Micropropagation and In Vitro Inflorescence of Pentas lanceolata

Author(s):  
Kitti Bodhipadma ◽  
Sompoch Noichinda ◽  
Chutikarn Tangtivaporn ◽  
Saowaros Phanomchai ◽  
David W. M. Leung

In this study, different concentrations of 6-benzyladenine (BA) on in vitro shoot and inflorescence inductions of P. lanceolata were investigated. The in vivo and in vitro floral characteristics of this plant were also compared. Nodal explants of P. lanceolata were cultured vertically with the cut ends inserted into semi-solid Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1, 2, 4, and 8 mg L–1 BA. The results showed that the explants formed the highest numbers of shoots even when cultured in MS basal medium without any addition of BA, while the shoots formed in the explants cultured in MS medium supplemented with 1 mg L–1 BA were the longest. No inflorescence was found in the shoots cultured in MS medium supplemented with 8 mg L–1 BA, while the highest percentage of inflorescence induction was found in the shoots cultured in the medium supplemented with 0.5 mg L–1 BA. The apperances of in vivo and in vitro flowers of P. lanceolata were the same in many aspects except that the number of flower/inflorescence formed was different. In addition, water accumulation was observed only inside the in vitro flowers. Water deposit in the long tubular structure of P. lanceolata flower could cause anther injury, suggesting that flowers developed in vitro may not always produce pollen.

2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis


2018 ◽  
Vol 30 (2) ◽  
pp. 283-294 ◽  
Author(s):  
Mani Manokari ◽  
Mahipal S. Shekhawat

Abstract The present study reports an efficient in vitro propagation system for Turnera ulmifolia using nodal segments as explants. Turnera ulmifolia (Passifloraceae) is an important garden plant with multipotent medicinal values. Effective shoot proliferation was achieved on agar gelled MS medium (Murashige and Skoog, 1962). The maximum number of shoots (8.3 ± 0.57) per initial explant was obtained on MS medium supplemented with 8.88 mM of 6-benzylaminopurine (BAP) and 0.54 mM of α-naphthalene acetic acid (NAA). The highest number of shoots (59.5 ± 2.10) proliferated on semi-solid MS medium (with agar) augmented with 2.22 mM of BAP and 2.32 mM of kinetin (Kin) along with 0.54 mM of NAA. Longer (4-5 cm) and healthy shoots were rooted (12.0 ± 0.10 roots per shoot) on half-strength MS medium fortified with 9.84 mM of indole-3 butyric acid (IBA). The in vitro regenerated plantlets were hardened in the greenhouse and transferred to the field. Significant developmental changes were observed in the foliar micromorphology of in vitro raised plantlets when these were transferred to the field. The stomatal index was gradually reduced (26.72 to 21.25) in the leaves from in vitro to field environments. But, vein-islets and veinlet terminations (13.4 and 7.6) were increased (39.7 and 18.4) respectively from in vitro to in vivo grown plants. Simple, unicellular, less frequent and underdeveloped trichomes were observed with the leaves of in vitro plants but fully developed trichomes recorded in the field transferred plants. The study could help in understanding the response and adaptation of tissue culture raised plantlets towards changed environmental conditions.


Author(s):  
Ajay Bhardwaj ◽  
T. Pradeepkumar ◽  
C. Varun Roch

A micropropagation protocol for parthenocarpic gynoecious cucumber reduces the burden of producing the seeds for each generation and their maintenance in-vivo. Thus an experiment was conducted in order to regenerate the plants in-vitro to check their performance after micropropagation. The micropropagation resulted in maximum shoot initiation (100%) from seedling excised cotyledonary explants with half strength MS medium supplemented with 0.5 mg/l IAA and 2 mg/l BAP along with half strength MS medium supplemented with 0.25 mg/l IAA for rooting and from stem nodal explants with Full MS + 1.5 mg/l IAA + 2 mg/l BAP media whereas half strength MS media without any hormones resulted in rooting and in both cases there were in-vitro flowers and change in their sex expression while grown in in-vivo conditions. On an average 61.11 and 48.15 percent survival was recorded from the plants regenerated through cotyledonary explants and stem nodal explants respectively. Out of five survived plants from regenerated parthenocarpic genotype CS 131 three showed monoecious sex expression and two exhibited gynoecious (parthenocarpic) sex expression. Mixed response of sex expression was evident in the regenerated parthenocarpic and gynoecious genotypes.


2012 ◽  
Vol 7 (4) ◽  
pp. 698-707 ◽  
Author(s):  
Ely Zayova ◽  
Ira Stancheva ◽  
Maria Geneva ◽  
Maria Petrova ◽  
Rumiana Vasilevska-Ivanova

AbstractAn effective in vitro protocol for rapid clonal propagation of Echinacea purpurea (L.) Moench through tissue culture was described. The in vitro propagation procedure consisted of four stages: 1) an initial stage - obtaining seedlings on Murashige and Skoog (MS) basal medium with 0.1 mg L−1 6-benzylaminopurine, 0.1 mg L−1 α-naphthalene acetic acid and 0.2 mg L−1 gibberellic acid; 2) a propagation stage — shoot formation on MS medium supplemented with 1 mg L−1 6-benzylaminopurine alone resulted in 9.8 shoots per explant and in combination with 0.1 mg L−1 α-naphthalene acetic acid resulted in 16.2 shoots per explant; 3) rooting stage — shoot rooting on half strength MS medium with 0.1 mg L−1 indole-3-butyric acid resulted in 90% rooted microplants; 4) ex vitro acclimatization of plants. The mix of peat and perlite was the most suitable planting substrate for hardening and ensured high survival frequency of propagated plants. Significant higher levels were observed regarding water-soluble and lipid-soluble antioxidant capacities (expressed as equivalents of ascorbate and α-tocopherol) and total pnenols content in extracts of Echinaceae flowers derived from in vitro propagated plants and adapted to field conditions in comparison with traditionally cultivated plants.


2020 ◽  
Vol 12 (1) ◽  
pp. 74-89
Author(s):  
Michael S. AKINROPO ◽  
Benjamin E. AYISIRE ◽  
Ejeoghene R. OGBIMI

This study was conducted to investigate the in vitro callus induction and rapid shoot regeneration potential in Enterolobium cyclocarpum, a plant native to central Mexico but widely introduced into Africa. The leaf, stem and nodal explants of E. cyclocarpum were cultured on full strength Murashige and Skoog (MS) medium supplemented with different concentrations of Cytokinins - Benzyladenine (BA) and/or Kinetin and Auxins - Naphthalene acetic acid (NAA) and/or 2,4-Dichlorophenoxylacetic acid (2,4-D) each alone and in combination.  The leaf explants did not respond to these treatments.  The Nodal explants were best for caulogenesis, while the explant responses were in the order- nodal > stem > cotyledon for callogenesis in MS medium supplemented with BA and/or Kin combined with NAA and/or 2,4-D. The varied combinations induced white compact callus.  The highest callus production was observed on MS medium supplemented with 2.7 µM NAA + 2.2 µM BA and 5.4 µM NAA alone.  Nodal and cotyledon explants developed callus and multiple shoots on MS supplemented with a combination of cytokinin (BA and/or Kin.) and auxin (NAA and/or 2,4-D). The maximum number of 3.98 ± 0.37 and 2.1±0.11 shoots/explants were recorded for nodal and cotyledon explants on MS medium supplemented with a combination of 8.8 µM BA+2.7 µM NAA and 2.2µM BA+2.7 µM NAA respectively.  On the basal medium, 10% of the excised shoots rooted successfully. Thus, this in vitro method can be exploited for conservation and mass propagation of this fast timber yielding tree and also utilized for embryogenesis studies.


2017 ◽  
Vol 6 (2) ◽  
pp. 5230 ◽  
Author(s):  
Miss. Bhawna ◽  
Santosh Kumar* ◽  
Rajeev Kumar

Justicia adhatoda L. (Family: Acanthaceae) has been exploited in India because of rich source of polyphenolic compounds which are used in stress related therapy. The Pharmacognostical study includes preliminary phytochemical screening of alkaloids, phenols, flavonoids and saponins which were confirmed in both in vivo grown garden plants as well as in vitro generated plantlets of J adhatoda. Explants inoculated on MS medium supplemented with different combination of auxins and cytokinins were successfully employed; combination of NAA (0.5µg/ml) and BA (2.0µg/ml) for nodal explants was found to be effective concentration to obtain plant regenerants from nodal explants. Rooting in the regenerants was also successfully initiated on transfer into half MS basal medium, supplemented with IBA (1.0 µg/ml). Addition of ADS (0.5µg/ml) in the above combinations of phytohormones induced multiple shoot from nodal explants. After hardening for 35 days in highly humidified acclimatized room, nearly 70 % of rooted plantlets survived in the field. Phytochemical screening revealed the presence of alkaloids, phenolic compounds, flavonoids and saponins in the methanol and aqueous extracts of both in vivo grown garden plants and in vitro genereated plantlet samples. Further analysis of the extracts on Thin Layer Chromatography and spectrophotometric estimation confirmed the presence of vasicine alkaloids in these extract samples with marked difference in quantity.


2018 ◽  
Vol 24 (4) ◽  
pp. 361-370
Author(s):  
Julián Guariniello ◽  
Jésica Iannicelli ◽  
Patricia Angélica Peralta ◽  
Alejandro Salvio Escandón

Achyrocline satureioides is a shrub native from South America. In popular medicine it is used in infusions such as digestive, carminative, antispasmodic, eupeptic and emmenagogue. However, its main use is as an ingredient in the liquor industry. Commercial exploitation is carried out through the collection of natural populations in an unsustainable way. The micropropagation of A. satureioides will allow its massive propagation and it will settle a base for its domestication. For this, a clone denominated as M1-5 was first propagated by cuttings. Subsequently, nodal segments obtained from young stems were disinfected by a standard method and cultured on MS medium. These shoots were used as a source of explants for subsequent assays. For its in vitro establishment MS medium and WPM were tested. Once the culture was established, the responses of the explants to increasing concentrations of 6-benzylaminopurine (BAP) (0.0; 0.5; 2.5 and 5.0 μM) with and without 0.05 μM α-naphthalene acetic acid (NAA) on WPM as basal medium were studied during 35 days. The proliferation of buds, the presence of callus and the number and length of the roots were evaluated. All of the “macela” cuttings in vivo propagated rooted and developed satisfactorily under the conditions tested. The application of 5.0 μM BAP alone generated the best multiplication rate, so it was selected as the multiplication medium. De novo shoots rooted spontaneously and finally, transferred to the greenhouse. Here in it was possible to establish a micropropagation protocol not only for the production of plantlets of selected clones but also for the application of biotechnological tools in the development of A. satureioides germplasm.


2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari

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.


2011 ◽  
Vol 39 (1) ◽  
pp. 84 ◽  
Author(s):  
Kantamaht KANCHANAPOOM ◽  
Suttinee JINGJIT ◽  
Kamnoon KANCHANAPOOM

A protocol for the regeneration of Gypsophila paniculata L. using nodal explants from 2-month-old field grown plants was established. The induction of multiple shoots was best obtained on Murashige and Skoog (MS) medium supplemented with 13.3 μM BA. Callus growth was observed on MS medium containing 44.3 μM BA. Calluses were transferred to MS medium supplemented with 2, 4-D (4.5, 13.5, 22.6 μM), NAA (5.3, 16.1, 26.8 μM) or BA (4.4, 13.3, 22.1 μM) for 2 months to induce shoot formation. After 6 weeks of initial culture, multiple shoots were regenerated from calluses cultured on MS medium supplemented with 13.3 μM BA. All regenerated shoots produced roots on 16.1 μM NAA containing MS medium within 4 weeks. Rooted plantlets were hardened and established in pots at 100% survival. For induction of in vitro flowering, regenerated shoots could be induced to flower efficiently when cultured on MS medium containing 13.3 μM BA and 50 g/l sucrose.


HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.


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