Rapid Micropropagation Protocol of Mucuna pruriens var. utilis using Cotyledonary Node Explant: A Cultivated Medicinal Edible Legume

2021 ◽  
Author(s):  
Sanjay Kumar Madkami ◽  
Arpita Moharana ◽  
Durga Prasad Barik
Keyword(s):  
Seed Germination ◽  
Large Scale ◽  
Shoot Multiplication ◽  
Cotyledonary Node ◽  
Production Method ◽  
Commercial Production ◽  
Mucuna Pruriens ◽  
Garden Soil ◽  
Ms Medium

Background: The presence of L-dopa coupled with rich protein and amino acid marked Mucuna pruriens var. utilis as an important under-utilised legume. Therefore, it is useful to develop a method for large-scale multiplication for commercial production. Method: Tissue culture technology is successfully utilized in propagation of plants with poor and uncertain response to conventional propagation. Murashige and Skoog’s (MS) medium without any Plant Growth Regulators (PGRs) was used for seed germination and with PGRs for shoot and root multiplication.Result: Highest 95% seed germination was found in fresh seeds at 7-8 days of culture. Shoot multiplication percentage was found to be 100% with highest c.a. 21.1 shoots with an average 4.8 cm shoot length on MS + BAP 1.5 mg L-1 per 10 days old cotyledonary node explant. A total c.a. 144 shoots were harvested after 3rd harvest of the mother cotyledonary node with two whole cotyledons at day 70. Rooting was best induced in in vitro derived shoots on MS medium without any PGRs and plantlets were acclimatized in sand and soil (1:1) and established in pot with garden soil. 

In vitro propagation of Camellia fascicularis: a plant species with extremely small populations

2020 ◽  
Vol 100 (2) ◽  
pp. 202-208
Author(s):  
Mengting Wang ◽  
Guiliang Zhang ◽  
Peiyao Xin ◽  
Yun Liu ◽  
Bin Li ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Large Scale ◽  
In Vitro Regeneration ◽  
Germplasm Conservation ◽  
Shoot Multiplication ◽  
Shoot Tips ◽  
Naphthalene Acetic Acid ◽  
Multiplication Rate ◽  
Ms Medium

Camellia fascicularis is an endangered evergreen ornamental plant with pale yellow flowers. An efficient and reproducible in vitro regeneration method is required for its large-scale propagation and germplasm conservation. In this study, one axillary bud per nodal stem was obtained from C. fascicularis cultured on Murashige & Skoog (MS) medium containing 0.1 mg L−1 indole-3-acetic acid (IAA) combined with 1.0 mg L−1 6-benzylaminopurine (BA). Axillary buds from the stem segments were transferred to modified woody plant medium (WPM) supplemented with 3.0 mg L−1 BA in combination with 0.3 mg L−1 IAA for multiplication, thereby resulting in a high shoot multiplication rate of 6.8. Multiple shoots were divided into nodal stems and shoot tips and were induced to root. The shoot tips were induced to root by culturing on one-half MS medium supplemented with 2.0 mg L−1 indole-3-butyric acid (IBA) in combination with 0.3 mg L−1 α-naphthalene acetic acid (NAA), which resulted in 76.0% rooting efficiency with 2.3 roots per shoot. The optimal hormone ratio for inducing rooting of nodal stems was 1.0 mg L−1 IBA in combination with 2.0 mg L−1 NAA, which resulted in 72.7% rooting efficiency with 1.7 roots per nodal stem. These two rooted plantlets were successfully acclimatized and established in a greenhouse.

scholarly journals Influence of Additives on Enhanced In vitro Shoot Multiplication of Orthosiphon aristatus (Blume) Miq.

2013 ◽  
Vol 5 (3) ◽  
pp. 338-345 ◽  
Author(s):  
Swarna JAYAKUMAR ◽  
Ravindhran RAMALINGAM
Keyword(s):  
Large Scale ◽  
Shoot Multiplication ◽  
Ms Medium ◽  
Morphological Variations ◽  
Different Types ◽  
Mother Plants ◽  
Half Strength ◽  
Different Parts ◽  
Number Of Shoots

Orthosiphon aristatus is a valuable medicinal plant and different parts of the plant are pharmaceutically used for the treatment of various diseases. The present study was designed to develop an efficient protocol for micropropagation of O. aristatus from nodal explants and to study the influence of additives on the enhancement of the number of shoots per explant. Among the different types of additives used, 10% coconut water and 30 mg/L glutamine added to Murashige and Skoog’s (MS) medium supplemented with 1.0 mg/L 6-benzyl amino purine (BAP) and 0.5 mg/L kinetin (KIN) was found to be most effective. Maximum number of shoots (44.07 ± 0.38) with 100% shooting response and shoot length of 7.47 ± 0.10 cm was recorded. In vitro rooting of the microshoots was achieved on half-strength MS medium containing 0.5 mg/L indole-3-butyric acid (IBA), producing an average of 30.27 ± 0.36 roots and 6.02 ± 0.20 cm root length. The rooted shoots were acclimatized with 100% survival rate on coco pith: soil (3:1) planting substrate and was successfully transferred to field conditions. The hardened plants exhibited homogeneity and no morphological variations were observed among the regenerants and the mother plants. Thus, the procedure described is a quick and reliable method which could be applied for efficient large-scale propagation, genetic transformation assays and secondary metabolite production.

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 of Camellia oleifera Abel. Using Hypocotyl, Cotyledonary Node, and Radicle Explants

HortScience ◽  
2016 ◽  
Vol 51 (4) ◽  
pp. 416-421 ◽  
Author(s):  
Ze Li ◽  
Xiaofeng Tan ◽  
Zhiming Liu ◽  
Qing Lin ◽  
Lin Zhang ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Cotyledonary Node ◽  
Direct Organogenesis ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Camellia Oleifera ◽  
Cotyledonary Nodes

Camellia oleifera Abel. is one of four major woody oil plants in the world. The objective of the current study was to evaluate the effect of different plant growth regulators (PGRs) and concentrations on direct organogenesis using cotyledonary nodes, hypocotyls, and radicle explants. High induction frequency of adventitious shoots were obtained from cotyledonary nodes, hypocotyls, and radicle explants (85.2%, 73.6%, and 41.0%, respectively) when cultured on half-strength Murashige and Skoog (1/2 MS) medium containing 2.0 mg·L−1 6-benzylaminopurine (BA) and 0.1 mg·L−1 indole-3-acetic acid (IAA). Microshoots from cotyledonary nodes, hypocotyls, and radicle explants were then transferred to 1/2 MS medium containing 2.0 mg·L−1 BA and 0.05 mg·L−1 indole-3-butyric acid (IBA) for shoot multiplication, resulting in 6.9 shoots per explant. The shoots were transferred to Woody Plant Medium (WPM) supplemented with various α-naphthalene acetic acid (NAA) and gibberellic acid (GA3) for shoot elongation. The mean length of shoots and the number of leaves per shoot were 3.7 and 6.6 cm, respectively, in WPM supplemented with 0.5 mg·L−1 NAA and 3.0 mg·L−1 GA3. The highest rooting of shoots (90.2%) or the number of roots per shoot (7.2) was obtained when elongated microshoots were transferred to 1/2 MS medium supplemented with 3.5% perlite, 1.0 mg·L−1 IBA and 2.0 mg·L−1 NAA. The rooted plantlets were successfully acclimatized in the greenhouse with a survival rate of 90.0%. The in vitro plant regeneration procedure described in this study is beneficial for mass propagation and improvement of C. oleifera through genetic engineering.

scholarly journals Chromosome stability of in vitro propagated Cucurbita cultivars

2019 ◽  
Vol 3 (3) ◽  
pp. 25
Author(s):  
Buse Dursun ◽  
Ahu Altınkut Uncuoğlu ◽  
Yıldız Aydin
Keyword(s):  
Chromosome Number ◽  
Cucurbita Pepo ◽  
Multiple Shoots ◽  
Cotyledonary Node ◽  
Garden Soil ◽  
Ms Medium ◽  
Ploidy Levels ◽  
Root Regeneration ◽  
Cucurbitaceae Family

Cucurbita pepo L. which is a member of Cucurbitaceae family, is a one-year plant with herbaceous stems, broad leaves and superficial scattered roots. Monoic flower structure in the Cucurbitaceae family and the differences in the maturing time of male and female organs in flowers cause an increase in the foreign fertilization rate. Therefore, there may be positive or negative changes in the existing characteristics of the species. Micro-propagation method can be performed in pumpkin species for clonal propagation, but genetic stability after tissue culture is an important consideration. Chromosome number and morphology are primary cytogenetic parameters that must remain stable after in vitro propagation. We analyzed plants of different hybrid pumpkin genotypes cultivated in our country (Ardendo, Angelina, Torpido, Roni, Sena Hanım) cytogenetically in order to determinate their stability level. Cotyledon nodes, nodes, shoot apex, hypocotyl and internode explants were prepared from the 4-week old C. pepo seedlings by making a horizontal slice through the hypocotyl region. Cotyledonary node explants produced multiple shoots and callus regeneration in MS medium in the presence of N6-benzylamino-purine BA 1 mg/L in Torpido genotype. Elongated shoots were excised from shoot clumps and transferred to rooting medium. The best root generation was obtained from Torpido and the most successful explant type for root regeneration was determined in cotyledon nodes as in shoot regeneration and MS medium without plant growth regulator showed the best root regeneration. The rooted plants were hardened in small pot containing standardized garden soil, well developed plant transferred to greenhouse. The chromosome number and karyotype analysis were determined in control and in vitro propagated Cucurbita pepo L. plants and ploidy levels were confirmed to be 2n = 40.

scholarly journals Develope Micro clonal -propagation protocol for Oxytenanthera abyssinica A.Rich. Munro to large scale micro-propagation

2020 ◽  
Author(s):  
Adugnaw Admas ◽  
Berhane Kidane ◽  
Melaku Admasu ◽  
Tesaka Misga
Keyword(s):  
Large Scale ◽  
Shoot Multiplication ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Shoot Initiation ◽  
Oxytenanthera Abyssinica ◽  
Micro Propagation ◽  
Viable Seeds ◽  
Propagation Methods

ABSTRACTIn Ethiopia, Oxytenanthera abyssinica A.Rich. Munro has varies economic importance. However, conventional propagation methods of O. abyssinica are generally inefficient due to their low multiplication rate, time consuming, labor intensive, and too costly. The objective of this study was to develop a protocol for mass micropropagation of O. abyssinica through seed culture. Murashige and Skoog (MS) medium augmented with 6-Benzylaminopurine (BAP) was used for shoot initiation and multiplication. For in vitro rooting, MS medium supplemented with 3-Indole –butric acid (IBA) was used.In shoot initiation experiment all viable seeds were proliferated in 5-7 days of culturing. In shoot multiplication at 0.004 g/L BAP was Sucssefuly shoot multiplied, also best root responding were found at 0.005 g/l IBA.The present optimized protocol enables for any acters who needs large numbers of low land bamboo seedling for industery, small and micro enterprize or for reafforestation programms.

scholarly journals In Vitro Seed and Clonal Propagation of the Mediterranean Aromatic and Medicinal Plant Teucrium capitatum

HortScience ◽  
2016 ◽  
Vol 51 (4) ◽  
pp. 403-411 ◽  
Author(s):  
Maria Papafotiou ◽  
Aekaterini N. Martini
Keyword(s):  
Seed Germination ◽  
Shoot Multiplication ◽  
Culture Conditions ◽  
Wild Plants ◽  
Ms Medium ◽  
Continuous Darkness ◽  
Half Strength ◽  
Concentrated Sulfuric Acid ◽  
And Storage

The effect of various pretreatments, culture conditions, and storage time on in vitro germination of seeds, as well as the effect of explant origin and plant growth regulators on in vitro propagation of Teucrium capitatum L. (Teucrium polium sp. capitatum Arcang., Lamiaceae) were examined. Seeds, collected from native plants and stored at room temperature for 3, 7, and 12 months, were cultured for germination in vitro in petri dishes with solid half-strength (½) Murashige and Skoog (1962) growth medium (MS) at 5, 10, 15, 20, 25, and 30 °C, and 16 hours light or continuous darkness. Pretreatments, such as cold stratification, scarification with sandpaper, dipping in concentrated sulfuric acid (H2SO4 > 95%), or dipping in boiling water were tested. Seeds without any pretreatment germinated at lower than 10%. Dipping in concentrated H2SO4 for 15 or 20 minutes was the most effective pretreatment, but still seed germination achieved was low (36%). Seeds preserved their germination capacity for at least 1 year, and germinated satisfactorily at a wide temperature range, from 15 to 25 °C (optimum), while photoperiod did not affect seed germination. Explants excised from in vitro-grown seedlings were established in vitro on MS medium with 1.0 mg·L−1 6-benzyladenine (BA) at much higher rates (≈90%) compared with those collected from plants grown from cuttings in a greenhouse (25%), while explants collected from adult wild plants failed to do so. Explants from seedlings showed strong variability in their response; those excised from branched seedlings formed shoots at significantly higher percentage (90%) at the establishment stage (cultured on MS medium with 0.5–2.0 mg·L−1 BA) than explants excised from unbranched seedlings (36% to 43%), while during subcultures on MS medium with 1.0 mg·L−1 BA, explants from branched seedlings also showed higher multiplication rates than those from unbranched ones. BA at 0.5–2.0 mg·L−1 induced shoot multiplication during both establishment and multiplication stages (7–8 and 14–15 shoots per explant, respectively), while kinetin and 6-γ-γ-(dimethylallylamino)-purine (2iP) were less effective than BA, and zeatin the least appropriate. Microshoot rooting was enhanced by 1-week culture on (½) MS medium with 1.0–4.0 mg·L−1 indole-3-butyric acid (IBA), followed by transfer to auxin-free, (½) MS medium (93% to 98%, 7–8 roots per microshoot), compared with culture on the same medium continuously for 5 weeks (69% to 80%, 5–6 roots per microshoot) or at lower IBA concentrations. Plantlets were acclimatized to ex vitro conditions at 98% on a peat–perlite (1:1, v/v) mixture.

scholarly journals Shoot Multiplication and Callus Induction of Labisia pumila var. alata as Influenced by Different Plant Growth Regulators Treatments and Its Polyphenolic Activities Compared with the Wild Plant

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3229
Author(s):  
Mat Yunus Najhah ◽  
Hawa Z. E. Jaafar ◽  
Jaafar Juju Nakasha ◽  
Mansor Hakiman
Keyword(s):  
Callus Induction ◽  
Antioxidant Activities ◽  
Shoot Multiplication ◽  
Wild Plants ◽  
Wild Plant ◽  
Total Phenolic ◽  
Nodal Segment ◽  
Ms Medium ◽  
In Vitro Plantlets

This study aims to investigate whether the in vitro-cultured L. pumila var. alata has higher antioxidant activity than its wild plant. An 8-week-old L. pumila var. alata nodal segment and leaf explants were cultured onto Murashige and Skoog (MS) medium supplemented with various cytokinins (zeatin, kinetin, and 6-benzylaminopurine (BAP)) for shoot multiplication and auxins (2,4-dichlorophenoxyacetic acid (2,4-D) and picloram) for callus induction, respectively. The results showed that 2 mg/L zeatin produced the optimal results for shoot and leaf development, and 0.5 mg/L 2,4-D produced the highest callus induction results (60%). After this, 0.5 mg/L 2,4-D was combined with 0.25 mg/L cytokinins and supplemented to the MS medium. The optimal results for callus induction (100%) with yellowish to greenish and compact texture were obtained using 0.5 mg/L 2,4-D combined with 0.25 mg/L zeatin. Leaves obtained from in vitro plantlets and wild plants as well as callus were extracted and analyzed for their antioxidant activities (DPPH and FRAP methods) and polyphenolic properties (total flavonoid and total phenolic content). When compared with leaf extracts of in vitro plantlets and wild plants of L. pumila var. alata, the callus extract displayed significantly higher antioxidant activities and total phenolic and flavonoid content. Hence, callus culture potentially can be adapted for antioxidant and polyphenolic production to satisfy pharmaceutical and nutraceutical needs while conserving wild L. pumila var. alata.

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 Micropropagation of the Medicinal Plant Physalis angulata L.

2016 ◽  
Vol 8 (2) ◽  
pp. 161-163
Author(s):  
Owk ANIEL KUMAR ◽  
Songa RAMESH ◽  
Sape SUBBA TATA
Keyword(s):  
Large Scale ◽  
Leaf Disc ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Medicinal Herb ◽  
Root Induction ◽  
Survival Percentage ◽  
Physalis Angulata

Physalis angulata L. is an important medicinal herb. An efficient direct adventitious plant regeneration protocol was developed for large scale propagation using leaf disc as explants. The explants were cultured on MS basal medium supplemented with 0.25-3.0 mg/L 6-benzyl amino purine (BAP) for primary shoot proliferation. Inclusion of indole-3-acetic acid (IAA) and gibberellic acid (GA3) in the culture medium along with BAP promoted a higher rate of shoot multiplication. The maximum number of shoots was produced in MS + BAP (1.0 mg/L) + IAA (0.5 mg/L) + GA3 (0.20 mg/L) after the third subculture. An average of 152.8 ± 0.40 shoots were produced from each leaf disc. For root induction the shootlets were transferred to MS medium supplemented with different concentrations of indole-3-butyric acid (IBA). The highest percentage of root induction was observed in 1.0 mg/L (IBA). Rooted plants were successfully established in the soil after hardening. The survival percentage of rooted plants on soil was found to be 85%. This result will facilitate the conservation and propagation of the important medicinal herb Physalis angulata L.

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