Characterization of copy numbers of 16S rDNA and 16S rRNA of Candidatus Liberibacter asiaticus and the implication in detection in planta using quantitative PCR
Identifieur interne : 001E02 ( Main/Exploration ); précédent : 001E01; suivant : 001E03Characterization of copy numbers of 16S rDNA and 16S rRNA of Candidatus Liberibacter asiaticus and the implication in detection in planta using quantitative PCR
Auteurs : Jeong-Soon Kim [États-Unis] ; Nian Wang [États-Unis]Source :
- BMC Research Notes [ 1756-0500 ] ; 2009.
Abstract
Citrus Huanglongbing (HLB) is one of the most devastating diseases on citrus and is associated with
We investigated the copy numbers of the 16S rDNA and 16S rRNA of the HLB pathogen and the implication of improving the diagnosis of HLB for early detection using Quantitative PCR. We compared the detection of HLB with different Quantitative PCR based methods with primers/probe targeting either 16S rDNA, beta-operon DNA, 16S rRNA, or beta-operon RNA. The 16S rDNA copy number of
Our result indicates that
Url:
DOI: 10.1186/1756-0500-2-37
PubMed: 19284534
PubMed Central: 2663771
Affiliations:
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Liberibacter asiaticus and the implication in detection <italic>in planta </italic>
using quantitative PCR</title>
<author><name sortKey="Kim, Jeong Soon" sort="Kim, Jeong Soon" uniqKey="Kim J" first="Jeong-Soon" last="Kim">Jeong-Soon Kim</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<wicri:noRegion>FL 33850</wicri:noRegion>
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<author><name sortKey="Wang, Nian" sort="Wang, Nian" uniqKey="Wang N" first="Nian" last="Wang">Nian Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA</nlm:aff>
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<wicri:regionArea>Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850</wicri:regionArea>
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<affiliation wicri:level="1"><nlm:aff id="I2">Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611, USA</nlm:aff>
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<wicri:regionArea>Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611</wicri:regionArea>
<wicri:noRegion>FL 32611</wicri:noRegion>
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Liberibacter asiaticus and the implication in detection <italic>in planta </italic>
using quantitative PCR</title>
<author><name sortKey="Kim, Jeong Soon" sort="Kim, Jeong Soon" uniqKey="Kim J" first="Jeong-Soon" last="Kim">Jeong-Soon Kim</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<author><name sortKey="Wang, Nian" sort="Wang, Nian" uniqKey="Wang N" first="Nian" last="Wang">Nian Wang</name>
<affiliation wicri:level="1"><nlm:aff id="I1">Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA</nlm:aff>
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<wicri:regionArea>Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611</wicri:regionArea>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>Citrus Huanglongbing (HLB) is one of the most devastating diseases on citrus and is associated with <italic>Candidatus </italic>
Liberibacter spp.. The pathogens are phloem limited and have not been cultured <italic>in vitro</italic>
. The current management strategy of HLB is to remove infected citrus trees and reduce psyllid populations with insecticides to prevent the spreading. This strategy requires sensitive and reliable diagnostic methods for early detection.</p>
</sec>
<sec><title>Results</title>
<p>We investigated the copy numbers of the 16S rDNA and 16S rRNA of the HLB pathogen and the implication of improving the diagnosis of HLB for early detection using Quantitative PCR. We compared the detection of HLB with different Quantitative PCR based methods with primers/probe targeting either 16S rDNA, beta-operon DNA, 16S rRNA, or beta-operon RNA. The 16S rDNA copy number of <italic>Ca</italic>
. Liberibacter asiaticus was estimated to be three times of that of the beta-operon region, thus allowing detection of lower titer of <italic>Ca</italic>
. L. asiaticus. Quantitative reverse transcriptional PCR (QRT-PCR) indicated that the 16S rRNA averaged 7.83 times more than that of 16S rDNA for the same samples. Dilution analysis also indicates that QRT-PCR targeting 16S rRNA is 10 time more sensitive than QPCR targeting 16S rDNA. Thus QRT-PCR was able to increase the sensitivity of detection by targeting 16S rRNA.</p>
</sec>
<sec><title>Conclusion</title>
<p>Our result indicates that <italic>Candidatus </italic>
Liberibacter asiaticus contains three copies of 16S rDNA. The copy number of 16S rRNA of <italic>Ca</italic>
. L. asiaticus <italic>in planta </italic>
averaged about 7.8 times of 16S rDNA for the same set of samples tested in this study. Detection sensitivity of HLB could be improved through the following approaches: using 16S rDNA based primers/probe in the QPCR assays; and using QRT-PCR assays targeting 16S rRNA.</p>
</sec>
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