Feasibility of the new method for orchid in vitro rooting using liquid and chemical sterilized culture medium under different sucrose concentration

Abstract The in vitro propagation of orchids is the only commercial large scale technique to obtain healthy and high quality plantlets with clonal origin. The use of new technologies in plant tissue culture systems could lead to efficiency increases and costs reduction of micropropagation systems. The main actual micropropagation system is based on semi-solid culture media solidified using agar, followed by sterilization using autoclaving, and cultivation under photomixotrophic conditions using sucrose as main source of energy to plant in vitro culture. We proposed in this study the use of new micropropagation system using chemical sterilized liquid medium using polyurethane foam as support and LED source of light in rooting stage of Miltassia ‘Shelobie Tolkien’. Thus, the objective of this research was to test different concentrations of sucrose, comparing the conventional semi-solid agar-based culture medium (control) and the use of liquid medium with polyurethane foam support. The following sucrose concentrations were used: 0, 7.5, 15, 22.5 and 30 g L-1. The experiment was conducted in a 2 × 5 factorial, in a completely randomized design with ten replications each over a total period of 105 days of cultivation. The chemical sterilization using ClO2 showed 100% of decontamination in all treatments. The use of liquid media with polyurethane foam showed better results than plants cultivated in agar medium, and can be used for replace agar-based for orchid in vitro rooting.

Download Full-text

Temporary immersion biorreators: efficient technique for the propagation of the ‘Pircinque’ strawberry

Revista Brasileira de Fruticultura ◽

10.1590/0100-29452019102 ◽

2019 ◽

Vol 41 (1) ◽

Author(s):

Samila Silva Camargo ◽

Leo Rufato ◽

Maicon Magro ◽

André Luiz Kulkamp de Souza

Keyword(s):

Liquid Medium ◽

Culture Medium ◽

Culture Media ◽

Time Factors ◽

Efficient Technique ◽

Control Treatment ◽

Solid Culture ◽

Temporary Immersion ◽

Solid Culture Medium

Abstract The in vitro propagation technique via temporary immersion bioreactors is a tool that, through the culture in a liquid medium, allows an increase in the efficiency of seedling production. Several researches with the strawberry crop have shown greater efficiency of the system compared to the conventional process of micropropagation in solid medium. In this sense, the objective herein was to establish a protocol of multiplication and rooting of the ‘Pircinque’ strawberry, in temporary immersion bioreactors. Two distinct and independent studies were carried out, characterized by the multiplication and rooting stages of strawberry explants, newly introduced and registered in Brazil. Two culture media (MS and KNOP) were studied and, as a control treatment, the growth of the explants in solid culture medium was evaluated with the addition of 5 g L-1 of agar. Different immersion times of the culture medium were explored: five or eight times a day, for 15 minutes. The study was composed of the culture medium and immersion time factors, as well as the control (solid) treatment. It was verified that the use of temporary immersion bioreactors system is an efficient technique for the multiplication and rooting of explants of strawberry cv. Pircinque, when compared to the conventional method of micropropagation with the use of solid culture medium, making it possible to optimize the production of seedlings in biofactories. The MS liquid medium, in contact with explants of ‘Pircinque’ strawberry five times a day, increased the growth of the aerial part and the root system.

Download Full-text

Effect of pH, Sucrose Concentrations and Medium States on in vitro Root-ing of Pineapple (Ananas comosus (L) Merr) cv Queen

AL-MUKHTAR JOURNAL OF SCIENCES ◽

10.54172/mjsc.v36i2.54 ◽

2021 ◽

Vol 36 (2) ◽

pp. 135-147

Author(s):

Abdelhamid M. Hamad

Keyword(s):

Liquid Medium ◽

In Vitro Rooting ◽

The Other ◽

Ananas Comosus ◽

Ph Adjustment ◽

Rooting Medium ◽

Important Approach ◽

Semi Solid ◽

Medium Cost

The effect of medium states (liquid, semi solid, solid), pH (5.0, 5.5, 6.0, 6.5) and sucrose concentrations (10, 20, 30, 40 g/l) on in vitro rooting of pineapple cultured in full strength MS enriched with IBA at 0.5 mg/l were investigated. According to average overall sucrose concentrations, overall pH adjustments, and at each combination of equal sucrose and pH, liquid medium was always super than solid and semisolid. The tallest plantlets (66 to 71 mm) obtained in liquid medium enriched with sucrose at 10 and 20 g/l both adjusted to pH 6.0; sucrose at 20 g/l and adjusted to pH 6.5 and sucrose at 30 g/l and adjusted to pH 5.0. All of the above combinations except sucrose at 20 and pH 6.0 resulted in 100% rooting. Sucrose at 30 g/l and pH 5.0 resulted in two times more (11 roots per shoot) and three times longer roots (39 mm) than the other treatments (5 roots each about 14 mm long). Each rooting parameter had different optimum combinations of medium state, sucrose and pH adjustment. For any parameter, proper pH adjustment could reduce the optimal sucrose enrichment from 30 to 20 and even to 10 g/l. Hence, pH adjustment is suggested as an important approach for reduction of in vitro rooting medium cost.

Download Full-text

In vitro Dendrobium nobile plant growth and rooting in different sucrose concentrations

Horticultura Brasileira ◽

10.1590/s0102-05362004000400023 ◽

2004 ◽

Vol 22 (4) ◽

pp. 780-783 ◽

Cited By ~ 18

Author(s):

Ricardo T. de Faria ◽

Fabiana N. Rodrigues ◽

Luciana do V.R. Oliveira ◽

Cláudio Müller

Keyword(s):

Plant Growth ◽

Culture Medium ◽

Culture Media ◽

Sucrose Concentration ◽

Ms Medium ◽

In Vitro Growth ◽

Dendrobium Nobile ◽

Modified Ms Medium ◽

Dendrobium Nobile Lindl

Sucrose is a very important component in in vitro culture media, serving as a source of carbon and energy. In this paper, the rooting and in vitro growth of Dendrobium nobile Lindl (Orchidaceae) were studied using different sucrose concentrations (0 g L-1; 5 g L-1; 10 g L-1; 20 g L-1; 30 g L-1 and 60 g L-1), in a modified MS medium containing half the regular concentration of macronutrients at pH 5.8. Greater increases in plant height (4.21±0.6 cm) and high seedling multiplication (1:4) were observed in the 60 g L-1 sucrose treatment, even without the addition of plant hormones. Sucrose concentration in the culture medium did not influence in vitro plant rooting.

Download Full-text

Influence of sucrose concentration in the culture medium on the condition of the photosynthetic apparatus of grapes cultured in vitro

BIO Web of Conferences ◽

10.1051/bioconf/20202504003 ◽

2020 ◽

Vol 25 ◽

pp. 04003

Author(s):

Maria Sundyreva ◽

Anton Rebrov ◽

Alisa Mishko

Keyword(s):

Culture Medium ◽

Culture Media ◽

Sucrose Concentration ◽

Photosynthetic Apparatus ◽

Photosynthetic Parameters ◽

Photosynthetic Genes ◽

Genes Expression ◽

Oxidative Processes ◽

Stress Formation

An influence of different sucrose concentrations in the culture media on the photosynthetic parameters, photosynthetic apparatus related genes expression, oxidative processes and acclimation of grape plants cultured in vitro was examined in this article. An increase of the sucrose concentration in the culture media resulted in a reduced expression of several photosynthetic genes. The most effective functioning of the photosynthetic apparatus was discovered by a decreased amount of surcose in culture media. An increase of the sucrose concentration in the culture media disrupts pigments synthesis, particularly carotenoids, which can be a cause of the secondary oxidative stress formation and grape plants growth reduction during acclimation.

Download Full-text

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

Agrologia ◽

10.30598/a.v1i1.293 ◽

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.

Download Full-text

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

Polysaccharides ◽

10.3390/polysaccharides2020032 ◽

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.

Download Full-text

Modeling and Optimizing a New Culture Medium for In Vitro Rooting of G×N15 Prunus Rootstock using Artificial Neural Network-Genetic Algorithm

Scientific Reports ◽

10.1038/s41598-018-27858-4 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 16

Author(s):

Mohammad Mehdi Arab ◽

Abbas Yadollahi ◽

Maliheh Eftekhari ◽

Hamed Ahmadi ◽

Mohammad Akbari ◽

...

Keyword(s):

Neural Network ◽

Genetic Algorithm ◽

Artificial Neural Network ◽

Culture Medium ◽

In Vitro Rooting ◽

Artificial Neural

Download Full-text

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

Biology of Reproduction ◽

10.1093/biolre/ioab095 ◽

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.

Download Full-text

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

Nativa ◽

10.31413/nativa.v6i1.4731 ◽

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:

Download Full-text

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

Genetika ◽

10.2298/gensr0602129r ◽

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.

Download Full-text