogenetic tree, the representative isolate was placed within same clade of B. cinerea. Based on the morphological characteristics and analysis of rDNA ITS sequence data, the fungus was identified as B. cinerea.

New Records of Fungi Isolated from Indoor Air of Greenhouse Used for Shiitake Cultivation in Korea (표고 재배사 실내 공기에서 분리한 국내 미기록 진균)

  • Kwon, Hyuk Woo;Yun, Yeo Hong;Kim, Jun Young;Kim, Seong Hwan;Ko, Han Kyu
    • The Korean Journal of Mycology
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    • v.43 no.1
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    • pp.58-63
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    • 2015
  • Mold contamination is one of the detrimental factors affecting sawdust media-based shiitake cultivation in greenhouses. During mold monitoring of indoor air of greenhouses, several fungi were isolated. Among them, Aspergillus pulverulentus and Cosmospora butyri were found to be new in Korea and Lecanicillium psalliotae and L. antillanum were known in Korea without taxonomic validation. In this study the morphological characteristics and phylogenetic analysis based on the internal transcribed spacer (ITS) rDNA region or ${\beta}$-tubulin gene of the four identified species were described.

Unrecorded fungi isolated from indoor air of cultivation houses used for field test of a newly bred domestic shiitake cultivar (표고 현장적응 시험 버섯 재배사내 공기에서 검출한 국내 미기록 진균 보고)

  • Ahn, Geum Ran;Ahn, Hong Seok;Kwon, Hyuk Woo;Ko, Han Gyu;Kim, Seong Hwan
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.168-173
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    • 2016
  • Four fungal species, during indoor air monitoring for fungi that possibly affect the field testing of a newly bred shiitake cultivar in cultivation houses located in Cheongyang, Chungnam Province and Jangheung, Jeonnam Province. Of these species, two are known to be plant pathogens and the other two are saprobes but potent contaminators in the mushroom cultivation environment. This study reports the morphological characteristics and phylogenetic relationships of these four species based on nucleotide sequences of the internal transcribed spacer (ITS) and 18S rDNA region, including their known information.

Complete Chloroplast Genome Sequence of Korean Endermic Species, Pseudostellaria longipedicellata

  • Kim, Yongsung;Heo, Kyeong-In;Lee, Sangtae;Park, Jongsun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.40-40
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    • 2018
  • Pseudostellaria Pax (Caryophyllaceae) is a small genus distributed in temperate region. It consists of 25 species presenting high diversity in Asia. Pseudostellaria longipedicellata S. Lee, K. Heo & S. C. Kim was first announced as new species in 2012. Morphological characters of P. longipedicellata are closely related to those of Psedusotellaria palibiniana and Psedusotellaria okmotoi. These are distinguished from P. longipedicellata by shorter pedicel and puberulent pedicels, respectively and by being distributed allopatically between P. longipedicellata and rest of species. The complete chloroplast genome of P. longipedicellata was successfully rescued from raw reads generated by HiSeq2000. Its total length is 149,626 bp consisting of four regions: large single copy (LSC) region (81,292 bp), small single copy (SSC) region (16,984bp), and inverted repeats (IRs; 25,765 bp per each). It contained 126 genes (81 coding DNA sequence (CDS), eight rRNAs, and 37 tRNAs); 18 genes (seven CDS, four rRNAs, and seven tRNAs) are duplicated in inverted repeat regions. The overall GC content of P. longipedicellata is 36.5% and in the LSC, SSC, and IR regions were 34.3%, 29.3%, and 42.4%, respectively. Based on phylogenetic analysis of chloroplast genomes of P. longipedicellata and relatives species presents clear phylogenetic positions of Pseudostellaria genus. This chloroplast genome will be an important sequence resources for further researches of Pseudostellaria genus.

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Isolation and Molecular Characterization of a New CRT Binding Factor Gene from Capsella bursa-pastoris

  • Wang, Xinglong;Liu, Li;Liu, Sixiu;Sun, Xiaoqing;Deng, Zhongxiang;Pi, Yan;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.37 no.5
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    • pp.538-545
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    • 2004
  • A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.

Occurrence of Sclerotium Rot of Cucumber Caused by Sclerotium rolfsii (Sclerotium rolfsii에 의한 오이 흰비단병 발생)

  • Kwon, Jin-Hyeuk;Lee, Sang-Dae;Choi, Okryun;Shen, Shun-Shan;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.229-232
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    • 2013
  • Sclerotium rot of cucumber (Cucumis sativus L.) occurred at the experimental field of Gyeongsangnam-do Agricultural Research and Extension Services in July 2012. The typical symptoms included wilt, rot, and water-soaking on stems and fruits and severely infected plants eventually died. White mycelial mats spread over lesions, and then sclerotia were formed on fruit and near soil line. The sclerotia were globoid in shape, white to brown in color and 1-3 mm in size and the hyphal width was 4-8 ${\mu}m$. The optimum temperature for mycelial growth and sclerotia formation on PDA was $30^{\circ}C$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. For further identification, the complete internal transcribed spacer (ITS) rDNA region was amplified and sequenced. On the basis of mycological characteristics, ITS rDNA region comparison, and pathogenicity to host plants, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report of sclerotium rot on cucumber caused by S. rolfsii in Korea.

Ribosomal Intergenic Spacer 1 Based Characterization of Button Mushroom (Agaricus bisporus) Strains

  • Kwon, Hyuk Woo;Choi, Min Ah;Kim, Dae Wook;Oh, Youn-Lee;Hyun, Min Woo;Kong, Won-Sik;Kim, Seong Hwan
    • Mycobiology
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    • v.44 no.4
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    • pp.314-318
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    • 2016
  • Breeding the button mushroom requires genetic information about its strains. This study was undertaken to genetically characterize four domestically bred button mushroom strains (Saea, Saejung, Saedo, Saeyeon cultivars) and to assess the possibility of using the intergenic spacer 1 (IGS1) region of rDNA as a genetically variable region in the genetic characterization. For the experiment, 34 strains of Agaricus bisporus, two strains of A. bitorquis, and one strain of A. silvaticus, from 17 countries were used. Nucleotide sequence analysis of IGS1 rDNA in these 37 Agaricus strains confirmed that genetic variations exist, not only among the four domestic strains, but also between the four domestic strains and foreign strains. Crossing two different haploid strains of A. bisporus seems to generate genetic variation in the IGS1 region in their off-spring haploid strains. Phylogenetic analysis based on the IGS1 sequence revealed all A. bisporus strains could be differentiated from A. silvaticus and A. bitorquis strains. Five genetic groups were resolved among A. bisporus strains. Saejung and Saeyeon cultivars formed a separate genetic group. Our results suggest that IGS1 could be complementarily applied in the polymorphism analysis of button mushroom.