The peptide AC 2 isolated from Bacillus-treated Trapa japonica fruit extract rescues DHT (dihydrotestosterone)-treated human dermal papilla cells and mediates mTORC1 signaling for autophagy and apoptosis suppression.

The Trapa japonica fruit is a natural plant growing in ponds with its roots in the mud. It has long been used as a home remedy for many diseases; however, a major problem with this kind of natural extract is the multicomponents-multitargets for diseases. Such problems make it difficult to identify the mechanism of action. Another problem is quality control and consistency. The aim of this research was to isolate a single bioactive compound (peptide) derived from the Trapa japonica fruit. The research was conducted with various experimental techniques, such as fermentation and liquid chromatography, to isolate a peptide. We isolated the AC 2 peptide from Trapa japonica fruit and found it to be promising on human dermal papilla cells. Dihydrotestosterone (DHT) stresses human dermal papilla cells and is a major cause of hair loss resulting from hormones and environmental factors. The purpose of this research was to develop an understanding of the mechanism by which the AC 2 peptide rescues dihydrotestosterone (DHT)-treated human dermal papilla cells. We explored the effects of the AC 2 peptide on the cell biological functions of human dermal papilla cells (HDPs). HDPs were treated with the AC 2 peptide and DHT. Then, a cytotoxicity assay, flow cytometry, Western blot, immunoprecipitation, and 3D cell culture for immunohistochemistry were conducted to investigate the mTORC1 pathway and suppression of autophagy and apoptosis. In addition, we also synthesized the AC2 peptide as an alternative to the expensive and difficult isolation and purification procedures and confirmed its potential in biomedical applications. We also validated the effects of the synthetic AC2 peptide as well as the isolated and purified AC2 peptide and established their similarity. Although extensive research has been carried out on natural extracts, few single studies have isolated and separated a bioactive peptide (single compound).

Characterization of Colletotrichum Species Causing Anthracnose of Pomegranate in the Southeastern United States.

Anthracnose fruit rot and leaf blight caused by Colletotrichum species are important diseases of pomegranate in the southeastern United States. In this study, 26 isolates from pomegranate were identified based on pathological and molecular characterization. Isolates were identified to species based on multilocus sequence analysis with the internal transcribed spacer region, glyceraldehyde-3-phosphate dehydrogenase, ß-tubulin, and chitin synthase genomic genes. Pomegranate isolates grouped within the C. acutatum and C. gloeosporioides species complexes, with more than 73% belonging to the latter group. Three species were identified within the C. acutatum species complex (C. nymphaeae [n = 5], C. fioriniae [n = 1], and C. simmondsii [n = 1]), and three other species were identified within the C. gloeosporioides species complex (C. theobromicola [n = 11], C. siamense [n = 6], and C. gloeosporioides [n = 2]). Inoculations of pomegranate fruit showed that isolates from the C. acutatum species complex were more aggressive than isolates from the C. gloeosporioides species complex. Interestingly, opposite results were observed when leaves of rooted pomegranate cuttings were inoculated. In addition, Colletotrichum isolates from pomegranate, strawberry, blueberry, mango, and citrus were cross-pathogenic when inoculated to fruit. This is the first study identifying six different species of Colletotrichum causing pomegranate leaf blight and fruit anthracnose in the southeastern United States and the potential cross-pathogenic capability of pomegranate isolates to other commercially important crops.

Green approach to phytopathogen: Characterization of lytic bacteriophages of Pseudomonas sp., an etiology of the bacterial blight of pomegranate.

Two morphologically different bacteriophages were isolated from the river and soil samples from various locations of Maharashtra, India against the phytopathogen Pseudomonas sp. that was recently reported to cause a new bacterial blight of pomegranate. Both the phages belonged to the order Caudovirales representing the families Siphoviridae (vB_Psp.S_PRɸL2) and Myoviridae (vB_Psp.M_SSɸL8). The multiplicity of infection ranged from 0.01 to 0.1, phage adsorption rate from 39% to 66%, latent period from 10 to 20 min with a burst size of 24-85 phage particles per infected host cell. The genome size of phages PRɸL2 and SSɸL8 was approximately 25.403 kb and 29.877 kb respectively. Restriction digestion pattern of phage genomic DNA was carried out for phage PRɸL2, Eco RI resulted in two bands and Hind III resulted in three bands while for phage SSɸL8, both Eco RI and Hind III each resulted in three bands. SDS-PAGE protein profile showed six bands for PRɸL2 and nine bands for SSɸL8 of different proteins. Phages showed high pH stability over a range of 4-9, temperature stability over a range of 4-50 °C and UV radiation showed a reduction up to 89.36% for PRɸL2 and 96% for SSɸL8. In short, the present research work discusses for the first time in-detailed characterization of phages of a phytopathogen Pseudomonas sp. from Maharashtra, India, which can be further efficiently used for biological control of the causative agent of a new bacterial blight disease of pomegranate.

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).

Nanomaterials for the control of bacterial blight disease in pomegranate: quo vadis?

Bacterial blight, caused by Xanthomonas axonopodis pv. punicae, Xap is a serious threat to commercially successful pomegranate (Punica granatum L) crop. Owing to the non-availability of disease-resistant varieties of pomegranate, integrated disease management involving change of season, adequate nutrition, and preventive sprays of bactericides is used to control Xap. We undertook a systematic study to assess the efficacy of metal-based nanomaterials (Cu, CuO, ZnO, CaO, MgO) for the control of Xap. The antimicrobial effectiveness was in the order Cu > ZnO > MgO > CuO with MIC (minimum inhibitory concentration) 2.5, 20, 190, 200, and 1600 µg/ml. A time-to-kill curve indicated that Cu nanoparticles (CuNPs) killed Xap cells within 30 min at 2.5 µg/ml. Under controlled conditions (polyhouse), foliar application of CuNPs (400 µg/ml) resulted in ~ 90 and ~ 15% disease reduction in 6-month-old infected plants at early (disease severity 10%) and established (disease severity 40%) stages of infection, respectively. In a subsequent field study on severely infected 7-year-old plants, applications of nanoparticles reduced the disease incidence by ~ 20% as compared to untreated control. Microscopic observations revealed that CuNPs reduced the bacterial colonization of the leaf surface. Anti-Xap activity of foliar applied CuNPs was on par with conventionally used copper oxychloride (3000 µg/ml) albeit at 8-fold reduced copper concentration. Thus, early disease detection and application of effective dosage of copper nanoparticles can indeed help the farmer in achieving rapid infection control. Further studies on use of combinations of nanoparticles for management of bacterial blight are warranted.

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.

Factors Affecting the Response of Pomegranate Fruit to Alternaria alternata, the Causal Agent of Heart Rot.

Heart rot, caused by Alternaria alternata, is a major pomegranate disease that impacts production worldwide; most fruits in orchards are colonized by A. alternata but, nevertheless, symptoms are apparent on only a small proportion of the colonized fruits. During the years of our previous research it was noticed that within individual orchards, the incidence of pomegranate fruits exhibiting heart rot symptoms was related to the visual appearance of the trees: trees that appeared visually frail bore more diseased fruits than robust trees. Furthermore, it was noticed that the disease responses of different pomegranate accessions and possibly of different variants of the same cultivar varied markedly. The specific objectives of the present study were: (i) to characterize the relationship between the visual appearance of pomegranate plants or individual stems and the incidence of heart rot and their vulnerability to heart rot; and (ii) to elucidate factors affecting the response of pomegranate fruit to A. alternata. Analysis of heart rot incidence in four orchards in 2014 revealed large differences in heart rot incidence among trees growing side by side in the same orchard; these differences were related to the visual appearance of the pomegranate trees. There were significant differences among germination rates of A. alternata spores in juice prepared from asymptomatic fruits originating from these trees, and comparable differences were found among the acidity levels (pH) of the juices. These differences may reflect differences among the physiological responses of pomegranate trees to heart rot. Fruits collected from the pomegranate collection located in Newe Ya'ar, which comprised 95 accessions in 2015 and 110 accessions in 2016, were also examined. There were differences among the acidity levels (pH) of the juices produced from these fruits and among the germination rates of A. alternata spores in the juices. These differences may reflect variances among the genetic responses of pomegranate accessions to heart rot. Results of studying the relationship between the acidity levels of pomegranate juice and the germination rates of A. alternata spores supported the hypothesis that, apart from pH, a compound(s) present in the juice regulates the germination of A. alternata spores in the juice.

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.

Nocardioides sonneratiae sp. nov., an endophytic actinomycete isolated from a branch of Sonneratia apetala.

A Gram-staining-positive, aerobic non-spore-forming and short-rod-shaped endophytic actinomycete was isolated from a branch of Sonneratia apetala, designated strain BGMRC0092T and investigated in detail data to determine its taxonomic position. On the basis of the 16S rRNA gene sequence analysis, the results of the phylogenetic analyses indicated that BGMRC0092T was most closely related to Nocardioides alpinus Cr7-14T (96.9 %), Nocardioides oleivorans DSM16090T (96.4 %) and Nocardiodes exalbidus RC825T (96.3 %). The predominant cellular fatty acids of BGMRC0092T were iso-C16 : 0 and C18 : 1ω8c. The major menaquinone was MK-8(H4). The diagnostic diamino acid in the cell-wall peptidoglycan was ll-2,6-diaminopimelic acid. The predominant cell-wall sugars were composed of galactose, mannose, rhamnose and xylose. The polar lipid pattern contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, one unknown phospholipid and three unknown polar lipids. The DNA G+C content was 69.3 mol%. On the basis of phenotypic and chemotaxonomic characteristics and the results of phylogenetic analysis, BGMRC0092T represents a novel species of the genus Nocardioides, for which the name Nocardioides sonneratiae sp. nov. is proposed. The type strain is Nocardioides sonneratiae BGMRC0092T (=KCTC 39565T=NBRC 110251 T=DSM 100390T).

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.

Impacts of pulsed electric field and heat treatment on quality and sensory properties and microbial inactivation of pomegranate juice.

Synergistic effects of pulsed electric field+mild heat on quality properties of pomegranate juice were modeled using the best-fit multiple (non-) linear regression models with inactivation kinetics parameters of Escherichia coli O157:H7 and Staphylococcus aureus. No significant difference was detected between the control and the treated samples in terms of pH; °Brix; total antioxidant capacity; total monomeric anthocyanin content; total ascorbic acid concentration; and the sensory properties of flavor, taste, aftertaste, and overall acceptance ( p > 0.05). An exposure of 65.3 J and 40 ℃ caused an increase on conductivity; titratable acidity; L*, a*, and b* values; and a decrease of browning index, total phenolic content, total antioxidant capacity, total monomeric anthocyanin content, total ascorbic acid concentration, and in the sensory properties of color and sourness in pomegranate juice. The goodness-of-fit for the best-fit multiple (non-) linear regression models in descending order belonged to E. coli O157:H7 (92.98%), S. aureus (84.06%), color a* (83.9%), titratable acidity (81.3%), color L* (78.5%), color b* (78.3%), conductivity (74.8%), total phenolic content (74.1%), and total ascorbic acid concentration (64.74%), respectively. De and ze values for E. coli O157:H7 and S. aureus ranged from 105.64 to 1093.25 and from 79.18 to 1057.73 µs with 27.39 and 30.80 J, consequently.

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.

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.

Influence of Fermentation with Different Lactic Acid Bacteria and in Vitro Digestion on the Biotransformation of Phenolic Compounds in Fermented Pomegranate Juices.

This study describes the effect of fermentation and the impact of simulated gastrointestinal digestion (SGD) of four fermented pomegranate juices with different lactic acid bacteria (LAB) on the biotransformation of phenolic compounds. The changes of the antioxidant capacity (AOC) and of LAB growth and survival in different fermented juices were also studied. Two new phenolic derivatives (catechin and α-punicalagin) were identified only in fermented juices. During SGD, the AOC increased together with the phenolic derivatives concentration mainly in the juices fermented with Lactobacillus. These derivatives were formed due to the LAB metabolism of the ellagitannins, epicatechin, and catechin after fermentation and during SGD. The FRAP assay performance might be associated with the degradation and biotransformation of catechin. The fermented pomegranate juices with these LAB increased the bioaccessibility of phenolic compounds, ensuring the survival of LAB after SGD, suggesting a possible prebiotic effect of phenolic compounds on LAB.

Development of a nested-PCR assay for the rapid detection of Pilidiella granati in pomegranate fruit.

Pilidiella granati, a causal agent of twig blight and crown rot of pomegranate, is an emerging threat that may cause severe risk to the pomegranate industry in the future. Development of a rapid assay for the timely and accurate detection of P. granati will be helpful in the active surveillance and management of the disease caused by this pathogen. In this study, a nested PCR method was established for the detection of P. granati. Comparative analysis of genetic diversity within 5.8S rDNA internal transcribed spacer (ITS) sequences of P. granati and 21 other selected fungal species was performed to design species-specific primers (S1 and S2). This primer pair successfully amplified a 450 bp product exclusively from the genomic DNA of P. granati. The developed method can detect 10 pg genomic DNA of the pathogen in about 6 h. This technique was successfully applied to detect the natural infection of P. granati in the pomegranate fruit. The designed protocol is rapid and precise with a high degree of sensitivity.

Polyketides with Immunosuppressive Activities from Mangrove Endophytic Fungus Penicillium sp. ZJ-SY2.

Nine polyketides, including two new benzophenone derivatives, peniphenone (1) and methyl peniphenone (2), along with seven known xanthones (3-9) were obtained from mangrove endophytic fungus Penicillium sp. ZJ-SY2 isolated from the leaves of Sonneratia apetala. Their structures were elucidated on the basis of MS, 1D, and 2D NMR data. Compounds 1, 3, 5, and 7 showed potent immunosuppressive activity with IC50 values ranging from 5.9 to 9.3 µg/mL.

XopN-T3SS effector of Xanthomonas axonopodis pv. punicae localizes to the plasma membrane and modulates ROS accumulation events during blight pathogenesis in pomegranate.

Bacterial blight caused by Xanthomonas axonopodis pv. punicae (Xap) is a major disease of pomegranate. Xap secretes effector proteins via type III secretion system (T3SS) to suppress pathogen-associated molecular pattern (PAMP)-triggered plant immunity (PTI). Previously we reported that XopN, a conserved effector of Xap, modulate in planta bacterial growth, and blight disease. In continuation to that here we report the deletion of XopN from Xap caused higher accumulation of reactive oxygen species (ROS) including H2O2 and O2-. We quantitatively assessed the higher accumulation of H2O2 in pomegranate leaves infiltrated with Xap ΔxopN compared to Xap wild-type. We analysed that 1.5 to 3.3 fold increase in transcript expression of ROS and flg22-inducible genes, namely FRK1, GST1, WRKY29, PR1, PR2 and PR5 in Arabidopsis when challenged with Xap ΔxopN; contrary, the up-regulation of all the genes were compromised when challenged with either Xap wild-type or Xap ΔxopN+xopN. Further, we demonstrated the plasma-membrane based localization of XopN protein both in its natural and experimental hosts. All together, the present study suggested that XopN-T3SS effector of Xap gets localized in the plasma membrane and suppresses ROS-mediated early defense responses during blight pathogenesis in pomegranate.

[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.