Bacillus capparidis sp. nov., an endophytic bacterium isolated from roots of Capparis spinosa L.

A novel endophytic bacterium, designated strain EGI 6500252T, was isolated from the surface-sterilized roots of a medicinal plant (Capparis spinosa L.) collected from Urumqi city, Xinjiang, north-west China. Cells were Gram-stain-positive, non-motile, aerobic, catalase- and oxidase-positive, rod-shaped and did not display spore formation. Strain EGI 6500252T grew at 10-40 °C (optimum 25-30 °C), at pH 6.0-8.0 (optimum pH 7.0) and in the presence of 0-10 % (w/v) NaCl (optimum 0-3 %). The major cellular fatty acids (>10 %) were identified as iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and summed feature 4. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unknown phospholipids, one unknown glycolipid and one unknown lipid. The dominant isoprenoid quinone was menaquinone 7 (MK-7). The DNA G+C content was 39.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain EGI 6500252T belonged to the genus Bacillus, and exhibited a highest 16S rRNA gene sequence similarity (96.2 %) that was lower than the suggested threshold (97.0 %) for separating bacterial species. On the basis of the phylogenetic analysis, chemotaxonomic data and physiological characteristics, strain EGI 6500252T represents a novel species of the genus Bacillus, for which the name Bacillus capparidis sp. nov. is proposed. The type strain is EGI 6500252T (=CGMCC 1.12820T=KCTC 33514T).

Streptomyces capparidis sp. nov., a novel endophytic actinobacterium isolated from fruits of Capparis spinosa L.

A novel endophytic actinobacterial strain, designated EGI 6500195T, was isolated from fruits of Capparis spinosa. Growth occurred at 10-45 °C (optimum 30 °C), at pH 6-8 (optimum pH 7) and in the presence of 0-1 % (w/v) NaCl. Strain EGI 6500195T shared highest 16S rRNA gene sequence similarity (97.74 %) with Streptomyces vitaminophilus DSM 41686T and less than 97 % sequence similarity with other members of the genus Streptomyces. The diagnostic amino acid in the peptidoglycan was ll-diaminopimelic acid. Whole-cell hydrolysates contained glucose, ribose, fructose and mannose. The predominant menaquinones were MK-9(H6) and MK-9(H8). The polar lipid profile of strain EGI 6500195T included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylcholine, three unknown phospholipids, an unknown aminophospholipid and an unknown aminolipid. The cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0, iso-C16 : 0, anteiso-C17 : 1ω9c, summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B) and iso-C17 : 1ω9c. The DNA G+C content of strain EGI 6500195T was 74.1 mol%. The level of DNA-DNA relatedness between strain EGI 6500195T and Streptomyces. vitaminophilus DSM 41686T was 14.1±3.5 %. On the basis of the phenotypic, phylogenetic, chemotaxonomic and DNA-DNA hybridization data, strain EGI 6500195T represents a novel species of the genus Streptomyces, for which the name Streptomyces capparidis sp. nov. is proposed. The type strain is EGI 6500195T (=DSM 42145T=JCM 30089T).

The Multifarious PGPR Serratia marcescens CDP-13 Augments Induced Systemic Resistance and Enhanced Salinity Tolerance of Wheat (Triticum aestivum L.).

The present study demonstrates the plant growth promoting (PGP) potential of a bacterial isolate CDP-13 isolated from 'Capparis decidua' plant, and its ability to protect plants from the deleterious effect of biotic and abiotic stressors. Based on 16S rRNA gene sequence analysis, the isolate was identified as Serratia marcescens. Among the PGP traits, the isolate was found to be positive for ACC deaminase activity, phosphate solubilization, production of siderophore, indole acetic acid production, nitrogen fixation, and ammonia production. CDP-13 showed growth at an increased salt (NaCl) concentration of up to 6%, indicating its potential to survive and associate with plants growing in saline soil. The inoculation of S. marcescens enhanced the growth of wheat plant under salinity stress (150-200 mM). It significantly reduced inhibition of plant growth (15 to 85%) caused by salt stressors. Application of CDP-13 also modulated concentration (20 to 75%) of different osmoprotectants (proline, malondialdehyde, total soluble sugar, total protein content, and indole acetic acid) in plants suggesting its role in enabling plants to tolerate salt stressors. In addition, bacterial inoculation also reduced the disease severity caused by fungal infection, which illustrated its ability to confer induced systemic resistance (ISR) in host plants. Treatment of wheat plants with the test organism caused alteration in anti-oxidative enzymes activities (Superoxide dismutase, Catalase, and Peroxidase) under various salinity levels, and therefore minimizes the salinity-induced oxidative damages to the plants. Colonization efficiency of strain CDP-13 was confirmed by CFU count, epi-fluorescence microscopy, and ERIC-PCR-based DNA fingerprinting approach. Hence, the study indicates that bacterium CDP-13 enhances plant growth, and has potential for the amelioration of salinity stress in wheat plants. Likewise, the results also provide insights into biotechnological approaches to using PGPR as an alternative to chemicals and pesticides.

Actividades citotóxicas y antibacterianas de las raíces de Capparis zeylanica Linn / Antibacterial and Cytotoxic Activities of Capparis zeylanica Linn Roots

Se analizaron extractos crudos y un ácido graso, ácido octadec-7-en-5-ynoic (1), de la corteza de la raíz de Cappariszeylanica Linn. (familia de las Capparidaceae) para observar sus actividades antibacterianas frente a la bacteria Gram positiva y Gram negativa. Entre los extractos crudos, el extracto de cloroformo mostró una buena actividad frente a todos los organismos de prueba. El ácido graso (1) aislado del extracto de cloroformo mostró actividades antibacterianas frente a todos los organismos de prueba, a excepción de E. coli. Las actividades se compararon con un antibiótico estándar: la kanamicina. Las concentraciones inhibitorias mínimas (CIH) de 1, determinadas mediante la técnica de dilución en serie, fueron 64 mg/ml frente a Bacillus subtilis y Shigella dysenteriae. Las actividades citotóxicas del extracto crudo y del ácido graso (1) se observaron mediante el bioensayo de gambas en salmuera yel valor de LC50 del compuesto fue 6,27 mg/ml (AU)

Crude extracts and a fatty acid, octadec-7-en-5-ynoic acid (1), from the root bark of Capparis zeylanica Linn. (Fam. Capparidaceae) were screened for their antibacterial activities against Gram positive and Gram negative bacteria. Among the crude extracts, chloroform extract showed good activity against all test organisms. The fatty acid (1) isolated from chloroform extract exhibited antibacterial activities against test organisms except E. coli. The activities were compared to a standard antibiotic- kanamycin. The minimum inhibitory concentrations (MICs) of 1, determined byserial dilution technique, were found to be 64 mg/ml against Bacillus subtilis and Shigella dysenteriae. The cytotoxic activities of crude extract and fatty acid (1) were observed by brine shrimp biassay and LC50 value of the compound was found to be 6.27 mg/ml (AU)

Safety and potential risks of enterococci isolated from traditional fermented capers.

A collection of 17 enterococci isolates obtained from fermentations of capers (the fruits of Capparis sp.) were investigated for incidence of known virulence determinants, antibiotic resistance and production of biogenic amines. Molecular identification revealed the presence of Enterococcus faecium (nine isolates), Enterococcus faecalis (4), E. avium (3) and Enterococcus casseliflavus/flavescens (1). Alpha-haemolytic activity was detected in two E. avium and one E. faecalis isolates, and beta-haemolytic activity was detected in E. casseliflavus/flavescens. The haemolytic component cylB was detected by PCR amplification in three non-haemolytic isolates and in E. casseliflavus/flavescens. The collagen adhesin ace gene and the endocarditis associated antigen gene efaA(fm) were detected in two isolates each. Genes encoding sex pheromone precursors (cpd, cob, ccf) were detected in E. faecalis and E. casseliflavus/flavescens. Other presumed virulence genes (agg, gelE, cylM, cylA and efaA(fs)) were not detected. All isolates were resistant to rifampicin, erythromycin and ciprofloxacin, and some were also resistant to quinupristin/dalfopristin, tetracycline, levofloxacin, gentamicin and streptomycin. Vancomycin resistance was not detected. Tyrosine decarboxylation was detected in all E. faecium isolates. Given the high resistance of enterococci to environmental conditions, and their implication in opportunistic infections, the incidence of potential virulent enterococci in foods (especially those of a higher risk-like home-made foods) should be carefully studied.

Plasmid profile patterns and properties of pediococci isolated from caper fermentations.

A collection comprising 14 isolates of Pediococcus pentosaceus and one Pediococcus acidilactici from the fermentation of caper fruits was studied. All isolates showed very similar fermentation profiles and produced a limited number of exoenzymes. All isolates carried large plasmids of diverse sizes between 20 and 55 kb, while some also contained smaller plasmids between 10 and 16 kb. Cluster analysis of plasmid profiles revealed four main groups with various degrees of similarities. All amino acid decarboxylation tests were negative, suggesting that pediococci are not involved in generation of biogenic amines. None of the isolates showed hemolytic activity. Antimicrobial resistance tests revealed that all isolates were sensitive to 11 different antimicrobials while being resistant to ciprofloxacin (MIC > or =2 mg/liter) and intrinsically resistant to vancomycin (MIC > or =16 mg/liter) and teicoplanin (MIC > or =16 mg/liter).

Microbiological study of lactic acid fermentation of Caper berries by molecular and culture-dependent methods.

Fermentation of capers (the fruits of Capparis sp.) was studied by molecular and culture-independent methods. A lactic acid fermentation occurred following immersion of caper berries in water, resulting in fast acidification and development of the organoleptic properties typical of this fermented food. A collection of 133 isolates obtained at different times of fermentation was reduced to 75 after randomly amplified polymorphic DNA (RAPD)-PCR analysis. Isolates were identified by PCR or 16S rRNA gene sequencing as Lactobacillus plantarum (37 isolates), Lactobacillus paraplantarum (1 isolate), Lactobacillus pentosus (5 isolates), Lactobacillus brevis (9 isolates), Lactobacillus fermentum (6 isolates), Pediococcus pentosaceus (14 isolates), Pediococcus acidilactici (1 isolate), and Enterococcus faecium (2 isolates). Cluster analysis of RAPD-PCR patterns revealed a high degree of diversity among lactobacilli (with four major groups and five subgroups), while pediococci clustered in two closely related groups. A culture-independent analysis of fermentation samples by temporal temperature gradient electrophoresis (TTGE) also indicated that L. plantarum is the predominant species in this fermentation, in agreement with culture-dependent results. The distribution of L. brevis and L. fermentum in samples was also determined by TTGE, but identification of Pediococcus at the species level was not possible. TTGE also allowed a more precise estimation of the distribution of E. faecium, and the detection of Enterococcus casseliflavus (which was not revealed by the culture-dependent analysis). Results from this study indicate that complementary data from molecular and culture-dependent analysis provide a more accurate determination of the microbial community dynamics during caper fermentation.

Resistance to antimicrobial agents in lactobacilli isolated from caper fermentations.

A collection of lactobacilli comprising species of Lactobacillus plantarum (43 isolates), Lactobacillus brevis (9 isolates) and Lactobacillus fermentum (6 isolates) obtained from spontaneous fermentations of capers (the fruits of Capparis spinosa) were investigated for resistance to antimicrobial agents. All isolates were resistant to vancomycin and teicoplanin (MIC > 16 microg/ml). Resistance to ciprofloxacin (MIC > 2 microg/ml) was detected in all isolates of L. brevis and L. fermentum as well as in most isolates of L. plantarum, whilst resistance to levofloxacin showed a much lower incidence. Among L. plantarum and L. brevis isolates, low levels of resistance to tetracycline and/or nitrofurantoin were detected. Higher resistance levels were also detected in some isolates. Resistance to penicillin and rifampicin were also detected among L. plantarum isolates. All isolates were sensitive to ampicillin, erythromycin, chloramphenicol, gentamicin, streptomycin, and quinupristin/dalfopristin.

Fungal endophytes from the three-leaved caper, Crataeva magna (Lour.) DC. (Capparidaceae).

Fungal endophytes were isolated from Crataeva magna, a medicinal plant growing along the streams and rivers, constituting riparian vegetation in Karnataka, southern India. Fresh bark and twig pieces were used for the isolation using standard methods. Ninety-six endophytic fungal isolates were isolated from 800 bark and twig segments. Mitosporic fungi represented as a major group (85%) followed by zygomycetes (10%) and ascomycetes (5%). Bark samples contained more endophytes than twig samples. Verticillium, Nigrospora oryzae and Fusarium verticilloides were the dominant fungal endophytes.