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

Molecules ◽  
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.

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

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
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.

scholarly journals Diazotrophic bacteria in the growth of micropropagated ornamental pineapple

2019 ◽  
Vol 13 (2) ◽  
pp. 269-278
Author(s):  
Adriano Bortolotti Silva ◽  
Ligiane Aparecida Florentino ◽  
Dalvana De Sousa Pereira ◽  
Paulo Roberto Correa Landgraf ◽  
Ana Carolina Rodrigues Alves ◽  
...  
Keyword(s):  
Shoot Growth ◽  
Ex Vivo ◽  
Control Treatment ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Ex Vitro ◽  
Diazotrophic Bacteria ◽  
Bacterial Strains ◽  
Indole 3 Acetic Acid

Ornamental pineapple is a hardy plant with significant landscaping value. Tissue culture of plants is viable for producing plants with a high phytosanitary quality. However, one of the difficulties with this cultivar is the acclimatization process, which is slow and can cause losses. The objective of the present study was to verify the potential of inoculation with diazotrophic bacteria for in vitro and ex vivo growth of ornamental pineapple. A group of diazotrophic bacterial strains selected at the Universidade José do Rosário Vellano (UNIFENAS) was prioritized in this study, and the treatments included bacterial strains UNIFENAS (100-13, 100-60, 100-68, 100-153, 100-167 and 100-198). These strains were evaluated in terms of their capacity to produce indole 3-acetic acid. Subsequently, plants were cultivated in a medium composed of MS medium salts (1/4), adding 1 mL of the bacterial strain. In the control treatment, the plants were maintained in 2 mL of MS medium. 7 days after inoculation, the plants were transplanted into the MS, where they were maintained for 30 days. After in vitro cultivation, the plants were transferred to pots containing commercial Plantmax® substrate and maintained under these conditions for 60 days. The diazotrophic bacteria were able to synthesize auxins, and their inoculation promoted greater growth in vitro and ex vitro in the plants. In the acclimatization phase, the plants inoculated with UNIFENAS strains (100-60, 100-68 and 100-153) promoted a higher shoot growth, chlorophyll content and nitrate reductase enzyme activity.

scholarly journals In vitro culture of jambu with different growth regulators

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.

scholarly journals Nitrogen assimilation in the bromeliad Ananas comosus var. ananassoides (Baker) Coppens & F.Leal grown in vitro with different sources of inorganic nitrogen

Hoehnea ◽  
2020 ◽  
Vol 47 ◽  
Author(s):  
Priscila Primo Andrade Silva ◽  
Ivomar Aparecido Medina ◽  
Jorge Luiz Marx Young ◽  
Vívian Tamaki
Keyword(s):  
Nitrogen Assimilation ◽  
Inorganic Nitrogen ◽  
Ananas Comosus ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Ex Vitro ◽  
N Assimilation ◽  
Modified Ms Medium ◽  
Different Sources

ABSTRACT Ananas comosus var. ananassoides (Baker) Coppens & F.Leal is a native ornamental bromeliad of the endangered biome Cerrado. Therefore, approaches aimed at the preservation of this species, such as in vitro cultivation and micropropagation are needed. Nitrogen (N) is absorbed by plants, mainly as NO3- and/or NH4+, and assimilated into amino acids. The aim of this work was to evaluate the N assimilation in this bromeliad. Plants were grown in vitro for seven months in modified MS medium with 15, 30, 60, and 90 mM of N as NO3-, NH4+ or NH4NO3, and then transferred to ex vitro conditions for acclimatization. Plants grown with NH4+ had high mortality. During acclimatization plants cultivated with 30, 60, and 90 mM of N as NH4NO3 showed higher biomass. With regard to N assimilation, GS and NR showed the highest activity in plants cultivated with NH4NO3, whereas plants cultivated with NH4+ had the highest GDH activity. Consequently, in vitro and ex vitro cultivation of this species with 60 mM N as NH4NO3 is recommended.

scholarly journals Sealing and explant types on the mangaba micropropagation

2012 ◽  
Vol 36 (4) ◽  
pp. 406-414 ◽  
Author(s):  
Aline de Jesus Sá ◽  
Ana da Silva Lédo ◽  
Carlos Alberto da Silva Lédo ◽  
Moacir Pasqual ◽  
Ana Veruska Cruz da Silva ◽  
...  
Keyword(s):  
In Vitro Propagation ◽  
Northeastern Brazil ◽  
Nodal Segments ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Botanical Variety ◽  
Pvc Film ◽  
Establishment Phase ◽  
The Ideal

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.

scholarly journals Utilization of Aseptic Seedling Explants for In vitro Propagation of Indian Red Wood

2013 ◽  
Vol 5 (4) ◽  
pp. 518-523 ◽  
Author(s):  
Kishore Kumar CHIRUVELLA ◽  
Arifullah MOHAMMED ◽  
Rama Gopal GHANTA
Keyword(s):  
Large Scale ◽  
Ex Situ Conservation ◽  
Cotyledonary Node ◽  
Multiple Shoot ◽  
Shoot Tip ◽  
Ex Situ ◽  
Nodal Segments ◽  
Ms Medium ◽  
Shoot Differentiation

Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae) was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS) medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1) supported highest average number and maximum multiple shoot differentiation (16.6). In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.

scholarly journals In Vitro Regeneration, Ex Vitro Rooting and Foliar Stoma Studies of Pseudostellaria heterophylla (Miq.) Pax

Agronomy ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 949
Author(s):  
Fengyun Wang ◽  
Xiaowei Xin ◽  
Hao Wei ◽  
Xiaohui Qiu ◽  
Boling Liu
Keyword(s):  
Stomatal Density ◽  
In Vitro Regeneration ◽  
Wild Plants ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Ex Vitro ◽  
Pseudostellaria Heterophylla ◽  
Stomatal Apparatus

Pseudostellaria heterophylla, in the family Caryophyllaceae, is an important Chinese medicinal plant commonly used to treat various diseases in children and valued for its ornamental properties. In this study, nodal segments were obtained from wild plants and used as explants to develop an efficient micropropagation protocol for this species. Murashige and Skoog (MS) medium supplemented with 1.5 mg·L−1 6-benzyladenine (6-BA) was the most suitable medium for inducing axillary buds and enhancing their growth, and MS medium containing 0.1 mg·L−1 indole-3-butyric acid (IBA) was the most effective for inducing in vitro rooting. To reduce labor, time, and cost, microshoots were rooted under ex vitro conditions. Pretreatments of the shoots with 100 mg·L−1 naphthaleneacetic acid (NAA) for 1 min ensured successful rooting in 86.7% of shoots. Comparison of the leaf microstructure between in vitro- and ex vitro-rooted plantlets revealed abnormal stomatal apparatus in the former. The stomatal apparatus of ex vitro plantlets were normal, although the stomatal density was reduced, which indicated that these plantlets were more likely to be able to adapt to environmental conditions in the field. We identified the optimal medium for P. heterophylla multiplication with respect to increased rooting efficiency of micropropagated shoots under ex vitro conditions. This results presented here will be helpful for agricultural cultivation of P. heterophylla.

scholarly journals Development of in vitro culture establishment conditions and micropropagation of grapevine rootstock cultivar ‘Ruggeri-140’

2021 ◽  
Vol 39 ◽  
pp. 03002
Author(s):  
Gayane Melyan ◽  
Andranik Barsegyan ◽  
Narek Sahakyan ◽  
Kima Dangyan ◽  
Yuri Martirosyan
Keyword(s):  
In Vitro Culture ◽  
Shoot Proliferation ◽  
Culture Conditions ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Resistant Rootstock

Optimization of in vitro culture conditions of grapevine phylloxera-resistant rootstock cultivar ‘Ruggeri-140’(Vitisberlandieri x Vitisrupestris) was carried out. Among the different sterilization treatments, maximum aseptic cultures were obtained for both explants apical tips and nodal segments when treated with Ca(ClO)2 at concentration of 1.5 % for 10 minutes plus 70 % ethanol for 30 s (T7). The maximum shoot proliferation was observed both in apical and nodal meristems cultured on MS medium supplemented with 1.0 mg/l BAP. MS/2 medium containing 1.0 mg/l indole-3-butric acid (IBA) gave the highest rooting percentage (100%) with the highest mean number and length of roots. The ex vitro survival of rooted micro shoots was 75.0%.

scholarly journals Efficient In Vitro Propagation by Ex Vitro Rooting Methods of Artemisia absinthium L., an Ethnobotanically Important Plant

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Anthropogenic Activities ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Ex Vitro ◽  
Artemisia Absinthium ◽  
Genetic Stock ◽  
In The Wild ◽  
Important Medicinal Plant

Artemisia absinthium is an important medicinal plant. Owing to the increasing anthropogenic activities and demand from the pharmaceutical industry, this plant species is overexploited; thereby this endangered its genetic stock in the wild. Therefore, it is urgently needed to develop nonconventional methods for conservation of A. absinthium. Nodal segments obtained from the field grown 2-month-old plants were used as explants. Murashige and Skoog (MS) medium containing 0.5 mg/L 6-benzylaminopurine (BAP) and 0.25 mg/L kinetin (Kn) were reported to be optimum for induction of shoots (6.0 ± 0.52 shoots per explant). The shoots were multiplied by repeated transfer of original explants and by subculturing of in vitro raised shoots on MS medium augmented with 1.0 mg/L each of BAP and Kn and 0.1 mg/L α-naphthaleneacetic acid (NAA). All in vitro regenerated shoots (100%) were rooted (4.4 ± 0.35 roots) on one-fourth strength MS medium supplemented with 2.0 mg/L indole-3 butyric acid (IBA). Cent percentage shoots rooted ex vitro on sterile Soilrite under the greenhouse conditions when the shoots were treated with 200 mg/L of IBA for 5 min. Plantlets rooted in vitro and ex vitro were acclimatized successfully in the greenhouse and exhibited 87% and 95% survival rate.

scholarly journals In vitro propagation and cytological analysis of Sophora mollis Royle: an endangered medicinal shrub

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Aakriti Bhandari ◽  
Harminder Singh ◽  
Amber Srivastava ◽  
Puneet Kumar ◽  
G. S. Panwar ◽  
...  
Keyword(s):  
Chromosome Number ◽  
Average Length ◽  
Germplasm Conservation ◽  
Basic Chromosome Number ◽  
Shoot Tip ◽  
Ex Situ ◽  
Ms Medium ◽  
Basic Chromosome ◽  
Shoot Tip Explants

Abstract Background Sophora mollis Royle (family Fabaceae, subfamily-Papilionaceae) is a multipurpose legume distributed in plains and foothills of the North-West Himalaya to Nepal and is facing high risk of extinction due to habitat loss and exploitation by the local people for its fuel and fodder values. Therefore, the present study was conducted to standardize a micropropagation protocol for Sophora mollis by using shoot tip explants and to study the meiotic chromosome count in the species. Results Multiple shoots were induced in shoot tip explants of Sophora mollis in Murashige and Skoog medium supplemented with different concentrations of cytokinins alone (BAP, TDZ, and Kinetin) and in combination with varying concentrations of NAA. MS medium supplemented with BAP (8.9 μM) was observed to be the optimal medium for multiple shoot induction and maximum 25.32 shoots per explant was obtained with average length of 4.5 ± 0.8 cm. In vitro developed shoots were transferred onto rooting media supplemented with different concentrations of auxin (IAA, IBA, and NAA). Maximum 86% rooting was observed in half-strength MS medium supplemented with 21.20 μM NAA with an average of 21.26 roots per culture. In vitro raised plantlets were adapted to greenhouse for better acclimatization and 60% plants were successfully transferred to the open environment. Based on the chromosome counts available from the literature and the current study, the species tend to show a basic chromosome number of x = 9. Conclusion The micropropagation protocol standardized can be helpful for the ex situ mass multiplication and germplasm conservation of the endangered species. Moreover, the ex situ conservation approach will be helpful in actively bridging the gap between ex situ and in situ approaches through the reintroduction of species in the wild. The cytological studies revealed the basic chromosome number x = 9 of the species.

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