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Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular fluid of infected sugarcane plants

Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular... DNA of Leifsonia xyli subsp. xyli (Lxx), the causal agent of ratoon stunting disease of sugarcane, was detected in the fibrovascular fluid of sugarcane plants using random amplified polymorphic DNA PCR-based amplification using two 10-mer oligonucleotide primers. The primers OPC-02 and OPC-11 produced Lxx-specific markers of approximately 800 bp and 1000 bp, respectively. A cloned DNA fragment from the 800 bp PCR product (pSKC2-800) hybridised to a single genomic DNA fragment from Lxx when used as a probe in Southern hybridisation. This cloned fragment did not hybridise to L. xyli subsp. cynodontis (Lxc), or L. xyli-like bacteria isolated from grasses in Australia, indicating the usefulness of this DNA fragment as a specif ic probe for Lxx. A cloned fragment from the 1000 bp PCR product (pSKC11-1000) hybridised to three genomic fragments in Lxx isolates, one genomic fragment in two of the four isolates of L. xyli-like bacteria, and in two of the four isolates of Lxc isolated from the USA. These results indicate that L. xyli-like bacteria are more likely to be related to Lxc than Lxx. These probes did not hybrid ise to the DNA from strains of the species of Clavibacter, Rathayibacter, Acidovorax, Ralstonia, Pseudomonas and Xanthomonas tested. Two oligonucleotide primers (21-mer) designed from the pSKC2-800 sequences specifically amplified template DNA from Lxx and detected as few as 5 × 104 cells/mL in fibrovascular fluid from sugarcane plants infected with Lxx. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Australasian Plant Pathology Springer Journals

Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular fluid of infected sugarcane plants

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Publisher
Springer Journals
Copyright
Copyright © 2003 by Australasian Plant Pathology Society
Subject
Life Sciences; Plant Pathology; Plant Sciences; Agriculture; Entomology; Ecology
ISSN
0815-3191
eISSN
1448-6032
DOI
10.1071/AP03036
Publisher site
See Article on Publisher Site

Abstract

DNA of Leifsonia xyli subsp. xyli (Lxx), the causal agent of ratoon stunting disease of sugarcane, was detected in the fibrovascular fluid of sugarcane plants using random amplified polymorphic DNA PCR-based amplification using two 10-mer oligonucleotide primers. The primers OPC-02 and OPC-11 produced Lxx-specific markers of approximately 800 bp and 1000 bp, respectively. A cloned DNA fragment from the 800 bp PCR product (pSKC2-800) hybridised to a single genomic DNA fragment from Lxx when used as a probe in Southern hybridisation. This cloned fragment did not hybridise to L. xyli subsp. cynodontis (Lxc), or L. xyli-like bacteria isolated from grasses in Australia, indicating the usefulness of this DNA fragment as a specif ic probe for Lxx. A cloned fragment from the 1000 bp PCR product (pSKC11-1000) hybridised to three genomic fragments in Lxx isolates, one genomic fragment in two of the four isolates of L. xyli-like bacteria, and in two of the four isolates of Lxc isolated from the USA. These results indicate that L. xyli-like bacteria are more likely to be related to Lxc than Lxx. These probes did not hybrid ise to the DNA from strains of the species of Clavibacter, Rathayibacter, Acidovorax, Ralstonia, Pseudomonas and Xanthomonas tested. Two oligonucleotide primers (21-mer) designed from the pSKC2-800 sequences specifically amplified template DNA from Lxx and detected as few as 5 × 104 cells/mL in fibrovascular fluid from sugarcane plants infected with Lxx.

Journal

Australasian Plant Pathology – Springer Journals

Published: Jan 28, 2011

References

  • Transposon mutagenesis of Pseudomonas solanacearum: isolation of Tn5-induced avirulent mutants
    Boucher, CA; Barberis, PA; Trigalet, PA; Demery, DA
  • Development and application of cloned DNA probes for Clavibacter xyli subsp. xyli, the causal agent of sugarcane ratoon stunting
    Chung, CH; Lin, CP; Chen, CT
  • Sugarcane germplasm conservation and exchange
    Croft, BJ; Smith, GR
  • Detection of the RSD-associated bacterium by serologically specif ic electron microscopy
    Damann, KE; Derrick, KS; Gillaspie, AG; Fontenot, DB; Kao, J
  • Comparison of diagnostic techniques for determining incidence of ratoon stunting disease of sugarcane in Florida
    Davis, MJ; Dean, JL
  • Ratoon stunting disease of sugarcane: Isolation of the causal organism
    Davis, MJ; Gillaspie, AG; Harris, RW; Lawson, RH
  • RNA from the yeast transposable element Ty l has both ends in the direct repeats, a structure similar to retrovirus RNA
    Elder, RT; Loh, EY; Davis, RW
  • Sensitive and specific detection of Clavibacrer xyli subsp. xyli, causal agent of ratoon stunting disease of sugarcane, with a polymerase chain reaction-based assay
    Fegan, M; Croft, BJ; Teakle, DS; Hayward, AC; Smith, GR
  • Ratoon stunting disease: serology
    Gillaspie, AG
  • Limitations of ELISA for detection of the RSD-associated bacterium in sugarcane and sudangrass
    Gillaspie, AG; Harris, RW
  • Diseases of sugarcane: major diseases
    Gillaspie, AG; Teakle, DS
  • Immunofluorescent diagnosis of ratoon stunting disease
    Harris, RW; Gillaspie, AG
  • A method for characterizing Pseudomonas solanacearum
    Hayward, AC
  • Primers are decisive for sensitivity of PCR
    He, Q; Marjamaki, M; Soini, H; Mertsola, J; Viljanen, MK
  • Laboratory protocols: CIMMYT Applied Molecular Genetics Laboratory
    Hoisington, D; Khairallah, M; Gonzalez de Leon, D
  • Some further developments in the study of ratoon stunting disease in Queensland
    Hughes, CG; Steindl, DRL
  • Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products
    Marchuk, D; Drumm, M; Saulino, A; Collins, FS
  • Leifsonia xyli-like bacteria are endophytes of grasses in eastern Australia
    Mills, L; Leaman, TM; Taghavi, SM; Shackel, L; Dominiak, BC; Taylor, PWJ; Fegan, M; Teakle, DS
  • A polymerase chain reaction protocol for the detection of Clavibacter xyli subsp. xyli, the causal bacterium of sugarcane ratoon stunting disease
    Pan, Y-B; Grisham, MP; Burner, DM; Damann, KE; Wei, Q
  • An improved method of xylem-sap extraction using positive pressure for the rapid diagnosis of ratoon stunting disease
    Richardson, SR
  • Clavibacter toxicus sp. nov., the bacterium responsible for annual ryegrass toxicity in Australia
    Riley, IT; Ophel, KM
  • Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues
    Rogers, SO; Bendich, AJ
  • Molecular cloning: a laboratory manual
    Sambrook, J; Fritsch, EF; Maniatis, T
  • Ratoon stunting disease
    Steindl, DRL
  • Harvester transmission of leaf scald and ratoon stunting disease, Demonstration and evaluation of methods of decontamination
    Taylor, PWJ; Ryan, CC; Birch, RG
  • The effect of high temperature on the sugar cane ratoon stunting bacterium, Clavibacter xyli subsp
    Teakle, DS; Ryan, CC