Sealing and explant types on the mangaba micropropagation

In micropropagation, especially for mangaba tree botanical variety of Northeastern Brazil, limiting aspects such as ethylene accumulation in the cultivation flask and loss of vigor in subcultures have been observed. This study was aimed at assessing the technical and scientific knowledge of the in vitro propagation of botanical mangaba tree variety and at improving the micropropagation protocol, establishing the in vitro cultivation time, the best type of flask sealing and explant at different micropropagation stages. For the establishment phase and for the first and second subcultures, the MS medium with 3% sucrose and 0.6% agar, supplemented with 1 mg L-1 IAA and 1 mg L-1 BA was used. Evaluations were performed at 30, 50 and 65 days of in vitro cultivation. The best types of flask sealing for the establishment phase were the PVC film and Para-film® and for the first subculture the Para-film® seal. In the second subculture the PVC film and Para-film® seals promoted the best growth. The median and basal nodal segments presented the best performance in the first subculture. No significant effect of explant type was observed in the second subculture. The ideal subculture interval in the establishment phase and the first and second subcultures is 50 days.

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In vitro propagation of muskmelon (Cucumis melo L.) from nodal segments, shoot tips and cotyledonary nodes

Rajshahi University Journal of Life & Earth and Agricultural Sciences ◽

10.3329/rujleas.v41i0.21627 ◽

2015 ◽

Vol 41 ◽

pp. 71-77

Author(s):

S. Parvin ◽

M. Kausar ◽

M. Enamul Haque ◽

M. Khalekuzzaman ◽

B. Sikdar ◽

...

Keyword(s):

Cucumis Melo ◽

In Vitro Propagation ◽

Multiple Shoots ◽

Multiple Shoot ◽

Shoot Tips ◽

Nodal Segments ◽

Ms Medium ◽

Cotyledonary Nodes ◽

Cucumis Melo L

A rapid and efficient protocol is outlined for in vitro propagation of muskmelon(Cucumis melo L.) Shoot tips, nodal segments and cotyledonary nodes from invitro grown seedlings were used as explants. The explants were inoculated on MS medium fortified with different combinations and concentrations of growthregulators viz., BAP, NAA, GA3 and IBA for multiple shoot regeneration.Effective result was found on MS medium supplemented with 2.0 mg/l BAP, inwhich 90% and 70% cultures induced multiple shoots from nodal segments andshoot tip explants, respectively. Whereas, 70% cultures of cotyledonary nodeswere found to induced shoots on MS medium with 1.5 mg/l BAP + 0.1 mg/l GA3. In vitro regenerated shoots were subcultured on half strength MS mediumsupplemented with different concentrations of IBA and NAA for successful rootinduction and the effective result (up to 70%) was found in medium with 1 mg/lIBA. Well rooted in vitro grown plantlets were acclimatized in sandy soil, whereas 70% plantlets survived

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In vitro culture of jambu with different growth regulators

Horticultura Brasileira ◽

10.1590/s0102-053620200204 ◽

2020 ◽

Vol 38 (2) ◽

pp. 134-138

Author(s):

Susan P Almeida ◽

Joanne MM Souza ◽

Andredy MT Amorim ◽

Sérgio AL de Gusmão ◽

Rodrigo ORM Souza ◽

...

Keyword(s):

Growth Regulators ◽

Callus Induction ◽

Nodal Segments ◽

Root Production ◽

In Vitro Cultivation ◽

Average Height ◽

Ms Medium ◽

Fresh Matter ◽

Friable Callus

ABSTRACT The aim of this study was to establish the best concentrations of growth regulators for in vitro cultivation of jambu for a subsequent elaboration of an efficient micropropagation protocol. After sterilized, the seeds were inoculated on different media (MS, ½MS and water-agar) for in vitro germination. Nodal segments of in vitro germinated jambu seedlings were used as explants in the micropropagation with different concentrations of 6-benzylaminopurine (BAP) (0.0; 0.125; 0.25; 0.50 and 0.75 mg L-1) and callus induction with 2.4- dichlorophenoxyacetic acid (2.4-D) (0.0; 0.25; 0.50; 0.75 and 1.0 mg L-1) on Murashige & Skoog’s (MS) medium. The highest germination rates were obtained on MS medium with better seedling development and greater height (3.7cm). In micropropagation, the best treatment was obtained on 0.125 BAP (T2), with an average of 2.2 sprouts/explant, average height of 2.4 cm and vigorous sprouts. In callus induction, all treatments with 2.4-D had developed friable calluses in 30 days and using doses of 0.25 and 0.50 mg L-1 provided greater fresh matter. The induction of friable callus and the root production occur without supplementation of exogenous growth regulator.

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In Vitro Multiplication and NMR Fingerprinting of Rare Veronica caucasica M. Bieb

Molecules ◽

10.3390/molecules26195888 ◽

2021 ◽

Vol 26 (19) ◽

pp. 5888

Author(s):

Desislava I. Mantovska ◽

Miroslava K. Zhiponova ◽

Milen I. Georgiev ◽

Tsvetinka Grozdanova ◽

Dessislava Gerginova ◽

...

Keyword(s):

Ex Situ ◽

Nodal Segments ◽

Cultivated Plants ◽

In Vitro Cultivation ◽

Ms Medium ◽

Ex Vitro ◽

Biotechnological Approach ◽

Iridoid Glucosides ◽

Development In Vitro

Micropropagation of rare Veronica caucasica M. Bieb. was achieved by successful in vitro cultivation of mono-nodal segments on MS medium supplemented with 1.0 mg L–1 6-benzylaminopurine (BA) and then transferring the regenerated plants on hormone free basal MS medium for root development. In vitro multiplicated plants were successively acclimated in a growth chamber and a greenhouse with 92% survival. The number of plastid pigments and the total phenolics content in in vitro cultivated and ex vitro adapted plants were unchanged, and no accumulation of reactive oxygen species (ROS) was detected by staining with 3-3′-diaminobenzidine (DAB) and 2′,7′-dichlorofluorescein diacetate (DCF-DA). Nuclear Magnetic Resonance (NMR) fingerprinting allowed for the identification of the major alterations in metabolome of V. caucasica plants during the process of ex situ conservation. Iridoid glucosides such as verproside, aucubin and catalpol were characteristic for in vitro cultivated plants, while in ex vitro acclimated plants phenolic acid–protocatechuic acid and caffeic acid appeared dominant. The successful initiation of in vitro and ex vitro cultures is an alternative biotechnological approach for the preservation of V. caucasica and would allow for further studies of the biosynthetic potential of the species and the selection of lines with a high content of pharmaceutically valuable molecules and nutraceuticals.

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Micropropagation, seed propagation and germplasm bank of Mandevilla velutina (Mart.) Woodson

Scientia Agricola ◽

10.1590/s0103-90162007000300008 ◽

2007 ◽

Vol 64 (3) ◽

pp. 263-268 ◽

Cited By ~ 14

Author(s):

Ronaldo Biondo ◽

Ana Valéria Souza ◽

Bianca Waléria Bertoni ◽

Andreimar Martins Soares ◽

Suzelei Castro França ◽

...

Keyword(s):

Medicinal Plant ◽

Plant Species ◽

In Vitro Propagation ◽

Nodal Segments ◽

Medicinal Plant Species ◽

Soil Conditions ◽

Ms Medium ◽

Germplasm Bank ◽

Seed Propagation

Mandevilla velutina (Mart.) Woodson (Apocynaceae) is a medicinal plant species with antivenom properties, native from Brazilian Savanna regions (Cerrado), which due to overexploitation and habitat deforestation is in danger of extinction. As an initiative for conserving this endangered but economically important plant species, a micropropagation protocol was developed and genotypes were stored in the Germplasm Bank "Cerrado In vitro". For the in vitro propagation of M. velutina, nodal segments were inoculated on Murashige and Skoog (MS) medium supplemented with different concentrations of BA, Zeatin, 2ip, DTT and TDZ. Best multiplication ratio was achieved when to the medium 0.44 µM BA, ranging 1: 6.7, were added. Plantlets cultured on MS/2 medium supplemented with 26.85 µM NAA rooted successfully (50.5%). Although rooted and un-rooted plantlets acclimatized to soil conditions, great losses were observed within un-rooted plantlets, while the rooted presented 100 % survival. It was possible to maintain 43% of the M. velutina germplasm under healthy conditions for six months, with no subcultures, using the MS medium supplemented with 2% sucrose, 13.8 mM spermidine, 2% sorbitol and 2% dextrose.

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Plant Regeneration from Leaf Explants of the Medicinal Herb Wedelia chinensis

Horticulturae ◽

10.3390/horticulturae7100407 ◽

2021 ◽

Vol 7 (10) ◽

pp. 407

Author(s):

Yung-Ting Tsai ◽

Kin-Ying To

Keyword(s):

Plant Regeneration ◽

In Vitro Propagation ◽

Liver Toxicity ◽

Leaf Explants ◽

Basal Medium ◽

Nodal Segments ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Average Percentage

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.

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In vitro propagation from nodal segments of Lippia origanoides (chemotype A)

Ciência Rural ◽

10.1590/0103-8478cr20210323 ◽

2022 ◽

Vol 52 (7) ◽

Author(s):

Julián Sebastián Ramírez Moreno ◽

Sergio Andrés Vega Porras ◽

Roosevelt Humberto Escobar ◽

Elena E. Stashenko ◽

Jorge Luis Fuentes Lorenzo ◽

...

Keyword(s):

Plant Growth ◽

Gibberellic Acid ◽

Growth Regulators ◽

In Vitro Propagation ◽

Callus Formation ◽

Nodal Segments ◽

Ms Medium ◽

Shoot Number ◽

Murashige And Skoog Medium

ABSTRACT: This research described an efficient micropropagation protocol for Lippia origanoides (Verbenaceae). Sterile seeds were used to obtain germinated seedlings in Murashige and Skoog medium (MS) supplemented with sucrose and agar. The nodal segments obtained from seedlings were grown on MS medium supplemented with different concentrations of gibberellic acid (GA), benzylaminopurine (BAP) and 1-naphthalenacetic acid (NAA) with BAP. The callus induction, shoots length, shoots number and root length, were analyzed. The treatments showed high percentage of callus formation at 0.5 to 1.5 mg L-1 of BAP alone or in combination with NAA (0.1 mg L-1). The highest value of shoot number per nodal segments was obtained at 1.5 mg L-1 of BAP (4.3 ± 0.8). The obtained plantlets were better rooted in vitro in the absence of plant growth regulators (PGRs) and they showed acclimatization rate of 90%. We reported a protocol for in vitro propagation and acclimatization of L. origanoides for A chemotypes from Colombia.

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Multiplicação de anador (Justicia pectoralis) in vitro

Revista Verde de Agroecologia e Desenvolvimento Sustentável ◽

10.18378/rvads.v11i3.3986 ◽

2016 ◽

Vol 11 (3) ◽

pp. 159 ◽

Cited By ~ 1

Author(s):

Rômulo Magno Oliveira Freitas ◽

Narjara Walessa Nogueira ◽

Sidney Carlos Praxedes

Keyword(s):

Plant Material ◽

In Vitro Propagation ◽

Culture Medium ◽

Culture Media ◽

Statistical Design ◽

The Other ◽

Nodal Segments ◽

Ms Medium ◽

Number Of Leaves

O trabalho teve como objetivo desenvolver um protocolo de micropropagação de segmentos nodais de anador (Justicia pectoralis). Para isso foram realizados dois experimentos. O delineamento estatístico utilizado foi o inteiramente casualizado, com 15 repetições. Os segmentos de J. pectoralis, após desinfestados, foram cultivados em meio MS durante 30 dias. No primeiro experimento, esse material foi repicado em três meios de cultura (MS, WPM e B5) e após 77 dias foram avaliados comprimento de plântula, número de raízes, número de folhas e o número de segmentos nodais. Para o segundo experimento foram testadas duas citocininas (BAP e Cinetina) nas seguintes dosagens 0,0; 0,5; 5,0 e 20,0 mM. Aos 60 dias após a repicagem foram avaliadas as seguintes características: números de folhas, número de raízes e número de explantes por planta. O meio MS foi o que apresentou maior comprimento de plântula. As demais variáveis não diferiram entre os meios utilizados. Por isso o meio MS foi utilizado para o segundo experimento onde se verificou que a utilização de BAP proporcionou maior número de folhas e de explantes quando submetido à concentração de 20 mM. Dessa forma, para multiplicação de seg Justicia pectoralis, recomenda-se a utilização de meio MS com adição de 20mM de BAP.In vitro propagation of Justicia pectoralisAbstract: The study aimed to establish a micropropagation protocol for Justicia pectoralis nodal segments. Two experiments were conducted. The statistical design was the completely randomized with 15 repetitions. After disinfestation, the segments of J. pectoralis were inoculated in the MS culture medium for 30 days. In the first experiment, the plant material was transferred to three culture media (MS, WPM and B5). The length of seedlings, number of roots, number of leaves, and number of nodal segments were evaluated at 77 days after transferring. In the second experiment two cytokinins (BAP and Kinetin) were tested in the following concentrations: 0.0; 0.5; 5.0 and 20.0 mM. At 60 days after transplanting the number of leaves, number of roots and number of explants per plant were evaluated. The MS medium induced the highest length of seedlings, but there was no effect for the other variables. Therefore, this medium was used for the second experiment, when it was found that BAP induced a larger number of leaves and explants when applied at 20 mM. Therefore, for multiplying J. pectoralis nodal segments we recommend the use of MS medium with 20 mM BAP.

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Micropropagation of Pyracantha coccinea

HortScience ◽

10.21273/hortsci11301-16 ◽

2017 ◽

Vol 52 (2) ◽

pp. 271-273

Author(s):

Chao Dong ◽

Xue Li ◽

Yue Xi ◽

Zong-Ming Cheng

Keyword(s):

In Vitro Propagation ◽

Basal Medium ◽

Ornamental Plant ◽

Axillary Buds ◽

Nodal Segments ◽

Ms Medium ◽

Southeast Europe ◽

Rooting Medium ◽

Pyracantha Coccinea

Pyracantha coccinea is a thorny evergreen shrub native to southeast Europe to southeast Asia. It is a popular ornamental plant because of its showy bright red fruits and small white flowers. However, in vitro vegetative propagation of P. coccinea has not been studied. Nodal segments with one or two axillary buds (1 to 1.5 cm in length) were cut and disinfected in a solution of 0.1% (v/v) mercuric chloride (HgCl2) for 5 minutes, and proliferated on Murashige and Skoog (MS) basal medium supplemented with various concentrations 6-benzylaminopurine (6-BA). After 4 weeks, newly formed shoots were transferred to proliferation and rooting media containing various concentrations of indole-3-butyric acid (IBA). Establishment of axillary buds was significantly better with an establishing rate of 67% on basal MS medium augmented with 6.6 µm 6-BA. The best medium for proliferation of shoots was three-fourth basal MS supplemented with 1.5 µm IBA, with a proliferation rate of 3.4 axillary bud. The optimum rooting medium was one-fourth MS basal medium containing 93 µm IBA. Rooting of shoots was as much as 77%. Rooted plantlets were transferred to pots containing vermiculite:perlite:peat (6:1:2) and acclimatized to ambient greenhouse conditions with a 95% survival rate. This protocol can be used for in vitro propagation of P. coccinea.

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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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In Vitro Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey

10.5772/intechopen.94378 ◽

2020 ◽

Author(s):

Nurşen Çördük ◽

Cüneyt Aki

Keyword(s):

Acetic Acid ◽

Medicinal Plant ◽

In Vitro Propagation ◽

Leaf Explants ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Northwestern Turkey ◽

Helen Of Troy ◽

Endemic Medicinal Plant

Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regeneration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber.

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