scholarly journals In vitro Propagation and Comparative Phytochemical Analysis of Wild Plant and Micropropagated Cleome rutidosperma DC.

Author(s):  
Deventhiran M. ◽  
John Wyson W. ◽  
Sheik Noor Mohamed M. ◽  
Jaikumar K. ◽  
Saravanan P. ◽  
...  

Plants are widely used by all sections of the society either as folk medicines or as pharmaceutical preparation of modern medicine. In vitro propagation of plants holds great promise for conservation and enhancement of valuable medicinal plants. Cleome rutidosperma has been used in indian ayurvedic medicine for the treatment of a wide number of health disorders. The present study deals with the influence of different plant growth regulators (PGR) including kinetin (Kin), 6- Benzylaminopurine (BAP) and 2, 4-Dichlorophenoxyacetic acid (2,4-D) on the growth of plant and the identification and comparison of bioactive constituents of wild and in situ propagated C. rutidosperma plant using Gas Chromatography - Mass Spectrometry analysis (GC-MS). Nodal segments used as explants were cultured on Murashige and Skoog's medium (MS) supplied with different concentrations of PGRs. Multiple shoot generation was achieved after 28 days of incubation. The GC-MS analysis showed the presence of ten compounds of micropropagated and seven compounds of wild plants were identified. The result concluded that various concentration of PGR had a significant role in in vitro regeneration of plant and showed that the phytoconstituents of micropropagated plant is comparatively higher than that of wild plant.

Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3229
Author(s):  
Mat Yunus Najhah ◽  
Hawa Z. E. Jaafar ◽  
Jaafar Juju Nakasha ◽  
Mansor Hakiman

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.


Author(s):  
Asmaa Abdelsalam ◽  
Ehab Mahran ◽  
Kamal Chowdhury ◽  
Arezue Boroujerdi

Abstract Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.


Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1657
Author(s):  
Nqobile P. Hlophe ◽  
Adeyemi O. Aremu ◽  
Karel Doležal ◽  
Johannes Van Staden ◽  
Jeffrey F. Finnie

In Africa and Asia, members of the genus Brachystelma are well-known for their diverse uses, especially their medicinal and nutritional values. However, the use of many Brachystelma species as a valuable resource is generally accompanied by the concern of over-exploitation attributed to their slow growth and general small size. The aim of the current study was to establish efficient micropropagation protocols for three Brachystelma species, namely Brachystelma ngomense (endangered), Brachystelma pulchellum (vulnerable) and Brachystelma pygmaeum (least concern), as a means of ensuring their conservation and survival. This was achieved using nodal segments (~10 mm in length) as the source of explants in the presence of different concentrations of three cytokinins (CK) namely N6-benzyladenine (BA), isopentenyladenine (iP) and meta-topolin riboside (mTR), over a period of 6 weeks. The highest (25 µM) concentration of cytokinin treatments typically resulted in significantly higher shoot proliferation. However, each species differed in its response to specific CK: the optimal concentrations were 25 µM mTR, 25 µM iP and 25 µM BA for Brachystelma ngomense, Brachystelma pulchellum and Brachystelma pygmaeum, respectively. During the in vitro propagation, both Brachystelma ngomense and Brachystelma pygmaeum rooted poorly while regenerated Brachystelma pulchellum generally lacked roots regardless of the CK treatments. Following pulsing (dipping) treatment of in vitro-regenerated shoots with indole-3-butyric acid (IBA), acclimatization of all three Brachystelma species remained extremely limited due to poor rooting ex vitro. To the best of our knowledge, the current protocols provide the first successful report for these Brachystelma species. However, further research remains essential to enhance the efficiency of the devised protocol.


Author(s):  
S. Parvin ◽  
M. Kausar ◽  
M. Enamul Haque ◽  
M. Khalekuzzaman ◽  
B. Sikdar ◽  
...  

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


2012 ◽  
Vol 7 (5) ◽  
pp. 931-940 ◽  
Author(s):  
Dagmar Skálová ◽  
Božena Navrátilová ◽  
Lenka Richterová ◽  
Michal Knit ◽  
Michal Sochor ◽  
...  

AbstractMany populations of high-mountainous relic dioecious willows in Central Europe only consist of female individuals and are thus limited in their reproductive potential. We completed micropropagation experiments with shoot apexes and nodal segments of common and endangered willow (Salix) species, which can help to reintroduce autochthonous genotypes to their natural sites. Until recently, cultivation of green young shoot apexes of S. alba and S. lapponum showed the highest percentage of regeneration. We successfully applied the two-times-sterilisation due to high contamination of natural explants. The OK medium was the most efficient culture medium. In vitro propagation of willows with unisexual catkins, anther and ovule cultures were tested and optimised. Isolated anthers were cultivated on selected media and then microcallus and calluses of S. caprea and calluses of S. viminalis were formed on the A medium. Among various tested and optimised media for the ovule culture, the CP medium was the most efficient one. In this case, only the microcalluses of S. viminalis were observed. We developed biotechnological procedures that can be useful in conserving fragmented populations of high-mountainous willows.


2000 ◽  
Vol 5 (1) ◽  
pp. 15
Author(s):  
A. S. AI-Wasel

Shoot multiplication of a putative variant of Catharanthus roseus (L.) G. Don, was achieved in vitro using shoot tips and nodal segments as explants. The addition of growth regulators to establishment medium stimulated bud breaking and shoot elongation. The maximum shoot multiplication (15.1 shoots/microshoot) and the longest shoots (7.0 cm) occurred on Murashige and Skoog medium (MS) containing 1.0 mg L-1 of N6-Benzyladenine (BA) and a- Naphthalene acetic acid (NAA). All microshoots formed roots and normal root morphology occurred on half strength MS salt supplied with 0.5 mg L-1 NAA or Indole-B-Butyric acid (IBA). Rooted microshoots (95 %) were successfully transferred to soil.


2007 ◽  
Vol 64 (3) ◽  
pp. 263-268 ◽  
Author(s):  
Ronaldo Biondo ◽  
Ana Valéria Souza ◽  
Bianca Waléria Bertoni ◽  
Andreimar Martins Soares ◽  
Suzelei Castro França ◽  
...  

Mandevilla velutina (Mart.) Woodson (Apocynaceae) is a medicinal plant species with antivenom properties, native from Brazilian Savanna regions (Cerrado), which due to overexploitation and habitat deforestation is in danger of extinction. As an initiative for conserving this endangered but economically important plant species, a micropropagation protocol was developed and genotypes were stored in the Germplasm Bank "Cerrado In vitro". For the in vitro propagation of M. velutina, nodal segments were inoculated on Murashige and Skoog (MS) medium supplemented with different concentrations of BA, Zeatin, 2ip, DTT and TDZ. Best multiplication ratio was achieved when to the medium 0.44 µM BA, ranging 1: 6.7, were added. Plantlets cultured on MS/2 medium supplemented with 26.85 µM NAA rooted successfully (50.5%). Although rooted and un-rooted plantlets acclimatized to soil conditions, great losses were observed within un-rooted plantlets, while the rooted presented 100 % survival. It was possible to maintain 43% of the M. velutina germplasm under healthy conditions for six months, with no subcultures, using the MS medium supplemented with 2% sucrose, 13.8 mM spermidine, 2% sorbitol and 2% dextrose.


2013 ◽  
Vol 61 (1) ◽  
pp. 67 ◽  
Author(s):  
Semiha Erişen ◽  
Zeynep Öncel

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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