HybProbes-based real-time PCR assay for specific identification of Streptomyces scabies and Streptomyces europaeiscabiei, the potato common scab pathogens.

UNLABELLED: The aim of this study was to develop and validate a HybProbes-based real-time PCR assay targeting the trpB gene for specific identification of Streptomyces scabies and Streptomyces europaeiscabiei. Four primer pairs and a fluorescent probe were designed and evaluated for specificity in identifying S. scabies and Streptomyces europaeiscabiei, the potato common scab pathogens. The specificity of the HybProbes-based real-time PCR assay was evaluated using 46 bacterial strains, 23 Streptomyces strains and 23 non-Streptomyces bacterial species. Specific and strong fluorescence signals were detected from all nine strains of S. scabies and Streptomyces europaeiscabiei. No fluorescence signal was detected from 14 strains of other Streptomyces species and all non-Streptomyces strains. The identification was corroborated by the melting curve analysis that was performed immediately after the amplification step. Eight of the nine S. scabies and S. europaeiscabiei strains exhibited a unique melting peak, at Tm of 69·1°C while one strain, Warba-6, had a melt peak at Tm of 65·4°C. This difference in Tm peaks could be attributed to a guanine to cytosine mutation in strain Warba-6 at the region spanning the donor HybProbe. The reported HybProbes assay provides a more specific tool for accurate identification of S. scabies and S. europaeiscabiei strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports a novel assay based on HybProbes chemistry for rapid and accurate identification of the potato common scab pathogens. Since the HybProbes chemistry requires two probes for positive identification, the assay is considered to be more specific than conventional PCR or TaqMan real-time PCR. The developed assay would be a useful tool with great potential in early diagnosis and detection of common scab pathogens of potatoes in infected plants or for surveillance of potatoes grown in soil environment.

Ecological and mechanistic insights into the direct and indirect antimicrobial properties of Bacillus subtilis lipopeptides on plant pathogens.

Members of the genus Bacillus produce a wide variety of antimicrobial compounds. Cyclic lipopeptides (CLP) produced by Bacillus subtilis strains have been shown to protect host plants from a numbers of pathogens. The representative families of these CLP (surfactins, fengycins, and iturins) share a polypeptide ring linked to a lipid tail of varying length. CLP provide plant protection through a variety of unique mechanisms. Members of the surfactin and fengycin families elicit induced systemic resistance in certain host plants, and they also function by directly affecting the biological membranes of bacterial and fungal pathogens, mainly resulting in membrane pore formation. Specific pore forming mechanisms differ between CLP families, causing differential activities. CLP also may aid in enhanced B. subtilis colonization of the plant environment in addition to potentially preventing the adhesion of competitive microorganisms. Several recent studies have highlighted the control of plant pathogens by CLP-producing B. subtilis strains. Strong ecological advantages through multifaceted activities of CLP provide these strains with immense promise in controlling pathogens in a variety of plant ecosystems.

A physical and transcript map of the MCOLN1 gene region on human chromosome 19p13.3-p13.2.

Mutations in MCOLN1 have been found to cause mucolipidosis type IV (MLIV; MIM 252650), a rare autosomal recessive lysosomal storage disorder found primarily in the Ashkenazi Jewish population. As a part of the successful cloning of MCOLN1, we constructed a 1.4-Mb physical map containing 14 BACs and 4 cosmids that encompasses the region surrounding MCOLN1 on human chromosome 19p13.3-p13.2-a region to which linkage or association has been reported for multiple diseases. Here we detail the precise physical mapping of 28 expressed sequence tags that represent unique UniGene clusters, of which 15 are known genes. We present a detailed transcript map of the MCOLN1 gene region that includes the genes KIAA0521, neuropathy target esterase (NTE), a novel zinc finger gene, and two novel transcripts in addition to MCOLN1. We also report the identification of eight new polymorphic markers between D19S406 and D19S912, which allowed us to pinpoint the location of MCOLN1 by haplotype analysis and which will facilitate future fine-mapping in this region. Additionally, we briefly describe the correlation between the observed haplotypes and the mutations found in MCOLN1. The complete 14-marker haplotypes of non-Jewish disease chromosomes, which are crucial for the genetic diagnosis of MLIV in the non-Jewish population, are presented here for the first time.

Mucolipidosis type IV is caused by mutations in a gene encoding a novel transient receptor potential channel.

Mucolipidosis type IV (MLIV) is a developmental neurodegenerative disorder characterized by severe neurologic and ophthalmologic abnormalities. The MLIV gene, ML4 (MCOLN1), has recently been localized to chromosome 19p13.2-13.3 by genetic linkage. Here we report the cloning of a novel transient receptor potential cation channel gene and show that this gene is mutated in patients with the disorder. ML4 encodes a protein, which we propose to call mucolipin, which has six predicted transmembrane domains and is a member of the polycystin II subfamily of the Drosophila transient receptor potential gene family. The role of a potential receptor-stimulated cation channel defect in the pathogenesis of mucolipidosis IV is discussed.

[Contents of the transversal foramen of the seventh cervical vertebra]. / Le contenu du trou transversaire de la septième vertèbre cervicale.

The purpose of our study was to evaluate the individual variations of the foramen sometimes has the same dimensions as the foramen of the other cervical vertebrae, but it can also be smaller, or absent. In cases where the foramen is present in the seventh cervical vertebra, vascular or nerve structures (or both) can be occasionally observed within. When the transverse foramina are found to contain none of the above structures, what then is to be found within this space? According to our results, the transverse foramina of the seventh cervical vertebra almost universally contain vascular and nerve branches as well as fibrous and adipose tissue. This report concerns the results of the macroscopic and microscopic studies concerning the contents of the transverse foramina of the seventh cervical vertebra.

[The inter-metacarpo-phalangeal spaces]. / Etude des espaces inter-métacarpo-phalangiens.

The study of the inter-metacarpophalangeal spaces on 20 human bodies raises the question of the existence of bursae at this level, in line with recent studies (BOSSEY, MIDY) suggesting that such bursae are present at the level of the foot. Careful dissection and histological studies revealed characteristics compatible with both of the following hypotheses: either there are bursae not described until now in anatomy textbooks, or these are virtual spaces limited by a cul-de-sac of the aponeuroses of the hand in the 2nd, 3rd and 4th spaces. Tissue analysis that could determine more specifically the nature of the limiting membrane of these spaces, as well as pathological studies demonstrating the occurrence of common anomalies of bursae in these spaces, are required in order to conclude definitively on the nature of the inter-metecarpophalangeal spaces.

A comparative study of the spaces between the metacarpal and metatarsal heads.

The study of the spaces between the metacarpal heads on 23 cadavers, raises the question of the existence of bursae at this level. This is in line with recent studies suggesting that such bursae are present at this level in the foot. Careful dissection and histological studies revealed characteristics compatible with both of the following hypotheses: either the existence of bursae not described until now in anatomy text-books, or spaces limited by a cul-de-sac of the aponeuroses of the hand in the 2nd, 3rd and 4th digits. Tissue analyses that could determine more specifically the nature of the limiting membrane of these spaces, as well as pathological studies demonstrating the occurrence of common anomalies of bursae in these spaces, are required in order to conclude definitively the nature of the spaces between the metacarpal heads. The gross anatomical and histological characteristics of the spaces between the metacarpal and metatarsal heads and of their limiting membranes were compared and found to be analogous in our series.