Publications

4468

Conventional PCR and two real‐time PCR (RTi‐PCR) methods were developed and compared using the primer pairs CQULA03F/CQULA03R and CQULA04F/CQULA04R, and TaqMan probe CQULAP1 designed from a species‐specific sequence of the rplJ/rplL ribosomal protein gene, for diagnosis of citrus huanglongbing (HLB) disease in southern China. The specificity and sensitivity of the three protocols for detecting ‘ Candidatus Liberibacter asiaticus’ in total DNA extracts of midribs collected from infected citrus leaves with symptoms in Guangxi municipality, Jiangxi Province and Zhejiang Province, were tested. Sensitivities using extracted total DNA (measured as copy number, CN per µ L of recombinant plasmid solution) were 439·0 (1·30 × 10 5 CN µ L −1 ), 4·39 (1·30 × 10 3 CN µ L −1 ) and 0·44 fg µ L −1 (1·30 × 10 2 CN µ L −1 ) for conventional PCR, TaqMan and SYBR Green I (SGI) RTi‐PCR, respectively. SGI RTi‐PCR was the most sensitive, but its specificity needed to be confirmed by running a melt‐curve assay. The TaqMan RTi‐PCR assay was rapid and had the greatest specificity. Concerning the correlation of PCR detection results with the various HLB symptoms, uneven mottling of leaves had the highest positive rate (96·50%), indicating that leaf mottling was the most reliable symptom for field surveys. Dynamic analysis results from the TaqMan assays showed that the titre (CN) g −1 citrus tissue of ‘ Ca. L. asiaticus’ was highest between October and December (threshold cycle ( C t ) average = 29·3, CN = 3·35 × 10 7 ) and lowest between March and May ( C t average = 32·0, CN = 5·10 × 10 6 ) in 2004 and 2005. The optimized molecular‐based assays should prove useful for presymptom diagnosis of HLB disease, monitoring and identification of ‘ Ca. L. asiaticus’, and field epidemic regulation.

Summary Twelve strains of filamentous bacteria morphologically identified as ‘ Microthrix parvicella ’ were isolated from industrial activated sludge wastewater treatment plants. 16S rRNA gene sequences analysis showed that these strains were all closely related to ‘ Candidatus Microthrix parvicella’. Six of them, however, had a 16S rRNA gene similarity of only 95.7% and 96.7% to ‘ Candidatus Microthrix parvicella’ suggesting the presence of a new species. The name ‘ Candidatus Microthrix calida’ is proposed for this new microorganism. The physiological properties of these six isolates supported the description of a new taxon. The ‘ Candidatus Microthrix calida’ strains produced thin filaments (0.3–0.7 µm diameter), they did not grow on the media supporting the growth of ‘ Candidatus Microthrix parvicella’ and could be cultivated at higher temperature (up to 36.5°C). Preliminary data on substrate uptake were obtained by microautoradiography on pure culture. Two new fluorescence in situ hybridization (FISH) probes, Mpa‐T1‐1260 specific for ‘ Candidatus Microthrix calida’ and Mpa‐all‐1410 targeting both Microthrix species, were designed. The presence of Microthrix spp. was investigated in 114 activated sludge plants. ‘ Microthrix parvicella ’ morphotype was detected in 23% of the analysed samples and FISH analysis revealed that ‘ Candidatus Microthrix calida’ was present in 5% of them. The remaining ‘ M. parvicella ’ filaments were positive with probe Mpa‐all‐1410 but could not all be identified as ‘ Candidatus Microthrix parvicella’ suggesting the presence of more hitherto undescribed biodiversity within this morphotype.

The purpose of this announcement is to effect the valid publication of the following new names and new combinations under the procedure described in the Bacteriological Code (1990 Revision). Authors and other individuals wishing to have new names and/or combinations included in future lists should send three copies of the pertinent reprint or photocopies thereof to the IJSEM Editorial Office for confirmation that all of the other requirements for valid publication have been met. It is also a requirement of IJSEM and the ICSP that authors of new species, new subspecies and new combinations provide evidence that types are deposited in two recognized culture collections in two different countries (i.e. documents certifying deposition and availability of type strains). It should be noted that the date of valid publication of these new names and combinations is the date of publication of this list, not the date of the original publication of the names and combinations. The authors of the new names and combinations are as given below, and these authors' names will be included in the author index of the present issue and in the volume author index. Inclusion of a name on these lists validates the publication of the name and thereby makes it available in bacteriological nomenclature. The inclusion of a name on this list is not to be construed as taxonomic acceptance of the taxon to which the name is applied. Indeed, some of these names may, in time, be shown to be synonyms, or the organisms may be transferred to another genus, thus necessitating the creation of a new combination.