scholarly journals Micropropagation of Rudbeckia hirta L. from seedling explants

2005 ◽  
Vol 11 (2) ◽  
pp. 105-108
Author(s):  
P. Szarvas ◽  
A. Zsila-André ◽  
Z. Kováts ◽  
M. G. Fári
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Axillary Meristem ◽  
Surface Disinfection ◽  
Shoot Bud ◽  
Rudbeckia Hirta ◽  
Seedling Explants ◽  
In Vitro Micropropagation ◽  
Special Value

We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L seedlings. We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps. First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times. For the propagation of seedling meristems, the combination of half-strenght solid Murashige and Skoog (1962) culture medium containing 10 mg/1 of kinetin or 2 mg/I of kinetin + 0.1 mg/1 of 2iP proved to be the most suitable. The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/1 of IAA. After four weeks of incubation we obtained elongated shoots which we separated and inoculated into a new culture medium and we obtained elongated roots. The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation. By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respect of breeding.

scholarly journals Micropropagation of Rudbeckia hirta L. from seedling explants

2006 ◽  
pp. 53-59
Author(s):  
Pál Szarvas ◽  
Anikó Zsila-André ◽  
Zoltán Kovács ◽  
Miklós Gábor Fári
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Surface Disinfection ◽  
Shoot Bud ◽  
Rudbeckia Hirta ◽  
Seedling Explants ◽  
In Vitro Micropropagation ◽  
Special Value ◽  
Half Strength

We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L seedlings. We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps. First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times. For the propagation of seedling meristems, the combination of half-strength solid Murashige and Skoog (1962) culture medium containing 10 mg/l of kinetin and 2 mg/l of kinetin + 0.1 mg/l of 2iP proved to be the most suitable. The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/l of IAA. After four weeks of incubation, we obtained elongated shoots, which we separated and inoculated into a new culture medium and from which we obtained elongated roots. The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation. By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respectt to breeding.

scholarly journals Optimization Studies of Culture Media for In-Vitro Clonal Micropropagation of New Grape Varieties

2021 ◽  
Author(s):  
Abdulmalik Batukaev ◽  
Eliza Sobralieva ◽  
Diana Palaeva
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Mineral Salts ◽  
Additional Increase ◽  
Shoot Bud ◽  
Clonal Micropropagation ◽  
Aseptic Culture ◽  
Grape Varieties

This article describes the effect of Gautheret, White, Heller and Murashige & Skoog mineral salts during in-vitro clonal micropropagation of new grape varieties. The optimal mineral compositions of the culture medium that support the in-vitro regeneration of isolated grape explants were identified. The grapes that were studied were the Bart and Augustine varieties. Primary grape explants were cultivated for 30 days in a non-transplanted culture. Increased regenerative activity was observed in the Murashige & Skoog and White media. Increased haemogenesis occurred and shoots regenerated. The addition of cytokinin 6-BAP to the medium for obtaining aseptic culture led to an increase in the frequency of shoot-bud production by 5 to 6 times, depending on the type of medium. Combining 6-BAP with the auxin NAA provided an additional increase in the frequency of shoot-bud production, but to a lesser extent. Adding growth regulators to the culture medium also reduced the frequency of explant necrosis. Keywords: grapes, mineral salts, culture medium, microclonal propagation, in-vitro, cytokinins, auxins

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.

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 In vitro growth of Cattleya nobilior Rchb. f.: culture media, sealing systems and irradiance

2021 ◽  
Vol 51 ◽  
Author(s):  
Kaliana Gottschalk de Freitas ◽  
José Carlos Sorgato ◽  
Jackeline Schultz Soares ◽  
Luan Marlon Ribeiro
Keyword(s):  
Plant Height ◽  
Natural Ventilation ◽  
Culture Medium ◽  
Culture Media ◽  
Ventilation System ◽  
Light Condition ◽  
Leaf Length ◽  
In Vitro Growth ◽  
Randomized Design

ABSTRACT Some factors may influence the in vitro culture of native orchids, especially the culture medium, sealing system and light condition. This study aimed to evaluate the addition of banana pulp and peel in the formulation of culture media, sealing system and irradiance, in the in vitro growth of Cattleya nobilior Rchb. f. A completely randomized design was used in a factorial scheme with five culture media [Murashige & Skoog supplemented or not with banana pulp or peel (100 or 200g L-1)], two light conditions [3,000 K LED lamps (43 µmol m-2 s-1 or 86 µmol m-2 s-1)] and two sealing systems (natural and conventional ventilation), with six replicates of one vial each. The plant height, diameter of the largest pseudobulb, length of the largest root and of the largest leaf, fresh weight and number of leaves, roots and shoots were evaluated. The use of both the banana pulp and peel as additives in the culture medium formulation promoted the C. nobilior growth. The natural ventilation system increased the plant height, pseudobulb diameter and leaf length, while the conventional system promoted tillering.

scholarly journals Types and concentrations of cytokinins in the micropropagation of Anthurium maricense

2018 ◽  
Vol 12 (2) ◽  
pp. 117
Author(s):  
Cecília Moreira Serafim ◽  
Arlene Santisteban Campos ◽  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cecília Ribeiro de Castro ◽  
Ana Cristina Portugal Pinto de Carvalho
Keyword(s):  
Light Intensity ◽  
Culture Medium ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Viable Option ◽  
Growth Room ◽  
In Vitro Induction

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.

scholarly journals Effective medium for in vitro sprouting of the buds and multiplication of the plantlets, and identification of the fungal contaminant associated with the explants of Gnetum

2021 ◽  
Vol 16 (2) ◽  
pp. 001-013
Author(s):  
Abwe Mercy Ngone ◽  
Lawrence Monah Ndam ◽  
Rita Mungfu Njilar ◽  
Doungous Oumar ◽  
Thomas Eku Njock
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Fungal Growth ◽  
Growth Media ◽  
Fungal Contamination ◽  
Surface Sterilization ◽  
Pure Cultures ◽  
Nodal Cuttings ◽  
Baseline Information

Plant tissue culture requires the optimization of growth media. Gnetum, known locally in Cameroon as “Eru” is an indigenous gymnospermous vegetable with diverse medicinal, nutritional, cultural and socio-economic values. This resource is over-exploited and expected to neighboring countries, resulting to increased scarcity in the forest. Preliminary work on the in vitro culture of nodal cuttings was faced by the problem of fungal contamination. It was therefore necessary to isolate and identify the fungal contaminant, optimize the surface sterilization of field material and compose an appropriate medium for sprouting. Pure cultures of the fungus were obtained and grown on Potato Dextrose Agar (PDA) and Sabouraud Dextrose Agar (SDA). The identification was based on the appearance of the fungal growth on plates and also on the microscopic view. This was affected by the use of keys. Gnetum explants were disinfected with the various concentrations of disinfectants, preceded in some instances by pre-treatments, as well as incorporating fungicides in the culture medium. Two different culture media were employed: the Woody Plant Medium (WPM) and the Murashige and Skoog (MS) based establishment medium (Y-1). Gnetum was found to live in association with a complex of Microsporum species. The level of contamination of cultures was reduced from 100% to 40% when pre-treated before disinfection and even lower to 10% by incorporating fungicides in the medium. Sprouting was observed in WPM. This study provides baseline information on the in vitro propagation of Gnetum and thus opens up avenues for more research to be carried out in this field.

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