Diversity and characteristics of culturable endophytic bacteria from Passiflora edulis seeds.
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ABSTRACT:
SUBMITTER: Ishida A
PROVIDER: S-EPMC8364935 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Bamboos, regarded as therapeutic agents in ethnomedicine, have been used to inhibit inflammation and enhance natural immunity for a long time in Asia, and there are many bamboo associated fungi with medical and edible value. In the present study, a total of 350 fungal strains were isolated from the uncommon moso bamboo (Phyllostachys edulis) seeds for the first time. The molecular diversity of these endophytic fungi was investigated and bioactive compound producers were screened for the first time. All the fungal endophytes were categorized into 69 morphotypes according to culturable characteristics and their internal transcriber spacer (ITS) regions were analyzed by BLAST search with the NCBI database. The fungal isolates showed high diversity and were divided in Ascomycota (98.0%) and Basidiomycota (2.0%), including at least 19 genera in nine orders. Four particular genera were considered to be newly recorded bambusicolous fungi, including Leptosphaerulina, Simplicillium, Sebacina and an unknown genus in Basidiomycetes. Furthermore, inhibitory effects against clinical pathogens and phytopathogens were screened preliminarily and strains B09 (Cladosporium sp.), B34 (Curvularia sp.), B35 (undefined genus 1), B38 (Penicillium sp.) and zzz816 (Shiraia sp.) displayed broad-spectrum activity against clinical bacteria and yeasts by the agar diffusion method. The crude extracts of isolates B09, B34, B35, B38 and zzz816 under submerged fermentation, also demonstrated various levels of bioactivities against bambusicolous pathogenic fungi. This study is the first report on the antimicrobial activity of endophytic fungi associated with moso bamboo seeds, and the results show that they could be exploited as a potential source of bioactive compounds and plant defense activators. In addition, it is the first time that strains of Shiraia sp. have been isolated and cultured from moso bamboo seeds, and one of them (zzz816) could produce hypocrellin A at high yield, which is significantly different from the other strains published.
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Project description:BackgroundIntragenomic variability in 16S rDNA is a limiting factor for taxonomic and diversity characterization of Bacteria, and studies on its occurrence in natural/environmental populations are scarce. In this work, direct DNA amplicon sequencing coupled with frequent-cutter restriction analysis allowed detection of intragenomic 16S rDNA variation in culturable endophytic bacteria from cacao seeds in a fast and attractive manner.MethodsTotal genomic DNA from 65 bacterial strains was extracted and the 16S rDNA hyper variable V5-V9 regions were amplified for enzyme digestion and direct Sanger-type sequencing. The resulting electropherograms were visually inspected and compared to the corresponding AluI-restriction profiles, as well as to complete genome sequences in databases. Restriction analysis were employed to substitute the need of amplicon cloning and re-sequencing. A specifically improved polyacrylamide-gradient electrophoresis allowed to resolve 5-bp differences in restriction fragment sizes. Chi-square analysis on 2 × 2 contingency table tested for the independence between the 'number of AluI bands' and 'type of eletropherogram'.ResultsTwo types of electropherograms were obtained: unique template, with single peaks per base (clean chromatograms), and heterogeneous template, with various levels of multiple peaks per base (mixed chromatograms). Statistics revealed significant interaction between number of restriction fragments and type of electropherogram for the same amplicons: clean or mixed ones associated to ≤5 or ≥6 bands, respectively. The mixed-template pattern combined with the AluI-restriction profiles indicated a high proportion of 49% of the culturable endophytes from a tropical environment showing evidence of intragenomic 16S rDNA heterogeneity.ConclusionThe approach presented here was useful for a rapid, first-tier detection of intragenomic variation in culturable isolates, which can be applied in studies of other natural populations; a preliminary view of intragenomic heterogeneity levels can complement culture-dependent and -independent methods. Consequences of these findings in taxonomic and diversity studies in complex bacterial communities are discussed.
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Project description:Dendrobium is known for its pharmacological actions including anti-cancer effect, anti-fatigue effect, gastric ulcer protective effect, and so on. At present, only studies on endophytic fungi of Dendrobium affecting the metabolites of host plants have been reported, very little research has been done on endophytic bacteria. In this study, we have demonstrated the great diversity of endophytic bacteria in 6 Dendrobium samples from different origins and cultivars. According to the results of the culture-independent method, the endophytic bacterial community in Dendrobium stems showed obvious different in the 6 samples and was influenced by origin and cultivar. Some bacteria including Ralstonia, Comamonas and Lelliottia were first detected in Dendrobium in this study. Based on the culture-dependent method, a total of 165 cultivable endophytic bacteria isolates were isolated from the sterilized Dendrobium stems, and were classified into 43 species according to the 16S rRNA gene sequence analysis. Moreover, 14 of the 43 strains showed antimicrobial activity against phytopathogen using the Kirby-Bauer method. Strain NA-HTong-7 (Bacillus megaterium, 99.12%) showed the highest antimicrobial activity. This study was the first comprehensive study on endophytic bacteria of Dendrobium from different origins and cultivars, which provides new insights into the endophytic bacteria from Dendrobium.
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Project description:Animal studies have shown the beneficial effects of piceatannol on metabolic health; however, there is a lack of human studies designed to examine these effects. The objective of this study was to investigate the effects of piceatannol on metabolic health in humans. This randomized, placebo-controlled study was conducted on 39 subjects, including 10 overweight men and 9 overweight women (BMI ? 25), as well as 10 non-overweight men and 10 non-overweight women (BMI < 25). Subjects received piceatannol (20 mg/day) or placebo capsules for eight weeks in a random order. The primary outcome was the effect of piceatannol on glucose-metabolism, including insulin sensitivity. The secondary outcomes were the effects on other parameters, including blood pressure (BP), heart rate (HR), endothelial function, lipids, inflammation, oxidative stress, mood status, and Sirt1 and phospho-AMP-activated kinase (p-AMPK) expression in isolated peripheral blood mononuclear cells (PBMNCs). Supplementation with piceatannol in overweight men reduced serum insulin levels, HOMA-IR, BP and HR. Other groups, including non-overweight men, as well as overweight and non-overweight women, showed no beneficial effects on insulin sensitivity, BP and HR. Furthermore, piceatannol is not associated with other data, including body weight (BW), body composition, endothelial function, lipids, inflammation, oxidative stress, mood status, and Sirt1/p-AMPK expression in PBMNCs. In conclusion, supplementation with piceatannol can improve metabolic health, including insulin sensitivity, BP and HR, in overweight men.
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Project description:Endophytes colonizing plant tissue play an essential role in plant growth, development, stress tolerance and plant protection from soil-borne diseases. In this study, we report the diversity of cultivable endophytic bacteria associated with marigold (Calendula officinalis L.) by using 16S rRNA gene analysis and their plant beneficial properties. A total of 42 bacterial isolates were obtained from plant tissues of marigold. They belonged to the genera Pantoea, Enterobacter, Pseudomonas, Achromobacter, Xanthomonas, Rathayibacter, Agrobacterium, Pseudoxanthomonas, and Beijerinckia. Among the bacterial strains, P. kilonensis FRT12, and P. rhizosphaerae FST5 showed moderate or vigorous inhibition against three tested plant pathogenic fungi, F. culmorum, F. solani and R. solani. They also demonstrated the capability to produce hydrolytic enzymes and indole-3-acetic acid (IAA). Five out of 16 isolates significantly stimulated shoot and root growth of marigold in a pot experiment. The present study reveals that more than half of the bacterial isolates associated with marigold (C. officinalis L.) provided antifungal activity against one or more plant pathogenic fungi. Our findings suggest that medicinal plants with antimicrobial activity could be a source for selecting microbes with antagonistic activity against fungal plant pathogens or with plant growth stimulating potential. These isolates might be considered as promising candidates for the improvement of plant health.
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Project description:Healthy cotton samples were collected and 93 endophytic fungal strains were isolated: 23 strains from the roots and 70 strains from the stems. Morphological characterization and ITS sequence analysis were used for the identification of these isolates. The results showed that the 93 strains including 20 species were highly diverse in terms of their taxonomy. Simpson's and Shannon's diversity indices were 0.915 and 3.848, respectively. Fusarium and Alternaria were the two dominant genera, constituting 19.4% of the total strains. Then, 72 spore-producing strains were tested for the suppression of cotton Verticillium wilt (CVW) caused by Verticillium dahliae in a greenhouse. Five strains exhibited effective suppression of CVW with average efficacy values higher than 50%. One of the effective strains, namely, Fusarium proliferatum 10R-7, was selected for the investigation of the role of fusaric acid, a secondary metabolite of strain 10R-7, in the suppression of V. dahliae and CVW. The results showed that F. proliferatum 10R-7 could produce fusaric acid, and this metabolite exhibited 100% inhibition of mycelial growth of V. dahliae at concentrations higher than 20 μg/ml. However, fusaric acid at 2.5 to 80 μg/ml was not effective in the suppression of CVW, compared with the control treatment with V. dahliae alone. F. proliferatum 10R-7 was labeled with green fluorescent protein (GFP), and the GFP-tagged strain was found to be able to colonize inside the taproots of cotton, suggesting that F. proliferatum 10R-7 is a true endophyte of cotton and endophytic colonization may play a role in the suppression of infection of cotton by V. dahliae.
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Project description:Purple passion fruit, Passiflora edulis Sims f. edulis, is an important semi-perennial, fruit bearing vine originating in South America that produces a commercial tropical juice pulp for international and national consumption. Within the round purple passion fruit are a large number of membranous seed sacs each containing individual seeds. Little is known about the seed anatomy of the commercial passion fruit, differences between wild collected and commercial types, and its effect on seedling germination. Therefore, our main objective for this study was to analyze the seed anatomy variability of different germplasm as well as the effect on viability and germination of the seeds of this species. Germplasm was evaluated from three sources: (1) commercial cultivars grown in current production areas, (2) genebank accession from the national seed bank, and (3) landraces collected across different high and mid-elevation sites of the Andean region. A total of 12 morphometric descriptors related to seed anatomy were evaluated on the 56 genotypes, of which three were most informative: Angle to the vertex which is related to the shape of the seed, the thickness of the tegument and the horizontal length; separating the seed according to its source of origin. Germination was found to be positively correlated with the number (r = 0.789) and depth (r = 0.854) of seed pitting. Seeds of the commercial cultivars had more seed pits and higher germination compared to seeds of landraces or genebank accessions showing a possible effect of domestication on the crop. Interestingly, passion fruits often germinate during the rainy season as escaped or wild seedlings especially in the disturbed landscapes of coffee plantations, so some dormancy is needed but faster germination is needed for intensive cultivation. Harnessing passion fruit diversity would be useful as the semi-domesticated landraces have valuable adaptation characteristics to combine with rapid germination selected in the commercial cultivars. The variability of seed pitting with cultivars more pitted than landraces possibly resulting in faster germination may indicate that purple passion fruit is still undergoing a process of selection and domestication for this trait.
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Project description:Flavonoids play a key role as a secondary antioxidant defense system against different biotic and abiotic stresses, and also act as coloring compounds in various fruiting plants. In this study, fruit samples of purple (Passiflora edulis f. edulis) and yellow (Passiflora edulis f. flavicarpa) passion fruit were collected at five developmental stages (i.e., fruitlet, green, veraison, maturation, and ripening stage) from an orchard located at Nanping, Fujian, China. The contents of flavonoid, anthocyanin, proanthocyanin, and their metabolites were determined using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS), activities of key enzymes involved in flavonoid metabolism were measured, and expression profiling of related genes was done using quantitative real-time PCR (qRT-PCR). The results revealed that total flavonoids, anthocyanins, and procyanidins were found to be increased in the fruit peel of both cultivars with fruit maturity. Total flavonoids, anthocyanins, procyanidins, flavonoid metabolites (i.e., rutin, luteolin, and quercetin), and anthocyanin metabolites (i.e., cyanidin-3-O-glucoside chloride, peonidin-3-O-glucoside, and pelargonidin-3-O-glucoside) were found abundant in the peel of purple passion fruit, as compared to yellow passion fruit. Principle component analysis showed that the enzymes, i.e., C4H, 4CL, UFGT, and GST were maybe involved in the regulation of flavonoids metabolism in the peel of passion fruit cultivars. Meanwhile, PePAL4, Pe4CL2,3, PeCHS2, and PeGST7 may play an important role in flavonoid metabolism in fruit peel of the passion fruit. This study provides new insights for future elucidation of key mechanisms regulating flavonoids biosynthesis in passion fruit.
| S-EPMC8620868 | biostudies-literature
Project description:The conditions for the solid-liquid extraction of the antioxidant polyphenol compounds from yellow passion fruit seeds were optimized by response surface methodology with the following variables as the extraction parameters: extraction time (12.8-147.2 min), ethanol concentration (13-97%), and temperature (16.4-83.6 °C). The polyphenol content and antioxidant capacity, which were assessed by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, oxygen radical absorbance capacity, ?-carotene bleaching assay, and ferric reducing antioxidant power assay, were considered dependent variables. The association of the dependent variables was effective for explaining the effect of the independent variables within a determination coefficient (R2) range of 0.88-0.96. A moderate-to-strong correlation for the polyphenol content and antioxidant capacity by the investigated methods was established, and optimized conditions were employed to maximize this response. Extraction was carried out at 80 °C using 70% ethanol concentration for 30 min, which was the most efficient condition to obtain an extract with high concentrations of polyphenolic compounds (3.12 g gallic acid equivalent/100 g seed dry basis) and a strong antioxidant capacity. The stilbene piceatannol was the major compound identified by liquid chromatography-electrospray ionization-tandem mass spectrometry (3.68 g/100 g seed dry basis). These results reinforce that agro-industrial waste demonstrates potential as a source of bioactive compounds, with implications in human health as well as in food and chemical industries.
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Project description:The aims of this study were to isolate, identify and characterize culturable endophytic bacteria from chickpea (Cicer arietinum L.) roots grown in different soils. In addition, the effects of rhizobial inoculation, soil and stress on the functionality of those culturable endophytic bacterial communities were also investigated. Phylogenetic analysis based on partial 16S rRNA gene sequences revealed that the endophytic bacteria isolated in this work belong to the phyla Proteobacteria, Firmicutes and Actinobacteria, with Enterobacter and Pseudomonas being the most frequently observed genera. Production of indoleacetic acid and ammonia were the most widespread plant growth-promoting features, while antifungal activity was relatively rare among the isolates. Despite the fact that the majority of bacterial endophytes were salt- and Mn-tolerant, the isolates obtained from soil with Mn toxicity were generally more Mn-tolerant than those obtained from the same soil amended with dolomitic limestone. Several associations between an isolate's genus and specific plant growth-promoting mechanisms were observed. The data suggest that soil strongly impacts the Mn tolerance of endophytic bacterial communities present in chickpea roots while rhizobial inoculation induces significant changes in terms of isolates' plant growth-promoting abilities. In addition, this study also revealed chickpea-associated endophytic bacteria that could be exploited as sources with potential application in agriculture.
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