scholarly journals Micropropagation of Bioencapsulation and Ultrastructural Features of Sainfoin (Onobrychis viciifolia) GrownIn VivoandIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-12
Author(s):  
Sadegh Mohajer ◽  
Rosna Mat Taha ◽  
Minoo Mohajer ◽  
Arash Khorasani Esmaeili
Keyword(s):  
Water Conservation ◽  
Adventitious Shoots ◽  
Ms Medium ◽  
Onobrychis Viciifolia ◽  
Half Strength ◽  
Somatic Embryogenic ◽  
First Time ◽  
Indole 3 Acetic Acid

To explore the potential ofin vitrorapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifoliaScop. syn.Onobrychis sativaL.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0–2.0 mg L−1) and Indole-3-acetic acid (0–2.0 mg L−1) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1 : 1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques inOnobrychis viciifoliawere evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed inin vivoleaves compared toin vitroleaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) inin vivoand 0.121 (abaxial), 0.201 (adaxial) inin vitroleaves.

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 Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli
Keyword(s):  
New Media ◽  
Mineral Composition ◽  
Culture Media ◽  
Dry Weight ◽  
Defined Medium ◽  
Ms Medium ◽  
Media Formulation ◽  
Half Strength

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.

scholarly journals Phenotypic aberrations during micropropagation of Soymida febrifuga (Roxb.) Adr. Juss

2014 ◽  
Vol 6 (1) ◽  
pp. 99-104 ◽  
Author(s):  
Kishore Kumar CHIRUVELLA ◽  
Arifullah MOHAMMED ◽  
Rama Gopal GHANTA
Keyword(s):  
Seed Viability ◽  
Ms Medium ◽  
Stem Cuttings ◽  
Endemic Plant ◽  
Seedling Mortality ◽  
Root Regeneration ◽  
Half Strength ◽  
Shoot Necrosis

Like most of the medicinal plants Soymida febrifuga (Meliaceae) possess significance for its valuable secondary metabolites. Multiplication of this endemic plant is limited by difficulty in rooting of stem cuttings, high seedling mortality rates and low seed viability period. Hence efficient protocols for in vitro mass propagation has been established from field grown and aseptic seedlings explants. Strikingly, we observed aberrant structures such as vitrified shoots, faciated shoots, albino shoots as well shoot necrosis during its micropropagation. These phenotypic maladies were observed during organogenesis and rooting. Compared to other abnormalities, shoot necrosis nonetheless was frequent and pronounced leading to plant death. Shoots when subjected to rooting also displayed necrosis which was controlled by transferring to MS medium containing various concentrations and combinations of calcium levels, activated charcoal, glucose, fructose and auxins. Microshoots initiated roots on half strength MS medium with IBA and IAA individually or in combination within two weeks. MS half strength solid medium supplemented with CAN (556 mg l–1), CAP (1.0 mg l–1), IAA (2.0 mg l–1) and IBA (2.0 mg l–1) in combination was found to be more efficient in showing high frequency (95%) of root regeneration. Rooted plantlets were successfully hardened and 70-85% of regenerated plants were successfully acclimatized to natural environment. In vitro derived plantlets were morphologically similar to in vivo plants.

scholarly journals Improved micropropagation and foliar micromorphological studies in Turnera ulmifolia L. – An important medicinal plant

2018 ◽  
Vol 30 (2) ◽  
pp. 283-294 ◽  
Author(s):  
Mani Manokari ◽  
Mahipal S. Shekhawat
Keyword(s):  
Shoot Proliferation ◽  
Nodal Segments ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Half Strength ◽  
Turnera Ulmifolia ◽  
Semi Solid ◽  
Number Of Shoots

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.

scholarly journals Non-symbiotic Seed Germination and In vitro Plant Development of Pholidota articulata

2021 ◽  
Vol 15 ◽  
pp. 44-51
Author(s):  
R. Devendra Prasad ◽  
Shreeti Pradan ◽  
Mukti Ram Poudel ◽  
Bijaya Pant
Keyword(s):  
Acetic Acid ◽  
Seed Germination ◽  
Plant Production ◽  
Sustainable Utilization ◽  
Ms Medium ◽  
Natural Habitats ◽  
Half Strength ◽  
Indole 3 Acetic Acid ◽  
Symbiotic Seed Germination

Pholidota articulata is an epiphytic orchid mostly used in ornamental cut/pot flower and in traditional medicine. As it has high ornamental and medicinal values, its population from natural habitats is decreasing, therefore, it is listed in the Appendix-II of Convention on International Trade in Endangered Species (CITES). The objective of the present study is to obtain the in vitro plants of P. articulata from seed culture to conserve its germplasm. The in vitro seed germination was carried out in different strengths of Murashige and Skoog (MS) and Knudson C (KnC) medium supplemented with various plant hormones. On the half-strength of MS medium, seeds were started to germinate after 4 weeks of primary culture and they were developed into protocorms with first leaf primordium earlier than on the other medium. Therefore, in vitro developed protocorms were sub-cultured on the half-strength of MS medium supplemented with different concentrations of 6-benzylaminopurine (BAP), gibberellic acid (GA3) and α-naphtalene acetic acid (NAA). They were successfully developed into shoots on the 1.5 mg/l BAP supplemented half-strength of MS medium. Later, they were inoculated on the half-strength of MS medium supplemented with different concentration of α-napthalene acetic acid (NAA), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) for the root formation, where IBA supplemented medium was found effective for the development of roots. Thus, this study provides a reliable protocol for non-symbiotic seed germination and plant production, and reveals that seed-derived protocorms are good explants for the in vitro mass propagation for conservation and sustainable utilization in horticulture.

scholarly journals In vitro propagation of Nanorrhinum ramosissimum (Wall.) Betsche: A traditionally important medicinal plant

2021 ◽  
Vol 48 (3) ◽  
Author(s):  
Jyotsna Sharma ◽  
◽  
Anuja Koul ◽  
Savita Sharma ◽  
Raju Shankarayan ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Shoot Organogenesis ◽  
Germplasm Conservation ◽  
Ms Medium ◽  
In Vitro Plantlets ◽  
Half Strength ◽  
Shoot Tip Explants ◽  
Indole 3 Acetic Acid ◽  
Important Medicinal Plant

An efficient micropropagation protocol facilitates successful conservation and improvement of Nanorrhinum ramosissimum (Wall.) Betsche by biotechnological means. Shoot tip explants exhibited optimal organogenic response when inoculated on half-strength(1/2) Murashige and Skoog (MS) medium supplemented with kinetin (KN) and indole-3-acetic acid (IAA) (0.5 mg/L each). Shoot organogenesis was further enhanced when the multiplication medium was fortified with dextrose (1%) (2.6 shoots/explant; 7.9 cm shoot length). The regenerated shoots formed roots; however, the best rooting frequency (87%) was achieved on half-strength MS medium containing only IAA (0.5 mg/L). Four-week-old in vitro plantlets were acclimatized with 95% survival under greenhouse conditions. The regeneration protocol developed in this study can be utilized for germplasm conservation of this elite traditional medicinal plant.

scholarly journals Effects of N-Benzyl -9-(2-tetrahydropyranyl and Indole –3-Acetic Acid In Vitro Culture of Bauhinia purpurea L.

2019 ◽  
pp. 238-242
Author(s):  
Belai Meeta Suwal Singh
Keyword(s):  
Analytical Procedure ◽  
Multiple Shoots ◽  
Nodal Explants ◽  
Leguminous Plant ◽  
Ms Medium ◽  
Half Strength ◽  
Bauhinia Purpurea ◽  
Mature Seeds ◽  
Indole 3 Acetic Acid

<p>Bauhinia purpurea L. is a leguminous plant moderate sized tree with multipurpose value. It is distributed in sub-Himalayan tracts. It has been cultivated in the plain region up to the elevation of 1350 m. Mature seeds of Bauhinia purpurea L. were cultured on half strength Murashige and Skoog (1962) (MS) medium. Nodal explants obtained from germinated seedlings were cultured on MS medium containing 0.5 M BAP produced multiple shoots which were used for experimental purposes. Nodal explants obtained from cultured were subcultured on different concentrations of N-Benzyl -9-(2-tetrahydropyranyl) (BPA) and Indole-3acetic acid (IAA). The best proliferation of nodes and shoots were observed on the MS medium supplemented with 0.5 M BPA and 0.1 M IAA. After 8 weeks of culture, the propagated plants were acclimatized and transferred to the sand box containing 1:1 soil and sand. Well rooted plants were then established in the field. All the data collected were worked out statistically with SPSS, a system of analytical procedure.</p>

scholarly journals Micropropagation, Micromorphological Studies, andIn VitroFlowering inRungia pectinataL.

Scientifica ◽  
2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari ◽  
C. P. Ravindran
Keyword(s):  
Tissue Culture ◽  
Shoot Multiplication ◽  
Developmental Changes ◽  
Bud Break ◽  
Ms Medium ◽  
Half Strength ◽  
Indole 3 Acetic Acid ◽  
Culture Protocol ◽  
Important Medicinal Plant

A tissue culture protocol was developed for an important medicinal plantRungia pectinataL. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2solution. Murashige and Skoog (MS) medium was used to establish the cultures ofR. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L−16-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L−1each of BAP and kinetin (Kin) + 0.1 mg L−1indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L−1indole-3 butyric acid (IBA).In vitroflowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots underin vitroconditions.

scholarly journals Direct Organogenesis from Rhizome Explants in Marsilea quadrifolia L.: A Threatened Fern Species

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Root Induction ◽  
Ms Medium ◽  
Full Strength ◽  
Marsilea Quadrifolia ◽  
Fern Species ◽  
Half Strength

An efficient micropropagation protocol has been developed for Marsilea quadrifolia L. through direct organogenesis. The mature rhizomes were used as explants and successfully sterilized using 0.1% HgCl2 for the establishment of cultures. The multiple shoots were differentiated from the explants on Murashige and Skoog (MS) medium augmented with 6-benzylaminopurin (BAP). Full strength MS medium was reported to be effective for the induction of sporophytes from the rhizomes after four weeks of inoculation. Maximum response (96%) with average of 6.2 shoots (2.72 cm length) was achieved on full strength of MS medium augmented with 0.5 mg/L BAP in culture initiation experiments. The cultures were further proliferated in clusters (79.0±0.37 shoots per explant) with stunted growth on half strength MS medium supplemented with 0.25 mg/L BAP after four weeks. These stunted shoots were elongated (5.30 cm long) on half MS medium devoid of growth hormones. Root induction and proliferation (3.0–4.0 cm long) were observed after 4th subculture of sporophytes on hormone-free half strength MS medium. The rooted plantlets were hardened in the fern house for 4-5 weeks and transferred to the field with 92% survival rate. There were no observable differences in between in vivo grown and in vitro propagated plantlets in the field.

scholarly journals In-vitro Flowering and in-vivo Sex Expression of Micropropagated Parthenocarpic Gynoecious Cucumber

2020 ◽  
pp. 17-24
Author(s):  
Ajay Bhardwaj ◽  
T. Pradeepkumar ◽  
C. Varun Roch
Keyword(s):  
Sex Expression ◽  
Nodal Explants ◽  
Ms Medium ◽  
Percent Survival ◽  
Shoot Initiation ◽  
Mixed Response ◽  
Cotyledonary Explants ◽  
Half Strength

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.

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