Molecular Identification and Classification of a Phytoplasma Associated with Phyllody of Strawberry Fruit in Maryland

Several phytoplasmas have been reported to be associated with phyllody of strawberry fruit, including clover yellow edge, clover proliferation, clover phyllody, eastern and western aster yellows, STRAWB2, strawberry multicipita, and Mexican periwinkle virescence phytoplasmas. Plant symptoms in addition to phyllody may include chlorosis, virescence, stunting, or crown proliferation. In this report we describe a new phytoplasma in association with strawberry leafy fruit (SLF) disease in Maryland. Diseased plants exhibited fruit phyllody, floral virescence, leaf chlorosis, and plant stunting. Phytoplasmal 16S rDNA was amplified from SLF diseased plants by using the polymerase chain reaction (PCR) primed by primer pair P1/P7 and was reamplified in nested PCR primed by primer pair R16F2n/R2 (F2n/R2) as previously described (1). These results indicated the presence of a phytoplasma, designated SLF phytoplasma. Identification of SLF phytoplasma was accomplished by restriction fragment length polymorphism (RFLP) analysis of DNA amplified in PCR primed by F2n/R2, using endonuclease enzyme digestion with AluI, HhaI, KpnI, HaeIII, MseI, HpaII, RsaI, and Sau3AI. Phytoplasma classification was done according to the system of Lee et al. (2). RFLP analyses of rDNA amplified in three separate PCRs gave identical patterns. On the basis of collective RFLP patterns of the amplified 16S rDNA, the SLF phytoplasma was classified as a member of group 16SrIII (group III, X-disease phytoplasma group). The HhaI RFLP pattern of SLF 16S rDNA differed from that of the apparently close relative, clover yellow edge (CYE) phytoplasma, and all other phytoplasmas previously described in group III. Based on these results, SLF phytoplasma was classified in a new subgroup, designated subgroup K (III-K), within group III. The 1.2 kbp DNA product of PCR primed by primer pair F2n/R2 was sequenced, and the sequence deposited in GenBank under Accession No. AF 274876. Results from putative restriction site analysis of the sequence were in agreement with the results from actual enzymatic RFLP analysis of rDNA amplified from phylloid strawberry fruit. Although the sequence similarity between the 1.2-kbp fragment from the 16S rDNA of SLF phytoplasma and that of CYE phytoplasma was 99.9%, the Hha1 RFLP pattern of SLF rDNA supports the conclusion that the SLF phytoplasma may be closely related to, but is distinct from, CYE and other strains that are classified in group III. These findings contribute knowledge about the diversity of phytoplasmas affiliated with group III and the diversity of phytoplasmas associated with diseases in strawberry. References: (1) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (2) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

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Molecular Identification of a Phytoplasma Associated with Witches'-Broom Disease of Black Raspberry in Oregon and Its Classification in Group 16SrIII, New Subgroup Q

Plant Disease ◽

10.1094/pdis.2001.85.10.1121a ◽

2001 ◽

Vol 85 (10) ◽

pp. 1121-1121 ◽

Cited By ~ 7

Author(s):

R. E. Davis ◽

E. L. Dally ◽

R. H. Converse

Keyword(s):

Nucleotide Sequence ◽

16S Rdna ◽

Sequence Data ◽

Sequence Similarity ◽

Rflp Analysis ◽

Close Relative ◽

Black Raspberry ◽

Rflp Pattern ◽

Nucleotide Sequence Data ◽

Group Iii

Plants of Rubus occidentalis (black raspberry) ‘Munger’ exhibiting symptoms of black raspberry witches'-broom (BRWB) disease were observed in commercial fields in Oregon (1). Symptoms were often severe, leading to death of infected plants, and a phytoplasma (mycoplasmalike bodies) was observed in ultrathin sections of diseased plants (1). In the current work, the association of phytoplasma with BRWB was assessed using the polymerase chain reaction (PCR) for specific amplification of phytoplasmal rDNA. DNA template for use in the PCR was extracted from plants as described elsewhere (2). Phytoplasmal 16S rDNA was amplified from diseased black raspberry plants in PCR primed by primer pair P1/P7 and reamplified in nested PCR primed by primer pair R16F2n/R2 (F2n/R2) by a method described previously (2). These results indicated the presence of a phytoplasma, designated BRWB phytoplasma, in the diseased plants. Identification of BRWB phytoplasma was accomplished by restriction fragment length polymorphism (RFLP) analysis of DNA amplified in PCR primed by F2n/R2. Phytoplasma classification was done according to the system of Lee et al. (3). On the basis of collective RFLP patterns of the amplified 16S rDNA, the BRWB phytoplasma was classified as a member of group 16SrIII (group III, X-disease phytoplasma group). The HhaI RFLP pattern of BRWB 16S rDNA differed from that of its close relative, clover yellow edge (CYE) phytoplasma. The RsaI RFLP pattern of BRWB rDNA differed from that of rDNA from all phytoplasmas previously described in group III. Based on these results, BRWB phytoplasma was classified in a new subgroup, designated subgroup Q (III-Q) within group III. The 1.8 kbp DNA product of PCR primed by primer pair P1/P7 was cloned and its nucleotide sequence determined. The sequence was deposited in GenBank under Accession no. AF302841. Results from putative restriction site analysis of the cloned and sequenced rDNA were in excellent agreement with the results from enzymatic RFLP analysis of uncloned rDNA amplified from BRWB diseased black raspberry. Sequence similarity between the 1.8 kbp rDNA of BRWB phytoplasma and that of CYE phytoplasma was 99.4%. The nucleotide sequence data support the conclusion that the BRWB phytoplasma is related to, but is distinct from, other strains that are classified in group III. These findings contribute knowledge about the diversity of phytoplasmas affiliated with group III and provide information to aid the diagnosis of BRWB disease. References: (1) R. H. Converse et al. Plant Dis. 66:949, 1982. (2) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (3) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

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A Sida sp. Is a New Host for “Candidatus Phytoplasma brasiliense” in Brazil

Plant Disease ◽

10.1094/pdis-09-10-0647 ◽

2011 ◽

Vol 95 (3) ◽

pp. 363-363 ◽

Cited By ~ 3

Author(s):

B. Eckstein ◽

J. C. Barbosa ◽

J. A. M. Rezende ◽

I. P. Bedendo

Keyword(s):

16S Rdna ◽

Sequence Similarity ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Sao Paulo ◽

São Paulo ◽

New Host ◽

Nucleotide Sequence Alignment ◽

Pcr Products

Sida is a genus of flowering herbs in the family Malvaceae, which includes several species that are weeds in Brazil. Plants of a Sida sp. exhibiting symptoms characterized by stunting, chlorosis, small leaves, and witches'-broom, indicative of infection by phytoplasmas, were found in a field previously cultivated with tomato, located in the region of Campinas, State of São Paulo, in December 2008. To demonstrate the presence of phytoplasmas in diseased tissues, DNA was extracted from shoots and leaves from three symptomatic and eight asymptomatic plants. Nested PCR was performed using primers P1/Tint followed by primer pair R16F2n/R16R2 (1). DNA fragments of 1.2 kb, corresponding to 16S rDNA, were amplified only for DNA from two symptomatic samples. Phytoplasma identification was initially carried out by restriction fragment length polymorphism (RFLP) analysis through digesting the PCR products with the restriction enzymes AluI, HhaI, HaeIII, HpaII, MseI, and RsaI. The two phytoplasma isolates found to be infecting a Sida sp. showed identical RFLP patterns, which were indistinguishable from the phytoplasma previously reported in association with hibiscus (Hibiscus rosa-sinensis) witches'-broom in Brazil (2). Nucleotide sequence alignment revealed that 16S rDNA of both phytoplasma isolates found in a Sida sp. (GenBank Accession No. HQ230579) shared 99.9% sequence similarity with 16S rDNA from hibiscus witches'-broom phytoplasma (HibWB) (GenBank Accession No. AF147708). HibWB is the representative of the 16SrXV group and it was proposed as a putative species nominated “Candidatus Phytoplasma brasiliense” (2). The disease is frequently observed in hibiscus plants used as ornamentals in the states of São Paulo (4) and Rio de Janeiro (2). “Ca. Phytoplasma brasiliense” has only been reported in Brazil to be infecting hibiscus (2,4) and periwinkle (Catharanthus roseus) (3). The presence of a phytoplasma belonging to group 16SrXV in a Sida sp. expands its natural host range. The role of this weed as a potential source of inoculum for crops should be investigated. References: (1) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) H. G. Montano et al. Int. J. Syst. Evol. Microbiol. 51:1109, 2001. (3) H. G. Montano et al. Plant Dis. 85:1209, 2001. (4) E. G. Silva et al. Summa Phytopathol. 35:234, 2009.

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First Report of Oat as Host of a Phytoplasma Belonging to Group 16SrI, Subgroup A

Plant Disease ◽

10.1094/pdis.2002.86.4.443b ◽

2002 ◽

Vol 86 (4) ◽

pp. 443-443 ◽

Cited By ~ 7

Author(s):

R. Jomantiene ◽

R. E. Davis ◽

A. Alminaite ◽

D. Valiunas ◽

R. Jasinskaite

Keyword(s):

16S Rdna ◽

Plant Species ◽

Nested Pcr ◽

Sequence Similarity ◽

Intergenic Spacer ◽

Rflp Analysis ◽

23S Rrna ◽

Universal Primer ◽

Intergenic Spacer Region ◽

Group I

Diseased plants of oat (Avena sativa L.) exhibiting abnormal proliferation of spikelets were observed in the field in Raseniai, Lithuania. The possible association of a phytoplasma with the disease, termed oat proliferation (OatP), was determined using polymerase chain reaction (PCR) for amplification of phytoplasmal ribosomal (r) RNA gene (rDNA) sequences from template DNA extracted from the diseased oats. DNA extractions and nested PCRs were conducted as previously described (2). In the nested PCRs, the first reaction was primed by phytoplasma-universal primer pair P1/P7, and the second (nested) PCR was primed by primer pair R16F2n/R16R2 (F2n/R2). Phytoplasmal rDNA was amplified in the nested PCR, indicating that the plants contained a phytoplasma, designated oat proliferation (OatP) phytoplasma. The OatP phytoplasma was identified and classified according to the system of Lee et al. (2) through restriction fragment length polymorphism (RFLP) analysis of 16S rDNA amplified in the PCR primed by F2n/R2. On the basis of collective RFLP patterns of the 16S rDNA, the OatP phytoplasma was classified as a member of group 16SrI (group I, aster yellows phytoplasma group). The RFLP patterns of the 16S rDNA were indistinguishable from those of 16S rDNA from tomato big bud (BB) phytoplasma and other phytoplasmas classified in group I, subgroup A (subgroup I-A, tomato big bud phytoplasma subgroup). The 1.8-kbp rDNA product of PCR primed by primer pair P1/P7 was cloned, and its nucleotide sequence was determined. The sequence was deposited in GenBank under Accession No. AF453416. Results from putative restriction site analysis of the cloned and sequenced rDNA were in excellent agreement with the results from enzymatic RFLP analysis of uncloned rDNA from OatP-diseased oat plants. Sequence similarity between the 1.8-kbp rDNA of OatP phytoplasma and that of BB phytoplasma (GenBank No. AF222064) was 99.2%; 9 of the 14 base changes were in the 16S-23S rRNA intergenic spacer region. The base differences in rDNA may signal that the OatP and BB phytoplasmas are mutually distinct in their biologies. Phytoplasmas classified in subgroup I-A have previously been reported in a broad range of plant species in North America and Europe, although there are no previous definitive reports of oat as a host of a subgroup I-A phytoplasma (3,4). In 1977, Fedotina (1) reported electron microscopy of a mycoplasma-like organism (phytoplasma) in pseudorosette-diseased oat plants in Siberia, but the identity of that phytoplasma remains unknown. Subgroup I-A phytoplasma strains are geographically widespread and have been found in numerous plant species (3,4). The discovery reported here, of a subgroup I-A phytoplasma in diseased oats in Lithuania, provokes questions concerning possible impacts of this phytoplasma on oat cultivation in central Europe and other regions. References: (1) V. L. Fedotina. Arch. Phytopathol. Pflanzenschutz 13:177, 1977. (2) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (3) C. Marcone et al. Int. J. Syst. Evol. Microbiol. 50:1703, 2000. (4) D. Valiunas et al. Plant Dis. 85:804, 2001.

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Single and Mixed Phytoplasma Infections in Phyllody- and Dwarf-Diseased Clover Plants in Lithuania

Plant Disease ◽

10.1094/pdis.2000.84.10.1061 ◽

2000 ◽

Vol 84 (10) ◽

pp. 1061-1066 ◽

Cited By ~ 20

Author(s):

Juozas B. Staniulis ◽

Robert E. Davis ◽

Rasa Jomantiene ◽

Audrone Kalvelyte ◽

Ellen L. Dally

Keyword(s):

16S Rdna ◽

Fragment Length Polymorphism ◽

Rflp Analysis ◽

Chain Reactions ◽

Polymerase Chain Reactions ◽

Restriction Sites ◽

Polymerase Chain ◽

Dwarf Disease ◽

Clover Phyllody

Naturally diseased plants of clover (Trifolium spp.) exhibiting symptoms of clover phyllody (virescence and phyllody of flowers) or of clover dwarf (abnormally small leaves, shortened internodes, proliferation of shoots, and dwarf growth habit) were observed in fields in Lithuania. Phytoplasma group-specific polymerase chain reactions (PCRs) and restriction fragment length polymorphism (RFLP) analysis of 16S rDNA revealed that the plants were infected by two mutually distinct phytoplasmas. Clover phyllody-diseased plants were infected by a subgroup 16SrI-C (subgroup I-C) phytoplasma (CPh-L) related to clover phyllody (CPh-C) phytoplasma in Canada. Clover dwarf-diseased plants were infected by both CPh-L and a phytoplasma (CYE-L) related to clover yellow edge (CYE-C) phytoplasma (subgroup 16SrIII-B = III-B) in Canada. A 1.8-kbp fragment of rRNA operon from CYE-L was amplified, cloned, and sequenced, and putative restriction sites mapped. This sequence shared high similarity (99.7%) with that of CYE-C and exhibited no differences from CYE-C in RFLP patterns of 16S rDNA; therefore, we tentatively classified CYE-L in subgroup 16SrIII-B (type strain, CYE = CYE-C phytoplasma) of the X-disease phytoplasma group. These findings extend the known geographical ranges of subgroup I-C and subgroup III-B taxa to the region of northern Europe including Lithuania and suggest a role of the subgroup III-B phytoplasma in clover dwarf disease.

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021 Strawberry Fruit Phyllody Caused by Phytoplasmas

HortScience ◽

10.21273/hortsci.35.3.391d ◽

2000 ◽

Vol 35 (3) ◽

pp. 391D-391

Author(s):

R. Jomantiene ◽

J.L. Maas ◽

E.L. Dally ◽

R.E. Davis

Keyword(s):

16S Rrna ◽

Flower Initiation ◽

Strawberry Fruit ◽

Group Vi ◽

Vegetative Organs ◽

Aster Yellows ◽

Group Iii ◽

Group I ◽

Leaf Chlorosis ◽

Phytoplasma Infection

Strawberry fruit phyllody, production of leaves and other vegetative organs from fruit tissue around achenes, has been ascribed to physiological causes due to temperature conditions during transplant cold storage, plant response to changing seasonal conditions at flower initiation time, and to phytoplasma infection. In examination of phylloid fruits from different strawberry clones and from different locations and sources, we found four distinct phytoplasmas associated with phyllody of strawberry fruit: strawberry multicipita (SM) phytoplasma (16S rRNA group VI, subgroup B), STRAWB2 phytoplasma (16S rRNA group I, subgroup K), clover yellow edge phytoplasma (16S rRNA group III, subgroup A), and a new group III phytoplasma. The SM and STRAWB2 phytoplasmas were detected in plants with phylloid fruit that also exhibited stunting and crown proliferation (SM phytoplasma) or stunting and leaf chlorosis (STRAWB2 phytoplasma). In no instances did we fail to detect phytoplasmas in phylloid fruit. To our knowledge, this is the first report to associate strawberry fruit phyllody with the presence of these phytoplasmas and to report that phytoplasmas other than those belonging to 16S rRNA group I (aster yellows group) can also be associated with strawberry fruit phyllody.

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First Report of a Group 16SrI, Subgroup B, Phytoplasma in Diseased Epilobium hirsutum in the Region of Tallin, Estonia

Plant Disease ◽

10.1094/pdis.2002.86.10.1177a ◽

2002 ◽

Vol 86 (10) ◽

pp. 1177-1177 ◽

Cited By ~ 1

Author(s):

A. Alminaite ◽

R. E. Davis ◽

D. Valiunas ◽

R. Jomantiene

Keyword(s):

16S Rdna ◽

Sequence Similarity ◽

Intergenic Spacer ◽

Rflp Analysis ◽

Nested Polymerase Chain Reaction ◽

Universal Primer ◽

Aster Yellows ◽

First Report ◽

Intergenic Spacer Region ◽

Aster Yellows Phytoplasma

Symptoms of phyllody of flowers and general plant yellowing indicating possible phytoplasma infection were observed in diseased plants of hairy willow-weed (Epilobium hirsutum L., family Onagraceae) growing in a meadow at Harku Village near Tallin, Estonia. DNA was extracted from diseased E. hirsutum using a Genomic DNA Purification Kit (Fermentas AB, Vilnius, Lithuania) and used as a template in nested polymerase chain reaction (PCR). Ribosomal (r) DNA was initially amplified in PCR primed by phytoplasma universal primer pair P1/P7 (4) and reamplified in PCR primed by nested primer pair 16SF2n/16SR2 (F2n/R2) (1) as previously described (2). Products of 1.8 kbp and 1.2 kbp were obtained in PCR primed P1/P7 and F2n/R2, respectively, from all four symptomatic plants examined. These data indicated that the diseased E. hirsutum plants were infected by a phytoplasma, termed epilobium phyllody (EpPh) phytoplasma. The 16S rDNA amplified in PCR primed by nested primer pair F2n/R2 was subjected to restriction fragment length polymorphism (RFLP) analysis using restriction endonucleases AluI, MseI, HpaI, HpaII, HhaI, RsaI, HinfI, and HaeIII (Fermentas AB). On the basis of the collective RFLP profiles, EpPh phytoplasma was classified in group 16SrI (aster yellows phytoplasma group), subgroup B (aster yellows phytoplasma subgroup), according to the phytoplasma classification scheme of Lee et al. (3). The 1.8-kbp rDNA product of P1/P7-primed PCR, which included 16S rDNA, 16S-23S intergenic spacer region, and the 5′ -end of 23S rDNA, was cloned in Escherichia coli using the TOPO TA Cloning Kit (Invitrogen, Carlsbad, Ca) according to manufacturer's instructions and sequenced. The sequence was deposited in the GenBank database as Accession No. AY101386. This nucleotide sequence shared 99.8% sequence similarity with a comparable rDNA sequence (GenBank Accession No. AF322644) of aster yellows phytoplasma AY1, a known subgroup 16SrI-B strain. The EpPh phytoplasma sequence was highly similar (99.9%) to operons rrnA (GenBank Accession No. AY102274) and rrnB (GenBank Accession No. AY102273) from Valeriana yellows (ValY) phytoplasma infecting Valeriana officinalis plants in Lithuania. ValY phytoplasma was found to exhibit rRNA interoperon sequence heterogeneity (D. Valiunas, unpublished data). To our knowledge, this is the first report to reveal E. hirsutum as a host of phytoplasma and to demonstrate the occurrence of a plant pathogenic mollicute in the northern Baltic region. References: (1) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (2) R. Jomantiene et al. HortScience 33:1069, 1998. (3) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (4) B. Schneider et al. Phlogenetic classification of plant pathogenic mycoplasma-like organisms or phytoplasmas. Page 369 in: Molecular and Diagnostic Procedures in Mycoplasmology, Vol 1, R. Razin, and J. G. Tully eds. Academic Press, San Diego, 1995.

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First Report of Aster Yellows-Related Subgroup I-A Phytoplasma Strains in Carrot, Phlox, Sea-Lavender, Aconitum, and Hyacinth in Lithuania

Plant Disease ◽

10.1094/pdis.2001.85.7.804c ◽

2001 ◽

Vol 85 (7) ◽

pp. 804-804 ◽

Cited By ~ 10

Author(s):

D. Valiunas ◽

A. Alminaite ◽

J. Staniulis ◽

R. Jomantiene ◽

R. E. Davis

Keyword(s):

16S Rdna ◽

Nested Pcr ◽

Sequence Similarity ◽

Rflp Analysis ◽

Plant Host ◽

Aster Yellows ◽

Mature Leaves ◽

Young Leaves ◽

Sea Lavender ◽

The Baltic

Phytoplasma strains that belong to group 16SrI (aster yellows phytoplasma group), subgroup A (I-A, North American tomato big bud phytoplasma subgroup) were discovered in diverse plant species in Lithuania. Plants in which the strains were found exhibited symptoms characteristic of infections by phytoplasma. Carrot (Daucus sativus) with carrot proliferation disease exhibited symptoms of proliferation of the crown, chlorosis of young leaves, and reddening of mature leaves. Diseased phlox (Phlox paniculata) exhibited symptoms of virescence and leaf chlorosis. Diseased sea-lavender (Limonium sinuatum) exhibited abnormal proliferation of shoots, chlorosis of young leaves, reddening of mature leaves, and degeneration of flowers. Diseased hyacinth (Hyacinthus orientalis) exhibited chlorosis of leaves and undeveloped flowers. Diseased Aconitum sp. exhibited proliferation of shoots. Phytoplasma-characteristic ribosomal (r) DNA was detected in the plants by use of the polymerase chain reaction (PCR). The rDNA was amplified in PCR primed by primer pair P1/P7 and reamplified in nested PCR primed by primer pair R16F2n/R16R2 (F2n/R2), as previously described (1). The phytoplasmas were classified through restriction fragment length polymorphism (RFLP) analysis of 16S rDNA, amplified in the nested PCR primed by F2n/R2, using single endonuclease enzyme digestion with AluI, MseI, KpnI, HhaI, HaeIII, HpaI, HpaII, RsaI, HinfI, TaqI, and Sau3AI. Collective RFLP patterns indicated that all detected phytoplasma strains were affiliated with subgroup I-A. The 16S rDNA amplified from the phytoplasma (CarrP phytoplasma) in diseased carrot was cloned in Escherichia coli, sequenced, and the sequence deposited in the GenBank data library (GenBank accession no. AF291682). The 16S rDNAs of CarrP and tomato big bud (GenBank acc. no. AF222064) phytoplasmas shared 99.8% nucleotide sequence similarity. Phytoplasmas belonging to group 16SrIII (3), group 16SrV (D. Valiunas, unpublished data), and subgroup I-C in group 16SrI (2,3) occur in Lithuania. This report records the first finding of a subgroup I-A phytoplasma in the Baltic region and expands the known plant host range of this phytoplasma subgroup. References: (1) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (2) Jomantiene et al. Phytopathology 90:S39, 2000. (3) Staniulis et al. Plant Dis. 84:1061, 2000.

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Bacteria diversity in paddy field soil by 16S rDNA-RFLP analysis in Ningxia

Biodiversity Science ◽

10.3724/sp.j.1003.2008.08118 ◽

2008 ◽

Vol 16 (6) ◽

pp. 586 ◽

Cited By ~ 2

Author(s):

Zhang Jianping ◽

Dong Naiyuan ◽

Yu Haobin ◽

Zhou Yongjun ◽

Lu Yongliang ◽

...

Keyword(s):

16S Rdna ◽

Paddy Field ◽

Rflp Analysis ◽

Field Soil ◽

Bacteria Diversity ◽

Paddy Field Soil

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Human intestinal spirochetosis diagnosed with colonoscopy and analysis of partial 16S rDNA sequences of involved spirochetes

Animal Health Research Reviews ◽

10.1079/ahrr200121 ◽

2001 ◽

Vol 2 (1) ◽

pp. 111-116 ◽

Cited By ~ 10

Author(s):

Wolfgang Kraatz ◽

Ulf Thunberg ◽

Bertil Pettersson ◽

Claes Fellström

Keyword(s):

16S Rdna ◽

Type Strain ◽

Sequence Similarity ◽

Rdna Sequences ◽

Intestinal Spirochetosis ◽

16S Rdna Sequences ◽

Pcr Products ◽

Human Intestinal Spirochetosis ◽

Type Strains ◽

Rrna Sequences

AbstractDNA was extracted from colonic biopsies of 33 patients with and three without evidence of intestinal spirochetosis (IS) in the large bowel. The biopsies were subjected to PCR. A pair of primers, generating a 207 bp fragment, were designed to detect specifically the 16S rDNA gene ofBrachyspira. PCR products of the expected size were obtained from 33 samples with histologic evidence of IS. The PCR amplicons were used for sequencing. The sequences obtained were aligned to the corresponding 16S rRNA sequences of five type strains ofBrachyspira. The sequences of 23 PCR products were 99–100% identical with the correspond-ingB.aalborgitype strain sequence. Two cases showed 99–100% sequence similarity with the type strain ofB.pilosicoliP43/6/78. Six cases could not be referred to any of the known species ofBrachyspira. Two PCR products gave incomplete sequences.

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16S rDNA Genotyping of Lactic Acid Bacteria Using PCR-RFLP Analysis

Nippon Shokuhin Kagaku Kogaku Kaishi ◽

10.3136/nskkk.64.355 ◽

2017 ◽

Vol 64 (7) ◽

pp. 355-364

Author(s):

Hideyuki Tamakawa ◽

Yoshihito Ito

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

16S Rdna ◽

Rflp Analysis ◽

Pcr Rflp

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