Vernonia condensata Baker: an alternative for large-scale seedling production

ABSTRACT: The increasing use of Vernonia condensata Baker highlights the importance of developing strategies to reduce the impact of exploitation on nature reserves. The aim of this study was to establish a micropropagation protocol to produce homogenous plants with high phytosanitary quality. Apical, nodal, and internodal segments of plants grown in the field were used for in vitro growth. The segments were disinfected in sodium hypochlorite solution (1.0 and 2.0%) for 15 and 30 minutes and then transferred to Petri dishes containing MS culture medium for 30 days. A completely randomized factorial experiment (3 x 2 x 2) with five replicates was designed. After this period, a completely randomized in vitro multiplication experiment was carried out with six treatments (BAP - 0.0; 0.5; 1.0; 1.5; 2.0; 2.5 mg L-1) and six replicates. The shoots obtained in the best treatment were transferred to flasks with rooting medium (MS, MS/2 or MS/4). The experiment was completely randomized with 12 replicates. Microplants were acclimatized in polyethylene terephthalate (PET) bottles filled with autoclaved topsoil. Our results showed that 40.0% of the nodal segments (immersed in 1.0% sodium hypochlorite for 30 minutes) were adequately disinfected and survived. In the in vitro multiplication experiment, the 0.5 mg L-1 concentration of BAP yielded the highest number of shoots and the best vegetative growth. With regard to the assessed characteristics, MS/4 was the best rooting medium, with 100% survival during acclimatization. This study showed that V. condensata in vitro culture might produce 32,000 seedlings in 7 months.

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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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In vitro culture and diversity of endophytic fungi in Bambusa oldhamii

Pesquisa Agropecuária Tropical ◽

10.1590/1983-40632019v4953760 ◽

2019 ◽

Vol 49 ◽

Cited By ~ 1

Author(s):

Ana Paula de Azevedo Pasqualini ◽

Mariely Cristine dos Santos ◽

Bruno Francisco Sant´Anna-Santos ◽

Hugo Pacheco de Freitas Fraga ◽

Marguerite Quoirin

Keyword(s):

In Vitro Culture ◽

Molecular Identification ◽

Endophytic Fungi ◽

Large Scale ◽

Culture Medium ◽

Current Knowledge ◽

Nodal Segments ◽

Scale Production ◽

Bambusa Oldhamii

ABSTRACT Bambusa oldhamii Munro is a fast-growing species of woody bamboo with strong commercial appeal. In Brazil, the use of this species is limited, mainly due to the low availability of seedlings for commercial plantations. Micropropagation is a technique used for the large scale production of seedlings, but protocols for the establishment of aseptic cultures are hampered by the presence of endophytic contamination. This study aimed to develop an in vitro establishment protocol for B. oldhamii, as well as to make the molecular identification of fungi associated with the explants used. Nodal segments of adult plants grown in the field were used as explants. This material was submitted to two experiments carried out to evaluate the effect of 6-benzylaminopurine (BAP) on multiplication and of Plant Preservative Mixture (PPMTM) as a disinfectant. In the first one, 10 µM, 15 µM or 20 µM of BAP were combined with 1 mL L-1, 2 mL L-1 or 3 mL L-1 of PPMTM; while the second one used 0 µM, 2.5 µM, 5 µM or 7.5 µM of BAP with 4 mL L-1 of PPMTM, both added to MS culture medium. After 21 days of culture, the use of 4 mL L-1 of PPMTM inhibited the bacterial growth and reduced fungal contamination. The addition of BAP to the culture medium above 10 µM inhibited the formation and growth of new shoots, while additions of less than 7,5 µM had no effect. The molecular identification of the endophytic fungi isolated during the in vitro culture indicated the presence of numerous fungal species, increasing the current knowledge about the diversity of fungi associated with bamboo.

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Effect of different carbon sources on the in vitro multiplication of Annona sp.

Ciência e Agrotecnologia ◽

10.1590/s1413-70542011005000002 ◽

2011 ◽

Vol 35 (3) ◽

pp. 487-493 ◽

Cited By ~ 5

Author(s):

José Raniere Ferreira de Santana ◽

Renato Paiva ◽

Ana Valéria de Souza ◽

Lenaldo Muniz de Oliveira

Keyword(s):

Culture Medium ◽

Carbon Sources ◽

Dry Weight ◽

Nodal Segments ◽

Seedling Production ◽

In Vitro Multiplication ◽

Bud Induction ◽

Significant Difference ◽

Annona Glabra

The Annonaceae family comprises approximately 2.300 species, some with significant commercial value. Although commercial plantations have suffered due to problems related to seedling production. As micropropagation is a viable technique for seedling production, the present work evaluated the effects of different carbon sources on in vitro bud induction in five Annonaceae species. Nodal segments obtained from plants of the Annona glabra, A. cauliflora, A. coriacea, A. bahiensis and Rollinia silvatica species were inoculated into solid WPM culture medium with 8.87 μM BAP, 0.86 mM of benomyl, and 87.64 mM of the following carbon sources: glucose, sucrose, fructose, galactose, sorbitol and maltose. We evaluated the buds number, the length and weight of the largest bud, the number of expanded leaves per bud, the length of the largest leaf and the dry matter of the buds. No significant difference was observed among the different carbon sources used in terms of the number of produced buds; however, the length of the largest bud, the number of expanded leaves, the length of the largest leaf, and dry weight of the buds presented significant difference according to the studied speciesas well as the carbon sources used, with the lowest value being obtained with sorbitol. The results obtained here indicated that, except for sorbitol, any of the carbohydrates tested could be used in the in vitro multiplication protocols for A. bahiensis, A. cauliflora, A. coriacea, A. glabra and R. silvatica.

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In-vitro callus induction and multiplication of inter-nodal explants in plants Dicoma tomentosa and Alhagi maurorum

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v9i4-a.3455 ◽

2019 ◽

Vol 9 (4-A) ◽

pp. 212-219 ◽

Cited By ~ 1

Author(s):

Shilpa Dhaniya ◽

Suman Kumari Parihar

Keyword(s):

Medicinal Plants ◽

Culture Medium ◽

Raw Material ◽

Nodal Segments ◽

Ms Medium ◽

In Vitro Growth ◽

Primary Metabolites ◽

Shoot Initiation ◽

Rooting Medium

Dicoma tomentosa and Alhagi maurorum are the two medicinal plants with fast in-vitro growth. Both the plants have high economic values. Both the plants were investigated on nodal segments and on leaves. The plants were cultured in five different conditions of medium ranging from MS1- MS5. The hormones were used in these mediums in different concentrations. BAP, NAA, Kinetin, and 2,4 D were use. The MS medium in combination with BAP (2.0 and 2.0mg/ml) with NAA 0.1 mg/ml with kinetin 0.25 mg/ml with 2-4 D were taken, where BAP 1 mg/ml with 2 mg/ml of NAA, BAP 2 mg/ml with 0.5 mg/ml of NAA showed better results with callus growth and root-shoot initiation. The best rooting medium found was MS medium supplemented with IAA and IBA 0.5mg/ each. The culture medium was used in different concentrations for estimation of primary metabolites. Maximum protein and lipid percentage were noticed in leaves of both the plants. It can be concluded that both the studied plants have high medicinal importance and can be used as raw material for industry. Keywords: - Dicoma tomentosa; Alhagi maurorum; Plant hormones; MS media.

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Multiplicação in vitro de Caesalpinia pyramidalis (Leguminosae)

SITIENTIBUS série Ciências Biológicas ◽

10.13102/scb320 ◽

2013 ◽

Vol 13 ◽

Cited By ~ 1

Author(s):

Tecla Dos Santos Silva ◽

Cristina Ferreira Nepomuceno ◽

Bárbara Paula dos Santos Borges ◽

Bruno Freitas Matos Alvim ◽

José Raniere Ferreira De Santana

Keyword(s):

Plant Growth ◽

Plant Growth Regulators ◽

Growth Regulators ◽

Culture Medium ◽

Dry Mass ◽

Shoot Length ◽

Nodal Segments ◽

In Vitro Multiplication ◽

Different Types

Caesalpinia pyramidalis is a species endemic to the Caatinga and known popularly as catingueira, which is widely used by local people, mainly for its timber and medicinal and fodder properties. This study investigated the effects of different types and concentrations of plant growth regulators on the in vitro multiplication of C. pyramidalis. In the first experiment, nodal segments were inoculated in media containing different combinations (0.0–8.0 µM) of BAP and NAA. In the second experiment, nodal segments wereinoculated in media containing different types (KIN, BAP and TDZ) and concentrations (0.0–16μM) of cytokinins. We used a WPM medium supplemented with 87.64 mM sucrose and solidified with 7.0 g L-1 agar. After 45 days, the highest number of shoots, leaf number, shoot length and dry mass of shoots were obtained when nodal segments were inoculated into a culture medium without plant growth regulators.

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Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽

10.3390/ani11051414 ◽

2021 ◽

Vol 11 (5) ◽

pp. 1414

Author(s):

Josep M. Cambra ◽

Emilio A. Martinez ◽

Heriberto Rodriguez-Martinez ◽

Maria A. Gil ◽

Cristina Cuello

Keyword(s):

Culture Medium ◽

Embryo Quality ◽

Transcriptional Profiling ◽

Defined Medium ◽

Platelet Factor ◽

Defined Media ◽

Defined Culture ◽

The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

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In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

Acta Botanica Croatica ◽

10.1515/botcro-2017-0012 ◽

2018 ◽

Vol 77 (1) ◽

pp. 80-87 ◽

Cited By ~ 3

Author(s):

Mahipal S. Shekhawat ◽

M. Manokari

Keyword(s):

Medicinal Plant ◽

Large Scale ◽

Nodal Segments ◽

Ms Medium ◽

Ex Vitro ◽

Culture Bottle ◽

Hybanthus Enneaspermus ◽

Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

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Microfluidic deep mutational scanning of the human executioner caspases reveals differences in structure and regulation

Cell Death Discovery ◽

10.1038/s41420-021-00799-0 ◽

2022 ◽

Vol 8 (1) ◽

Author(s):

Hridindu Roychowdury ◽

Philip A. Romero

Keyword(s):

Large Scale ◽

Cysteine Proteases ◽

Amino Acid Substitutions ◽

Substrate Preferences ◽

Functional Differences ◽

Caspase Family ◽

Executioner Caspases ◽

The Impact ◽

Human Caspase

AbstractThe human caspase family comprises 12 cysteine proteases that are centrally involved in cell death and inflammation responses. The members of this family have conserved sequences and structures, highly similar enzymatic activities and substrate preferences, and overlapping physiological roles. In this paper, we present a deep mutational scan of the executioner caspases CASP3 and CASP7 to dissect differences in their structure, function, and regulation. Our approach leverages high-throughput microfluidic screening to analyze hundreds of thousands of caspase variants in tightly controlled in vitro reactions. The resulting data provides a large-scale and unbiased view of the impact of amino acid substitutions on the proteolytic activity of CASP3 and CASP7. We use this data to pinpoint key functional differences between CASP3 and CASP7, including a secondary internal cleavage site, CASP7 Q196 that is not present in CASP3. Our results will open avenues for inquiry in caspase function and regulation that could potentially inform the development of future caspase-specific therapeutics.

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Clonal bamboo production based on in vitro culture

Bioscience Journal ◽

10.14393/bj-v36n4a2020-48169 ◽

2020 ◽

Vol 36 (4) ◽

Author(s):

Anatálya dos Santos Ribeiro ◽

Alexssandra Jéssica Rondon de Figueiredo ◽

Gabriela Cristina Rech Tormen ◽

André Luís Lopes da Silva ◽

Wellington Ferreira Campos ◽

...

Keyword(s):

Activated Carbon ◽

Large Scale ◽

Culture Medium ◽

Clonal Plant ◽

Adventitious Rooting ◽

Clonal Plants ◽

Ex Vitro ◽

Bambusa Vulgaris ◽

In Vitro Establishment

Bamboo species are an alternative for the composition of forest plantations. However, their potential has not been explored due to the hard time in producing large-scale clonal plants. Thus, the aim this work was to evaluate the in vitro establishment, bud multiplication and ex vitro rooting of Bambusa vulgaris. The first experiment tested different systemic and contact fungicide solutions, based on exposure time, during the establishment phase. Established explants were subjected to evaluation of residual fungicide effect on subcultures during the multiplication and elongation phases. The second experiment evaluated the influence of activated carbon on ex vitro survival and on adventitious rooting. Explant immersion in liquid culture medium added with 1.0 mL of fungicide for 120 hours has favored the in vitro establishment and reduced fungal contamination. On the other hand, it favored the shoot emission of shoots per explant during the multiplication phase. Both rooting induction culture medium and mini-incubator system use were effective in enabling adventitious root formation. The presence of activated carbon in the rooting induction culture medium resulted in a higher clonal plant survival rate.

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Types and concentrations of cytokinins in the micropropagation of Anthurium maricense

Revista Agro mbiente On-line ◽

10.18227/1982-8470ragro.v12i2.4671 ◽

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.

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