AJMR- 20 September, 2012 Issue

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Volume 6 Number 36 20 September, 2012

African Journal of

Microbiology ResearchISSN 1996-0808


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University of Mauritius

Reduit

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Luki Subehi

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International Journal of Medicine and Medical SciencesAfrican Journal of Microbiology Research

ARTICLES

Research Articles

Conventional and molecular characterization of Trichophyton rubrum 6502

Farzad Aala, Rosimah Nulit, Umi Kalsom Yusuf and Sassan Rezaie

Inhibitory effect of some plant extracts on clinical isolates of Staphylococcus

aureus 6517

Rajaa Milyani and Nahed Ashy

Role of CSE1034 in bacterial lipids and polysaccharides involved in biofilm

formation: a comparison with other drugs 6525

Chaudhary Manu and Anurag Payasi

Analytical specificity and sensitivity determination of 16SrRNA gene based

diagnostic polymerase chain reaction (PCR) for molecular detection of

Coxiella burnetii 6532

Mohammad Soleimani, Keivan Majidzadeh A., Amirhossein Mohseni and

Mohammad Khalili

Effect of Bacillus cereus Br on bacterial community and gossypol content

during fermentation in cottonseed meal 6537

Xin Wang, Jiang-wu Tang, Xiao-hong Yao, Yi-fei Wu, Hong Sun and Yao-xing Xu

Identification and characterization of a fungal strain with lignin and cellulose

hydrolysis activities 6545Ran Jin, Hongdong Liao, Xuanming Liu, Mang Zheng, Xianqiu Xiong, Xinwu Liu,

Liyong Zhang and Yonghua Zhu

Table of Contents: Volume 6 Number 36 20 September, 2012


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ARTICLES

Influence of ciprofloxacin on glioma cell line GL26: A new application for

an old antibiotic

Abdolreza Esmaeilzadeh, Massoumeh Ebtekar, Alireza Biglari and

Zuhair Mohammad Hassan 4891

Identification of microbial diversity in caecal content of broiler chicken

S. Nathiya, G. Dhinakar Raj, A. Rajasekar, D. Vijayalakshmi and T. Devasena 4897

Microbial quality of some non-sterile pharmaceutical products sourced

from some retail pharmacies in Lagos, Nigeria

Adeola Anifowoshe R., Opara Morrison I. and Adeleye Isaac A. 4903

Molecular detection of adhesins genes and biofilm formation in methicillin

resistant Staphylococcus aureus

Karima BEKIR, Omayma HADDAD, Mohammed GRISSA, Kamel CHAIEB,

Amina BAKHROUF and Salem IBRAHIM ELGARSSDI 4908

Amylase production by moderately halophilic Bacillus cereus in solid

state fermentation

P. Vijayabaskar, D. Jayalakshmi and T. Shankar 4918

Networking clusters and sequence characteristics of clustered regularly

interspaced short palindromic repeats (CRISPR) direct repeats and their

evolutionary comparison with cas1 genes in lactic acid bacteria

Kaibo Deng, Fei Liu, Chuntao Gu and Guicheng Huo 4927

Table of Content: Volume 6 Number 23 21 June, 2012

ARTICLES

DNA viral infections and transient bone marrow failure in southern Iran 6551

Kambiz Bagheri, Mohammad Hossein Karimi, Ramin Yaghobi,

Behnam Mohammadi, Mehdi Dehghani and Padideh Ebadi

Survival of microorganisms in high pressure treated minced meat during

chilled storage and at pH and temperature mimicking gastrointestinal tract 6558

Sami Bulut

Efficacy and toxicity of neutralizers against disinfectants and antiseptics

used in vaccine production facility 6565

Norhan S. Sheraba, Aymen S. Yassin, Aly Fahmy and Magdy A. Amin

Effects of essential oil extracted from Citrullus colocynthis (CCT) seeds on

growth of phytopathogenic bacteria 6572

Zahra Setayesh Mehr, Nima Sanadgol and LeylaVafadar Ghasemi

Development and evaluation of a novel TaqMan fluorescence probe-based

real-time reverse transcriptase polymerase chain reaction assay for

detection and quantification of West Nile virus 6576

Lijun Shi, Huiqiong Yin, Jingang Zhang, Zhan-zhong Zhao and Gang Li

Microbial water quality in the upper Olifants River catchment: Implications

for health 6580

W. J. le Roux, L. M. Schaefer and B. Genthe

Partial characterization of a bacteriocin produced by Lactobacillus alivarius

isolated from oral cavity of desert foxes 6589

Aly E. Abo-Amer and Mohammed Y. Shobrak

Table of Contents: Volume 6 Number 36 20 September, 2012


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African Journal of Microbiology Research Vol. 6(36), pp. 6502-6516, 20 September, 2012 Available online at http://www.academicjournals.org/AJMRDOI: 10.5897/AJMR10.736ISSN 1996-0808 ©2012 Academic Journals

Full Length Research Paper

Conventional and molecular characterization ofTr ichophy ton rubrum

Farzad Aala1*, Rosimah Nulit2, Umi Kalsom Yusuf 2 and Sassan Rezaie3

1Department of Medical Mycology and Parasitology, School of Medicine, Kurdistan University of Medical Sciences,

Sanandaj, Iran.2Department of Biology, Faculty of Science; Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.

3Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical

Sciences, Tehran, Iran.

Accepted 10 September, 2012

Different studies illustrated that Trichophyton rubrum , among all species of Trichophyton , is the mostprevalent and consequently the most important genus. T. rub rum as a worldwide filamentous pathogenfungus can infect human keratinized tissue (skin, nails and rarely hair), and causes dermatophytosis.Researchers use two general methods for the identification of dermatophytes namely, conventionalmethods on the basis of phenotype variations and molecular methods on the basis of moleculardifferences. Due to some limitations in traditional methods, in the recent years, molecular biologicalmethods are regarded as useful in the exact and rapid recognition of dermatophytes. The present studyidentified nine clinical isolates and one ATCC as a control strain of T. rubrum by using bothconventional and molecular methods. The molecular systematics method was used to elucidate geneticdiversity among strains of T. rubru m and within Trichophyton species. Morphological characteristics ofall colonies T. rubru m quite varies among each other; we revealed that that conventional methods aregenerally prolonged and may be indecisive. However, molecular studies based on internal transcribed

spacer (ITS) sequencing provides a very accurate result, which is more than 96% the similarity of T.rubrum among all isolates, and more than 90% similarity within Trichophyton spp.

Key words: Trichophyton rubrum, conventional method, internal transcribed spacer (ITS) regions, identification,dermatophytes.

INTRODUCTION

Trichophyton rubrum is one of the most commonlyencountered dermatophytes that infect human keratinizedtissue such as skin, nails and possibly hair. Thispathogen causes well-characterized superficial infections,and also produces skin infections in unusual parts of thebody in immunodepressed patients (Cervelatti et al.,2004). Nearly 80% of onychomycosis due to T . rubrum and 90% of the chronic dermatophyte infections arecaused mostly by T. rubrum, this pathogen developedmechanisms to avoid or suppress cell- mediatedimmunity ((Baeza et al., 2006; Baeza et al., 2007).

*Corresponding author. E-mail: [email protected]. Tel:

+98-9197544944.

Researchers use two general methods for thelaboratory identification of various species ofdermatophytes: a) identification on the basis ofphenotype differences (conventional methods) and bIdentification on the basis of molecular differences. Fagget al. (2001) mentioned that identification odermatophyte species by conventional methods requiresthe examination of colony, particularly with the method oslide culture and microscopic morphological structuresMorphological and physiological features are dynamic. Asa matter of fact, outside factors such as temperaturevariation, medium and chemotherapy, can greatlyinfluence the phenotypic characteristics andconsequently can make the identification more difficult.

Molecular biological methods, in the recent years, areregarded as useful in the exact and rapid recognition o


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dermatophytes. Sequencing of the Internal TranscribedSpacer (ITS) region of the ribosomal DNA, Sequencing ofprotein-encoding genes, Restriction Fragment LenghtPolymorphism (PCR-RFLP) analysis of mitochondrialDNA, Polymerase Chain Reaction (PCR); Random

Amplification of Polymorphic DNA (RAPD), Arbitrarily

Primed PCR [AP-PCR], and PCR fingerprinting are allinstances of molecular techniques which have broughtprominent advance in differentiating between species andstrains (Faggi et al., 2001; Kanbe et al., 2003; Girgis etal., 2006; Yoshida et al., 2006; Li et al., 2007). In therecent years, quite a few molecular studies have beenconducted on the internal transcribed spacer (ITS) regionof the rRNA gene. Sequencing analysis of the ITSregions is considered as a useful tool for phylogeneticdelineation and the identification of dermatophytes(Yoshida et al., 2006; Li et al., 2007).

Even though about 80 to 90% of all isolated are T.rubrum (Brasch and Hipler, 2008), has been isolated toidentify the morphological similarity and the variabilityamong this species, but only a few study has been doneabout the genetic relationship of Trichophyton.

The aim of this work is to identify ten clinical isolates ofT. rubrum by using both conventional methods andmolecular method based on universal fungal primerswhich are internal transcribed spacer 1 (ITS1). T. rubrum(ATCC-10218) was used as a control strain. Themolecular systematics method was used to elucidategenetic diversity among strains of T. rubrum and withinTrichophyton species.

MATERIALS AND METHODS

Isolates

Nine isolates of T. rubrum which are T. rubrum (1138), T. rubrum(1059), T. rubrum (1164), T. rubrum (1208), T. rubrum (1160), T.rubrum (1008), T. rubrum (1298), T. rubrum (1044) and T. rubrum(2970), were obtained from the culture collection of clinical isolatespreserved at the laboratory of Medical Mycology Department inTehran University of Medical Sciences, Iran for study; and T.rubrum (ATCC-10218) was used as a control strain. All clinicalisolates were kept in sterile saline (0.85%) v/v NaCl at 4°C untilrequired for bioassays.

Conventional method

All isolates of T. rubrum were cultured on Sabouraud dextrose agarmedia (Difco Laboratories, Detroit, Michigan) at 28°C for 14 days.Then, slide cultures of isolates were prepared and identified underlight microscope (Carl Zeiss, Germany).

Molecular method

All isolates of T. rubrum maintained on Sabouraud’s dextrose agarmedium and stored at 4°C. Then fungus was cultured in Sabourauddextrose broth, and incubated at 28°C for 14 days. 200 to 300 mgof mycelia was harvested and centrifuged at 1600×g for 10 min,then washed twice with ice-cold sterile phosphate buffered saline

Aala et al. 6503

(PBS) and finally stored at −70°C.

DNA extraction

Fungal genomic DNA from T. rubrum was isolated according toRezaie et al. (2000) with slight modification. 200 to 300 mg omycelia was ground with liquid nitrogen to powder form. 500 μl oDNA extraction buffer (50 mM Tris-HCl pH 8.0), 50 mM EDTA, 25 μ20% SDS, and 10 μl of proteinase-K, was added and mixed gentlyThen, incubated at 65°C for 60 min and centrifuged at 3000×g for15 min. 25 μl Rnase H (10 mg/ml) was added to supernatant andincubated again at 37°C for 30 min. Then mixed with 500 μl ophenol:chloroform:isoamyl alcohol (25:24:1) and and centrifuge a10000×g for 10 min and the supernatant were collected andtransferred to new steril eppendorff tubes. Then mixed again with500 μl of chloroform:isoamyl alcohol (24:1) and centrifuge a10000×g for 10 min, and the supernatant were collected andtransferred to new steril eppendorff tubes. DNA was precipitated byadding 500 μl isopropanol and 30 μl 3 M sodium acetate followedby centrifugation at 15000×g for 30 min and the supernatant wasdiscarded. DNA pellet was rinsed twice or more with 200 μl of 70%cold ethanol and centrifuged at 10000×g for 10 min. The pellet was

air-dry and resuspended DNA pellet in 30 μl of distilled water a37°C for 60 min and stored at -20°C.

PCR amplification

Internal transcribed spacer 1 and 4 (ITS1 and ITS4) (AIT-BiotechSingapore) were designed as ITS1 forward primer is 5’-TCC GTAGGT GAA CCT GC-3’ and the ITS4 reverse primer 5’-TCC TCCGCT TAT TGA TAT G-3’ (Shehata et al., 2008; Yang et al ., 2008 ).

PCR reaction mixtures were prepared in a 25 μl volumecontaining 2.5 μl of 10× reaction buffer, 1.5 μl of 25 mM MgCl2, 0.5μl of 10 mM dNTPs, 0.5 μl of 0.2 mM of each ITS 1 primer and ITS4 primer, 0.5 μl of genomic DNA and 0.5 μl of 1 U Go Taq DNApolymerase (Promega Corporation, USA), and 18.5 μl of distiledwater. PCR reactions were carried out on a thermal cycler (MJ

Research. Inc. USA) with the following conditions: 1 cycle in aninitial step of 94°C for 5 min and then subjected to 30 cyclesconsisting of denaturation at 94°C for 30 s, annealing at 55°C for 40s, and extention at 72°C for 40 s. After the last cycle, this wasfollowed by a final extention step at 72°C for 10 min. Then, 5 μl oPCR product was loaded on 1% agarose in 1X Tris – Acetic Acid –EDTA buffer and stained with 0.5 mg/ml ethidium bromide at 80 Vfor 40 min and visualised with UV transilluminator (Alpha InnotechUSA), compared with a standard DNA size marker; 100 bp DNAladder (Fermenats, USA), and photographed in UV light.

PCR purification

DNA PCR products were purified according to the QIAquick PCR

Purification Kit (Qiagen, Germany) and send for sequencing (1

s

Laboratories, Seri Kembangan, Malaysia).

RESULTS AND DISCUSSION

Morphological characteristics of colonies T. rubrum

This study used both conventional and molecularmethods to diagnose ten isolates of T. rubrum. Studiesrevealed that colonies characterization of all isolatesquite varies among each other. Of these isolates, isolate

http://en.wikipedia.org/wiki/Trichophyton_rubrumhttp://en.wikipedia.org/wiki/Trichophyton_rubrumhttp://en.wikipedia.org/wiki/Trichophyton_rubrumhttp://en.wikipedia.org/wiki/Trichophyton_rubrum


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6504 Afr. J. Microbiol. Res.

numbers 1138 and 1059 are white and cottony or fluffybut isolates number 1164, 1160, 1008, and 1298 arecream, flat and downy, but the others are cream with acarmine and woolly or granular type (isolates numbers1208, 1044, 2970 and 10218) (Figure 1). Themicroscopic features of the isolates also varies, which are

macroconidia and microconidia of isolates numbers,1138, 1008, 1298, 1044, 2970 and 10218 more abundantthan isolates number 1059, 1164, 1208, and 1160.However, the shape of the macroconidia andmicroconidia of all isolates are almost similar, which iscyclindrical to cigar shaped (Figure 1).

Isolation, identification and molecularcharacterization of ITS1 of T. rubrum

Figure 2 showed that ITS1 of all isolates T. rubrum hadbeen amplifed and then were isolated and sequenced.The length of nucleotides sequence of all isolates are notsimilar which is T. rubrum (1138) 658 bp, T. rubrum (1059) 715 bp, T. rubrum (1164) 722 bp, T. rubrum (1208) 713 bp, T. rubrum (1160) 614 bp, T. rubrum (1160) 614 bp, T. rubrum (1008) 719 bp, T. rubrum (1298) 668 bp, T. rubrum (1044) 658 bp, T. rubrum (2970) 660 bp and T. rubrum (ATCC-10218) 633 bp.Nucleotide sequence of all isolates of T. rubrum and

ATCC-10218 are shown in Figure 3. Previous studies byRakeman et al. (2005) and Shehata et al. (2008) alsorevealed that the universal fungal primers amplified theITS regions (ITS1-5.8S-ITS2) of the ribosomal DNAnearly 690 bp for T. rubrum isolates.

Nucleotide sequence of ten isolates of T. rubrum

shown in Figure 3. All nucleotide sequences of T. rubrum isolates were analyzed using online software CLUSTALW(www.Pir.geogetown.edu/pirwww/search/multialn.shtm)to reveal the similarities among isolates. Figure 4 showedthat the similarities among nine isolates of T. rubrum arehigher, which is more than 96% identities.

Nucleotide sequence of isolates T. rubrum wereanalyzed using online software CLUSTALW (www.Pir.geogetown.edu/pirwww/search/multialn.shtm) toreveal the similarities among isolates T. rubrum and otherspecies of Trichophyton which are Trichophytonraubitschekii strain NOMH 789 (GenBank accession no.

AF170469), T. rubrum strain UAMH 8547 (GenBank

accession no. AF170471), T. kanei (GenBank accessionno. AF170460), T. rubrum strain WM 06.348 (GenBankaccession no. EF568093), T. rubrum strain 05-287-3929(GenBank accession no. EU200395), T. rubrum 5.8SrRNA (GenBank accession no. AJ270808), T.soudanense strain UAMH 8548 (GenBank accession no.

AF170474), T. rubrum strain NCPF 295 (GenBankaccession no. EU181449), T. megninii strain ATCC12106 (GenBank accession no. AF170464), and T.rubrum strain ATCC 28188 (GenBank accession no. AF170472). The similarities of all isolates of T. rubrum and

other species of Trichopthyon is also higher than 90% asshown in Figure 5, CLUSTAL 2.0.12 multiple sequencealignment.

DISCUSSION

Traditional method such as investigation of macroscopicand microscopic features of cultures of fungi had beenapplied since early 19

th century. However, these methods

seem to be difficult to amplify due to the polymorphicfeature of these characters, besides increased bydifferences in media compounds, temperature variationsand other variables of cultivation. Furthermore, in somecases, the dermatophytes fail to make reproductiveorganization in culture (sterile mycelia) that makes iimpossible for final identification (Malinovschi et al.2009). Besides that, conventional method is often difficuldue to abnormal microscopic or macroscopic morphology(Li et al., 2008). Currently, molecular studies becomecrucial and necessary for identification of pathogenicfungi (Borman et al., 2008; Malinovschi et al., 2009). Theinternal transcribed spacer (ITS) regions of the fungaribosomal DNA (rDNA) had been used as one oftechniques for species identification becuase it is fasteraccurate species determination, specific, and are lessfeasible to be affected by exterior effects such astemperature changes and chemotherapy (Girgis et al.2006; Kong et al., 2008). Studies revealed thamorphological characteristics of colonies of all isolates Trubrum are similar to T. rubrum isolated from tinea cruristinea pedis, and tinea capitis of human (Graser et al.2000). Colonies of T. rubrum are fluffy to cotonny and

white to cream in colour. Macroconidia are sparse orabundant and microconidia are present in all isolates.

In this studies, the length of ITS1 of all isolates is abou690 bp, 10 clinical isolates of T. rubrum were collectedfrom the Clinical Mycology Laboratory at WestmeadHospital, Sydney, and the Women’s and Children’sHospital, Adelaide, Australia also have almost the samelength of ITS1, which is 666 bp (Kong et al., 2008)Consequently, the results of our study are in agreementwith these studies and showed that molecular methodbased on ITS sequencing is a reliable and useful methodfor the identification of dermatophytes as well as foconfirmation of diagnosis of the conventional methods.

In this study, the molecular method was also used toclarify genetic diversity among strains of T. rubrum andwithin Trichophyton species. The results of this studyregarding nucleotide sequence of isolates of T. rubrumdemonstrated that the similarities among ten isolates oT. rubrum are more than 96% identities. It also showedthe similarities among ten isolates of T. rubrum and tenisolates of other genus of Trichophyton are higher than90%. The results of this study are in agreement withGraser et al. (2000) who showed that the Trichophytonspecies are supported by high similarities with value o


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Aala et al. 6505

Figure 1. The colonies and microscopy of 10 isolates of T. rubrum with (macroconidia and microconidia) × 400.

more than 86% among isolates of T. rubrum and isolatesof other genus of Trichophyton. Our results are also in

agreement with Li et al. (2008), who revealed thapercentage identity of Trichophyton species with


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Figure 2. PCR amplification of isolates of T. rubrum on 1% agarose gelelectrophoresis. T. rubrum ATCC-10218 as positive control strain also showed DNAamplification at 690 bp.

> T. rubrum (1138)50 NNNNGGGAGAGCGTAAGTGGGCTGCCACTATAGAGGACCGGACATTCCAT

100 CAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACC150 TCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTC

200 CGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGA

250 CAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGC

300 AAGCACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATC

350 GATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCG

400 TGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGG

450 GCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGAT

500 GGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCA

550 GTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAG

600 CGCCCTCAGGACCGGCCGCCTGGCCCCAATCTTTATATATATATATATC

650 TTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCAT

658 ATCAAAAG

> T. rubrum (1059)50 NCCAGTAACCGTAGGTGACCTGCGCATATCAATAAGCGGAGGACTCCGTG

100 GGTGAGCATACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACCTCACC

150 CGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCCGGCG

200 GGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACAGAC

250 ACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCACAGA

300 CAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGA

350 AGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTGAAT

400 CATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGCATG

450 CCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGGACG

500 ACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGTGGC

550 CAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCGCCC

600 TCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTTTTCA

650 GGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAAT

700 AAGCCGGAGGAAGGGGGGGCCCCCCATAGGGCCCCCCCGCTCTCTTTTTG

715 GGGAAGCAAAATGGG

> T. rubrum (1164)50 CNNNNNAGACCGTACGTTGGCTGCGCATATCAGATAACCGGACATGACAT

100 CGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACCT

150 CACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCC

200 GGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACA

250 GACACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCAA

300 GCACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGA

350 TGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTG

400 AATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGC

450 ATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGG

500 ACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGT

550 GGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCG

600 CCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTT

650 TTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATAT

700 CAAAAGGGGGGAGGAAGAGGGGGGCCCCCCATAGGGGCCCCCCCCTTTTT

722 TTTTGGGGTAGCGAGAAGGGGG

Figure 3. Nucleotide sequences of 9 isolates of T. rubrum and ATCC-

10218. Nucleotide sequence numbering is shown on the left.


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> T. rubrum (1208)50 TNNGCAGACGTACGTGGGCTGCGAATATCAGGAAGCGACATGACTTCGGG

100 GGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACCTCACC

150 CGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCCGGCG

200 GGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACAGACA

250 CCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCAAGCAC

300 AATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGAA

350 GAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTGAATC

400 ATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGCATGC

450 CTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGGACGA

500 CCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGTGGCC

550 AGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCGCCCTC

600 AGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTTTTCAGG

650 TTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAATAA

700 GCCGGGAGGAAGGGGGGGCCCCCCAAAATGCCCCCCCCTCTCTTTTTGGG

713 GGGGAGAGCGGGG

> T. rubrum (1160)50 NNNNNAAGAATCGTAAGTGACCTGCGCATATCAATAAGCGGAGGATCCGT

100 AGGTGAACCTGCGCGTATCAATAAGCGGAGGACATTCTTGTCTACCTCAC

150 CCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCCGGC

200 GGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACAGAC

250 ACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCAAGCA

300 CAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGA

350 AGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTGAAT

400 CATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGCATG

450 CCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGGACG

500 ACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGTGGC

550 CAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCGCCC

600 TCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTTTTC

650 AGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCAA

614 TAAGCGGGGAGGAA

> T. rubrum (1008)50 NACNAAGAGCCGTAGGTGACCTGCGCATATCAATAAGCGAGAGGACTCCG

100 TAGGTGAACCTGCGTGTATCGGCCGTACGCCCACATTCTTGTCTACCTCA

150 CCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCCGG

200 CGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACAGA

250 CACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCAAGC

300 ACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATG

350 AAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTGAA

400 TCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGCAT

450 GCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGGAC

500 GACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGTGG

550 CCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCGCC

600 CTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTTTT

650 CAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCA

700 ATAAGCCGGAGGAAGGGGGCCCCGAAGAGGAGCCACCCCCCTCAGGGTGT

719 GTGAAACAAACGGCGGGCC

> T. rubrum (1298)50 NNACNNAGTATCGTAGGTGACCTGCGCATATCAATAAGCGGAGGATTCCG

100 TAGGTGAACCTGCGCATATCAATAAGCGGAGGATTCCGTTGGTTACCTCG

150 CCCGGTTGCCTCGGCGGGGCGCGCTCCCCCTGCCAGGGAGAGCCGTCCGG

200 CGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACAGA

250 CACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAGCAAGC

300 ACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATG

350 AAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCCGTGAA

400 TCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGGGGCAT

450 GCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGGAC

500 GACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGTGG

550 CCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCGCC

600 CTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCTTTT

650 CAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATATCA

668 ATAAGCGGAGGAA

>

T. rubrum (1044) 50 NNNANCGGGACAGCCGTAGTGGGCTGCGCATATCAGATAACGCGGAGATT

100 ACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTCT

150 ACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCC

200 GTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGG

250 ACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTTAG

300 CAAGCACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCAT

350 CGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCC

400 GTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGGGG

450 GGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGA

500 TGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCA

550 GTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAG

600 CGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATC

650 TTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCAT

658 ATCAAAAG

Figure 3. Cotnd.


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> T. rubrum (2970) 50 ANCGGACAGCCGTAGTGGCCTGCGACATATCAGATAACGCGGAGAGGACT

100 TCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACC

150 TCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTC

200 CGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACA





450 ATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGG500 ACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGT

550 GGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAG

600 CGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATATATCT

650 TTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAGCATA

660 TCAAAAGCGG

> T. rubrum (ATCC-10218)

50 NGGGACCGCCGTAGTGGCCTGCGACATATCAGATAACGCGGAGAGGACTT

100 CGGGGGTGAGCATACGTGCGCCGGCCGTACGCCCCCATTCTTGTCTACCT

150 CACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGCCGTCC

200 GGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGAGGACA





450 ATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGTGATGG

500 ACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAAGCAGT

550 GGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATTCAGCG

600 CCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATCGCGATATATCTT633 GGCAGGTTGACCTCGGATCAGGTAGGGATACGT

Figure 3. Cotnd.

T. rubrum (1138) --NNNNGGGAGAGCGTAAGTGGGCTGCCA-CTAT-AGAGGAC-CGGACAT 50

T. rubrum (1164) ---CNNNNNAGACCGTACGTTGGCTGCGC-ATATCAGATAAC-CGGACAT 50

T. rubrum (1208) ----TNNGCAGA-CGTACGTGGGCTGCGA-ATATCAGGAAGC---GACAT 50

T. rubrum (1044) NNNANCGGGACAGCCGTAGTGGGCTGCGC-ATATCAGATAACGCGGAGAT 50

T. rubrum (ATCC-10218) -----NGGGACCGCCGTAGTGGCCTGCGACATATCAGATAACGCGGAGAG 50

T. rubrum (2970) ----ANCGGACAGCCGTAGTGGCCTGCGACATATCAGATAACGCGGAGAG 50

T. rubrum (1059) ----NCCAGTAACCGTAGGTGACCTGCGC-ATATCAATAAGC----GGAG 50

T. rubrum (1160) --NNNNNAAGAATCGTAAGTGACCTGCGC-ATATCAATAAGC---G-GAG 50

T. rubrum (1298) --NNACNNAGTATCGTAGGTGACCTGCGC-ATATCAATAAGC---G-GAG 50

T. rubrum (1008) ---NACNAAGAGCCGTAGGTGACCTGCGC-ATATCAATAAGC---GAGAG 50

* ** **** *** * * *

T. rubrum (1138) TCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100T. rubrum (1164) GACATCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (1208) GACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (1044) TACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (ATCC-10218) GACTTCGGGGGTGAGCATACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (2970) GACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (1059) GACTCCGTGGGTGAGCATACGTGCGCCGGCCGTACGCCCCCATTCTTGTC 100

T. rubrum (1160) GAT-CCGTAGGTGAACCTGCGCGTATCAATAAGCGGAGGACATTCTTGTC 100

T. rubrum (1298) GATTCCGTAGGTGAACCTGCGCATATCAATAAGCGGAGGATTCCGTTGGT 100

T. rubrum (1008) GACTCCGTAGGTGAACCTGCGTGTATCGGCCGTACGCCCACATTCTTGTC 100

* ***** * ** * * ***

T. rubrum (1138) TACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGC 150




T. rubrum (ATCC-10218) TACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTGCCAGGGAGAGC 150




T. rubrum (1298) TACCTCGCCCGGTTGCCTCGGCGGGGCGCGCTCCCCCTGCCAGGGAGAGC 150


****** ******************************************

T. rubrum (1138) CGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGA 200




T. rubrum (ATCC-10218) CGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCGCGCCCGCCGGA 200






**************************************************

Figure 4. Comparison of nucleotide sequence between T. rubrum ITS1orthologues. Nucleotide sequences that are present in all ITS1 are shaded inblue colour. Nucleotide sequence numbering is shown on the right.


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Aala et al. 6509

T. rubrum (1138) GGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTT 250




T. rubrum (ATCC-10218) GGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAGTCTGAGCGTTT 250






**************************************************

T. rubrum (1138) AGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGC 300




T. rubrum (ATCC-10218) AGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGC 300






**************************************************

T. rubrum (1138) ATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATT 350




T. rubrum (ATCC-10218) ATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATT 350






**************************************************

T. rubrum (1138) CCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGG 400


T. rubrum (1208) CCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGG 400T. rubrum (1044) CCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGG 400

T. rubrum (ATCC-10218) CCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTCTGGCATTCCGG 400





**************************************************

T. rubrum (1138) GGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGT 450




T. rubrum (ATCC-10218) GGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGCCCGGCTTGTGT 450





**************************************************

T. rubrum (1138) GATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAA 500




T. rubrum (ATCC-10218) GATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAA 500

T. rubrum (2970) GATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAA 500T. rubrum (1059) GATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGACGCGCCCGAAAA 500




**************************************************

T. rubrum (1138) GCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATT 550




T. rubrum (ATCC-10218) GCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATGGGCAGCCAATT 550






**************************************************

T. rubrum (1138) CAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATATATATAT 600




T. rubrum (ATCC-10218) CAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTATATCGCGATAT 600






****************************************** ****

T. rubrum (1138) ATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGAACTTAAG 650




T. rubrum (ATCC-10218) ATCTTGGCAGGTTGACCTCGGATCAGGTAGGGATACGT------------ 650






***** *****************************

Figure 4. Contd.


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T. rubrum (1138) CATATCAAAAG--------------------------------------- 700

T. rubrum (1164) CATATCAAAAGGGGGGAGGAAGAGGGGGGCCCCCCATAGGGGCCCCCCCC 700

T. rubrum (1208) CATATCAATAAGCCGGGAGGAAGGGGGGGCCCCCCA-AAATGCCCCCCCC 700

T. rubrum (1044) CATATCAAAAG--------------------------------------- 700

T. rubrum (ATCC-10218) -------------------------------------------------- 700

T. rubrum (2970) CATATCAAAAGCGG------------------------------------ 700

T. rubrum (1059) CATATCAATAAGCCGG-AGGAAGGGGGGGCCCCCCATAGGGCCCCCCCGC 700

T. rubrum (1160) CATATCAATAAGCGGGGAGGAA---------------------------- 700

T. rubrum (1298) CATATCAATAAGCGGAGGAA------------------------------ 700

T. rubrum (1008) CATATCAATAAGCCGGAGGAAGGGGGCCCCGAAGAGGAGCCACCCCCCTC 700

T. rubrum (1138) --------------------------- 727

T. rubrum (1164) TTTTTTTTTGGGGTAGCGAGAAGGGGG 727

T. rubrum (1208) TCTCTTTTTGGGGGGGAGAGCGGGG-- 727

T. rubrum (1044) --------------------------- 727

T. rubrum (ATCC-10218) --------------------------- 727

T. rubrum (2970) --------------------------- 727

T. rubrum (1059) TCTCTTTTTGGGGAAGCAAAATGGG-- 727

T. rubrum (1160) --------------------------- 727

T. rubrum (1298) --------------------------- 727

T. rubrum (1008) AGGGTGTGTGAAACAAACGGCGGGCC- 727

Figure 4. Contd.

T. raubitschekii strain NOMH 789 CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50

T. megninii strain ATCC 12106 CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50

T. megninii strain ATCC 12106 CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50

T. rubrum strain UAMH 8547 CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50

T. rubrum strain ATCC 28188 CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50T. kanei CGCCCGTCGCTACTACCGATTGAATGGCTCAGTGAGGCCTTCGGACTGGC 50

T.rubrum 5.8S rRNA gene --------------------------------------------------

T.rubrum strain WM 06.348 --------------------------------------------------

T.rubrum strain NCPF 295 --------------------------------------------------

T.rubrum strain 05-287-3929 --------------------------------------------------

T. rubrum (1138) --------------------------------------------------

T. rubrum (1164) --------------------------------------------------

T. rubrum (1298) --------------------------------------------------

T. rubrum (1008) --------------------------------------------------

T. rubrum (1059) --------------------------------------------------

T. rubrum (1208) --------------------------------------------------

T. rubrum (1264) --------------------------------------------------

T. rubrum (2970) --------------------------------------------------

T. rubrum (ATCC-10218) --------------------------------------------------

T. rubrum (1044) --------------------------------------------------

T. raubitschekii strain NOMH 789 CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100

T. megninii strain ATCC 12106 CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100

T. saudanese UAMH 8548 CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100

T. rubrum strain UAMH 8547 CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100

T. rubrum strain ATCC 28188 CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100T. kanei CCAGGGAGGTTGGAAACGACCGCCCAGGGCCGGAAAGTTGGTCAAACTCG 100

T. rubrum 5.8S rRNA gene --------------------------------------------------

T. rubrum strain WM 06.348 --------------------------------------------------

T. rubrum strain NCPF 295 --------------------------------------------------

T. rubrum strain 05-287-3929 --------------------------------------------------

T. rubrum (1138) --------------------------------------------------

T. rubrum (1164) --------------------------------------------------

T. rubrum (1298) --------------------------------------------------

T. rubrum (1008) --------------------------------------------------

T. rubrum (1059) --------------------------------------------------

T. rubrum (1208) --------------------------------------------------

T. rubrum (1264) --------------------------------------------------

T. rubrum (2970) --------------------------------------------------

T. rubrum (ATCC-10218) --------------------------------------------------

T. rubrum (1044) --------------------------------------------------

T.raubitschekii strain NOMH 789 GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. megninii strain ATCC 12106 GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. saudanese UAMH 8548 GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. rubrum strain UAMH 8547 GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. rubrum strain ATCC 28188 GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. kanei GTCATTTAGAGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTG 150

T. rubrum 5.8S rRNA gene --------------------------ACAAGGTTTCCGTAGGTGAACCTG 24

T. rubrum strain WM 06.348 --------------------------------------------------

T. rubrum strain NCPF 295 ----------------------------------TCCGTAGGTGAACCTG 16

T. rubrum strain 05-287-3929 --------------------------------------------------

T. rubrum (1138) --------------------------------------------------

T. rubrum (1164) --------------------------------------------------

T. rubrum (1298) --------------------------------------------------

T. rubrum (1008) --------------------------------------------------

T. rubrum (1059) --------------------------------------------------

T. rubrum (1208) --------------------------------------------------

T. rubrum (1264) --------------------------------------------------

T. rubrum (2970) --------------------------------------------------

T. rubrum (ATCC-10218) --------------------------------------------------

T. rubrum (1044) --------------------------------------------------

Figure 5. Comparison of nucleotide sequence between T. rubrum ITS1

orthologues. Nucleotide sequences that are present in all ITS1 are shaded inblack colour. Nucleotide sequence numbering is shown on the right.


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Aala et al. 6511

T. raubitschekii strain NOMH 789 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. saudanese UAMH 8548 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. megninii strain ATCC 12106 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. rubrum strain UAMH 8547 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. rubrum strain ATCC 28188 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. kanei CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 198

T. rubrum 5.8S rRNA gene CGGAAGGATCATTAACGCGCNGGCCGGAGGCTGGCCCCC-CACGATAG-G 72

T. rubrum strain WM 06.348 ------GATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 42T. rubrum strain NCPF 295 CGGAAGGATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 64

T. rubrum strain 05-287-3929 ------GATCATTAACGCGCAGGCCGGAGGCTGGCCCCC-CACGATAG-G 42

T. rubrum (1138) -------------NNNNGGGAGAGCGTAAG-TGGGCTGC-CACTATAGAG 35

T. rubrum (1164) -------------NNNNNAAGAATCGTAAGTGACCTGCG-CATATCA-AT 35

T. rubrum (1298) -------------NNACNNAGTATCGTAGGTGACCTGCG-CATATCA-AT 35

T. rubrum (1008) --------------NACNAAGAGCCGTAGGTGACCTGCG-CATATCA-AT 34

T. rubrum (1059) ---------------NCCAGTAACCGTAGGTGACCTGCG-CATATCA-AT 33

T. rubrum (1208) ----------------TNNGCAGACGTACGTGGGCTGCG-AATATCA-GG 32

T. rubrum (1264) --------------CNNNNNAGACCGTACGTTGGCTGCG-CATATCAGAT 35

T. rubrum (2970) --------------ANCGGACAGCCGTA-GTGGCCTGCGACATATCAGAT 35

T. rubrum (ATCC-10218) ---------------NGGGACCGCCGTA-GTGGCCTGCGACATATCAGAT 34

T. rubrum (1044) ----------NNNANCGGGACAGCCGTA-GTGGGCTGCG-CATATCAGAT 38

** * * * *

T. raubitschekii strain NOMH 789 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. megninii strain ATCC 12106 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. saudanese UAMH 8548 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. rubrum strain UAMH 8547 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. rubrum strain ATCC 28188 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. kanei GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 246

T. rubrum 5.8S rRNA gene GA-CCG-ACGTTC-ATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 119

T. rubrum strain WM 06.348 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 90

T. rubrum strain NCPF 295 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 112

T. rubrum strain 05-287-3929 GA-CCG-ACGTTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 90

T. rubrum (1138) GA-CCGGACATTCCATCAGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 84

T. rubrum (1164) AA-GCG--GAGGAT-CCGTAGGTGAACCTGCGCGTATCAATAAGCGGAGG 81

T. rubrum (1298) AA-GCG--GAGGATTCCGTAGGTGAACCTGCGCATATCAATAAGCGGAGG 82

T. rubrum (1008) AA-GCGA-GAGGACTCCGTAGGTGAACCTGCGTGTATCGGCCGTACGCCC 82

T. rubrum (1059) AA-GCG--GAGGACTCCGTGGGTGAGCATACGTGCGCCGGCCGTACGCCC 80

T. rubrum (1208) AA-GCGA-CATGACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 80

T. rubrum (1264) AA-CCGGACATGACATCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 84

T. rubrum (2970) AACGCGGAGAGGACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 85

T. rubrum (ATCC-10218) AACGCGGAGAGGACTTCGGGGGTGAGCATACGTGCGCCGGCCGTACGCCC 84

T. rubrum (1044) AACGCGGAGATTACTTCGGGGGTGAGCAGACGTGCGCCGGCCGTACGCCC 88

* ** * ***** * ** * *

T. raubitschekii strain NOMH 789 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. megninii strain ATCC 12106 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. saudanese UAMH 8548 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. rubrum strain UAMH 8547 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. rubrum strain ATCC 28188 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. kanei CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 296

T. rubrum 5.8S rRNA gene C-ATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 168

T. rubrum strain WM 06.348 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 140

T. rubrum strain NCPF 295 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 162

T. rubrum strain 05-287-3929 CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 140

T. rubrum (1138) CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 134T. rubrum (1164) ACATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 131

T. rubrum (1298) ATTCCGTTGGTTACCTCGCCCGGTTGCCTCGGCGGGGCGCGCTCCCCCTG 132

T. rubrum (1008) ACATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 132

T. rubrum (1059) CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 130




T. rubrum (ATCC-10218) CCATTCTTGTCTACCTCACCCGGTTGCCTCGGCGGGCCGCGCTCCCCCTG 134


*** ****** ****************** *************

Figure 5. Contd.


20/110


T. raubitschekii strain NOMH 789 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 346

T. megninii strain ATCC 12106 CCAGAGAGAGCCGTCCGGCGGGCCTCTTCCGGGGGCCTCGAGCCGGACCG 346

T. saudanese UAMH 8548 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGGGCCTCGAGCCGGACCG 346

T. rubrum strain UAMH 8547 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 346

T. rubrum strain ATCC 28188 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 346

T. kanei CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 346

T. rubrum 5.8S rRNA gene CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 218

T. rubrum strain WM 06.348 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 190T. rubrum strain NCPF 295 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 212

T. rubrum strain 05-287-3929 CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 190

T. rubrum (1138) CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 184








T. rubrum (ATCC-10218) CCAGGGAGAGCCGTCCGGCGGGCCCCTTCTGGGAGCCTCGAGCCGGACCG 184


**** ******************* **** *** ****************

T. raubitschekii strain NOMH 789 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. megninii strain ATCC 12106 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. saudanese UAMH 8548 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. rubrum strain UAMH 8547 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. rubrum strain ATCC 28188 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. kanei CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 396

T. rubrum 5.8S rRNA gene CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 268

T. rubrum strain WM 06.348 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 240

T. rubrum strain NCPF 295 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 262

T. rubrum strain 05-287-3929 CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 240

T. rubrum (1138) CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 234





T. rubrum (1208) CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 230T. rubrum (1264) CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 234


T. rubrum (ATCC-10218) CGCCCGCCGGAGGACAGACACCAAGAAAAAATTCTCTGAAGAGCTGTCAG 234


**************************************************

T. raubitschekii strain NOMH 789 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. megninii strain ATCC 12106 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. saudanese UAMH 8548 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. rubrum strain UAMH 8547 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. rubrum strain ATCC 28188 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. kanei TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 446

T. rubrum 5.8S rRNA gene TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 318

T. rubrum strain WM 06.348 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 290

T. rubrum strain NCPF 295 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 312

T. rubrum strain 05-287-3929 TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 290T. rubrum (1138) TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 284

T. rubrum (1164) TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 281







T. rubrum (ATCC-10218) TCTGAGCGTTTAGCAAGCACAATCAGTTAAAACTTTCAACAACGGATCTC 284


**************************************************

Figure 5. Contd.


21/110

Aala et al. 6513

T.raubitschekii strain NOMH 789 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. megninii strain ATCC 12106 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. saudanese UAMH 8548 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. rubrum strain UAMH 8547 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. rubrum strain ATCC 28188 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. kanei TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 496

T. rubrum 5.8S rRNA gene TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 368

T. rubrum strain WM 06.348 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 340T. rubrum strain NCPF 295 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 362

T. rubrum strain 05-287-3929 TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 340

T. rubrum (1138) TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 334








T. rubrum (ATCC-10218) TTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGA 334


**************************************************

T. raubitschekii strain NOMH789 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. megninii strain ATCC 12106 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. saudanese UAMH 8548 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. rubrum strain UAMH 8547 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. rubrum strain ATCC 28188 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. kanei ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 546

T. rubrum 5.8S rRNA gene ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 418

T. rubrum strain WM 06.348 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 390

T. rubrum strain NCPF 295 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 412

T. rubrum strain 05-287-3929 ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 390

T. rubrum (1138) ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 384








T. rubrum (ATCC-10218) ATTGCAGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCTC 384


**************************************************

T. raubitschekii strain NOMH789 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. megninii strain ATCC 12106 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. saudanese UAMH 8548 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. rubrum strain UAMH 8547 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. rubrum strain ATCC 28188 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. kanei TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 596

T. rubrum 5.8S rRNA gene TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 468

T. rubrum strain WM 06.348 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 440

T. rubrum strain NCPF 295 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 462

T. rubrum strain 05-287-3929 TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 440T. rubrum (1138) TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 434

T. rubrum (1164) TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 431







T. rubrum (ATCC-10218) TGGCATTCCGGGGGGCATGCCTGTTCGAGCGTCATTTCAACCCCTCAAGC 434


**************************************************

Figure 5. Contd.


22/110


T. raubitschekii strain NOMH789 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. megninii strain ATCC 12106 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. saudanese UAMH 8548 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. rubrum strain UAMH 8547 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. rubrum strain ATCC 28188 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. kanei CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 646

T. rubrum 5.8S rRNA gene CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTC--TTCGGGGGCGGGAC 516

T. rubrum strain WM 06.348 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 490T. rubrum strain NCPF 295 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 512

T. rubrum strain 05-287-3929 CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 490

T. rubrum (1138) CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 484








T. rubrum (ATCC-10218) CCGGCTTGTGTGATGGACGACCGTCCGGCCCCTCCCTTCGGGGGCGGGAC 484


********************************** **************

T. raubitschekii strain NOMH789 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 696

T. megninii strain ATCC 12106 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTGGGCGAATG 696

T. saudanese UAMH 8548 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTGGGCGAATG 696

T. rubrum strain UAMH 8547 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 696

T. rubrum strain ATCC 28188 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 696

T. kanei GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 696

T. rubrum 5.8S rRNA gene GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 566

T. rubrum strain WM 06.348 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 540

T. rubrum strain NCPF 295 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 562

T. rubrum strain 05-287-3929 GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 540

T. rubrum (1138) GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 534








T. rubrum (ATCC-10218) GCGCCCGAAAAGCAGTGGCCAGGCCGCGATTCCGGCTTCCTAGGCGAATG 534


***************************************** ********

T. raubitschekii strain NOMH789 GGCAGCCAATTCAGCGCCCTCAGG-------------------------- 720

T. megninii strain ATCC 12106 GGCAGCCAAACCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 746

T. saudanese UAMH 8548 GGCAGCCAAACCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 746

T. rubrum strain UAMH 8547 GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 746

T. rubrum strain ATCC 28188 GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 746

T. kanei GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 746

T. rubrum 5.8S rRNA gene GGCAGCCAATTCAGCGCCCTCAGGACCGGCNGCCCTGGCCCCAATCTTTA 616

T. rubrum strain WM 06.348 GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 590

T. rubrum strain NCPF 295 GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 612

T. rubrum strain 05-287-3929 GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 590T. rubrum (1138) GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 584

T. rubrum (1164) GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 581







T. rubrum (ATCC-10218) GGCAGCCAATTCAGCGCCCTCAGGACCGGCCGCCCTGGCCCCAATCTTTA 584


********* *************

Figure 5. Contd.


23/110

Aala et al. 6515

T. raubitschekii strain NOMH789 --------------------------------------------------

T. megninii strain ATCC 12106 TATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCT 796

T. saudanese UAMH 8548 TATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCGCT 796

T. rubrum strain UAMH 8547 TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 796

T. rubrum strain ATCC 28188 TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 796

T. kanei TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 796

T. rubrum 5.8S rRNA gene TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 666

T. rubrum strain WM 06.348 TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 640

T. rubrum strain NCPF 295 TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 662

T. rubrum strain 05-287-3929 TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 640T. rubrum (1138) TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 634

T. rubrum (1164) TATATATATATATCTTTTCAGGTTGACCTCGGATCAGGTAGGGATACCCG 631







T. rubrum (ATCC-10218) TATCGCGATATATCTTGGCAGGTTGACCTCGGATCAG------------- 621


T.raubitschekii strain NOMH 789 --------------------------------------------------

T. megninii strain ATCC 12106 GAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTGCC 846

T. saudanese UAMH 8548 GAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTGCC 846

T. rubrum strain UAMH 8547 CTGAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTG 846

T. rubrum strain ATCC 28188 CTGAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTG 846

T. kanei CTGAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTG 846

T. rubrum 5.8S rRNA gene CTGAACTTAAGCATATCAATAAGCGGAGGAAAAGAAACCAACAGGGATTG 716

T. rubrum strain WM 06.348 CTGAACTTAA---------------------------------------- 650T. rubrum strain NCPF 295 CTGAACTTAAGCATATCAAT------------------------------ 682

T. rubrum strain 05-287-3929 CTGAACTTAAGCATATCAATAAGCGG------------------------ 666

T. rubrum (1138) CTGAACTTAAGCATATCAAAAG---------------------------- 656

T. rubrum (1164) CTGAACTTAAGCATATCAATAAGCGGGGAGGAA----------------- 664

T. rubrum (2970) -------------------------------------------------

T. rubrum (ATCC-10218) --------------------------

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AJMR- 20 September, 2012 Issue