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 ◽  
...  

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.

Scientifica ◽  
2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari ◽  
C. P. Ravindran

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.


Author(s):  
Ashu Pandey ◽  
Oshin Verma ◽  
Suresh Chand

Boerhaavia diffusa L., also known as santhi, or punarnava is an important medicinal plant, belonging to the family Nyctaginaceae. This species is said to be distributed throughout Malwa plateau in central India, as per ayurvedic literature, but due to extensive commercial exploitation, the species has become vulnerable. For callus induction, leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4- dichlorophenoxy acetic acid (2, 4-D 2.26 µM-9.04 µM) and N 6-benzyladenine (BA 1.11 µM-4.44 µM), either alone or in combinations. Calli formed within 10-12 days of culture, followed by shoots regeneration within 20-25 days. Direct organogenesis was achieved from nodal explants in MS media fortified with 2,4-D (2.26 µM-9.04 µM) along with BA (1.11 µM-4.44 µM) within 20 days. Multiple shooting was observed during subculture of in vitro regenerated shoots when 2,4-D was replaced with α-naphthalene acetic acid (NAA). Rooting was achieved in MS medium fortified with 2.85 µM IAA, within 7-10 days and also on half strength MS medium containing 2.85 µM Indole-3-acetic acid (IAA). For hardening, regenerated plants, with roots (3-4cm) were initially maintained on half-strength MS liquid medium (MS1/2) without growth regulators followed by quarter strength MS (MS1/4) liquid medium for 10 days. For acclimatization sterile mixture of soil, sand and manure (2:1:1) was used. Survival rate of regenerated plants was nearly 100%.


2021 ◽  
Vol 13 (3) ◽  
pp. 897-906
Author(s):  
Amit ◽  
Rajkumar ◽  
Narender Singh

The present investigation aimed to standardize efficient plant regeneration protocol through in vitro culture by using nodal segment for mass multiplication of Lawsonia inermis an economically important medicinal plant species. Mass multiplication of shoots induced on Murashige and Skoog (MS) medium supplemented with different growth regulators like auxins and cytokinins separately and in different combinations. The medium fortified with 6-Benzylaminopurine ( BAP) 1.0 mg/l + kinetin (KN) 1.5mg/l  explained best compared to all other combinations. In vitro raised plantlets were excised and transferred in half strength MS  medium supplemented with different growth regulators like Indole Butyric acid ( IBA)  and naphthalene acetic acid (NAA ) (0.5-3.0 mg/l) in an experiment that gave rise to rooting. The half strength of MS medium additive with IBA in separate and in different combinations with NAA concentrations (0.5-3.0 mg/l) supported root development. The best response of rooting was obtained on half MS medium fortified with 1.0 mg/l IBA. The regenerated plantlets were successfully transplanted to pots. Regenerants were transferred to the field conditions and recorded the survival rate.. Among all the carbon sources and gelling agents used, sucrose (3%) in combination with 0.8 per cent agar-agar has proved significantly better. Multiple shoots formation with longer shoots were achieved on medium with 1.0mg/l BAP and 1.5mg/l Kn. Thus, it is possible to develop a large number of plants of L. inermis through shoot bud regeneration which can cater for the need of pharmaceutical as well as other industries.


Author(s):  
Aakriti Bhandari ◽  
Harminder Singh ◽  
Amber Srivastava ◽  
Puneet Kumar ◽  
G. S. Panwar ◽  
...  

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.


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.


2014 ◽  
pp. 125-136
Author(s):  
Marija Markovic ◽  
Mihailo Grbic ◽  
Dragana Skocajic ◽  
Matilda Djukic

The effect of the concentration of MS salts and explant type on D. serotinus rooting and acclimatization was investigated in order to optimize a protocol for the micropropagation of this species. The obtained results showed that explant type as well as the concentration of MS salts had a significant effect on rooting, and the highest rooting rate (85-86,7%) was achieved when culturing single-node cuttings and terminal buds on a half-strength MS medium supplemented with 0,5 mgL-1 NAA. Nevertheless, mean number of roots per explant was higher on the MS media (15,3-18,6) than on the half-strength MS media (11,8-13,4). The best acclimatization rate was obtained in a 4:1 mixture of peat and sand (83,3-86,7%). The explant type from which in vitro plantlets developed had no effect on the acclimatization rate.


2015 ◽  
pp. 99-112
Author(s):  
Marija Markovic ◽  
Dragana Skocajic ◽  
Mihailo Grbic ◽  
Matilda Djukic ◽  
Dragica Obratov-Petkovic ◽  
...  

The aim of this study was to determine the possibility of micropropagation of the medicinal plant A. millefolium on half-strength MS medium and ex vitro rooting and acclimatization of the obtained microshoots in hydroculture in order to establish an efficient production method. Two explant types were used: basal and terminal cuttings, and better results were achieved when terminal cuttings were used. The development of shoots in the multiplication phase was successful with a regeneration percentage of 100%. Ex vitro rooting in a modified Hoagland nutrient solution was successful (83%), but the percentage of in vitro rooting on half-strength MS medium without hormones was higher (95%). However, bearing in mind that mass production of A. millefolium is more efficient when the phase of in vitro rooting is excluded, this method could be recommended for commercial propagation of this medicinal plant. It is necessary to conduct additional research in order to optimize the composition, EC and pH value of the hydroponic nutrient solution.


2014 ◽  
Vol 22 (1) ◽  
pp. 35-39 ◽  
Author(s):  
Mafatlal M. Kher ◽  
Dimpal Joshi ◽  
Sureshkumar Nekkala ◽  
M. Nataraj ◽  
Dharmesh P. Raykundaliya

AbstractPluchea lanceolata is an important medicinal plant of Asteraceae family known for its anti-arthritic and anti-inflammatory activity. A protocol was established for micropropagation of P. lanceolata using nodal explants. Nodal explants were inoculated onto Murashige and Skoog (1962) - MS medium supple–mented with 6-benzylaminopurine (BAP), kinetin (Kin), thidiazuron (TDZ) and 2iP (2-isopentenyladenine) at various concentrations (0.0, 0.5, 1.0, 1.5 and 2.0 mg·dm-3). The highest multiplication rate was obtained for nodal explants cultured on MS medium, supplemented with 0.5 mg·dm-3 thidiazuron (TDZ). In vitro raised shoots were successfully rooted on ½ mineral salt concentration of MS medium supplemented with 1.0 mg dm-3 IBA.


2021 ◽  
Author(s):  
Awadhesh Kumar Mishra ◽  
Kavindra Nath Tiwari ◽  
Pallavi Mishra ◽  
Sunil Kumar Mishra ◽  
Shailesh Kumar Tiwari

Abstract An efficient encapsulation and germplasm conservation protocol were developed for Nyctanthes arbor-tristis L. In this study the gel matrix containing three percent sodium alginate (SA) and 100 mM calcium chloride (CaCl2. 2H2O) was found best for the formation of encapsulated seeds from node explant of this economically valuable species. The viability of encapsulated seeds and shoot sprouting potential was optimized. Encapsulated seeds stored at 4ºC and 24 ºC maintained its viability up to 90 days and showed sprouting potential 42.89±6.04 and 33.53±7.15 percent respectively. Node explant maintain under slow growth condition up to 180 days on one-eighth (1/8th) strength MS medium supplemented with 0.5 percent sucrose found suitable to maintain high span viability percent (40.28±2.04) with average number of shoots/ node (1.61±0.28) and shoots length (1.12±0.32 cm) respectively. One-eighth (1/8th) strength MS medium supplemented with 0.5 percent sucrose and enriched with 0.5 mg/l abscisic acid (ABA) prolonged the viability up to 40.36±1.01 percent of explant. The best rooting response was achieved on half (½) strength MS medium enriched with 4 mg/l indole-3-acetic acid (IAA). The rooted plant shows 65 percent survivability in open field condition. The true-to-type clonal fidelity assessment of tissue culture recovered acclimated plants with start codon targeted (SCoT) primer profile shows same banding mobility patterns as with source parent mother plant. The maximum banding profile is monomorphic and consistent. Hence on this basis it confirmed the true-to-type clonal stability among them. The protocols display the novel method for conservation of this species under in-vitro condition and facilitate easy exchange of plant germplasm.


2021 ◽  
Vol 15 ◽  
pp. 44-51
Author(s):  
R. Devendra Prasad ◽  
Shreeti Pradan ◽  
Mukti Ram Poudel ◽  
Bijaya Pant

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.


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