scholarly journals Vertical Transmission of Diverse Cultivation-Recalcitrant Endophytic Bacteria Elucidated Using Watermelon Seed Embryos

2021 ◽  
Vol 12 ◽  
Author(s):  
Pious Thomas ◽  
Pramod Kumar Sahu
Keyword(s):  
Bacterial Diversity ◽  
Vertical Transmission ◽  
Endophytic Bacteria ◽  
Bacterial Colonization ◽  
Confocal Imaging ◽  
Plant Biology ◽  
Watermelon Seed ◽  
Tissue Homogenates ◽  
Embryo Sections

Seed transmission of endophytic microorganisms is a growing research area in plant biology and microbiology. We employed cultivation versus cultivation-independent approaches on excised embryos from watermelon seeds (6–12 months in storage) and on embryo-derived in vitro seedlings (EIVS) to assess the vertical transmission of endophytic bacteria. Surface-disinfected watermelon seeds bore abundant residual bacteria in the testa and perisperm tissues, predominantly Bacillus spp. propounding the essentiality of excluding all non-embryonic tissues for vertical transmission studies. Tissue homogenates from re-disinfected seed embryos displayed no cultivable bacteria during the 1-week monitoring. Bright-field live microscopy revealed abundant bacteria in tissue homogenates and in embryo sections as intracellular motile particles. Confocal imaging on embryo sections after SYTO-9 staining and eubacterial fluorescent in situ hybridization (FISH) endorsed enormous bacterial colonization. Quantitative Insights Into Microbial Ecology (QIIME)-based 16S rRNA V3–V4 taxonomic profiling excluding the preponderant chloroplast and mitochondrial sequences revealed a high bacterial diversity in watermelon seed embryos mainly Firmicutes barring spore formers followed by Proteobacteria, Bacteroidetes, and Actinobacteria, and other minor phyla. Embryo-base (comprising the radicle plus plumule parts) and embryo-cotyledon parts differed in bacterial profiles with the abundance of Firmicutes in the former and Proteobacteria dominance in the latter. EIVS displayed a higher bacterial diversity over seed embryos indicating the activation from the dormant stage of more organisms in seedlings or their better amenability to DNA techniques. It also indicated embryo-to-seedling bacterial transmission, varying taxonomic abundances for seed embryos and seedlings, and differing phylogenic profiles for root, hypocotyl, and cotyledon/shoot-tip tissues. Investigations on different watermelon cultivars confirmed the embryo transmission of diverse cultivation recalcitrant endophytic bacteria. Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes formed the core phyla across different cultivars with 80–90% similarity at genus to phylum levels. Conversely, freshly harvested seeds displayed a dominance of Proteobacteria. The findings revealed that dicot seeds such as in different watermelon cultivars come packaged with abundant and diverse vertical and seedling-transmissible cultivation recalcitrant endophytic bacteria with significant implications for plant biology.

scholarly journals Influence of native endophytic bacteria on the growth and bacterial crown rot tolerance of papaya (Carica papaya)

2021 ◽  
Author(s):  
Mark Paul Selda Rivarez ◽  
Elizabeth P. Parac ◽  
Shajara Fatima M. Dimasingkil ◽  
Eka Mirnia ◽  
Pablito M. Magdalita
Keyword(s):  
Plant Growth ◽  
Endophytic Bacteria ◽  
Plant Biomass ◽  
Growth Promotion ◽  
Bacterial Colonization ◽  
Fold Increase ◽  
Pathogen Resistance ◽  
Crown Rot ◽  
Native Plant

Abstract The native plant microbiome is composed of diverse communities that influence its overall health, with some species known to promote plant growth and pathogen resistance. Here, we show the antibacterial and growth promoting activities of autoclaved culture metabolites (ACMs) from native endophytic bacteria (NEB) in a papaya cultivar that is tolerant to bacterial crown rot (BCR) caused by Erwinia mallotivora . Initially, bacterial colonization in recovering tissues of this cultivar was observed before onset of tissue regeneration or 'regrowth'. We further isolated and characterized these bacteria and were able to identify two culturable stem NEB under genera Kosakonia (EBW), related to Enterobacter , and Sphingomonas (EBY). We also identified root NEB (BN, BS and BT) under genus Bacillus . Inhibition assays indicated that ACMs from these NEB promptly (18-30h) and efficiently inhibited (60-65%) E. mallotivora proliferation in vitro. Interestingly, when ACMs from BN and EBW were inoculated in surface-sterilized papaya seeds, germination was variably retarded (20-60% reduction) depending on plant genotype, but plant biomass accumulation was significantly stimulated, at around two-fold increase. Moreover, greenhouse experiments show that ACMs from all isolates, especially EBW, significantly reduced BCR incidence and severity in susceptible genotype, at around two-fold. In general, our observations of pathogen antagonism, plant growth promotion leading to disease reduction by ACMs of native endophytic bacteria suggested its contribution to increased fitness of papaya and tolerance against the (re)emerging BCR disease.

In vitro regeneration of watermelon seed segments

2015 ◽  
Vol 4 (8) ◽  
Author(s):  
M. Ameri ◽  
M. Lahouti ◽  
A. Bagheri ◽  
A. Sharifi ◽  
F. Keykha Akhar
Keyword(s):  
In Vitro Regeneration ◽  
Watermelon Seed

scholarly journals Use of electromagnetic stimulation on an Enterococcus faecalis biofilm on root canal treated teeth in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Beatriz H. D. Panariello ◽  
Justin K. Kindler ◽  
Kenneth J. Spolnik ◽  
Ygal Ehrlich ◽  
George J. Eckert ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Imaging ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Electromagnetic Stimulation ◽  
Root Canal Disinfection

AbstractRoot canal disinfection is of utmost importance in the success of the treatment, thus, a novel method for achieving root canal disinfection by electromagnetic waves, creating a synergistic reaction via electric and thermal energy, was created. To study electromagnetic stimulation (EMS) for the disinfection of root canal in vitro, single rooted teeth were instrumented with a 45.05 Wave One Gold reciprocating file. Specimens were sterilized and inoculated with Enterococcus faecalis ATCC 29,212, which grew for 15 days to form an established biofilm. Samples were treated with 6% sodium hypochlorite (NaOCl), 1.5% NaOCl 1.5% NaOCl with EMS, 0.9% saline with EMS or 0.9% saline. After treatments, the colony forming units (CFU) was determined. Data was analyzed by Wilcoxon Rank Sums Test (α = 0.05). One sample per group was scored and split for confocal laser scanning microscopy imaging. There was a significant effect with the use of NaOCl with or without EMS versus 0.9% saline with or without EMS (p = 0.012 and 0.003, respectively). CFUs were lower when using 0.9% saline with EMS versus 0.9% saline alone (p = 0.002). Confocal imaging confirmed CFU findings. EMS with saline has an antibiofilm effect against E. faecalis and can potentially be applied for endodontic disinfection.

Bacterial endosymbiont inhabiting Leucaena leucocephala leaves and their antioxidant and antidiabetic potential

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Sridevi Chigurupati ◽  
Shantini Vijayabalan ◽  
Kesavanarayanan Krishnan Selvarajan ◽  
Ahmad Alhowail ◽  
Fatema Kauser
Keyword(s):  
Free Radical ◽  
Crude Extract ◽  
Leucaena Leucocephala ◽  
Endophytic Bacteria ◽  
Radical Scavenging ◽  
16S Rrna Gene Sequencing ◽  
Free Radical Scavenging ◽  
Rrna Gene ◽  
Standard Drug

AbstractObjectivesResearch on endosymbionts is emerging globally and is considered as a potential source of bioactive phytochemicals. The present study examines the antioxidant and antidiabetic of the endophytic crude extract isolated from Leucaena leucocephala leaves.MethodsEndophytic bacteria were isolated from the leaves of L. leucocephala and 16S rRNA gene sequencing was used to establish their identity. The in vitro antioxidant effect of endophytic crude extract (LL) was evaluated using 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) free radical scavenging methods. The in vitro antidiabetic properties of LL were evaluated using α-amylase and α-glucosidase enzyme inhibition assay.ResultsThe isolated endophytic bacteria were identified as Cronobacter sakazakii. LL displayed potent free radical scavenging effect against ABTS and DPPH radicals with an inhibitory concentration 50% (IC50) value of 17.49 ± 0.06 and 11.3 ± 0.1 μg/mL respectively. LL exhibited α-amylase and α-glucosidase inhibition with an IC50 value of 23.3 ± 0.08 and 23.4 ± 0.1 μg/mL respectively compared to the standard drug (acarbose). Both glucose loaded normoglycemic rats and STZ induced diabetic rats treated with LL (200 mg/kg) exhibited a considerable reduction in blood glucose levels p<0.01 after 8 h of treatment when compared to normal and diabetic control rats respectively.ConclusionsThus, the study shows that LL has a wellspring of natural source of antioxidants, and antidiabetic agents and phytoconstituents present in endophytes could be the rich source for bioactive compounds.

scholarly journals DDEL-12. NANOPARTICLE DELIVERY OF DOXORUBICIN FOR THE TREATMENT OF DIFFUSE INTRINSIC PONTINE GLIOMA (DIPG)

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii286-iii286
Author(s):  
Caitlin Ung ◽  
Maria Tsoli ◽  
Jie Liu ◽  
Domenico Cassano ◽  
Dannielle Upton ◽  
...  
Keyword(s):  
Treatment Strategies ◽  
Pediatric Brain Tumors ◽  
Diffuse Intrinsic Pontine Glioma ◽  
Confocal Imaging ◽  
Cell Content ◽  
Potent Effect ◽  
Orthotopic Mouse Model ◽  
In Vivo Testing

Abstract DIPGs are the most aggressive pediatric brain tumors. Currently, the only treatment is irradiation but due to its palliative nature patients die within 12 months. Effective delivery of chemotherapy across the blood-brain barrier (BBB) has been a key challenge for the eradication of this disease. We have developed a novel gold nanoparticle functionalised with human serum albumin (Au-NP, 98.8 ±19 nm) for the delivery of doxorubicin. In this study, we evaluated the cytotoxic efficacy of doxorubicin delivered through gold nanoparticles (Au-NP-Dox). We found that DIPG neurospheres were equally sensitive to doxorubicin and Au-NP-Dox (at equimolar concentration) by alamar blue assay. Colony formation assays demonstrated a significantly more potent effect of Au-NP-Dox compared to doxorubicin alone, while the Au-NP had no effect. Furthermore, western blot analysis indicated increased apoptotic markers cleaved Parp, caspase 3/7 and phosphorylated H2AX in Au-NP-Dox treated DIPG neurospheres. Live cell content and confocal imaging demonstrated significantly higher uptake of Au-NP-Dox compared to doxorubicin alone. Treatment of a DIPG orthotopic mouse model with Au-NP-Dox showed no signs of toxicity with stable weights being maintained during treatment. However, in contrast to the above in vitro findings the in vivo study showed no anti-tumor effect possibly due to poor penetration of Au-NP-Dox into the brain. We are currently evaluating whether efficacy can be improved using measures to open the BBB transiently. This study highlights the need for rigorous in vivo testing of new treatment strategies before clinical translation to reduce the risk of administration of ineffective treatments.

scholarly journals The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr.

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1276
Author(s):  
Olga A. Aleynova ◽  
Andrey R. Suprun ◽  
Nikolay N. Nityagovsky ◽  
Alexandra S. Dubrovina ◽  
Konstantin V. Kiselev
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Endophytic Fungi ◽  
Endophytic Bacteria ◽  
Fungal Endophytes ◽  
Biomass Accumulation ◽  
Vitis Amurensis ◽  
Grape Cell ◽  
Stilbene Biosynthesis

Plant endophytes are known to alter the profile of secondary metabolites in plant hosts. In this study, we identified the main bacterial and fungal representatives of the wild grape Vitis amurensis Rupr. microbiome and investigated a cocultivation effect of the 14 endophytes and the V. amurensis cell suspension on biomass accumulation and stilbene biosynthesis. The cocultivation of the V. amurensis cell culture with the bacteria Agrobacterium sp., Bacillus sp., and Curtobacterium sp. for 2 weeks did not significantly affect the accumulation of cell culture fresh biomass. However, it was significantly inhibited by the bacteria Erwinia sp., Pantoea sp., Pseudomonas sp., and Xanthomonas sp. and fungi Alternaria sp., Biscogniauxia sp., Cladosporium sp., Didymella sp. 2, and Fusarium sp. Cocultivation of the grapevine cell suspension with the fungi Didymella sp. 1 and Trichoderma sp. resulted in cell death. The addition of endophytic bacteria increased the total stilbene content by 2.2–5.3 times, while the addition of endophytic fungi was more effective in inducing stilbene accumulation by 2.6–16.3 times. The highest content of stilbenes in the grapevine cells cocultured with endophytic fungi was 13.63 and 13.76 mg/g of the cell dry weight (DW) after cultivation with Biscogniauxia sp. and Didymella sp. 2, respectively. The highest content of stilbenes in the grapevine cells cocultured with endophytic bacteria was 4.49 mg/g DW after cultivation with Xanthomonas sp. The increase in stilbene production was due to a significant activation of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) gene expression. We also analyzed the sensitivity of the selected endophytes to eight antibiotics, fluconazole, and trans-resveratrol. The endophytic bacteria were sensitive to gentamicin and kanamycin, while all selected fungal strains were resistant to fluconazole with the exception of Cladosporium sp. All endophytes were tolerant of trans-resveratrol. This study showed that grape endophytes stimulate the production of stilbenes in grape cell suspension, which could further contribute to the generation of a new stimulator of stilbene biosynthesis in grapevine or grape cell cultures.

scholarly journals Unraveling the Metabolite Signature of Endophytic Bacillus velezensis Strain Showing Defense Response Towards Fusarium oxysporum

Agronomy ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 683
Author(s):  
Yang Cao ◽  
Wei Ding ◽  
Chang Liu
Keyword(s):  
Fusarium Oxysporum ◽  
Inhibitory Activity ◽  
Defense Response ◽  
Endophytic Bacteria ◽  
Rice Seedling ◽  
Seedling Blight ◽  
Bacillus Velezensis ◽  
Secondary Bile Acid ◽  
Pathogen Inhibition

Seedling blight, caused by the fungus Fusarium oxysporum, significantly lowers rice production globally. Earlier reports have opined that endophytic bacteria strains could be possible biocontrol agents, but the mechanistic actions involved are still unclear. Therefore, this study aimed to isolate the endophytic bacteria with high inhibitory activity and elucidate its possible mechanisms for inducing resistance by metabolomics. The results showed that mdj-36 had the strongest in vitro pathogen inhibition of F. oxysporum, while mdj-34 displayed the lowest inhibitory activity identified as Bacillus velezensis strains. Metabolomic analyses demonstrated that B. velezensis mdj-36 growth medium could produce higher organic acids, terpenes, and diterpene than B. velezensis mdj-34. Further investigation revealed that ‘secondary bile acid biosynthesis’ and ‘glycerophospholipid metabolism’ pathways played essential roles in defense response towards F. oxysporum. This study’s findings provide a credible theoretical basis for the possible use of the B. velezensis strain against rice seedling blight.

scholarly journals Antagonistic activity of endophytic bacteria isolated from weed plant against stem end rot pathogen of pitaya in Vietnam

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
The Anh Luu ◽  
Quyet Tien Phi ◽  
Thi Thu Hang Nguyen ◽  
Mai Van Dinh ◽  
Bich Ngoc Pham ◽  
...  
Keyword(s):  
Endophytic Bacteria ◽  
Indole Acetic Acid ◽  
Pathogenic Fungi ◽  
Antagonistic Activity ◽  
Biofilm Production ◽  
Weed Plant ◽  
Seedling Biomass ◽  
Rot Disease ◽  
Positive Results

Abstract Background Fungal stem end rot disease of pitaya caused by Alternaria alternata is one of the most destructive diseases in Binh Thuan province, Vietnam. This study aimed to assess the antagonistic effects of some endophytic bacteria isolated from the weed plant (Echinochloa colonum) against A. alternata. Results A total of 19 endophytic bacteria were isolated and 5 of them presented in vitro antagonistic activity against A. alternata. Of five, strain EC80 significantly inhibited the pathogenic growth with a mean inhibition diameter of 11.88 ± 0.08 mm, while the other four (C79, EC83, EC90, and EC97) showed a weak inhibition. Interestingly, the combination of EC79 and EC80 reduced more biomass of pathogenic fungi than the single one did. EC79 showed positive results for amylase, indole acetic acid (IAA), and biofilm production, whereas EC80 presented positive capabilities for IAA and biofilm production and a negative one for amylase production. In addition, the combined filtrate of EC79 and EC80 presented non-antifungal activity on biocontrol tests in vitro, indicating that bacteria cells played a role in defending against the pathogen. Moreover, both isolates EC79 and EC80 significantly increased seedling biomass than the control. Conclusions The results suggest that those two strains in combination had the potential to be used as a biocontrol agent against A. alternata. More studies should be done in the future to evaluate their efficiency under the field conditions.

Sign in / Sign up

Export Citation Format

Share Document