A novel plant-based-sea water culture media for in vitro cultivation and in situ recovery of the halophyte microbiome

Saleh, Mohamed Y.; Sarhan, Mohamed S.; Mourad, Elhussein F.; Hamza, M.; Abbas, Mohamed; Othman, Amal A.; Youssef, Hanan H.; Morsi, Ahmed; Youssef, Gehan H.; El-Tahan, Mahmoud; Amer, Wafaa; Fayez, M. B. E.; Ruppel, Silke; Hegazi, N. A.

doi:10.1016/j.jare.2017.06.007

Cited by 13 publications

(21 citation statements)

References 48 publications

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“…The lowest culturability was reported for artificial culture media (3.4–3.9%). These results not only provide more depth to our previous studies ( 18 , 32 , 41 , 43 , 52 ), but also offer an insight into the relevance of plant materials selected for the preparation of the plant-only-based culture medium and the respective plant sphere or niche under investigation. All tested plant materials used for culture medium preparation generally supported the good development and culturability of endophytes.…”

Section: Resultssupporting

confidence: 66%

“…The present study amplifies the spectrum of our previous findings on the sole use of plant-only-based culture media to support the in situ recovery and in vitro growth as well as biomass production of representative isolates of rhizobacteria ( 18 , 32 , 41 , 43 , 52 ). The plant materials tested are refuse of mowed turfgrass, cactus pads, and Egyptian berseem clover plants.…”

supporting

confidence: 70%

“…The use of suitable plant materials for the in vitro growth and biomass production of rhizobacteria, in the form of plant-only-based culture media, represents a novel approach in culturing technologies for examining the plant microbiome. Based on various plant materials, these culture media have proven to be ample and sustainable sources of nutrients ( 18 , 32 , 41 , 43 , 52 ). Further advances in this respect will contribute to the discovery of new members of rhizobacteria including promising biomass production needed for biotechnological applications in support of plant productivity and environment sustainability.…”

mentioning

confidence: 99%

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Plant Materials are Sustainable Substrates Supporting New Technologies of Plant-Only-Based Culture Media for <i>in vitro</i> Culturing of the Plant Microbiota

Mourad

Sarhan

Daanaa

et al. 2018

Microbes and environments

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In order to improve the culturability and biomass production of rhizobacteria, we previously introduced plant-only-based culture media. We herein attempted to widen the scope of plant materials suitable for the preparation of plant-only-based culture media. We chemically analyzed the refuse of turfgrass, cactus, and clover. They were sufficiently rich to support good in vitro growth by rhizobacteria isolates representing Proteobacteria and Firmicutes. They were also adequate and efficient to produce a cell biomass in liquid batch cultures. These culture media were as sufficient as artificial culture media for the cultivation and recovery of the in situ rhizobacteria of barley (Hordeum murinum L.). Based on culture-dependent (CFU plate counting) and culture-independent analyses (qPCR), mowed turfgrass, in particular, supported the highest culturable population of barley endophytes, representing >16% of the total bacterial number quantified with qPCR. This accurately reflected the endophytic community composition, in terms of diversity indices (S′, H′, and D′) based on PCR-DGGE, and clustered the plant culture media together with the qPCR root populations away from the artificial culture media. Despite the promiscuous nature of the plant materials tested to culture the plant microbiome, our results indicated that plant materials of a homologous nature to the tested host plant, at least at the family level, and/or of the same environment were more likely to be selected. Plant-only-based culture media require further refinements in order to provide selectivity for the in vitro growth of members of the plant microbiome, particularly difficult-to-culture bacteria. This will provide insights into their hidden roles in the environment and support future culturomic studies.

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Section: Resultssupporting

confidence: 66%

supporting

confidence: 70%

mentioning

confidence: 99%

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Plant Materials are Sustainable Substrates Supporting New Technologies of Plant-Only-Based Culture Media for <i>in vitro</i> Culturing of the Plant Microbiota

Mourad

Sarhan

Daanaa

et al. 2018

Microbes and environments

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“…To gain insights into their unknown functions and exploit their potentials, different approaches were recently introduced to culture such not-yet-cultured bacteria [2,3]. In this respect, the plantonly-based culture media were presented as natural culture media to replace myriad formulas of synthetic culture media, and strongly recommended to increase the cultivability of the plant microbiota [4][5][6][7][8][9][10]. To alleviate the stress of disproportionate nutrients, present in common culture media, we aimed at culturing maize and sunflower microbiota on the natural nutrients present in the plant inoculum itself, compared to highly diluted plant-only-based culture media and standard R2A.…”

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confidence: 99%

An inoculum-dependent culturing strategy (IDC) for the cultivation of environmental microbiomes and the isolation of novel endophytic Actinobacteria

et al. 2019

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The recent introduction of plant-only-based culture media enabled cultivation of not-yet-cultured bacteria that exceed 90% of the plant microbiota communities. Here, we further prove the competence and challenge of such culture media, and further introduce "the inoculum-dependent culturing strategy, IDC". The strategy depends on direct inoculating plant serial dilutions onto plain water agar plates, allowing bacteria to grow only on the expense of natural nutrients contained in the administered inoculum. Developed colonies are successively transferred/subcultured onto plant-only-based culture media, which contains natural nutrients very much alike to those found in the prepared plant inocula. Because of its simplicity, the method is recommended as a powerful tool in screening programs that require microbial isolation from a large number of diverse plants. Here, the method comfortably and successfully recovered several isolates of endophytic Actinobacteria represented by the six genera of Curtobacterium spp., Plantibacter spp., Agreia spp., Herbiconiux spp., Rhodococcus spp., and Nocardioides spp. Furthermore, two of the isolates are most likely novel species belonging to Agreia spp. and Herbiconiux spp.

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“…In view of our previous publications [32][33][34][35]37], we set up a number of preliminary experiments to assess the growth of Rhizobium leguminosarum and biomass production on the expense of increasing concentrations of dehydrated clover plant powder and boosting effects of supplementation with rich and available agro-byproducts of local molasses and glycerol in various concentrations. Then, the main experiments were designed to measure the suitability of supplemented concentrations of molasses and glycerol not only to enrich the plant material but also to formulate stable and consistent plant pellets.…”

Section: Experimental Designmentioning

confidence: 99%

Plant Pellets: A Compatible Vegan Feedstock for Preparation of Plant-Based Culture Media and Production of Value-Added Biomass of Rhizobia

et al. 2020

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Although plant-based culture media enhances in vitro cultivation of rhizobacteria, studies assessing their biomass potential for large-scale applications are lacking. Here, we advance plant pellets (PPs) as a novel technology to unlock the potential of such vegan culture media for biomass production of Rhizobium leguminosarum. PP formulations were based on mixtures of Egyptian clover powder and the agro-byproducts glycerol and molasses. These mixtures were either contained or not contained in teabags during culture media preparation. Metrics of biomass included colony forming units, optical density (OD600nm), and cell dry weight (DW). Biomass comparisons between culture media based on PPs and standard yeast extract mannitol (YEM) revealed that the following PPs composition, contained in teabags, cultivated rhizobia at levels comparable to YEM: 16 g clover powder, 5% molasses, and 0.8% glycerol. This PPs composition enabled shorter generation times of rhizobia (PP: 3.83 h, YEM: 4.28 h). Strikingly, PPs mixtures supplemented with 10% molasses and not contained in teabags promoted rhizobia without apparent lag phases and produced 25% greater DW than YEM. PPs potentiate the use of dehydrated vegan feedstocks for both plant microbiota cultivation and biomass production and appear as cost- and labor-effective tools, easy to handle and store for plant-based culture media preparation.

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A novel plant-based-sea water culture media for in vitro cultivation and in situ recovery of the halophyte microbiome

Abstract: Graphical abstract

Cited by 13 publications

References 48 publications

Plant Materials are Sustainable Substrates Supporting New Technologies of Plant-Only-Based Culture Media for <i>in vitro</i> Culturing of the Plant Microbiota

Plant Materials are Sustainable Substrates Supporting New Technologies of Plant-Only-Based Culture Media for <i>in vitro</i> Culturing of the Plant Microbiota

An inoculum-dependent culturing strategy (IDC) for the cultivation of environmental microbiomes and the isolation of novel endophytic Actinobacteria

Plant Pellets: A Compatible Vegan Feedstock for Preparation of Plant-Based Culture Media and Production of Value-Added Biomass of Rhizobia

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