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

Taylor, P. W. J., Petrasovits, L. A., Van der Velde, R., Birch, R. G., Croft, B. J., Fegan, M., Smith, G. R. and Brumbley, S. (2003) Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular fluid of infected sugarcane plants. Australasian Plant Pathology, 32 3: 367-375. doi:10.1071/AP03036


Author Taylor, P. W. J.
Petrasovits, L. A.
Van der Velde, R.
Birch, R. G.
Croft, B. J.
Fegan, M.
Smith, G. R.
Brumbley, S.
Title Development of PCR-based markers for detection of Leifsonia xyli subsp. xyli in fibrovascular fluid of infected sugarcane plants
Journal name Australasian Plant Pathology   Check publisher's open access policy
ISSN 0815-3191
1448-6032
Publication date 2003
Sub-type Article (original research)
DOI 10.1071/AP03036
Volume 32
Issue 3
Start page 367
End page 375
Total pages 9
Place of publication Collingwood
Publisher CSIRO Publishing
Collection year 2003
Language eng
Subject C1
270403 Plant Pathology
620000 - Plant Production and Plant Primary Products
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 specific 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 hybridise 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 x 10(4) cells/mL in fibrovascular fluid from sugarcane plants infected with Lxx.
Keyword Molecular Markers
Polymerase Chain Reaction
Ratoon Stunting Disease
Ratoon-stunting-disease
Pseudomonas-solanacearum
Causal Bacterium
Australia
Agent
Dna
Q-Index Code C1

 
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Created: Wed, 15 Aug 2007, 02:13:20 EST