scholarly journals In vitro germination, initial development and acclimatization of Cattleya nobilior Rchb. f. (Orchidaceae): an approach to curb the eventual endangerment of this exuberant, near-threatened Cerrado species

2021 ◽  
Vol 6 (2) ◽  
pp. 2167-2191
Author(s):  
Silene Lívia Aires de Oliveira ◽  
Kellen Lagares Ferreira Silva ◽  
Rafael José de Oliveira ◽  
Maíra Jéssica de Souza ◽  
Jaderson Roney Gomes de Oliveira ◽  
...  
Keyword(s):  
Large Scale ◽  
Culture Medium ◽  
Culture Media ◽  
Natural Environments ◽  
In Vitro Germination ◽  
Initial Development ◽  
Anatomical Characteristics ◽  
Soybean Straw ◽  
Leaf Anatomical Characteristics

ABSTRACT: The objective of the present study was to evaluate the effects of different culture media on the in vitro germination and initial development of Cattleya nobilior, and to acclimatize this species using soybean straw as a substrate component, aiming the production of plants at large scale to reintroduce it in natural environments. Anatomical characteristics associated with development during acclimatization are also depicted. The influence of Murashige and Skoog, Knudson, and Vacin and Went culture media on the in vitro germination and protocorm development were assessed. Acclimatization was accomplished using different proportions of soybean straw (SS) and Bioplant (BP) as substrates. Anatomical studies were conducted on the leaves of plants during acclimatization. KC was the most suitable culture medium for both germination and initial development of C. nobilior. Regarding acclimatization, it is recommended that a substrate composed of 60% SR and 40% BP be initially used. Upon transfer to shade-house conditions, plants should be grown in a mix of 40% SR: and 60% BP. The leaf anatomical characteristics observed are typical of epiphytic orchids adapted to water-poor environments, which indicates that SR did not negatively affect the development of the species. The in vitro propagation of C. nobilior as described herein is efficient for its multiplication for commercial and conservation purposes and SS can be used as an alternative component of the substrate for its acclimatization. KEYWORDS: Foliar anatomy, culture media, soybean straw.

scholarly journals Does soaking time during disinfestation affect germination rates in Dendrobium?

2020 ◽  
Vol 36 (1) ◽  
Author(s):  
Jose Carlos Sorgato ◽  
Jackeline Schultz Soares ◽  
Silvana de Paula Quintão Scalon ◽  
Suzana Targanski Sajovic Pereira ◽  
Débora De Freitas Brotto ◽  
...  
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Germination Rate ◽  
Sodium Hypochlorite Solution ◽  
In Vitro Germination ◽  
Soaking Time ◽  
Growth Room ◽  
Aseptic Conditions ◽  
Species Specific

Asymbiotic germination is considered an efficient and viable technique that can increase germination rates. The effect of type and concentration of disinfestants, and the exposure time to disinfestants may differ according to the plant species. Therefore, species-specific standardization of disinfestation agent and procedure is necessary to achieve optimal germination rates. The objective of this study was to determine a disinfestation methodology to increase in vitro germination rates and the early development of seedlings of Dendrobium nobile and Dendrobium phalaenopsis, using different times for seed disinfestation and different culture media. Seeds were disinfected by soaking in a 0.8% sodium hypochlorite solution for 5 or 15 min under aseptic conditions, after which seed suspensions were either washed with water or left unwashed. Next, they were seeded in culture flasks containing four different culture media (MS, ½MS, K, and VW). The flasks were then transferred to a growth room under controlled photoperiod and temperature, where they remained under an irradiance of 20 μmol m-2 s-1. Germination rates of the species were evaluated 45 days after placement in the culture flasks. A higher germination rate was observed when the seeds were triple washed, regardless of the culture medium or soaking time. Seed soaking disinfestation for 5 min is also recommended. MS and ½MS media were the most effective culture media in promoting in vitro germination of the species under study.  

scholarly journals In vitro germination of zygotic embryos of hybrid BRS Manicoré (E. guineensis X E. oleifera)

2016 ◽  
Vol 88 (3 suppl) ◽  
pp. 1841-1850
Author(s):  
KEILA A.P. BONETTI ◽  
MARGUERITE QUOIRIN ◽  
REGINA C. QUISEN ◽  
SUELEN C.S. LIMA
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Germination Rate ◽  
Phosphate Concentration ◽  
Zygotic Embryos ◽  
In Vitro Germination ◽  
N6 Medium ◽  
Whole Plants ◽  
Tetrazolium Test

ABSTRACT The interspecific oil palm hybrid BRS Manicoré (E. guineensis x E. oleifera) has superior agronomic characteristics. However, the germination rate is low (30%) and the process is slow when the seeds are sown in a conventional form. The purpose of this study was to optimize the in vitro germination of zygotic embryos of this hybrid comparing seed lots. The viability of zygotic embryos was evaluated by the tetrazolium test (0.075%) for 4 h. The embryos were cultured on MS and Y3 culture media, with and without the addition of NaH2PO4, as well as on MS, MS1/2 and N6 medium. In MS medium containing NaH2PO4, the germination rate was increased from 40 to 70% in comparison with the medium without sodium phosphate. The comparison between the culture media MS, MS 1/2, N6 and Y3 showed that 75% of zygotic embryos cultured in the Y3 medium formed whole plants (with roots and shoots defined), a higher percentage than embryos cultured on MS, MS 1/2 and N6 media (46, 35 and 17% respectively). In the same Y3 culture medium, the embryos were larger (36% ≥ 2 cm and 30% ≥ 5 cm) than in the other media. Results obtained by the tetrazolium test were similar to those of germination, showing the effect of the genotype of each seed lot. For the germination and development of plantlets it is essential to add NaH2PO4 to a culture medium containing no phosphate or with a low phosphate concentration.

scholarly journals A Chemically Defined, Xeno- and Blood-Free Culture Medium Sustains Increased Production of Small Extracellular Vesicles From Mesenchymal Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Aliosha I. Figueroa-Valdés ◽  
Catalina de la Fuente ◽  
Yessia Hidalgo ◽  
Ana María Vega-Letter ◽  
Rafael Tapia-Limonchi ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Large Scale ◽  
Culture Medium ◽  
Culture Media ◽  
Cell Phenotype ◽  
Target Cells ◽  
Scale Production ◽  
Paracrine Factors

Cell therapy is witnessing a notable shift toward cell-free treatments based on paracrine factors, in particular, towards small extracellular vesicles (sEV), that mimic the functional effect of the parental cells. While numerous sEV-based applications are currently in advanced preclinical stages, their promised translation depends on overcoming the manufacturing hurdles posed by the large-scale production of purified sEV. Unquestionably, the culture medium used with the parental cells plays a key role in the sEV’s secretion rate and content. An essential requisite is the use of a serum-, xeno-, and blood-free medium to meet the regulatory entity requirements of clinical-grade sEV’s production. Here, we evaluated OxiumTMEXO, a regulatory complying medium, with respect to production capacity and conservation of the EV’s characteristics and functionality and the parental cell’s phenotype and viability. A comparative study was established with standard DMEM and a commercially available culture medium developed specifically for sEV production. Under similar conditions, OxiumTMEXO displayed a three-fold increase of sEV secretion, with an enrichment of particles ranging between 51 and 200 nm. These results were obtained through direct quantification from the conditioned medium to avoid the isolation method’s interference and variability and were compared to the two culture media under evaluation. The higher yield obtained was consistent with several harvest time points (2, 4, and 6 days) and different cell sources, incluiding umbilical cord-, menstrual blood-derived mesenchymal stromal cells and fibroblasts. Additionally, the stem cell phenotype and viability of the parental cell remained unchanged. Furthermore, OxiumTMEXO-sEV showed a similar expression pattern of the vesicular markers CD63, CD9, and CD81, with respect to sEV derived from the other conditions. The in vitro internalization assays in different target cell types and the pharmacokinetic profile of intraperitoneally administered sEV in vivo indicated that the higher EV production rate did not affect the uptake kinetics or the systemic biodistribution in healthy mice. In conclusion, the OxiumTMEXO medium sustains an efficient and robust production of large quantities of sEV, conserving the classic functional properties of internalization into acceptor target cells and biodistribution in vivo, supplying the amount and quality of EVs for the development of cell-free therapies.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

Control of browning during seed germination of Meizotropis pellita: a rare and an endangered plant

2021 ◽  
Vol 16 (8) ◽  
pp. 118-120
Author(s):  
Renu Chandola ◽  
Anshulika Upadhyaya
Keyword(s):  
Seed Germination ◽  
Culture Media ◽  
Light Condition ◽  
Endangered Plant ◽  
In Vitro Germination ◽  
Light Conditions ◽  
Low Intensity ◽  
Browning Reaction ◽  
Threatened Plant Species

Meizotropis pellita is a rare, endangered and threatened plant species found in Patwadanger forest. In nature this species grows from rootstock in precise and sensitive habitats. However, phenolic compounds provoke browning reaction which is responsible for lethal browning in plant tissue culture and hindering in vitro germination. The aim of this study was to minimize the effect of phenolic compound during seed germination. We evaluate different antibrowning treatments in Meizotropis pellita seeds. The seeds were pretreated in Polyvinylpyrrolidone and then treated seeds were cultured on MS media and sterilized seeds were directly inoculated on PVP containing MS media. These culture media incubate under dark and light conditions at different durations respectively. Our results showed that addition of antibrowning agent, PVP inhibit onset of browning. Dark and light condition also played a crucial role on onset of browning. Delayed and low intensity of browning were observed in case of seeds that were inoculated on MS media containing PVP and were kept in dark. The present research offers a positive in vitro seed germination protocol for Meizotropis pellita conservation.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

scholarly journals MODIFICAÇÕES NO MEIO DE CULTURA, FOTOPERÍODO E TEMPO DE CULTIVO AFETAM O ALONGAMENTO E ENRAIZAMENTO IN VITRO DE BANANEIRA CV. PACOVAN

Nativa ◽  
2018 ◽  
Vol 6 (1) ◽  
pp. 27
Author(s):  
Marcos Vinícius Marques Pinheiro ◽  
Ana Cristina Portugal Pinto De Carvalho ◽  
Fabrina Bolzan Martins
Keyword(s):  
Plant Height ◽  
Culture Medium ◽  
Culture Media ◽  
Dry Weight ◽  
Salt Mixture ◽  
Musa Spp ◽  
Fresh Biomass ◽  
Shoot Dry Weight ◽  
Number Of Leaves

No intuito de elevar as taxas de sobrevivência durante a etapa de aclimatização e posterior plantio a campo, avaliou-se o enraizamento in vitro de bananeira cv. Pacovan, em diferentes concentrações de sais MS e de sacarose. Utilizou-se DIC, esquema fatorial (6x2x3), com seis meios de cultura [sendo três concentrações de nutrientes do meio MS (100%; 50% de macronutrientes; e 50% dos sais macro e micronutrientes), e duas concentrações de sacarose (1,5/3,0%)], dois fotoperíodos (12/16 h) e três tempos de cultivo (21, 28 ou 35 dias) e seis repetições/tratamento. Analisaram-se: altura da planta, número de folhas/planta, massas frescas e secas das partes aérea e radicular. Para altura da planta, massa fresca da parte aérea e radicular, o meio MS 50% dos sais + sacarose (1,5%) com fotoperíodo de 16 h e tempo de cultivo de 35 dias foi satisfatório. Para massa seca da parte aérea foi MS 50% de sais + sacarose (3%), e para massa seca da parte radicular, MS 100% + sacarose (3%) (em 12hs/28 dias e 16hs/21 dias). Para o alongamento/enraizamento in vitro da bananeira cv. Pacovan sugere-se MS 50% de sais (macro e micronutrientes), redução ou manutenção de sacarose (1,5 ou 3%) em 16h/35 dias de cultivo.Palavra-chave: Musa spp., propagação in vitro, sistema radicular. CHANGES IN CULTURE MEDIUM, PHOTOPERIOD AND TIME OF CULTIVATION AFFECT THE IN VITRO ELONGATION AND ROOTING OF BANANA CV. PACOVAN ABSTRACT:In order to achieve high rates of survival during the acclimatization and later planting in the field, was evaluated the in vitro of banana cv. Pacovan plants under different concentrations of sucrose and MS basal salt mixture. The experiment was assembled in a DIC, in 6x2x3, six different culture media [three different MS salt mixture concentrations (100%; 50% of macronutrients; and 50% of macro/micronutrients) and two sucrose concentrations (1.5/3%)], two photoperiods (12/16 hours) and three cultivation times (21, 28 or 35 days). Each treatment was composed by 6 replicates. Plant height, number of leaves/plant, fresh and dry weight of roots and shoots, were analyzed. Satisfactory results for plant height and shoot and root fresh biomass were observed in MS with macro/micronutrients (50%) + sucrose (3%), 16 hours/35 days. The highest values of shoot dry weight were observed in MS with macro/micronutrients (50%) + sucrose (3%); the highest root dry weight was achieved with MS 100% + sucrose (3%) (12hs/28 and 16hs/21 days). The suggested medium for the in vitro elongation and rooting stage of banana cv. Pacovan is the MS with 50% of salts (macro and micronutrients), reduction or maintenance of sucrose (1.5 or 3%) in 16h/35 days of cultivation.Keywords: Musa spp., in vitro propagation, root system. DOI:

scholarly journals In vitro answer of Bulgarian pepper (Capsicum annuum L.)

Genetika ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 129-136
Author(s):  
Velichka Rodeva ◽  
Stanislava Grozeva ◽  
Velichka Todorova
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Casein Hydrolysate ◽  
Basal Medium ◽  
Regeneration Ability ◽  
Stem Explants ◽  
Media Composition ◽  
Breeding Lines ◽  
High Level

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.

scholarly journals ACTIVATED CHARCOAL APPLICATION FOR THE MICROPROPAGATION OF Cattleya crispata (Thunb.) Van den Berg

Nativa ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 352-358
Author(s):  
Denys Matheus Santana Costa Souza ◽  
Sérgio Bruno Fernandes ◽  
Letícia Vaz Molinari ◽  
Maria Lopes Martins Avelar ◽  
Gilvano Ebling Brondani
Keyword(s):  
Endemic Species ◽  
Activated Charcoal ◽  
Culture Medium ◽  
Genetic Conservation ◽  
Campo Rupestre ◽  
In Vitro Germination ◽  
Germination Speed

 Micropropagation is an alternative for the genetic conservation and propagation of endemic species from “Campo Rupestre Ferruginoso”, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, seedling number, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of seedlings of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots.

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