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

Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1177-1177 ◽  
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.

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

Plant Disease ◽  
2002 ◽  
Vol 86 (4) ◽  
pp. 443-443 ◽  
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.

scholarly journals First Report of Aster Yellows Phytoplasma in Alfalfa

Plant Disease ◽  
1999 ◽  
Vol 83 (5) ◽  
pp. 488-488 ◽  
Author(s):  
R. D. Peters ◽  
M. E. Lee ◽  
C. R. Grau ◽  
S. J. Driscoll ◽  
R. M. Winberg ◽  
...  
Keyword(s):  
16S Rdna ◽  
Nested Pcr ◽  
Sequence Data ◽  
Sequence Similarity ◽  
Similarity Function ◽  
Sterile Water ◽  
Aster Yellows ◽  
First Report ◽  
Pcr Product ◽  
Aster Yellows Phytoplasma

Samples of alfalfa (Medicago sativa L.) leaves and stems showing symptoms of inter-veinal chlorosis and purpling, commonly associated with insect feeding, were collected from 8 sites in central and southern Wisconsin in autumn of 1998. Samples were frozen within 24 h of collection. Approximately 0.3 g of plant tissue from each sample was used for total DNA extraction according to the protocol of Zhang et al. (4), with minor modifications in grinding procedures and reagent volumes to optimize results. Nested polymerase chain reaction (PCR) was carried out by amplification of 16S rDNA with the universal primer pairs R16mF2/R16mR1 followed by R16F2n/R16R2 as described by Gunder-sen and Lee (1). Undiluted total sample DNA was used for the first amplification; PCR products were diluted (1:30) in sterile water prior to final amplification. Alfalfa DNA and sterile water were used as negative controls; DNA from phytoplasma causing X-disease in peach (CX) served as a positive control. Fragments of 16S rDNA from putative phytoplasmas amplified by PCR with the primer pair R16F2n/R16R2 were characterized by restriction endonuclease digestion (3). The resulting restriction fragment length polymorphism (RFLP) patterns were compared with patterns for known phytoplasmas described by Lee et al. (3). Products of nested PCR were also purified and sequenced with primers R16F2n/R16R2 and an automated DNA sequencer (ABI 377XL; C. Nicolet, Biotechnology Center, University of Wisconsin-Madison). Of 51 samples of alfalfa assessed, one sample from Evansville, WI, yielded a nested PCR product of the appropriate size (1.2 kb), indicating the presence of phytoplasma. Digestion of this product with various restriction enzymes produced RFLP patterns that were identical to those for phytoplasmas in the aster yellows phytoplasma subgroup 16SrI-A (3). Alignment of the DNA sequence of the nested PCR product from the positive sample with sequences found in the GenBank sequence data base (National Center for Biotechnology Information, Bethesda, MD) with the BLAST sequence similarity function confirmed this result. Although other phytoplasma strains (particularly those causing witches'-broom) have been reported to infect alfalfa (2), this is the first report of the presence of the aster yellows phytoplasma in the alfalfa crop. Vectors involved in transmission and the potential agronomic impacts of aster yellows phytoplasma in alfalfa are topics of current investigation. References: (1) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (2) A.-H. Khadhair et al. Microbiol. Res. 152:269, 1997. (3) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (4) Y.-P. Zhang et al. J. Virol. Methods 71:45, 1998.

scholarly journals First Report of an Aster Yellows Phytoplasma Associated with Cabbage in Southern Texas

Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 447-447 ◽  
Author(s):  
I.-M. Lee ◽  
R. A. Dane ◽  
M. C. Black ◽  
Noel Troxclair
Keyword(s):  
16S Rdna ◽  
Nested Pcr ◽  
Restriction Enzymes ◽  
Diagnostic Procedures ◽  
Acid Extraction ◽  
Insect Vector ◽  
Aster Yellows ◽  
First Report ◽  
Pcr Products ◽  
Aster Yellows Phytoplasma

In early spring 2000 carrot crops in southwestern Texas were severely infected by an outbreak of phyllody associated with aster yellows phytoplasma. Cabbage crops that had been planted adjacent to these carrot fields began to display previously unobserved symptoms characteristic of phytoplasma infection. Symptoms included purple discoloration in leaf veins and at the outer edges of leaves on cabbage heads. Proliferation of sprouts also occurred at the base of the stem and between leaf layers of some plants, and sprouts sometimes continued to proliferate on extended stems. About 5% of cabbage plants in the field exhibited these symptoms. Two symptomless and four symptomatic cabbage heads were collected in early April from one cabbage field. Veinal tissues were stripped from each sample and used for total nucleic acid extraction. To obtain specific and sufficient amount of PCR products for analysis, nested PCR was performed by using primer pairs (first with P1/P7 followed by R16F2n/R16R2) (1,2) universal for phytoplasma detection. A specific 16S rDNA fragment (about 1.2 kb) was strongly amplified from the four symptomatic but not from the two asymptomatic samples. The nested PCR products obtained from the four symptomatic samples were then analyzed by restriction fragment length polymorphism (RFLP) using the restriction enzymes MseI, HhaI, and HpaII, and the RFLP patterns were compared to the published patterns of known phytoplasmas (1). The resulting RFLP patterns were identical to those of a phytoplasma belonging to subgroup B of the aster yellows phytoplasma group (16SrI). These RFLP patterns were also evident in putative restriction sites observed in a 1.5 kbp nucleotide sequence of the 16S rDNA. This is the first report of aster yellows phytoplasma associated disease symptoms in cabbage in Texas. The occurrence of cabbage proliferation coincided with the presence of high populations of the insect vector, aster leafhopper. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) B. Schneider et al. 1995. Molecular and Diagnostic Procedures in Mycoplasmology, Vol. I. Academic Press, San Diego, CA.

scholarly journals First Report of Phytoplasmas in Soybean, Alfalfa, and Lupinus sp. in Lithuania

Plant Disease ◽  
2000 ◽  
Vol 84 (2) ◽  
pp. 198-198 ◽  
Author(s):  
R. Jomantiene ◽  
R. E. Davis ◽  
L. Antoniuk ◽  
J. Staniulis
Keyword(s):  
16S Rdna ◽  
Type Strain ◽  
Sequence Data ◽  
Sequence Similarity ◽  
Dna Amplification ◽  
Plant Host ◽  
Nucleotide Sequence Data ◽  
Aster Yellows ◽  
Veinal Necrosis ◽  
Aster Yellows Phytoplasma

Plants of cultivated soybean (Glycine max) and alfalfa (Medicago sativa) in Dotnuva and of wild Lupinus sp. in Ledakalnis, Lithuania, exhibited symptoms that suggested phytoplasmal infections. Soybean plants were of normal growth habit but exhibited veinal necrosis. Alfalfa and Lupinus plants exhibited stunting, abnormally small leaves, and witches'-broom symptoms. Diseases in the plants were termed soybean veinal necrosis (SVN), alfalfa stunt (AlfS), and Lupinus stunt (LupS), respectively. The presence of phytoplasmas in diseased plants was assessed using polymerase chain reaction (PCR) for amplification of phytoplasma-specific 16S rDNA. A phytoplasma-characteristic 1.2-kbp DNA fragment was amplified from all diseased plants but not from known healthy plants in nested PCRs in which the first DNA amplification was primed by primer pair P1/P7 and reamplification of DNA was primed by primer pair F2n/R2 (2,4). Products from the nested PCR primed by F2n/R2 were subjected to restriction fragment length polymorphism (RFLP) analysis, and the RFLP patterns obtained were compared with patterns previously published (1–4). On the basis of AluI, HaeIII, HhaI, HpaI, KpnI, MseI, and RsaI RFLP patterns, the SVN and LupS phytoplasmas were classified in group 16SrIII (peach X-disease phytoplasma group), subgroup B (III-B, type strain clover yellow edge phytoplasma), and the AlfS phytoplasma was classified in group 16SrI (aster yellows phytoplasma group), subgroup B (I-B, type strain aster yellows phytoplasma). Nucleotide sequences were determined for 16S rDNA fragments amplified from SVN and AlfS phytoplasmas in nested PCRs primed by F2n/R2. The sequences were deposited in GenBank under Accession nos. AF177383 for SVN and AF177384 for AlfS. Sequence similarity between the 16S rDNAs of SVN and Canadian clover yellow edge (strain CYE-C, GenBank Accession no. AF175304) phytoplasmas was 99.8%; sequence similarity between 16S rDNAs of AlfS and aster yellows (strain SAY, GenBank Accession no. M86340) phytoplasmas was 99.6%. The SVN phytoplasma 16S rDNA shared 100% sequence similarity with a 16S rDNA from the Lithuanian clover yellow edge (CYE-L, GenBank Accession no. AF173558) phytoplasma. The nucleotide sequence data supported the conclusion that the SVN and AlfS phytoplasmas were closely related to strains classified in subgroups III-B and I-B, respectively. Our findings extend the known geographic ranges of phytoplasma subgroups I-B and III-B to northern Europe, including Lithuania, and expand the known plant host ranges of these pathogens. References: (1) R. E. Davis et al. Int. J. Syst. Bacteriol. 47:262, 1997. (2) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (3) R. Jomantiene et al. HortScience 33:1069, 1998. (4) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

scholarly journals First Report of an Aster Yellows Subgroup 16SrI-B Phytoplasma Infecting Chayote in Costa Rica

Plant Disease ◽  
2002 ◽  
Vol 86 (3) ◽  
pp. 330-330 ◽  
Author(s):  
W. Villalobos ◽  
L. Moreira ◽  
C. Rivera ◽  
K. D. Bottner ◽  
I.-M. Lee
Keyword(s):  
Costa Rica ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Potential Threat ◽  
Nested Polymerase Chain Reaction ◽  
Aster Yellows ◽  
First Report ◽  
Rdna Sequences ◽  
Pcr Products ◽  
Production Area

An outbreak of a witches' broom disease affected approximately 20% of plants in several chayote (Sechium edule (Jacq.) Schwartz) fields in the commercial production area of the Ujarrás Valley, Cartago Province, Costa Rica during 2000 and 2001. Affected chayote plants exhibited symptoms, including basal proliferation with severe foliage reduction, aborted flowers, and deformed fruits, suggestive of phytoplasmal infection. Two other symptomatic cucurbit species growing near the chayote fields were also identified. These species were tacaco plants (S. tacaco (Pitt.) C. Jeffrey), an edible cucurbit for domestic marketing in Costa Rica, showing severe size reduction of leaves and fruits, and Rytidostylis carthaginensis (Jacq.) Kuntze, a weed in chayote and tacaco fields, exhibiting abnormal tendril proliferation. Plants were analyzed for phytoplasma infection by a nested polymerase chain reaction (PCR) assay, using the universal rRNA primer pair P1/P7 followed by R16F2n/R16R2 (2). Phytoplasmas were detected in all symptomatic samples (18 chayote, 6 tacaco, and 3 weed) tested but were undetectable in all asymptomatic samples (10 chayote, 6 tacaco, and 2 weed). Restriction fragment length polymorphism (RFLP) analysis of PCR products (16S rDNA sequences) by separate digestion with eight restriction enzymes (RsaI, HhaI, KpnI, BfaI, HaeIII, HpaII, AluI, MseI) revealed that a phytoplasma belonging to subgroup 16SrI-B in the aster yellows phytoplasma group (16SrI) was associated with chayote witches' broom (CWB). The same or very similar phytoplasmas were found in both symptomatic tacaco and R. carthaginensis plants. Phylogenetic analysis of 16SrDNA sequences also confirmed the CWB phytoplasma to be most similar to members of subgroup 16SrI-B. Similar diseases in chayote and other cucurbits have been reported in Brazil (3), Taiwan (1), and Mexico (4). The CWB phytoplasma differs from the phytoplasma (16SrIII-J subgroup) associated with chayote in Brazil. The identities of phytoplasmas associated with cucurbits in Taiwan and Mexico are unknown. The occurrence of an aster yellows group phytoplasma in chayote may pose a potential threat to continued production and exportation of this cash crop. To our knowledge, this is the first report of 16SrI-B subgroup phytoplasmas in naturally infected cucurbits in Costa Rica. References: (1) T. G. Chou et al. Plant Dis. Rep. 60:378, 1976. (2) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (3) H. G. Montano et al. Plant Dis. 84:429, 2000. (4) E. Olivas. Rev. Fitopatol. (Lima) 13:14, 1978.

scholarly journals Corn with Symptoms of Reddening: New Host of Stolbur Phytoplasma

Plant Disease ◽  
2006 ◽  
Vol 90 (10) ◽  
pp. 1313-1319 ◽  
Author(s):  
B. Duduk ◽  
A. Bertaccini
Keyword(s):  
16S Rdna ◽  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Gene Sequences ◽  
New Host ◽  
Intergenic Spacer Region ◽  
New Finding ◽  
Polymerase Chain ◽  
Stolbur Phytoplasma ◽  
Uncertain Etiology

Recurrent epiphytotic outbreaks of a disease of uncertain etiology known as reddening of corn (Zea mays) have occurred in some areas of Serbia during the last 50 years. Affected plants show early and abnormal ripening, dry precociously, and have poor, shriveled grains. Using molecular tools, phytoplasmas were detected in diseased plants and their identity was subsequently deduced as a subgroup 16SrXII-A strain by a variety of supporting assays involving restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified 16S rDNA and tuf gene sequences, selective amplification of phytoplasma DNA using primer pair G35p/m, similarity of 16-23S intergenic spacer region (SR) sequences, and similarity and phylogenetic analysis of 16S rDNA gene sequences. Presence of stolbur phytoplasmas in corn with reddening symptoms is a new finding not only for Serbia: it is the first report of stolbur phytoplasma in this species worldwide.

scholarly journals First Report of Barley as Host of a Phytoplasma Belonging to Group 16SrI, Subgroup B, and Ribosomal Protein Subgroup rpI-B in Lithuania

Plant Disease ◽  
2005 ◽  
Vol 89 (3) ◽  
pp. 339-339 ◽  
Author(s):  
L. Urbanaviciene ◽  
R. Jomantiene ◽  
R. E. Davis
Keyword(s):  
Ribosomal Protein ◽  
16S Rdna ◽  
Rflp Analysis ◽  
Gene Sequences ◽  
Universal Primer ◽  
Group I ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Template Dna ◽  
Aster Yellows Phytoplasma

Numerous diseased plants of barley (Hordeum vulgaris L.) exhibiting twisted, abnormally thin and yellowed awns, reduced spikelets, and general stunting and yellowing were observed in fields in the Vilnius and Kaisiadorys regions of Lithuania. The possible association of a phyto-plasma with the disease, termed barley deformation (BaDef), was assessed using polymerase chain reaction (PCR). Three phytoplasma universal primer pairs (P1/P7, R16F2n/R16R2, and rpF1/rpR1) (1,2,4) were employed to amplify ribosomal (r) RNA gene (rDNA) and ribosomal protein (rp) gene sequences. Template DNA extractions and PCR (direct and nested) were conducted as previously described (4). Although DNA was amplified in PCRs containing template extracted from diseased plants, no amplification was observed in PCRs containing DNA from symptomless plants sampled from the same fields. The BaDef phytoplasma was identified and classified according to Lee et al. (4) through restriction fragment length polymorphism (RFLP) analysis of 1.2-kbp 16S rDNA amplified in the PCR primed by primer pair R16F2n/R16R2 and analysis of the 1.2-kbp rp gene sequences amplified in PCR primed by primer pair rpF1/rpR1. On the basis of collective RFLP patterns of amplified 16S rDNA and rp gene sequences, the BaDef phytoplasma was classified as a member of group 16SrI (group I, aster yellows phytoplasma group), subgroup B (16SrI-B), and rp subgroup rpI-B. Ribosomal protein subgroup B was distinguished from other rp subgroups on the basis of the presence of a recognition site for HpaII. The 1.8-kbp rDNA product of PCR primed by P1/P7 and the 1.2-kbp rpF1/rpR1 PCR product were cloned and sequenced, and the sequences were deposited in GenBank under Accession No. AY734453 for the BaDef 16S rDNA and Accession No. AY735448 for the BaDef rp gene sequence. Previously, only oat proliferation (OatP) phytoplasma, a member of subgroup 16SrI-A, had been characterized in a cereal crop (Avena sativa L.) in Europe (3); BaDef is another phytoplasmal disease threatening cereal crops in the region. References: (1) S. Deng and D. Hiruki. J. Microbiol. Methods 14:53, 1991. (2) D. E. Gundersen and I. M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) R. Jomantiene et al. Plant Dis. 86:443, 2002. (4) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

scholarly journals First Report of Spirea Witches'-Broom Disease in China

Plant Disease ◽  
2007 ◽  
Vol 91 (5) ◽  
pp. 635-635 ◽  
Author(s):  
R. Gao ◽  
J. Wang ◽  
X. D. Li ◽  
X. P. Zhu ◽  
G. Z. Tian
Keyword(s):  
New York ◽  
16S Rdna ◽  
Rflp Analysis ◽  
Northern China ◽  
Aster Yellows ◽  
First Report ◽  
Taxonomy Group ◽  
Ctab Method ◽  
Broom Disease ◽  
Intensive Investigation

Bumald spirea (Spiarea bumalda Burv.) is an important ornamental tree widely grown in northern China. In August of 2006, spirea plants exhibiting symptoms of witches'-broom, stunting, yellowing, and shoot dieback were found at an incidence of 5 to 15% in Qingzhou City, Shandong Province, China. Total DNA was extracted separately from 0.1 g of phloem tissue from leaf midribs and stems of six symptomatic and six asymptomatic plants with a modified cetyltriethylammonium bromide (CTAB) method (3). Resulting DNA samples were analyzed for phytoplasma DNA by a nested PCR assay using phytoplasma universal 16S rDNA gene primer pairs R16mF2/R16mR1 and R16F2n/R16R2 (2). These primers amplified 1.5- and 1.2-kb products, respectively, from DNA of all symptomatic plants only. Restriction fragment length polymorphism (RFLP) analysis of the 1.2-kb 16S rDNA product using enzymes AluI, MseI, and HhaI indicated that all symptomatic plants contained a group 16SrI (aster yellows group) subgroup B (16SrI-B) phytoplasma strain (4). A 16S rDNA sequence derived from this strain (GenBank Accession No. EF176608) was most similar (99.8 and 99.6%) to those of severe aster yellows (GenBank Accession No. M86340) and Maryland aster yellows (GenBank Accession No. AF322644) phytoplasmas, respectively, thereby confirming strain identity based on RFLP analysis. A phytoplasma (Spiarea stunt phytoplasma, GenBank Accession No. AF190228), which belongs to X-disease group (16SrIII), was reported to infect spirea and probably be lethal to S. tomentosa in New York (1,4). The phytoplasma reported here shared low identity (90.8%) with Spiarea stunt phytoplasma, but also caused dieback of spirea shoots. The epidemiology and economic impact of this disease need further intensive investigation. To our knowledge, this is the first report of spirea witches'-broom disease and of its association with a subgroup 16SrI-B phytoplasma in China. References: (1) H. M. Griffiths et al. Can. J. Plant Pathol. 16:255, 1994. (2) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) Y. Qi et al. Biotechnol. Bull. 4:44, 2004. (4) The IRPCM Phytoplasma/Spiroplasma Working Team-Phytoplasma Taxonomy Group. Int. J. Syst. Bacteriol. 54:1243, 2004.

scholarly journals Detection of XIIA phytoplasma group on cultivar Zupljanka in Zupa vineyard region by RFLP analysis of 16s rDNA sequences

Genetika ◽  
2010 ◽  
Vol 42 (1) ◽  
pp. 145-153
Author(s):  
Dragana Josic ◽  
Slobodan Kuzmanovic ◽  
Sasa Stojanovic ◽  
Goran Aleksic ◽  
Snezana Pavlovic ◽  
...  
Keyword(s):  
16S Rdna ◽  
Direct Sequencing ◽  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Intergenic Spacer Region ◽  
Region Detection ◽  
Rdna Sequences ◽  
16S Rdna Sequences ◽  
Stolbur Phytoplasma

'Bois noir' (BN) is an important grapevine disease associated with phytoplasmas belonging to ribosomal subgroup 16SrXII-A. Phytoplasmas cause diseases in several hundred plant species. The number of infected cultivars is growing each year and it is important to follow the spreading of the phytoplasma in the different regions and identify which strains are present in specific regions on specific cultivars. Phytoplasmas are identified and classified based on direct sequencing of phytoplasma 16S rDNA or the 16S to 23S intergenic spacer region, but this approach is not always practical when a large number of unknown phytoplasmas is to be analyzed. Classification by RFLP analysis has provided a simple and rapid method that can be used to differentiate and identify a large number of unclarified phytoplasmas. Our objective was to investigate presence of phytoplasmas of 16SrXII-A group (Stolbur) in Zupa vineyard region. Detection was based on RFLP analysis of 16s rDNA sequences using four restriction enzymes: Tru1I, AluI, KpnI and TaqI. We identified phytoplasmas of XIIA group on two of three investigated cultivars (Zupljanka and Frankovka, but not on Plovdina) in the Zupa vineyard regions (Gornje Rataje and Tules locality). This is the first report of Stolbur phytoplasma on cv. Zupljanka in Zupa region.

scholarly journals First Report of Alfalfa Witches Broom Disease in Oman Caused by a Phytoplasma of the 16Sr II Group

Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1287-1287 ◽  
Author(s):  
A. J. Khan ◽  
K. M. Azam ◽  
M. L. Deadman ◽  
A. M. Al-Subhi ◽  
P. Jones
Keyword(s):  
Sweet Potato ◽  
Infected Plant ◽  
Rflp Analysis ◽  
Negative Control ◽  
Sultanate Of Oman ◽  
Nested Polymerase Chain Reaction ◽  
Aster Yellows ◽  
First Report ◽  
Pcr Products ◽  
Plant Pathol

Alfalfa (Medicago sativa L.) is a primary forage crop in the Sultanate of Oman. A new disease of alfalfa in Oman is characterized by proliferation of shoots and yellowing of leaves in 1- to 2-year-old plants and tillering of stems in 4- to 5-year-old plants. Annual losses due to this disease are estimated at more than US$ 23 million. Samples of healthy and infected alfalfa plants were collected from different regions. Total DNA was extracted according to Khadhair et al. (1), with minor modifications. Amplification of 16S rDNA was done using a nested polymerase chain reaction (PCR) approach with primers P1/P7 and R16F2n/R16R2. DNA from healthy leaves and sterile water was used as a negative control, while DNA from periwinkle infected with faba bean phyllody (16SrII-C), aster yellows (16SrI), tomato big bud (16SrII-D), sweet potato little leaf (16SrII-D), catharanthus phyllody (16SrVI), and sesame phyllody (16SrII-A) were used as positive controls and for restriction fragment length polymorphism (RFLP) comparisons. Nested 1.25-kb PCR products from infected plant samples were subjected to RFLP analysis with restriction endonucleases RsaI, AluI, HaeIII, HhaI, EcoRI, TaqI, Tru9I, and Sau3AI. The analysis showed that the alfalfa witches' broom phytoplasma (AWBP) belonged to the 16SrII group (peanut witches' broom) and that the AWBP was most similar to sweet potato little leaf (16SrII-D) but distinct from “Candidatus Phytoplasma aurantifolia,” the cause of lime witches' broom in Oman. Other phytoplasmas infecting alfalfa have been reported from Europe and North America (1,3), but they belong to the 16SrVI (clover phyllody) and 16SrI (aster yellows) groups. An alfalfa witches' broom reported from Italy (2) forms a separate grouping (4). To our knowledge, this is the first report of a phytoplasma from the peanut witches' broom group infecting alfalfa in the Sultanate of Oman. References: (1) A. H. Khadhair et al. Microbiol. Res. 152:259, 1997. (2) C. Marcone et al. J. Plant Pathol. 79:211, 1997. (3) R. D. Peters et al. Plant Dis. 83:488, 1999. (4) E. Seemuller et al. J. Plant Pathol. 80:3, 1998.

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