scholarly journals Diversity of Nematode Microbial Antagonists from Algeria Shows Occurrence of Nematotoxic Trichoderma spp.

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 941
Author(s):  
Nawal Benttoumi ◽  
Mariantonietta Colagiero ◽  
Samira Sellami ◽  
Houda Boureghda ◽  
Abdelaziz Keddad ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Fusarium Spp ◽  
Trichoderma Spp ◽  
Bacillus Spp ◽  
Phytoparasitic Nematodes ◽  
Meloidogyne Spp ◽  
Rrna Gene Sequencing

Fungi and bacteria associated to phytoparasitic nematodes Globodera rostochiensis and Meloidogyne spp. in Algeria were identified and characterized. Trichoderma spp. showed the highest prevalence in the cysts of G. rostochiensis. A number of isolates were identified through PCR amplification and the sequencing of the internal transcribed spacer (ITS)1-2 and Rpb2 gene regions. The most represented species were T. harzianum and T. afroharzianum. The latter and T. hirsutum were reported for the first time in Algeria. Fusarium spp., including F. oxysporum and F. solani, comprised a second group of fungi found in cysts. Taxa associated to females of Meloidogyne spp. included T. harzianum, Fusarium spp. and other hyphomycetes. To assess the efficacy of Trichoderma spp., two assays were carried out in vitro with the culture filtrates of two T. afroharzianum and T. harzianum isolates, to check their toxicity versus the second stage juveniles of M. incognita. After 24–48 h exposure, a mortality significantly higher than the control was observed for both filtrates at 1% dilutions. The TRI genes involved in the production of trichothecenes were also amplified with the PCR from some Trichoderma spp. isolates and sequenced, supporting a putative role in nematode toxicity. Bacteria isolated from the cysts of G. rostochiensis included Brucella, Rhizobium, Stenotrophomonas and Bacillus spp., identified through 16S rRNA gene sequencing. The potential of the microbial isolates identified and their mechanisms of action are discussed, as part of a sustainable nematode management strategy.

scholarly journals Bacterial Communities Associated with the Cycling of Non-Starch Polysaccharides and Phytate in Aquaponics Systems

Diversity ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 631
Author(s):  
Daniel Menezes-Blackburn ◽  
Nahad Al-Mahrouqi ◽  
Buthaina Al-Siyabi ◽  
Adhari Al-Kalbani ◽  
Ralf Greiner ◽  
...  
Keyword(s):  
Bacterial Communities ◽  
Pcr Amplification ◽  
Aquatic Organisms ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Culture Techniques ◽  
Operational Taxonomic Units ◽  
Catalytic Mechanisms ◽  
Rrna Gene Sequencing

Aquaponics are efficient systems that associate aquatic organisms’ production and plants by recirculating water and nutrients between aquaculture and hydroponic tanks. In this study, we characterised the bacterial communities in the freshwater aquaponics system that can mineralise polysaccharides and phytate by producing carbohydrate-degrading enzymes and phytases, by 16S rRNA gene sequencing and in vitro culture techniques. Around 20% of the operational taxonomic units (zOTUs) identified were previously reported to carry fibre-degrading enzyme putative genes, namely β-glucanase (1%), xylanase (5%), or cellulases (17%). Ten % of the zOTUs were previously reported to carry putative genes of phytases with different catalytic mechanisms, namely β-propeller (6%), histidine acid phytases (3%), and protein tyrosine phytase (<1%). Thirty-eight morphologically different bacteria were isolated from biofilms accumulated in fish and plant compartments, and identified to belong to the Bacilli class. Among these, 7 could produce xylanase, 8 produced β-glucanase, 14 produced cellulase, and 11 isolates could secrete amylases. In addition, Staphylococcus sp. and Rossellomorea sp. could produce consistent extracellular phytate-degrading activity. The PCR amplification of β-propeller genes both in environmental samples and in the isolates obtained showed that this is the most ecologically relevant phytase type in the aquaponics systems used. In summary, the aquaponics system is abundant with bacteria carrying enzymes responsible for plant-nutrient mineralisation.

scholarly journals Bacterial Communities Associated with the Cycling of Non-Starch Polysaccharides and Phytate in Aquaponics Systems

2021 ◽  
Author(s):  
Daniel Menezes-Blackburn ◽  
Nahad Al-Mahrouqi ◽  
Buthaina Al-Siyabi ◽  
Adhari Al-Kalbani ◽  
Ralf Greiner ◽  
...  
Keyword(s):  
Bacterial Communities ◽  
Pcr Amplification ◽  
Aquatic Organisms ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Beta Glucanase ◽  
Culture Techniques ◽  
Degrading Enzymes ◽  
Rrna Gene Sequencing

Aquaponics are efficient systems that associate aquatic organisms&rsquo; production and plants by recirculating water and nutrients between aquaculture and hydroponic tanks. In this study, we have characterised the bacterial communities in the fresh water aquaponics system that can mineralise polysaccharides and phytate by producing carbohydrate degrading enzymes and phytases, by 16S rRNA gene sequencing and in vitro culture techniques. Around 20% of the operational taxonomic units (OTUs) identified were previously reported to carry fibre-degrading enzymes putative genes, namely &beta;-glucanase (1%), xylanase (5%) or cellulases (17%). Ten % of the OTUs were previously reported to carry putative genes of phytases with different catalytic mechanisms, namely &beta;-propeller (6%), histidine acid phytases (3%) and protein tyrosine phytase (&amp;lt;1%). Thirty-eight morphologically different bacteria were isolated from biofilms accumulated in fish and plant compartments, and identified to belong to the Bacilli class. Among these, seven could produce xylanase, 8 produced &beta;-glucanase, 14 produced cellulase, and 11 isolates could secrete amylases. In addition, Staphylococcus sp. and Rossellomorea sp. could produce consistent extracellular phytate-degrading activity. The PCR amplification of &beta;-propeller genes both in environmental samples and in the isolates obtained showed that this is the most ecologically relevant phytase type in the aquaponics systems used. In summary, the aquaponics system is abundant with bacteria carrying enzymes responsible for plant-nutrient mineralisation.

scholarly journals The growth response of rice (Oryza sativa L. var. FARO 44) in vitro after inoculation with bacterial isolates from a typical ferruginous ultisol

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Musa Saheed Ibrahim ◽  
Beckley Ikhajiagbe
Keyword(s):  
16S Rrna ◽  
Bacillus Cereus ◽  
Proteus Mirabilis ◽  
Growth Response ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Rice Seedlings ◽  
Rrna Gene Sequencing

Abstract Background Rice forms a significant portion of food consumed in most household worldwide. Rice production has been hampered by soil factors such as ferruginousity which has limited phosphorus availability; an important mineral component for the growth and yield of rice. The presence of phosphate-solubilizing bacteria (PSB) in soils has been reported to enhance phosphate availability. In view of this, the present study employed three bacteria species (BCAC2, EMBF2 and BCAF1) that were previously isolated and proved P solubilization capacities as inocula to investigate the growth response of rice germinants in an in vitro setup. The bacteria isolates were first identified using 16S rRNA gene sequencing and then applied as inoculum. The inolula were prepared in three concentrations (10, 7.5 and 5.0 ml) following McFarland standard. Viable rice (var. FARO 44) seeds were sown in petri dishes and then inoculated with the three inocula at the different concentrations. The setup was studied for 28 days. Results 16S rRNA gene sequencing identified the isolates as: isolate BCAC2= Bacillus cereus strain GGBSU-1, isolate BCAF1= Proteus mirabilis strain TL14-1 and isolate EMBF2= Klebsiella variicola strain AUH-KAM-9. Significant improvement in rice germination, morphology, physiology and biomass parameters in the bacteria-inoculated setups was observed compared to the control. Germination percentage after 4 days was 100 % in the inoculated rice germinants compared to 65% in the control (NiS). Similarly, inoculation with the test isolates enhanced water-use efficiency by over 40%. The rice seedlings inoculated with Bacillus cereus strain GGBSU-1 (BiS) showed no signs of chlorosis and necrosis throughout the study period as against those inoculated with Proteus mirabilis strain TL14-1 (PiS) and Klebsiella variicola strain AUH-KAM-9 (KiS). Significant increase in chlorophyll-a, chlorophyll-b and alpha amylase was observed in the rice seedlings inoculated with BiS as against the NiS. Conclusion Inoculating rice seeds with Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 in an in vitro media significantly improved growth parameters of the test plant. Bacillus cereus strain GGBSU-1 showed higher efficiency due to a more improved growth properties observed.

scholarly journals Identification and Genomic Characterization of Pathogenic Bacillus altitudinis from Common Pear Trees in Morocco

Agronomy ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1344
Author(s):  
Naima Lemjiber ◽  
Khalid Naamani ◽  
Annabelle Merieau ◽  
Abdelhi Dihazi ◽  
Nawal Zhar ◽  
...  
Keyword(s):  
16S Rrna Gene Sequencing ◽  
Genomic Islands ◽  
Rrna Gene ◽  
In Planta ◽  
Bacterial Strains ◽  
Bacillus Altitudinis ◽  
Genes Encoding ◽  
Bacillus Spp ◽  
Pear Trees ◽  
Rrna Gene Sequencing

Bacterial burn is one of the major diseases affecting pear trees worldwide, with serious impacts on producers and economy. In Morocco, several pear trees (Pyrus communis) have shown leaf burns since 2015. To characterize the causal agent of this disease, we isolated fourteen bacterial strains from different parts of symptomatic pear trees (leaves, shoots, fruits and flowers) that were tested in planta for their pathogenicity on Louise bonne and Williams cultivars. The results showed necrotic lesions with a significant severity range from 47.63 to 57.77% on leaves of the Louise bonne cultivar inoculated with isolate B10, while the other bacterial isolates did not induce any disease symptom. 16S rRNA gene sequencing did not allow robust taxonomic discrimination of the incriminated isolate. Thus, we conducted whole-genome sequencing (WGS) and phylogenetic analyzes based on gyrA, gyrB and cdaA gene sequences, indicating that this isolate belongs to the Bacillus altitudinis species. This taxonomic classification was further confirmed by the Average Nucleotide Identity (ANI) and the in silico DNA-DNA hybridization (isDDH) analyzes compared to sixty-five Bacillus spp. type strains. The genome was mined for genes encoding carbohydrate-active enzymes (CAZymes) known to play a role in the vegetal tissue degradation. 177 candidates with functions that may support the in planta phytopathogenicity results were identified. To the best of our knowledge, this is the first data reporting B. altitudinis as agent of leaf burn in P. communis in Morocco. Our dataset will improve our knowledge on spread and pathogenicity of B. altitudinis genotypes that appears as emergent phytopathogenic agent, unveiling virulence factors and their genomic location (i.e., within genomic islands or the accessory genome) to induce trees disease.

scholarly journals Live Bacillus subtilis natto Promotes Rumen Fermentation by Modulating Rumen Microbiota In Vitro

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1519
Author(s):  
Meinan Chang ◽  
Fengtao Ma ◽  
Jingya Wei ◽  
Junhao Liu ◽  
Xuemei Nan ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
16S Rrna ◽  
Rumen Fluid ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rumen Fermentation ◽  
Rrna Gene ◽  
Bacillus Subtilis Natto ◽  
Rrna Gene Sequencing

Previous studies have shown that Bacillus subtilis natto affects rumen fermentation and rumen microbial community structure, which are limited to detect a few microbial abundances using traditional methods. However, the regulation of B. subtilis natto on rumen microorganisms and the mechanisms of microbiota that affect rumen fermentation is still unclear. This study explored the effects of live and autoclaved B. subtilis natto on ruminal microbial composition and diversity in vitro using 16S rRNA gene sequencing and the underlying mechanisms. Rumen fluid was collected, allocated to thirty-six bottles, and divided into three treatments: CTR, blank control group without B. subtilis natto; LBS, CTR with 109 cfu of live B. subtilis natto; and ABS, CTR with 109 cfu of autoclaved B. subtilis natto. The rumen fluid was collected after 0, 6, 12, and 24 h of fermentation, and pH, ammonia nitrogen (NH3-N), microbial protein (MCP), and volatile fatty acids (VFAs) were determined. The diversity and composition of rumen microbiota were assessed by 16S rRNA gene sequencing. The results revealed LBS affected the concentrations of NH3-N, MCP, and VFAs (p < 0.05), especially after 12 h, which might be attributed to changes in 18 genera. Whereas ABS only enhanced pH and NH3-N concentration compared with the CTR group (p < 0.05), which might be associated with changes in six genera. Supplementation with live B. subtilis natto improved ruminal NH3-N and propionate concentrations, indicating that live bacteria were better than autoclaved ones. This study advances our understanding of B. subtilis natto in promoting ruminal fermentation, providing a new perspective for the precise utilization of B. subtilis natto in dairy rations.

scholarly journals Findings from the microbiota of tooth apical periodontitis and the search for pathogen forgranulomatous inflammation

2021 ◽  
Author(s):  
Yuanyuan Wang ◽  
Hao Xu ◽  
Minghui Wei ◽  
Yuhong Wang ◽  
Wenzhe Wang ◽  
...  
Keyword(s):  
Granulomatous Inflammation ◽  
16S Rrna Gene Sequencing ◽  
Apical Periodontitis ◽  
Rrna Gene ◽  
Normal Microbiota ◽  
Rrna Gene Sequencing ◽  
Neisseria Subflava ◽  
Foot Pad

Abstract BackgroundOrofacial granulomatosis (OFG) is a granulomatous inflammation (GI) disease in maxillofacial region, the underlying cause of it remains unknown. Our previous study demonstrated that tooth apical periodontitis (AP) plays a significant role in the pathogenesis of OFG, we aimed here to characterize the AP bacterial signatures of OFG patients, and identify bacteria that may be important pathogens capable of inducing OFG.ResultsThe composition of AP microbiota in OFG cases and common AP controls was compared using 16S rRNA gene sequencing, the results showed a specific AP microbiota signature in OFG patients, characterized by domination of phyla Firmicutes and Proteobacteria , notably members of Streptococcus, Lactobacillus and Neisseria. To assess the pathogenicity of the potential pathogens in OFG, we isolated and successfully in vitro cultured Streptococcus, Lactobacillus casei, Neisseria subflava, Veillonella parvula and Actinomyces from OFG patients, and injected the clinical isolates into mice respectively. Ultimately, foot pad injection with N. subflava elicited granulomatous inflammation, and the virulence of N. subflava was verified based on Koch’s postulates.ConclusionsOur findings confirmed the role of bacteria in OFG, and first suggested that the component of the host normal microbiota, N. subflava is likely a pathogen for GI.

scholarly journals Personalized Bioconversion of Panax Notoginseng Saponins Mediated by Gut Microbiota Between Chinese-diet and Western-diet Healthy Subjects

2021 ◽  
Author(s):  
Li Wang ◽  
Man-Yun Chen ◽  
Li Shao ◽  
Wei Zhang ◽  
Xiang-Ping Li ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Lactobacillus Rhamnosus ◽  
16S Rrna Gene Sequencing ◽  
Panax Notoginseng ◽  
Rrna Gene ◽  
Bifidobacterium Adolescentis ◽  
Correlative Analysis ◽  
Rrna Gene Sequencing ◽  
The Relationship

Abstract Background: Panax notoginseng saponins (PNS) as the main effective substances from P. notoginseng with low bioavailability could be bio-converted by human gut microbiota. In our previous study, PNS metabolic variations mediated by gut microbiota have been observed between high fat, high protein (HF-HP)-diet and low fat, plant fiber-rich (LF-PF)-diet subjects. In this study, we aimed to correspondingly characterize the relationship between distinct gut microbiota profiles and PNS metabolites. Methods: Gut microbiota were collected from HF-HP and LF-PF healthy adults, respectively and profiled by 16S rRNA gene sequencing. PNS were incubated with gut microbiota in vitro. A LC-MS/MS method was developed to quantify the five main metabolites yields including ginsenoside F1 (GF1), ginsenoside Rh2 (GRh2), ginsenoside compound K (GC-K), protopanaxatriol (PPT) and protopanaxadiol (PPD). The selected microbial species, Bifidobacterium adolescentis and Lactobacillus rhamnosus, were employed to metabolize PNS for the corresponding metabolites.Results: The five main metabolites were significantly different between the two diet groups. Compared with HF-HP group, the microbial genus Blautia, Bifidobacterium, Clostridium, Corynebacterium, Dorea, Enhydrobacter, Lactobacillus, Roseburia, Ruminococcus, SMB53, Streptococcus, Treponema and Weissella were enriched in LF-PF group, while Phascolarctobacterium and Oscillospira were relatively decreased. Furthermore, Spearman’s correlative analysis revealed gut microbiota enriched in LF-PF and HF-HP groups were positively and negatively associated with PNS metabolites yields, respectively. Conclusions: Our data showed gut microbiota diversity led to the personalized bioconversion of PNS.

scholarly journals Morphological and Molecular Characterization of Trichoderma spp. from Rhizosphere Soil and Their Antagonistic Activity against Fusarium spp.

2021 ◽  
pp. 100-112
Author(s):  
Jaygendra Kumar ◽  
Mukesh Kumar ◽  
Akash Tomar ◽  
. Vaishali ◽  
Pushpendra Kumar ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Rhizosphere Soil ◽  
Pcr Amplification ◽  
Its Region ◽  
Culture Method ◽  
Dual Culture ◽  
Fusarium Spp ◽  
Trichoderma Spp ◽  
Trichoderma Species

Trichoderma species are well known for their biocontrol activity which colonize many soil and tuber-borne and foliage plant pathogens. In this study, 12 native isolates of Trichiderma spp were collected from various crop rhizosphere soil samples and characterized them phenotypically based on morphological and cultural features and genotypically based on sequence analysis of internal transcribed spacer (ITS) region-PCR amplification. The results obtained from phenotypic and genotypic observation revealed that isolates were belonged to five different species namely T. asperellum, T. harzianum, T. longibrachiatum, T. koningii and T. koningiopsis. All Trichoderma isolates produced ~600 bp amplicon and phylogenetic analysis revealed that all isolates were grouped with respective species. Further, the antagonistic potential of all the isolates was evaluated against Fusarium spp. following in vitro dual culture method. The results showed that isolates of T. harzianum exhibited maximum growth inhibition activity. The highest rate of inhibition was recorded with T. harzianum isolate TBT6 (87.1%) followed by TBT7 (82.2%), while the least inhibition was observed in T. longibrachiatum isolate TBT10 (59.7%) after 7 days of incubation. The antagonistic T. harzianum isolate TBT6 can be used for development of Trichoderma based bio-formulation and served as bio-control agent against Fusaium spp. under field conditions.

scholarly journals Analysis of Vaginal and Endometrial Microbiota Communities in Infertile Women With a History of Repeated Implantation Failure

2020 ◽  
Author(s):  
Takuhiko Ichiyama ◽  
Keiji Kuroda ◽  
Yoko Nagai ◽  
Daichi Urushiyama ◽  
Motoharu Ono ◽  
...  
Keyword(s):  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Implantation Failure ◽  
Repeated Implantation Failure ◽  
Infertile Women ◽  
Healthy Women ◽  
History Of ◽  
Rrna Gene Sequencing ◽  
Clinical Trials Registry

Abstract Background: Repeated implantation failure (RIF) is estimated to occur in 15%–20% of infertile women undergoing in vitro fertilization-embryo transfer (IVF-ET). Molecular identification recently confirmed that the uterine microbiota may have implications for reproductive and obstetrical outcomes. One hundred forty-five women who had been diagnosed with RIF were enrolled in the study. Twenty-one healthy women were also enrolled as controls. We investigated their vaginal and endometrial microbiotas using 16S rRNA gene sequencing and compared the microbiota profiles in the patients with RIF and controls.Results: The endometrial microbiotas had higher α-diversities than did the vaginal microbiotas (p<0.001 in both patients with RIF and healthy women). The microbiota profiles showed that vaginal and endometrial samples in patients with RIF had significantly higher levels of 5 and 14 bacterial genera, respectively, than those in healthy women. These genera included Atopobium, Gardnerella, Prevotella and Megasphaera. Vaginal Lactobacillus rates in patients with RIF were significantly lower at 76.4 ± 38.9% compared with those of the controls at 91.8 ± 22.7% (p=0.018), but endometrial Lactobacillus rates did not significantly differ between the RIF patients and controls (56.2 ± 36.4% and 58.8 ± 37.0%, respectively, p=0.79) Conclusions: Impaired microbiota communities containing specific bacteria in both the endometrium and vagina were associated with implantation failure. The Lactobacillus rate in the vagina, but not the endometrium, may be a biomarker for RIF.Trial registration: UMIN Clinical Trials Registry, UMIN000031731, Registered 15 March 2018; https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000036121

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