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 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.

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 In vitro regeneration protocol of Rauvolfia serpentina L.

2018 ◽  
Vol 53 (2) ◽  
pp. 133-138 ◽  
Author(s):  
S Khan ◽  
TA Banu ◽  
S Akter ◽  
B Goswami ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Root Induction ◽  
Ms Medium ◽  
Regeneration System ◽  
Rauvolfia Serpentina ◽  
Number Of Shoots

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13∙4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Bangladesh J. Sci. Ind. Res.53(2), 133-138, 2018

scholarly journals In Vitro Micropropagation of Orchid (Vanda tessellata L.) from Shoot Tip Explant

1970 ◽  
Vol 17 ◽  
pp. 139-144 ◽  
Author(s):  
MS Rahman ◽  
MF Hasan ◽  
R Das ◽  
MS Hossain ◽  
M Rahman
Keyword(s):  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Shoot Tip ◽  
Root Induction ◽  
Ms Medium ◽  
Culture Techniques

Context: Orchid produces a huge number of minute seeds but the seeds can not germinate easily in nature due to the lack of endosperm in the seeds is an incompatibility barrier that limits its propagation in nature. Objectives: To develop in vitro culture techniques for quick propagation of Vanda tessellate, a commercially important orchid species. Materials and Methods: Shoot tips were used as experimental materials. The explants were surface sterilized and the shoot tips were excised. The isolated shoot tips were cultured in MS medium supplemented with different concentration and combinations of auxin and cytokinin. Results: The combination of 1.5 mgl-1 NAA and 1.0 mgl-1 BAP was proved to be the best medium formulation for multiple shoot formation as well as maximum shoot elongation. The single shoots were isolated from the multiple shoots and subcultured in MS medium having NAA and IBA individually and in combinations for root induction. Maximum root induction was obtained in MS agarified medium having 0.5 mgl-1NAA and 1.0 mgl-1IBA. The well rooted plantlets were hardened successfully in the potting mixture containing coconut husk, perlite, charcoal, brick pieces in the ratio of 2:1:1:1 and eventually established under natural condition.Conclusion: An efficient regeneration protocol for micropropagation in V. tessellata through shoot tip culture has been established.Key words: Shoot tip; micropropagation; orchid.DOI: 10.3329/jbs.v17i0.7122J. bio-sci. 17: 139-144, 2009

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

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

scholarly journals Influence of the Genotype and TDZ on the in Vitro Regeneration of Hybrid Grapes

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 877B-877
Author(s):  
Maritza I. Tapia ◽  
Paul E Read
Keyword(s):  
In Vitro Regeneration ◽  
Shoot Proliferation ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Ms Medium ◽  
High Concentrations ◽  
Grape Cultivars

It has been previously demonstrated that thidiazuron (TDZ) enhanced the regeneration and multiple shoot proliferation of vinifera grape cultivars. To determine the effect of TDZ on the multiplication of hybrid grapes, in vitro nodal segments from cultivars Chancellor, Leon Millot, and Valiant were cultured on MS medium supplemented with 0, 0.01, 0.05, 0.1, 0.5, and 1.0 mg TDZ/liter. After 1 month, the higher percentage of rooted shoots was obtained from the explants cultured in medium containing the lowest concentration of TDZ (0.01 mg–liter–1) independent of the genotype. Multiple shoot proliferation was favored by high concentrations of TDZ (0.5 and 1.0 mg–liter–1). An average of 0.39 and 0.39 shoots, respectively, was obtained from `Chancellor' cultures, 0.56 and 0.59 from `Leon Millot', and 1.93 and 2.38 from `Valiant'. Vitrification and teratological structures were observed in all the cultures of the three genotypes, but less vitrification occurred in `Valiant' plantlets.

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 An efficient shoot regeneration system for medicinally important Elephantopus scaber Linn.

2015 ◽  
Vol 15 (2) ◽  
pp. 94-99 ◽  
Author(s):  
Jyothi Abraham ◽  
T. Dennis Thomas
Keyword(s):  
Age Groups ◽  
Cotyledonary Node ◽  
Multiple Shoot ◽  
Ms Medium ◽  
Shoot Induction ◽  
Regeneration System ◽  
Multiple Shoot Induction ◽  
Elephantopus Scaber ◽  
Cotyledonary Node Explants

An efficient protocol for the rapid micropropagation of medicinally important Elephantopus scaber has been standardized using cotyledonary node explants. Direct multiple shoot induction was observed when the cotyledonary node explants at various age groups were cultured on MS medium supplemented with various plant growth regulators. The highest shoot induction was obtained when the cotyledonary node explants from 20-day-old seedlings were cultured on MS medium supplemented with 1.5 mg L-1 TDZ and 0.5 mg L-1 NAA. On this medium, 98% of the cultures responded, with an average number of 33.7 shoots per explant. The highest frequency of rooting (100%) and mean number of roots (3.3 per shoot) were observed when the shoots were transferred to MS medium supplemented with 1.0 mg L-1 IBA. The plantlets raised in vitro were acclimatized and transferred to soil with a 92% success rate. The protocol described here may be utilized for multiplication and conservation of elite clones of E. scaber.

scholarly journals Efficient Micropropagation Protocol for the Conservation of the Endangered Aloe peglerae, an Ornamental and Medicinal Species

Plants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 506
Author(s):  
Nontobeko A. Hlatshwayo ◽  
Stephen O. Amoo ◽  
Joshua O. Olowoyo ◽  
Karel Doležal
Keyword(s):  
Conservation Status ◽  
Shoot Tip ◽  
Ex Situ ◽  
Rapid Decline ◽  
Ms Medium ◽  
Medicinal Species ◽  
Aromatic Cytokinin ◽  
African Plants ◽  
Over Exploitation

A number of Aloe species are facing an extremely high risk of extinction due to habitat loss and over-exploitation for medicinal and ornamental trade. The last global assessment of Aloe peglerae Schönland (in 2003) ranked its global conservation status as ‘endangered’ with a decreasing population trend. In the National Red List of South African Plants, the extremely rapid decline of this species has resulted in its conservation status being elevated from ‘endangered’ to ‘critically endangered’ based on recent or new field information. This dramatic decline necessitates the development of a simple, rapid and efficient micropropagation protocol as a conservation measure. An in vitro propagation protocol was therefore established with the regeneration of 12 shoots per shoot-tip explant within 8 weeks using Murashige and Skoog (MS) medium supplemented with 2.5 µM meta-topolin riboside (an aromatic cytokinin). The rooting of the shoots with a 100% frequency on half-strength MS medium without any plant growth resulted in additional six shoots produced per cultured shoot. The resultant plantlets were successfully acclimatized with a 100% survival frequency after 6 weeks. Overall, the developed protocol can result in the production of 3906 transplantable shoots that are ready for rooting per annum from a single shoot-tip explant. It is simple and efficient for seedling production in the ex situ cultivation and conservation of the endangered A. peglerae.

In vitro propagation of the threatened plant Sphaerophysa kotschyana (Fabaceae): inter simple-sequence-repeat (ISSR) analysis and salt tolerance of the regenerants

2013 ◽  
Vol 61 (1) ◽  
pp. 67 ◽  
Author(s):  
Semiha Erişen ◽  
Zeynep Öncel
Keyword(s):  
In Vitro Propagation ◽  
Ex Situ Conservation ◽  
Conservation Strategy ◽  
Ex Situ ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Shoot Height ◽  
Issr Analysis ◽  
Sphaerophysa Kotschyana

Sphaerophysa kotschyana is a threatened endemic species in Turkey and according to the Bern Convention, it is on the absolute preservation plant list. In vitro propagation methodologies were evaluated as an ex situ conservation strategy for this species. Nodal segments were cultured on Murashige and Skoog (MS) media with different cytokinins (benzyladenine, thidiazuron (TDZ) and zeatine), with or without auxin (α-naphthaleneacetic acid; NAA), to investigate shoot initiation. TDZ produced the highest number of shoots (11.0 shoots per explant) on MS medium at a concentration of 0.05 mg L–1. Rooting reached 100% when 0.5 mg L–1 NAA was combined with half strength MS and 1.5% sucrose and rooted plantlets were successfully acclimatised. Somaclonal variation of a mother plant and 10 regenerants was assessed using ISSR analysis. The same banding profiles were exhibited by all plants. In vitro response to salinity stress (NaCl) was also investigated in this halophytic species. Higher concentrations of NaCl negatively affected shoot multiplication, whereas shoot height was enhanced at 50 mM NaCl. These results suggest that the established protocol is an efficient and reliable system of in vitro propagation for ex situ conservation of S. kotschyana.

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