Characterization of ISR region and development of a PCR assay for rapid detection of the fish pathogen Tenacibaculum soleae

The aims of this work were to characterize the 16S-23S internal spacer region of the fish pathogen Tenacibaculum soleae and to develop a PCR assay for its identification and detection. All T.?soleae strains tested displayed a single internal spacer region class, containing tRNA(I) (le) and tRNA(A) (la) genes; nevertheless, a considerable intraspecific heterogeneity was observed. However, this region proved to be useful for differentiation of T.?soleae from related and non-related species. Species-specific primers were designed targeting the 16S rRNA gene and the internal spacer region region, yielding a 1555-bp fragment. Detection limit was of 1?pg DNA per reaction (相似文献   

Observations on the venous system of three species of Polypterus (Pisces)

The venous system of Polypterus exhibits general asymmetry. The features that characterize the venous system of this peculiarly African fish are the possession of many blood sinuses, the segmental drainage of blood, the continuation of the posterior cardinal veins as separated vessels and the occurrence of several anastomozes between the latter.
In the absence of any recent comprehensive work on the blood system of this fish, it was thought that investigation of the circulatory system of Polypterus would be valuable. In this paper attention is drawn to Budgettapos;s (1902) original work on a larva of Polypterus senegalus. Contrary to Budgettapos;s findings it is concluded that what he called the "interrenal" vein is in fact the right posterior cardinal vein. It is also found that paired posterior cardinal veins exist in all adults.     

The carriage population of Staphylococcus aureus from Mali is composed of a combination of pandemic clones and the divergent Panton-Valentine leukocidin-positive genotype ST152

Staphylococcus aureus is an important human pathogen, but it appears more commonly in asymptomatic colonization of the nasopharynx than in cases of invasive disease. Evidence concerning the global population structure of S. aureus is limited by the overrepresentation in the multilocus sequence testing database of disease isolates recovered from Western Europe, the Americas, Australia, and Japan. We address this by presenting data from the S. aureus carriage population in Mali, the first detailed characterization of asymptomatic carriage from an African population. These data confirm the pandemic spread of many of the common S. aureus clones in the carriage population. We also note the high frequency (approximately 24%) of a single divergent genotype, sequence type 152 (ST152), which has not previously been recovered from nasal carriage isolates but corresponds to a sporadic Panton-Valentine leukocidin (PVL)-positive, community-acquired methicillin-resistant S. aureus clone noted mostly in Central Europe. We show that 100% of the ST152 isolates recovered from nasal carriage samples in Mali are PVL positive and discuss implications relating to the emergence and spread of this virulent genotype.     

Divergent responses of different endothelial cell types to infection with Candida albicans and Staphylococcus aureus

Endothelial cells are important in the pathogenesis of bloodstream infections caused by Candida albicans and Staphylococcus aureus. Numerous investigations have used human umbilical vein endothelial cells (HUVECs) to study microbial-endothelial cell interactions in vitro. However, the use of HUVECs requires a constant supply of umbilical cords, and there are significant donor-to-donor variations in these endothelial cells. The use of an immortalized endothelial cell line would obviate such difficulties. One candidate in this regard is HMEC-1, an immortalized human dermal microvascular endothelial cell line. To determine if HMEC-1 cells are suitable for studying the interactions of C. albicans and S. aureus with endothelial cells in vitro, we compared the interactions of these organisms with HMEC-1 cells and HUVECs. We found that wild-type C. albicans had significantly reduced adherence to and invasion of HMEC-1 cells as compared to HUVECs. Although wild-type S. aureus adhered to and invaded HMEC-1 cells similarly to HUVECs, an agr mutant strain had significantly reduced invasion of HMEC-1 cells, but not HUVECs. Furthermore, HMEC-1 cells were less susceptible to damage induced by C. albicans, but more susceptible to damage caused by S. aureus. In addition, HMEC-1 cells secreted very little IL-8 in response to infection with either organism, whereas infection of HUVECs induced substantial IL-8 secretion. This weak IL-8 response was likely due to the anatomic site from which HMEC-1 cells were obtained because infection of primary human dermal microvascular endothelial cells with C. albicans and S. aureus also induced little increase in IL-8 production above basal levels. Thus, C. albicans and S. aureus interact with HMEC-1 cells in a substantially different manner than with HUVECs, and data obtained with one type of endothelial cell cannot necessarily be extrapolated to other types.     

Tinkering with the C-Function: A Molecular Frame for the Selection of Double Flowers in Cultivated Roses

Background

Roses have been cultivated for centuries and a number of varieties have been selected based on flower traits such as petal form, color, and number. Wild-type roses have five petals (simple flowers), whereas high numbers of petals (double flowers) are typical attributes of most of the cultivated roses. Here, we investigated the molecular mechanisms that could have been selected to control petal number in roses.

Methodology/Principal Findings

We have analyzed the expression of several candidate genes known to be involved in floral organ identity determination in roses from similar genetic backgrounds but exhibiting contrasting petal numbers per flower. We show that the rose ortholog of AGAMOUS (RhAG) is differentially expressed in double flowers as compared to simple flowers. In situ hybridization experiments confirm the differential expression of RhAG and demonstrate that in the double-flower roses, the expression domain of RhAG is restricted toward the center of the flower. Conversely, in simple-flower roses, RhAG expression domain is wider. We further show that the border of RhAG expression domain is labile, which allows the selection of rose flowers with increased petal number. Double-flower roses were selected independently in the two major regions for domestication, China and the peri-Mediterranean areas. Comparison of RhAG expression in the wild-type ancestors of cultivated roses and their descendants both in the European and Chinese lineages corroborates the correlation between the degree of restriction of RhAG expression domain and the number of petals. Our data suggests that a restriction of RhAG expression domain is the basis for selection of double flowers in both the Chinese and peri-Mediterranean centers of domestication.

Conclusions/Significance

We demonstrate that a shift in RhAG expression domain boundary occurred in rose hybrids, causing double-flower phenotype. This molecular event was selected independently during rose domestication in Europe/Middle East and in China.     

A natural recessive resistance gene against potato virus Y in pepper corresponds to the eukaryotic initiation factor 4E (eIF4E)

We show here that the pvr2 locus in pepper, conferring recessive resistance against strains of potato virus Y (PVY), corresponds to a eukaryotic initiation factor 4E (eIF4E) gene. RFLP analysis on the PVY-susceptible and resistant pepper cultivars, using an eIF4E cDNA from tobacco as probe, revealed perfect map co-segregation between a polymorphism in the eIF4E gene and the pvr2 alleles, pvr2(1) (resistant to PVY-0) and pvr2(2) (resistant to PVY-0 and 1). The cloned pepper eIF4E cDNA encoded a 228 amino acid polypeptide with 70-86% nucleotide sequence identity with other plant eIF4Es. The sequences of eIF4E protein from two PVY-susceptible cultivars were identical and differed from the eIF4E sequences of the two PVY-resistant cultivars Yolo Y (YY) (pvr2(1)) and FloridaVR2 (F) (pvr2(2)) at two amino acids, a mutation common to both resistant genotypes and a second mutation specific to each. Complementation experiments were used to show that the eIF4E gene corresponds to pvr2. Thus, potato virus X-mediated transient expression of eIF4E from susceptible cultivar Yolo Wonder (YW) in the resistant genotype YY resulted in loss of resistance to subsequent PVY-0 inoculation and transient expression of eIF4E from YY (resistant to PVY-0; susceptible to PVY-1) rendered genotype F susceptible to PVY-1. Several lines of evidence indicate that interaction between the potyvirus genome-linked protein (VPg) and eIF4E are important for virus infectivity, suggesting that the recessive resistance could be due to incompatibility between the VPg and eIF4E in the resistant genotype.     

Biological control of the wheat root rot caused by Fusarium graminearum using some PGPR strains in Saudi Arabia

The aim of this study was to evaluate the efficacy of selected bacterial strains against the wheat soil‐borne pathogen Fusarium graminearum under greenhouse conditions. The most potent isolates were 3 isolates out of 18 isolates, which have numbers 3, 9 and 10 with in vitro inhibition index 42.5%, 41.3% and 46.3% respectively. Isolates 3 and 10 were selected for the following experiments. Isolates 3 and 10 were identified as Bacillus subtilis MAA03 and Pseudomonas fluorescens MAA10, respectively according to International Identification Keys and, confirmed by using Biolog system and 16S rDNA where the strains exhibited more than 99.5% sequence identity. Their close taxonomic relationship was further documented by phenotypic similarities. The using of B. subtilis and P. fluorescens separately or in mixture as biocontrol agent against F. graminearum on wheat significantly increased the final germination percent, the mean daily germination and germination index of wheat cultivar, while the mean germination time was significantly decreased relative to infested control. The final infection percent, the mean daily infection and infection index were decreased significantly, while the mean infection time was significantly increased relative to infested control. The use of P. fluorescens as biocontrol agent was the most efficient than B. subtilis or in mixture and the best treatment was seed coating. The application of B. subtilis and P. fluorescens separately or in combination significantly affected the growth parameters of wheat cultivar Tabuki, the root length was significantly increased in seed coating and seed soaking treatments, while non‐significantly decreased in case of soil drench treatment relative to infested control. Shoot length was significantly decreased in case of seed coating treatment relative to infested control. The shoot fresh and dry weights were significantly increased in seed coating and seed soaking treatments relative to infested control. The root fresh and dry weights were significantly increased in seed coating and seed soaking treatments relative to infested control. The number of leaves was significantly increased in all treatments relative to infested control.     

Volatile Oils from the Aerial Parts of Eremophila maculata and Their Antimicrobial Activity

The essential oils isolated from the fresh flowers, fresh leaves, and both fresh and air‐dried stems of Eremophila maculata (Scrophulariaceae) were characterized by GC‐FID and GC/MS analyses. Sabinene was the major component in most of the oils, followed by limonene, α‐pinene, benzaldehyde, (Z)‐β‐ocimene, and spathulenol. The leaf and flower essential oils showed antibacterial and antifungal activity against five Gram‐positive and four Gram‐negative bacterial strains, multi‐resistant clinical isolates from patients, i.e., methicillin‐resistant Staphylococcus aureus (MRSA), as well as two yeasts. Minimum inhibitory concentrations (MICs) and minimum microbicidal concentrations (MMCs) were between 0.25 and 4 mg/ml.     

Bioerosion in the Miocene Reefs of the northwest Red Sea,Egypt

Macroborings provide detailed information on the bioerosion, accretion and palaeoenvironment of both modern and fossil reefs. Dolomitized reefal carbonates in the Um Mahara Formation exhibit an outstanding example of spatially distributed, well‐preserved bioerosion structures in tropical to subtropical syn‐rift Miocene reefs. Ten ichnospecies belonging to five ichnogenera are identified; three belonging to the bivalve‐boring ichnogenus Gastrochaenolites, three attributed to the sponge‐boring ichnogenus Entobia, and four ichnospecies assigned to three worm‐boring ichnogenera Trypanites, Maeandropolydora and Caulostrepsis. The distribution of the reported borings is strongly linked to the palaeo‐reef zones. Two distinctive ichnological boring assemblages are recognized. The Gastrochaenolites‐dominated assemblage reflects shallower‐marine conditions, under water depths of a few metres, mostly in back‐reef to patch‐reef zones of a back‐reef lagoon. The Entobia‐dominated assemblage signifies relatively deeper marine conditions, mostly in reef core of the fringing Miocene reefs. These ichnological assemblages are attributed herein to the Entobia sub‐ichnofacies of the Trypanites ichnofacies. This ichnofacies indicates boring in hard carbonate substrates (such as corals, rhodoliths, carbonate cements and hardgrounds) during periods of non‐sedimentation or reduced sediment input.