In vitro cultivation of Tamarindus indica L.:  explants obtention and contamination in culture medium

The tamarind tree (Tamarindus indica L.) is a common tree in tropical countries with a great exploitation potential due to its high nutritional value and important pharmaceutical characteristics, justifying its potential as a promising crop. The scarcity of scientific studies of the species, especially on propagation, hinders its availability and, consequently, the supply of the product in the market. The aim of this study was to verify the obtainment of nodal segments via sexual propagation and the in vitro establishment of sweet tamarind in MS culture medium (25, 50, 75 and 100% of salts) and with or without activated charcoal (2 g.L-1). The experiment was carried out in a completely randomized design in a 2 x 4 factorial scheme (presence and absence of activated carbon x salt concentrations), with 25 replicates, each replicate consisting of a test tube with an inoculated explant. According to the results, it is possible to conclude that from seedlings with 45 days after sowing, nodal segments of sweet tamarind are obtained for in vitro establishment. As a precursor of protocol for in vitro formation of healthy seedlings is indicated the use of MS culture medium with 75 % of the salts and added with 2 gL-1 of activated charcoal to reduce the contamination index.

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DESINFESTAÇÃO E MEIO DE CULTURA PARA O ESTABELECIMENTO In Vitro DE SEGMENTOS NODAIS DE Liquidambar styraciflua

FLORESTA ◽

10.5380/rf.v40i3.18916 ◽

2010 ◽

Vol 40 (3) ◽

Author(s):

Gilvano Ebling Brondani ◽

Fabricio Augusto Hansel ◽

Leonardo Ferreira Dutra ◽

Ivar Wendling

Keyword(s):

Sodium Hypochlorite ◽

Bacterial Contamination ◽

Culture Medium ◽

Morphological Characteristics ◽

Liquidambar Styraciflua ◽

Nodal Segments ◽

Hydroponic System ◽

Randomized Design ◽

In Vitro Establishment

O objetivo deste trabalho foi testar a desinfestação e meios de cultura para o estabelecimento in vitro de segmentos nodais de Liquidambar styraciflua L. Os explantes foram coletados de minicepas propagadas pelo processo de estaquia e manejadas em minijardim clonal sob sistema semi-hidropônico em leito de areia. O experimento foi conduzido em delineamento inteiramente casualizado no arranjo fatorial (3x2x2), sendo os fatores constituídos por três clones (L 26, L 35 e L 63), duas metodologias de desinfestação (A1 - hipoclorito de sódio (NaOCl) durante 10 minutos a 2,5% v/v de cloro ativo e A2 - explantes mergulhados durante 40 minutos em solução a base de benomyl à 1% p/v) e dois meios de cultura (WPM e MS), com quatro repetições. Os clones não diferiram em relação à assepsia e meio de cultura, obtendo-se média de 4% de explantes isentos de contaminação. Contudo, cerca de 80% dos explantes apresentaram contaminação bacteriana, indicando a necessidade do desenvolvimento de um protocolo de desinfestação mais eficiente. Embora tenham ocorrido poucas diferenças entre os meios de cultura testados, o meio de cultura MS apresentou superioridade em relação ao WPM para a maioria das características morfológicas estudadas.Palavras-chave: Micropropagação; assepsia; contaminação bacteriana; hipoclorito de sódio; benomyl. AbstractDisinfestation and culture medium for the in vitro establishment of Liquidambar styraciflua nodal segments. The objective of this research was to test the culture medium sterilization for the in vitro establishment of Liquidambar styraciflua L. nodal segments. Ministumps, from which the explants were collected, were propagated by cutting process and managed in clonal mini garden under semi-hydroponic system in a sand bed. The experiment was conducted in a completely randomized design under factorial arrangement (3x2x2); the factors were: three clones (L 26, L 35 and L 63), two sterilization methods (A1 - sodium hypochlorite (2.5% v/v of active chlorine) during 10 minutes; A2 - explants immersed during 40 minutes in a solution of benomyl 1% w/v) and two culture mediums (WPM and MS), with four replications. The clones did not differ in relation to the asepsis and culture medium. In average, 4% of the explants were free of contamination. However, almost 80% of the material presented bacterial contamination, what indicated the necessity of developing a more efficient sterilization protocol. Although only a little difference between the culture mediums had been observed, the MS medium showed superiority in relation to the WPM regarding the majority of the morphological characteristics studied.Keywords: Micropropagation; asepsis; bacterial contamination; sodium hypochlorite; benomyl.

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Zinc as promoter of growth and biochemical activity in basil cultivars under in vitro conditions

Scientia Agraria Paranaensis ◽

10.18188/sap.v18i4.21909 ◽

2019 ◽

pp. 342

Author(s):

Caroline Galego Comar ◽

Edinara Maria Barbosa ◽

Vanusa Souza Rocha Pereira ◽

Julliane Destro de Lima ◽

Thiago Teodoro Santana ◽

...

Keyword(s):

Antioxidant Activity ◽

Phenolic Compounds ◽

Culture Medium ◽

Total Phenolic ◽

In Vitro Cultivation ◽

Biochemical Activity ◽

Randomized Design ◽

Number Of Leaves ◽

Completely Randomized Design

In vitro cultivation of basil allows the manipulation of the concentration of certain micronutrients, commonly neglected by the micropropagation protocols. It is a plant of great economic importance for the cosmetic and pharmaceutical industry, due to the components present in its essential oil. In view of the above, the objective of this study was to evaluate zinc (Zn) concentrations in the micropropagation of basil, in addition to antioxidant activity and total phenolic compounds. Basil seeds, cultivars Manolo and Grecco Palla were oxygenated for 4 h, passed through asepsis and placed in test tubes with MS medium supplemented with 30 g L-1 sucrose and 6.5 g L-1 agar and pH adjusted to 5.8. The treatments were composed by the addition or not of 25 μM of zinc sulfate (ZnSO4) and arranged in a completely randomized design. The tubes containing the seeds and the culture medium were kept in a growth chamber for 90 days. The cultivar Manolo was more sensitive to the addition of ZnSO4 due to the increase in the number of leaves and in the antioxidant activity, however, the addition of this component in the culture medium did not influence the production of phenolic compounds or the activity of the antioxidant enzymes SOD, CAT and APX.

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In vitro establishment of pepper-rosmarin nodal segments

Horticultura Brasileira ◽

10.1590/s0102-05362008000200025 ◽

2008 ◽

Vol 26 (2) ◽

pp. 255-258 ◽

Cited By ~ 3

Author(s):

Arie F Blank ◽

Andréa S da Costa ◽

Maria de Fátima Arrigoni-Blank ◽

Aline B de Mendonça ◽

Ana da S Ledo

Keyword(s):

Essential Oil ◽

Medicinal Plant ◽

Growth Regulators ◽

Culture Medium ◽

Antimicrobial Properties ◽

Shoot Length ◽

Nodal Segments ◽

Lippia Sidoides ◽

In Vitro Establishment

Pepper-rosmarin (Lippia sidoides Cham.) is a native medicinal plant from the Brazilian biome Caatinga. Its high economical importance comes from the antimicrobial properties of thymol and carvacrol, both present in the pepper-rosmarin essential oil. Nodal segments of pepper-rosmarin were established in vitro to evaluate different growth regulators. We tested four concentrations of IBA (0.0; 0.01; 0.05; and 0.1 mg L-1) combined with six concentrations of BAP (0.0; 0.1; 0.5; 1.0; 2.0; and 4.0 mg L-1) in assay 1; five concentrations of NAA (0.0; 0.1; 0.25; 0.5; and 1.0 mg L-1) in assay 2; two concentrations of NAA (0.0 and 0.01 mg L-1) combined with five concentrations of kinetin (0.0; 0.25; 0.5; 1.0; and 2.0 mg L-1) in assay 3; five concentrations of IAA (0.0; 0.5; 1.0; 1.5; and 2.0 mg L-1) in assay 4; and five concentrations of GA3 (0.0; 0.1; 0.5; 1.0; and 1.5 mg L-1) in assay 5. The use of BAP, IBA, NAA and KIN did not influence the number of shoots. The addition of 0.1 mg L-1 of NAA at the culture medium resulted in larger shoot length per explant, while the concentration of 1.0 mg L-1 of kinetin promoted larger shoot length in general. GA3 did not affect the elongation of pepper-rosmarin shoots when cultivated in vitro.

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Effect of different culture tube caps and concentrations of activated charcoal and sucrose on in vitro growth and budding induction of Annona glabra L.

Ciência e Agrotecnologia ◽

10.1590/s1413-70542011000500008 ◽

2011 ◽

Vol 35 (5) ◽

pp. 916-923 ◽

Cited By ~ 5

Author(s):

José Raniere Ferreira de Santana ◽

Renato Paiva ◽

Ana Valéria de Souza ◽

Lenaldo Muniz de Oliveira

Keyword(s):

Activated Charcoal ◽

Dry Weight ◽

Test Tube ◽

Leaf Abscission ◽

Nodal Segments ◽

Bud Induction ◽

Pvc Film ◽

Annona Glabra ◽

In Vitro Induction

The present work evaluated the effects of different types of culture flask seals and varying concentrations of sucrose and activated charcoal on the in vitro induction and growth of buds of Annona glabra L.; an edible fruit-producing species popularly known as "araticum". Nodal segments obtained from A. glabra plants maintained in green houses were surface sterilized and inoculated into a WPM culture medium solidified with 7 g L-1 of agar and supplemented with sucrose (0.00; 29.21; 58.63 and 116.84 mM), activated charcoal (0.0 and 2.0 g L-1), and 250 mg L-1 benomyl. In addition to the varying concentrations of sucrose and activated charcoal, we evaluated the efficiency of two types of test tube seals: PVC film, and cotton plugs. All possible combinations of caps and nutrient media were tested with 4 repetitions with 5 tubes each, evaluating the number of buds, the percentage of explant responses, the number of expanded leaves per bud, the length of the largest leaves, leaf abscission, and the length and dry weight of the buds. The type of seal influenced organogenesis in nodal segments of A. glabra, and no bud induction was observed in the absence of sucrose. The largest number of expanded leaves were obtained when 58.42 mM of sucrose was used in tubes sealed with cotton plugs, and leaf abscission was halved in the presence of activated charcoal. The greatest bud length and dry weight were obtained in tubes sealed with cotton plugs and in the presence of activated charcoal.

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Solidifying agents and activated charcoal for in vitro culture of Solanum sessiliflorum

Pesquisa Agropecuária Brasileira ◽

10.1590/s0100-204x2017001100019 ◽

2017 ◽

Vol 52 (11) ◽

pp. 1123-1126 ◽

Cited By ~ 2

Author(s):

Filipe Almendagna Rodrigues ◽

Renata Alves Lara Silva Rezende ◽

Moacir Pasqual ◽

Maria Teresa Gomes Lopes

Keyword(s):

In Vitro Culture ◽

Activated Charcoal ◽

Root Length ◽

Culture Medium ◽

In Vitro Cultivation ◽

Fresh Matter ◽

Number Of Leaves

Abstract: The objective of this work was to evaluate the effects of the solidifying agents agar and phytagel and of activated charcoal on the in vitro cultivation of two maná cubiu (Solanum sessiliflorum) varieties: Thaís and Santa Luzia. The phytotechnical characteristics analyzed included number of leaves, number of roots, shoot and root length, and fresh matter of shoot and root. Regardless of the variety, phytagel was superior to agar as a culture medium. A greater number of leaves and longer shoots were observed in the Santa Luzia variety, in the absence of charcoal. The Thaís variety showed longer shoots and roots in the presence of charcoal.

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Control of contaminants during introduction and establishment of Bambusa vulgaris in vitro

Research in Biotechnology ◽

10.19071/rib.2016.v7.3056 ◽

2016 ◽

Vol 7 ◽

Cited By ~ 1

Author(s):

Gustavo Rubens Castro Torres ◽

Laureen Michelle Houllou ◽

Robson Antônio De Souza

Keyword(s):

Microbial Contamination ◽

Culture Medium ◽

Nodal Segments ◽

Fungal Contamination ◽

Liquid Culture Medium ◽

Randomized Design ◽

Bambusa Vulgaris ◽

Completely Randomized Design ◽

Pre Treatment

The aim of this work was to test techniques to reduce microbial contamination in the phases of introduction and establishment of the in vitro cultivation of Bambusa vulgaris through two experiments. The first experiment was carried out in a completely randomized design using a factorial arrangement (pre-treatment of nodal segments using or not a solution of Derosal 500 SC® and Chloramphenicol × culture medium with half or full concentration of salts × culture medium with presence or absence of sucrose × culture medium with presence or absence of Plant Preservative MixtureTM). In a second experiment, carried out in a completely randomized design, the effect of different fungicides associated to Chloramphenicol in a liquid culture medium was tested. It was possible to verify that the isolated effects of the pre-treatment by immersion of the nodal segments in a solution of 4 mL L-1 of Derosal 500 SC® and 200 mg L-1 of Chloramphenicol for 30 minutes and explants placed in a sucrose-free medium reduced fungal contamination. In the second experiment, the treatment that reduced fungal contamination corresponded to explants placed for seven days in a liquid medium with half the concentration of salts, sucrose-free, with 2 mL L-1 of Plant Preservative MixtureTM and with 4 mL L-1 of Derosal 500 SC® and 200 mg L-1 of Chloramphenicol.

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Chemical sterilization in in vitro propagation of Arundina bambusifolia Lindl. and Epidendrum ibaguense Kunth

Revista CERES ◽

10.1590/s0034-737x2013000400001 ◽

2013 ◽

Vol 60 (4) ◽

pp. 447-451 ◽

Cited By ~ 6

Author(s):

Donizetti Tomaz Rodrigues ◽

Roberto Ferreira Novais ◽

Víctor Hugo Alvarez Venegas ◽

José Maria Moreira Dias ◽

Wagner Campos Otoni ◽

...

Keyword(s):

Activated Charcoal ◽

Culture Medium ◽

Flow Chamber ◽

Nodal Segments ◽

Control Treatment ◽

Fresh Weight ◽

Production Methods ◽

Weight Yield ◽

Chemical Sterilization

There is a great demand for simpler and less costly laboratory techniques and for more accessible procedures for orchid breeders who do not have the necessary theoretical basis to use the traditional seed and clone production methods of orchids in vitro. The aim of this study was to assess the use of sodium hypochlorite (NaClO) as a decontaminant in the process of inoculating adult orchid explants of Arundina bambusifolia and Epidendrum ibaguenses. Solutions of NaClO (1.200, 2.400, 3.600, 4.800 and 6.000 mg L-1 - equivalent to 50, 100, 150, 200 and 250 mL L-1 of commercial bleach - CB) were sprayed on the explants (1.0 mL) and the culture medium (GB5), in the presence or absence of activated charcoal (2 g L-1). The explants used were nodal segments of field-grown adult plants. The procedures for inoculating the explants were conducted outside the laminar flow chamber (LFC), except for the control treatment (autoclaved medium and explant inoculation inside the LFC). The best results for fresh weight yield, height and number of shoots were obtained using NaClO in solution at 1.200 mg L-1 (equivalent to 50 mL L-1 commercial bleach) with activated charcoal in the culture medium. Fresh weight figures were 1.10 g/jar for Arundina bambusifolia and 0.16 g/jar for Epidendrum ibaguenses. Spraying the NaClO solutions controls the contamination of the culture medium already inoculated with the explants.

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In vitro behaviour of Aspasia variegata, an epiphytic orchid from the Brazilian Cerrado

Ciência Rural ◽

10.1590/s0103-84782013001200010 ◽

2013 ◽

Vol 43 (12) ◽

pp. 2178-2184 ◽

Cited By ~ 1

Author(s):

Vespasiano Borges de Paiva Neto ◽

Guilherme de Oliveira Campos ◽

Amanda Galdi Boaretto ◽

Monica Cristina Rezende Zuffo ◽

Mateus de Aguiar Torrezan ◽

...

Keyword(s):

Activated Charcoal ◽

Culture Medium ◽

Culture Media ◽

Survival Rates ◽

In Vitro Cultivation ◽

In Vitro Growth ◽

Mato Grosso ◽

Chlorophyll Contents ◽

Coconut Fibre

Aspasia variegata occurs naturally in the savanna of the Mato Grosso do Sul State, Brazil and it has been widely collected for its beautiful flowers. Additionally, its habitat has been greatly reduced and little or no investigation of its spread has been performed. Aiming to establish a protocol to obtain seedlings of the orchid A. variegata, different compositions of culture medium were tested to identify which one provided better in vitro growth and development and to assess the influence of these media in seedling acclimatisation. Thus, seeds obtained from mature capsules were inoculated in Knudson culture medium for 120 days until the protocorm stage. They were transferred to different culture media formulations, including MS and Knudson with half or full formulation, and 3.0 and 6.0g L-1 activated charcoal were added to them or not. After 180 days of protocorm inoculation, seedlings were evaluated for length of roots and shoots, number of roots and leaves, and chlorophyll contents. After that, seedlings were transferred to trays containing a mixture of Plantmax® and coconut fibre (1:1) for acclimatisation. Best results for the in vitro growth of A. variegata were obtained with the use of MS medium supplemented with 6.0g L-1 activated charcoal. Higher levels of chlorophyll were obtained, however, in treatments containing MS salts without activated charcoal presence, and lower levels in media containing Knudson salts with the presence of activated charcoal. The seedlings originated by higher chlorophyll levels during in vitro cultivation presented the highest survival rates and better development in the acclimatisation phase.

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Culture media for the multiplication of wild Manihot species

Ciência e Agrotecnologia ◽

10.1590/1413-70542018426024718 ◽

2018 ◽

Vol 42 (6) ◽

pp. 598-607 ◽

Cited By ~ 1

Author(s):

Jucieny Ferreira de Sá ◽

Emília dos Santos Sampaio ◽

Maria Inês de Souza Mendes ◽

Karen Cristina Fialho dos Santos ◽

Antônio da Silva Sousa ◽

...

Keyword(s):

Plant Height ◽

Disease Transmission ◽

Culture Media ◽

Test Tube ◽

Nodal Segments ◽

Randomized Design ◽

Lateral Bud ◽

Successive Generations ◽

Shoot Mass

ABSTRACT The cassava propagation system is slow and favors disease transmission through successive generations. Micropropagation is an alternative to overcome the aforementioned limitations, besides allowing the generation of a larger number of pest- and pathogen-free plants. Therefore, the aim of the present study is to investigate the effect of culture media on the multiplication in vitro of five wild Manihot species. The experiment followed a completely randomized design, at factorial arrangement 5 (wild Manihot species) x 6 (culture media), with 11 repetitions. Explants consisted in nodal segments (91 cm long and one lateral bud) of species Manihot flabellifolia, M. tristis, M. caerulescens, M. chlorosticta and M. jacobinensis, which were extracted in vitro from the collection of wild cassava species. One segment was placed in each test tube added with 10 mL of MS media 0.01, 17N, 12A3, 4E, 8S and WPM, and kept for 90 days in growth room under 30 μmol m-2 s-1irradiance, temperature 27 ± 1 °C and 16h photoperiod. Variables plant height (cm), number of green leaves, number of senescent leaves, number of shoots, number of microcuttings, fresh and dry shoot mass, fresh and dry root mass (mg) and callus mass (mg) were analyzed. Our results showed that the culture medium 12A3 was not responsive to any of the species; however, if one takes into consideration variables plant height and number of microcuttings, this medium can possibly be used in the micropropagation of other wild species belonging to genus Manihot.

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Colchicine and oryzalin effects on tetraploid induction and leaf anatomy of Solanum commersonii ssp.

Ciência Rural ◽

10.1590/0103-8478cr20150477 ◽

2016 ◽

Vol 46 (11) ◽

pp. 1973-1979 ◽

Cited By ~ 1

Author(s):

Lívia Gracielle Oliveira Tomé ◽

Adriano Bortolotti da Silva ◽

Cesar Augusto Brasil Pereira Pinto ◽

Lisete Chamma Davide ◽

Dalvana Sousa Pereira ◽

...

Keyword(s):

Flow Cytometry ◽

Leaf Anatomy ◽

Culture Medium ◽

Nodal Segments ◽

In Vitro Cultivation ◽

Solanum Commersonii ◽

Breeding Programs ◽

Genetic Breeding ◽

Equatorial Diameter

ABSTRACT: The aim of the present study was to evaluate the effect of the colchicine and oryzalin anti-mitotic substances on the induction of tetraploid plants and foliar anatomy of two diploids clones of Solanum commersonii ssp. Nodal segments of Solanum commersonii subsp. commersonii Dun. and Solanum commersonii subsp. malmeanum Bitt. were treated with colchicine (3.5, 5.0, and 6.5mM; 72h) or oryzalin (10, 30, and 50µM; 24h). After the treatment with anti-mitotic substances, nodal segments were inoculated in the MS culture medium and cultivated in vitro (60 days). After in vitro cultivation, the plants were transferred to vases with the substrate Plantmax(r) and kept in the greenhouse (45 days). Plant ploidy level was assessed by flow cytometry and leaf anatomy was assessed by anatomic cuts. An increase was observed in the polar and equatorial diameter of stomata ("gigas effect") of the Solanum commersonii subsp. commersonii (SCC) and Solanum commersonii subsp. malmeanum (SCM) clones, which was due to the use of the anti-mitotic substances. Treatments with colchicine (6.5mM; 72h) and oryzalin (50µM; 24h) caused death of the SCC plants cultured in vitro. In the others treatments of the SCC clone, the use of oryzalin and colchicine caused production of chimeric plants. The treatment of nodal segments of SCM with oryzalin (10-50µM; 24h) was effective on induction of tetraploid plants, which can be employed in genetic breeding programs in crossbreeding with cultured potato.

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