Fulvimarina endophytica sp. nov., a novel endophytic bacterium isolated from bark of Sonneratia caseolaris.

A Gram-negative, aerobic, short-rod-shaped, motile (with a terminal flagellum), non-spore-forming bacterium, designated strain 85T, was isolated from a surface-sterilized bark of Sonneratia caseolaris collected from Qinzhou in Guangxi, China and was analyzed using a polyphasic approach to determine its taxonomic position. Strain 85T grew optimally in the presence of 1-2% (w/v) NaCl at 30°C and pH 6.0-7.0. Phylogenetic analysis based on 16S rRNA gene sequence suggested that strain 85T belonged to the genus Fulvimarina and shared the highest 16S rRNA gene sequence similarity with Fulvimarina pelagi HTCC2506T (96.16%). The cell-wall peptidoglycan contained meso-diaminopimelic acid and ubiquinone Q-10 was the predominant respiratory lipoquinone. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unidentified amino lipid, three unidentified phospholipids and six unidentified lipids. The major fatty acid was C18:1ω7c. The DNA G+C content of strain 85T was 65.4 mol%, and the average nucleotide identity and estimated DDH values between strain 85T and the type strain of Fulvimarina pelagi HTCC2506T were 77.3% and 21.7%, respectively. Based on the phylogenetic, phenotypic, and chemotaxonomic analyses, strain 85T should be considered as a novel species of the genus Fulvimarina with the proposed name Fulvimarina endophytica sp. nov., and its type strain is 85T (= KCTC 62717T = CGMCC 1.13665T).

Cohnella endophytica sp. nov., a novel endophytic bacterium isolated from bark of Sonneratia apetala.

A Gram-stain-positive, aerobic, rod-shaped, endospore-forming bacterium, designated strain M2MS4P-1T, was isolated from surface-sterilized bark of Sonneratiaapetala sampled in Guangxi, China. The bacterium was characterized by a polyphasic approach to determine its taxonomic position. 16S rRNA gene sequence comparisons revealed that strain M2MS4P-1T belonged to the genus Cohnella and was most closely to Cohnella luojiensis HY-22RT (98.4 % similarity). The average nucleotide identity value and estimated DDH value between strain M2MS4P-1T and the type strain of C. luojiensis HY-22RT were 79.2 and 20.1 %, respectively. Neither substrate nor aerial mycelia were formed, and no diffusible pigments were observed on the media tested. Strain M2MS4P-1T grew in the pH range 6.0-9.0 (optimum, pH 7.0-8.0), at temperatures between 10-37 °C (30 °C) and in 0-1 % (w/v) NaCl (0 %). The predominant isoprenoid quinone in strain M2MS4P-1T was menaquinone-7. The major fatty acids were anteiso-C15 : 0 and iso-C16 : 0. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, lysyl-phosphatidylglycerol, four unidentified aminophospholipids and two unidentified phospholipids. The DNA G+C content was 51.5 mol%. According to the phylogenetic, phenotypic and chemotaxonomic evidence, strain M2MS4P-1T was clearly distinguishable from other species with validly published names in the genus Cohnella and should therefore be classified as a novel species, for which we suggest the name Cohnellaendophytica sp. nov. The type strain is M2MS4P-1T (=KCTC 43011T=CGMCC 1.13745T).

Improving the antifungal activity of clove essential oil encapsulated by chitosan nanoparticles.

Encapsulation of clove essential oil (CEO) by chitosan nanoparticles (ChNPs) was performed, using an emulsion-ionic gelation technique to improve the antifungal efficacy of CEO. The mass ratios of chitosan (Ch) to tripolyphosphate (TPP), 1:1, for unloaded ChNPs and 1:1:1 for Ch to TPP to CEO, for CEO-loaded ChNPs (CEO-ChNPs), were selected as optimum formulations based on dynamic light scattering and ultraviolet-visible spectroscopy. The presence of CEO in optimum CEO-ChNPs, was evidenced by Fourier transform infrared spectroscopy. Particle size distribution, of around 40 and 100 nm for the most optimum unloaded and oil-loaded ChNPs, was obtained by field emission-scanning electron microscopy. In vitro release studies of CEO-ChNPs revealed a controlled release during 56 days. The nano-encapsulated CEO demonstrated a superior performance against Aspergillus niger, isolated from spoiled pomegranate, compared with ChNPs and free oil. Therefore, this study revealed that CEO-ChNPs can be used as a promising natural fungicide in agriculture and food industry.

Effects of fludioxonil, propolis and black seed oil application on the postharvest quality of "Wonderful" pomegranate.

Pomegranate fruit consumption has increased rapidly throughout the world, mainly because of its medical and nutritive attributes. Thus, considerable commercial and scientific interest exists in prolonging its postharvest life with non-chemical applications as much as possible to meet the year-round demand for this fruit. The present work aimed to study the effects of black seed oil (0.1% and 0.5%), propolis (0.01% and 0.1%) and fludioxonil (0.06%), with and without modified atmosphere packaging (MAP), on the postharvest quality of pomegranate cv. Wonderful. Treated fruits were stored at 6.5±1 °C and 90-95% relative humidity for 150 days. The results indicated that both black seed oil and propolis treatments significantly influenced the maintenance of fruit weight and quality. At 150 days after storage, the fruit weight loss of the samples treated with MAP + 0.5% black seed oil, MAP + 0.1% propolis and MAP alone were found to be 5.5%, 6.3%, and 9.1%, respectively, whereas the weight loss of the untreated control fruits was 19.8%. Application of either 0.5% black seed oil or 0.1% propolis, especially when combined with MAP, was also effective in controlling gray mold development and slowing the occurrence of chilling injury.

Xanthomonas axonopodis pv. punicae uses XopL effector to suppress pomegranate immunity.

Xanthomonas axonopodis pv. punicae (Xap) causing bacterial blight is an important pathogen that incurs significant losses to the exportability of pomegranate. Xap uses the Xop TTSS-effector, via the type three secretion system, to suppress pomegranate immunity. Here, we investigate the role of XopL during blight pathogenesis. We observed that XopL is essential for its in planta growth and full virulence. Leaves inoculated with Xap ΔxopL produced restricted water-soaked lesions compared to those inoculated with wild-type Xap. XopL supports Xap for its sustained multiplication in pomegranate by suppressing the plant cell death (PCD) event. We further demonstrated that XopL suppresses immune responses, such as callose deposition and production of reactive oxygen species (ROS). RT-qPCR analysis revealed that immune responsive genes were upregulated when challenged with Xap ΔxopL, whereas upregulation of such genes was compromised in the complemented strain containing the xopL gene. The transiently expressed XopL::EYFP fusion protein was localized to the plasma membrane, indicating the possible site of its action. Altogether, this study highlights that XopL is an important TTSS-effector of Xap that suppresses plant immune responses, including PCD, presumably to support the multiplication of Xap for a sufficient time-period during blight disease development.

Flaxseed gum in combination with lemongrass essential oil as an effective edible coating for ready-to-eat pomegranate arils.

Flaxseed gum (FSG) in combination with lemongrass essential oil (LGEO) was investigated for coating of ready-to-eat pomegranate arils. FSG was used at 0.3% and 0.6% concentrations and with both concentrations LGEO was incorporated at levels of 0ppm, 200ppm, 500ppm and 800ppm. Changes in headspace gases, physicochemical, microbiological and sensory attributes of pomegranate arils stored at 5°C were studied on different days of analysis during the 12day storage period. Coatings containing LGEO were effective in reducing total plate count and yeast and mold populations. Increasing LGEO concentrations in the coatings resulted in more decline in microbial populations. Reduced weight loss occurred in coated samples as compared to uncoated (control) sample. Coated samples showed a gradual decrease in ripening index in contrast with control where a significantly higher decline was observed. Total soluble solids, pH and titratable acidity significantly varied over the storage period. Color change (ΔE) for control increased steeply over the storage time in comparison to coated samples. Furthermore, chroma decreased while as hue angle increased over time.

Acetylcholinesterase Inhibitory Meroterpenoid from a Mangrove Endophytic Fungus Aspergillus sp. 16-5c.

One new meroterpenoid, named 2-hydroacetoxydehydroaustin (1), together with nine known meroterpenoids, 11-acetoxyisoaustinone (2), isoaustinol (3), austin (4), austinol (5), acetoxydehydroaustin (6), dehydroaustin (7), dehydroaustinol (8), preaustinoid A2 (9), and 1,2-dihydro-acetoxydehydroaustin B (10), were isolated from the mangrove endophytic fungus, Aspergillus sp. 16-5c. These structures were characterized by spectroscopic analysis, further the absolute configurations of stereogenic carbons for Compounds 1, 3, 4, 6, 7, 8, 9, and 10 were determined by single crystal X-ray diffraction analysis using Cu Kα radiation. Moreover, the absolute configurations of stereogenic carbons for Known Compounds 3, 7, 8, and 9 are identified here for the first time. Compounds 3, 7, and 8 showed acetylcholinesterase (AchE) inhibitory activity with IC50 values of 2.50, 0.40, and 3.00 µM, respectively.

Swarm motility inhibitory and antioxidant activities of pomegranate peel processed under three drying conditions.

During processing of ready-to-eat fresh fruits, large amounts of peel and seeds are discarded as waste. Pomegranate (Punicagranatum) peels contain high amounts of bioactive compounds which inhibit migration of Salmonella on wet surfaces. The metabolic distribution of bioactives in pomegranate peel, inner membrane, and edible aril portion was investigated under three different drying conditions along with the anti-swarming activity against Citrobacter rodentium. Based on the multivariate analysis, 29 metabolites discriminated the pomegranate peel, inner membrane, and edible aril portion, as well as the three different drying methods. Punicalagins (∼38.6-50.3mg/g) were detected in higher quantities in all fractions as compared to ellagic acid (∼0.1-3.2mg/g) and punicalins (∼0-2.4mg/g). The bioactivity (antioxidant, anti-swarming) and phenolics content was significantly higher in peels than the edible aril portion. Natural anti-swarming agents from food waste may have promising potential for controlling food borne pathogens.

[Diversity and cytotoxic activity of endophytic bacteria isolated from Sonneratia apetala of Maowei Sea].

Objective: The purpose of this study was to study the distribution, diversity and cytotoxic activity of endophytic bacteria of Sonneratia apetala collected from Maowei Sea, Qinzhou city. Methods: The 16S rRNA gene sequencing and MTT were used to explore the diversity and cytotoxic activity of endophytic bacteria isolated from different organs and tissues of Sonneratia apetala. Results: Total of 38 isolates were obtained. The result of diversity analysis showed that these isolates could be phylogenetically classified into 21 genera and 12 families, based on their 16S rRNA gene sequencing. Of them 5 were potential new genera or new genus. Five strains (R74, R71, S92, S85 and S84) had cytotoxic activity against human liver carcinoma Hep G2 cell line. Conclusion: Endophytic bacteria of Sonneratia apetala are genetically diverse and most of them have abundant new bioactivities.

Inhibitory effect of pomegranate (Punica granatum L.) polyphenol extracts on the bacterial growth and survival of clinical isolates of pathogenic Staphylococcus aureus and Escherichia coli.

In the present study major polyphenols of pomegranate arils and peel by-products were extracted in 50% (v/v) aqueous ethanol, characterized and used in microbiological assays in order to test antimicrobial activity against clinically isolated human pathogenic microorganisms. Total concentration of polyphenols and in vitro antioxidant properties were determined by the Folin-Ciocalteu and DPPH methods, respectively. The most abundant bioactive molecules, including anthocyanins, catechins, tannins, gallic and ellagic acids were identified by RP-HPLC-DAD, also coupled to off-line matrix assisted laser desorption/ionization (MALDI-TOF) mass spectrometry (MS). The inhibitory spectrum of extracts against test microorganisms was assessed by the agar well-diffusion method. Data herein indicated that both pomegranate aril and peel extracts have an effective antimicrobial activity, as evidenced by the inhibitory effect on the bacterial growth of two important human pathogens, including Staphylococcus aureus and Escherichia coli, which are often involved in foodborne illness.

Characterization of bacterial knot disease caused by Pseudomonas savastanoi pv. savastanoi on pomegranate (Punica granatum L.) trees: a new host of the pathogen.

UNLABELLED: This study aimed to isolate and identify the causal organism causing hyperplastic outgrowths (knots) on stems and branches of pomegranate trees in the Eastern Mediterranean region of Turkey. Bacterial colonies were isolated from young knots on plates containing selective nutrient media. Biochemical tests, fatty acid analysis and PCR were performed to identify possible causal disease agent. Representative isolates were identified as Pseudomonas.pv.savastanoi (Psv) using biochemical tests, fatty acid profiling and PCR. Following inoculation of pomegranate plants (cv. hicaz) with bacterial suspensions, 25 of 54 bacterial isolates caused typical knots at the site of inoculation. PCR analysis, using specific primer for Psv, generated a single amplicon from all isolates. The similarity of the sequence of Turkish pomegranate isolate was 99% similar to the corresponding gene sequences of Psv in the databases. Based on symptoms, biochemical, molecular, pathogenicity tests and sequence analyses, the disease agent of knots observed on the pomegranate trees is Psv. To the best of our knowledge, this research has revealed pomegranate as a natural host of Psv, which extends the list of host plant species affected by the pathogen in the world and Turkey. SIGNIFICANCE AND IMPACT OF THE STUDY: Pomegranate trees were affected by the disease with outgrowths (galls or knot) disease. Currently, there is no published study on disease agent(s) causing the galls or knots on pomegranate trees in worldwide. Bacterial colonies were isolated from young knots. The causal agent of the knot Pseudomonas savastanoi pv.savastanoi (Psv) was identified based on symptoms, biochemical, molecular methods, pathogenicity tests and sequence analysis. To the best of our knowledge, this is the first report of Psv on pomegranate as a natural host, which extends the growing list of plant species affected by this bacterium in the world and Turkey.

Changes on indigenous microbiota, colour, bioactive compounds and antioxidant activity of pasteurised pomegranate juice.

Juices prepared from arils of 'Mollar' pomegranates were analysed for naturally occurring microorganisms, CIE Lab colour parameters, total phenols, anthocyanins and punicalagins, ellagic acid content and antioxidant capacity before and after low-, mild- and high-temperature pasteurisations (LTPs, MTPs and HTPs): 65, 80 and 90 °C for 30 or 60s. Mean aerobic plate count (APC), yeast and mold count (YMC), and lactic acid bacteria (LAB) for fresh juices were 5.7, 5.36 and 4.0 log CFU/mL, respectively. MTPs and HTPs were sufficiently effective to decrease APCs to nil or negligible levels. An increase in CIE a values and decrease in CIE b values were the characteristic colour changes in heat-treated juices. The effect of pasteurisations showed that total phenols, punicalagins and ellagic acid were not much affected by thermal processing. Total anthocyanin content and antioxidant capacity were substantially and significantly influenced by the heat treatment applied. A linear relationship was observed between Trolox equivalent antioxidant capacity (TEAC) values and total anthocyanins, suggesting that they contributed strongly to the antioxidant capacity of pomegranate juice.

Effect of chitosan coating on maintenance of aril quality, microbial population and PPO activity of pomegranate (Punica granatum L. cv. Tarom) at cold storage temperature.

BACKGROUND: Chitosan edible coating was used in an attempt to extend the storage life of pomegranate arils during 12 days at 4 °C. Prior to storage, treated arils were dipped in 0.25, 0.5 and 1% (w/v) chitosan aqueous solutions and 1% (v/v) acetic acid for 1 min, while control arils were dipped in distilled water with 1% (v/v) acetic acid. RESULTS: Chitosan coating inhibited bacterial and fungal growth on the surface of arils. The water content of arils coated with 0.5 and 1% chitosan was maintained during 12 days of storage. Chitosan reduced the increase in total soluble solids (TSS) and titratable acidity (TA) of arils during storage. The lowest TSS and TA were detected in arils coated with 0.5 and 1% chitosan, which maintained the highest TSS/TA ratio after 12 days of storage. In contrast, application of chitosan delayed the decrease in total phenolics, total anthocyanins and antioxidant capacity during storage. The results also showed that chitosan coating suppressed the monophenolase activity of polyphenol oxidase (PPO) with pyrogallol substrate and the diphenolase activity of PPO with dopamine hydrochloride substrate, but the diphenolase activity of PPO with pyrocatechol substrate increased during storage. CONCLUSION: The results suggest that chitosan coating has the potential to extend the storage life of pomegranate arils by reducing the microbial population on their surface.

The metabolites of mangrove endophytic fungus Zh6-B1 from the South China Sea.

Two new metabolites, 3R,5R-Sonnerlactone (1) and 3R,5S-Sonnerlactone (2), were isolated from the mangrove endophytic fungus Zh6-B1 obtained from the South China Sea. Their structures were elucidated by MS and NMR. The absolute configuration of compound 1 was determined by single-crystal X-ray analysis using Cu Kalpha radiation. The absolute configuration of compound 2 was determined by NOESY analysis and comparing circular dichroism spectroscopy with compound 1. The antiproliferative activity of compound 1 and 2 against the multi-drug resistant human oral floor carcinoma cells (KV) was evaluated.

Overall quality and shelf life of minimally processed and modified atmosphere packaged 'ready-to-eat' pomegranate arils.

Minimally processed ready-to-eat pomegranate arils have become popular due to their convenience, high value, unique sensory characteristics, and health benefits. The objective of this study was to monitor quality parameters and to extend the shelf life of ready-to-eat pomegranate arils packaged with modified atmospheres. Minimally processed pomegranate arils were packed in PP trays sealed with BOPP film under 4 atmospheres including low and super atmospheric oxygen. Packaged arils were stored at 5 degrees C for 18 d and monitored for internal atmosphere and quality attributes. Atmosphere equilibrium was reached for all MAP applications except for high oxygen. As a general trend, slight or no significant change was detected in chemical and physical attributes of pomegranate arils during cold storage. The aerobic mesophilic bacteria were in the range of 2.30 to 4.51 log CFU/g at the end of the storage, which did not affect the sensory quality. Overall, the pomegranate arils packed with air, nitrogen, and enriched oxygen kept quality attributes and were acceptable to sensory panelists on day 18; however, marketability period was limited to 15 d for the low oxygen atmosphere. PP trays sealed with BOPP film combined with either passive or active modified atmospheres and storage at 5 degrees C provided commercially acceptable arils for 18 d with high quality and convenience.