scholarly journals Comparative analyses of stevioside between fresh leaves and in-vitro derived callus tissue from Stevia rebaudiana Bert. using HPLC

2015 ◽  
Vol 49 (4) ◽  
pp. 199-204 ◽  
Author(s):  
S Mahmud ◽  
S Akter ◽  
IA Jahan ◽  
S Khan ◽  
A Khaleque ◽  
...  
Keyword(s):  
Retention Time ◽  
Callus Tissue ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Poor Performance ◽  
The Other ◽  
Ms Medium ◽  
Green Callus

A protocol was developed to produce large amount of callus in short a period of time from leaf explants of Stevia rebaudiana Bert. The highest amount of white callus was obtained on MS medium supplemented with 2.5 mg/l 2, 4-D and 0.5 mg/l BAP after 3 weeks of inoculating leaf segments. On the other hand, 0.5 mg/l BAP and 1.0 mg/l Kn exhibits poor performance towards callus formation while after using 1.0 mg/l Kn alone did not develop any callus. In this experiment, highest amount of green callus was obtained when MS medium supplemented with 2.5 mg/l NAA and 10% coconut water was used. An improved analytical method HPLC was applied to analyze stevioside extracted from the leaf and callus of Stevia rebaudiana. The stevioside in each sample were analyzed by comparing their retention times with those of the standards. The retention time (RT) of stevioside for leaves were found 14.96 and for callus 13.81 mins. The percentage of stevioside content from leaves and callus was 12.19% and 12.62% respectively DOI: http://dx.doi.org/10.3329/bjsir.v49i4.22621 Bangladesh J. Sci. Ind. Res. 49(4), 199-204, 2014

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

scholarly journals In vitro adventitious shoot regeneration from leaf explants of some apricot cultivars

2019 ◽  
Vol 43 ◽  
Author(s):  
Olga Vladimirovna Mitrofanova ◽  
Irina Vjacheslavovna Mitrofanova ◽  
Tatyana Nikolaevna Kuzmina ◽  
Nina Pavlovna Lesnikova-Sedoshenko ◽  
Sergey Vladimirovich Dolgov
Keyword(s):  
Callus Formation ◽  
System Development ◽  
Low Frequency ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Limiting Factor ◽  
Ms Medium ◽  
Regeneration System ◽  
Propagation Methods

ABSTRACT Apricot is one of the most valuable commercial fruits. In vitro propagation of apricot is very important for rapid multiplication of cultivars with desirable traits and production of cleaning up and virus-free plants. Low frequency of multiplication is the main limiting factor for traditional propagation methods. In this regard, the objective of our investigation was to study the morphogenetic capacity of apricot leaf explants of the promising cultivars ‘Iskorka Tavridy’, ‘Magister’ and ‘Bergeron’ for regeneration system development and solving some breeding questions. The source of explants was in vitro plants regenerated and cultured on QL medium. Leaves were maintained in the dark at 24±1 °C in thermostat for three-four weeks. Morphogenic callus and structures were mainly formed at the central and proximal parts of leaves on MS, QL and WPM media with 1.5 or 2.0 mg L-1 BAP and 1.5 or 2.0 mg L-1 IAA in different combinations, or TDZ (0.6 and 1.3 mg L-1). Callus with adventive buds was transferred to regeneration medium and placed into a growth chamber at 24±1 °C and 16-hour photoperiod with a light intensity of 37.5 μmol m-2 s-1. The best results were obtained when adaxial leaf surface was in contact with the culture medium. Frequency of leaf callus formation on MS medium with 1.5 mg L-1 BAP and 1.5 mg L-1 IAA was higher in the explants of ‘Iskorka Tavridy’ (80.0%) than in - ‘Bergeron’ (50.0%) and ‘Magister’ (36.7%). The best results of callogenesis for ‘Magister’ was obtained on MS medium with 1.3 mg L-1 TDZ (53.3%). Active microshoot regeneration in ‘Iskorka Tavridy’ cultivar was shown on MS medium with BAP and IAA and in ‘Magister’ cultivar - on MS medium with TDZ. Rhizogenesis was obtained on half strength MS medium with 2.0 mg L-1 IBA.

scholarly journals Assessment of the Effects of Newly Fabricated CaO, CuO, ZnO Nanoparticles on Callus Formation Maintainance of Alfalfa (Medicago Sativa L.) Under In Vitro Salt Stress

2021 ◽  
Author(s):  
Merve Simsek ◽  
Busra Yazicilar ◽  
Fatma Boke ◽  
Hayrunisa Nadaroglu ◽  
Azize Alayli ◽  
...  
Keyword(s):  
Salt Stress ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Nacl Stress ◽  
The Other ◽  
Antioxidant Enzyme Activities ◽  
Zno Nps ◽  
Induction Stage

Abstract Nanoparticules plays an important role in plant adaptation to abiotic stress, especially in response to salt stress. In this study, two alfalfa lines (Erzurum, and Muş) were used as the material for the response NaCl to CuO, ZnO and CaO nanoparticules (NPs). CaO is evident to be higher effective than CuO, ZnO in callus induction from leaf explants. The antioxidant enzyme activities were also determined in the callus cultures. The maximum activity in MDA analysis was observed from callus treated of 50 mM NaCl with 0.8 ppm CuO NPs. The callus induction stage without salt treatments indicated a best result in 0.8 ppm CaO NPs for H2O2 value compared to the other NPs. The callus induction stage without salt treatments indicated a best result in 0.8 ppm CaO NPs for POD value compared to the other NPs for POD activity. The best response in protein rate was obtained from callus induction stage and callus formation stage after 50 mM treatment NaCl with 0.8 ppm CuO. LSCM analysis evident that the NPs could migitate the negative effects of NaCl stress by the elimination of stress severity in callus cells. SEM analysis was supported the results obtained by LSCM analysis. Our findings suggest that CuO, CaO and ZnO NPs can offer a simple and effective method to protect alfalfa callus from NaCl stress severity.

scholarly journals In vitro CULTURE OF BIG-SAGE (Lantana camara L.) PLANT

2020 ◽  
Vol 19 (2) ◽  
pp. 67-73
Author(s):  
Majid Abdulhameed Ibrahim ◽  
Manal Zebari Sabty ◽  
Shaimaa Hussein Mussa
Keyword(s):  
Root Length ◽  
Root Formation ◽  
Callus Formation ◽  
Shoot Multiplication ◽  
Lantana Camara ◽  
The Other ◽  
Ms Medium ◽  
Formation Number ◽  
Number Of Shoots

The study was conducted to mass micropropagation of big sage (Lantana camara L.) plant by shoot multiplication technique. The treatments 2.22 and 2.66 µmol·L–1 BA gave the highest significant increase in the percentage of response to shoot multiplication and number of shoots per explant compared to the other treatments as reached 96.70% and 100.00% and 4.33 and 6.00 shoots, respectively. The results showed that these two treatments did not differ significantly between them. While the 1.33 µmol·L–1 BA gave the lowest values in the percentage of response to shoot multiplication and number of shoots per explant were 80.00% and 2.00 shoots per explant, respectively. The MS medium supplemented with 4.30 or 5.37 µmol·L–1 NAA gave a high response to root formation, number of roots per shoot and root length. While the MS medium supplemented with 6.44 or 7.52 µmol·L–1 NAA gave low values in these characteristics. The MS medium with 2.22 or 2.66 µmol·L–1 concentration of BA or 7.52 µmol·L–1 concentration of NAA recorded the highest significant increase in the percentage of response to callus formation. While the MS medium supplemented with 1.33 µmol·L–1 BA or 4.30 µmol·L–1 NAA gave less response to the callus formation.

scholarly journals Explants selection for in vitro propagation of Pachyrhizus erosus L.

2021 ◽  
Vol 13 (1) ◽  
pp. 10844
Author(s):  
Idowu A. OBISESAN ◽  
Ayobola M. A. SAKPERE ◽  
Bamidele J. AMUJOYEGBE ◽  
Michael S. AKINROPO
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Propagation ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stem Explants ◽  
Ms Medium ◽  
Friable Callus ◽  
Pachyrhizus Erosus ◽  
Regeneration Response

Pachyrhizus erosus tuber is rich in protein asides its agronomical value as a legume, but the seeds by which it is propagated have very low viability. This study established sterilization protocol and effect of various concentrations of auxins and cytokinins on callus production and shoot regeneration from explants of P. erosus. Explants and seeds were sterilized using sodiumhypochlorite (NaClO) solution (5, 10 and 15% v/v) for 5 and 10 mins. Nodal, stem and leaf explants from in vitro germinated P. erosus and tuber from field grown plant were sterilized and cultured on Murashige and Skoog (MS) medium (control) and MS combined with different concentrations of auxins (NAA and 2, 4-D) and cytokinin (BA and Kinetin) and the cultured explants were monitored in terms of degree of callus formation, morphology and colour of callus and also for shoot induction. The results showed that seeds of P. erosus sterilized with 10% NaClO solution for 10 mins and germinated in vitro is the best way of getting sterile nodal, stem and leaf explants for the in vitro propagation of the plant, while tuber explants could be sterilized with 15% NaClO for 10 minutes. Nodal explants inoculated in MS medium supplemented with 1.0 mg/L BA gave the highest shoot regeneration response, while stem explants inoculated on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L NAA also gave the highest amount of friable callus. The study concluded that in vitro germinated seeds were the best way of getting explant for P. erosus.

scholarly journals Efficient in vitro plant regeneration from leaf derived callus and genetic fidelity assessment of an endemic medicinal plant of Ranunculus wallichianus Wight & Arnn using RAPD and ISSR markers

2021 ◽  
Author(s):  
Srinivasan P ◽  
David Raja H ◽  
Tamilvanan R
Keyword(s):  
Callus Formation ◽  
Leaf Explants ◽  
Genetic Fidelity ◽  
Issr Markers ◽  
Ms Medium ◽  
Genetic Purity ◽  
Fidelity Assessment ◽  
Half Strength ◽  
Rapd And Issr

Abstract Ranunculus wallichianus is a medicinally important plant and an endemic species to Western Ghats of South India. An efficient and reliable indirect regeneration protocol system for R. wallichianus was developed from leaf explants in the present investigation. Leaf explants were cultured on both full-strength and half-strength MS (Murashige & Skoog) medium supplemented with different concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of 2,4-D and NAA. Among the different concentrations tested, the highest percentage of yellowish green compact nodular callus formation was observed on half-strength MS medium with 2.0 mg L− 1 of 2, 4-D. Then, the in vitro raised organogenic callus was cultured on half strength MS medium containing various concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of BA, KIN and TDZ with 0.5 mg L− 1 NAA and 10% CW for in vitro shoot regeneration. The highest percentage of regeneration response (97%) and maximum number of shoots formation (11.1 ± 0.13 shoots/culture with 9.2 ± 0.35 cm mean shoot length) were obtained from MS medium containing 2.5 mg L− 1 BA with 0.5 mg L− 1 NAA and 10% CW. The well elongated in vitro raised shoots were rooted in half strength MS medium with 2.5 mg L− 1 IBA + 250 mg L− 1 activated charcoal shows high frequency of root formation. The well rooted plantlets were successfully hardened and acclimatized with the survival rate of 94%. Clonal fidelity of in vitro raised plantlets was assessed by using DNA based RAPD and ISSR molecular markers. The total of 56 and 47 monomorphic bands were obtained from RAPD and ISSR markers respectively. This present in vitro propagation protocol system could be an effective for the conservation of R. wallichianus with their genetic purity and its further investigations.

scholarly journals In vitro cultivation and regeneration of Solanum melongena (L.) using stem, root and leaf explants

1970 ◽  
Vol 1 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Bishnu Pada Ray ◽  
Lutful Hassan ◽  
Smreeti Kana Sarker
Keyword(s):  
Key Words ◽  
Callus Induction ◽  
Callus Formation ◽  
Solanum Melongena ◽  
Leaf Explants ◽  
In Vitro Cultivation ◽  
Ms Medium

The treatment combinations was BAP (0, 2.0, 3.0 and 4.0 mg/L) and NAA (0, 0.1, 0.5, and 1.0 mg/L). The rate of callus formation varied in different treatments. The highest amount of callus (48.66%) was produced on MS medium containing 2.0 mg/l BAP and 0.5 mg/l NAA from stem and 8.2 days required for callus induction. The number of shoot regenerated through callus from stem containing 2.0 mg/l BAP and 0.5 mg/l NAA was 3.4 (23.287%) and days required for 38.8 days. Key words: Regeneration; BAP; NAA. Nepal Journal of Biotechnology. Jan. 2011, Vol. 1, No. 1 : 49-54

scholarly journals Protocorm-like bodies and plant regeneration from foliar explants of Coelogyne flaccida, a horticulturally and medicinally important endangered orchid of Eastern Himalaya

Lankesteriana ◽  
2015 ◽  
Vol 15 (2) ◽  
Author(s):  
Kaylan K. De ◽  
Sujit Sil
Keyword(s):  
Chromosome Number ◽  
Callus Tissue ◽  
Leaf Explants ◽  
Mass Scale ◽  
Plantlet Regeneration ◽  
Eastern Himalaya ◽  
Ms Medium ◽  
Young Leaves ◽  
Efficient Induction

An efficient induction of protocrom-like bodies (PLBs) and plantlet regeneration from the young leaves of in vitro grown seedlings of Coelogyne flaccida, an horticulturally and medicinally important endangered epiphytic orchid, was accomplished in order to develop mass-scale propagation. The young leaves (1.5 cm in length) from 110 days old aseptically germinated seedling were grown in vitro in Murashige and Skoog’s (MS) medium supplemented with different concentrations and combinations of NAA ( 0.5-2 mg/L), BAP (0.5-2 mg/L) and Kn ( 0.5-2 mg/L ). The explants produced protocorm-like bodies directly from the epidermal cells of leaf without the formation of intervening callus tissue within four weeks of culture. The highest number of plantlets regenerated through PLBs per explant after 15 weeks was 35-36 in presence of NAA (2mg/l) and Kn (2mg/l). Within 20-25 weeks individual plantlets produced 2-3 leaves and 2-3 roots. Chromosome number from all plants regenerated from leaf explants showed the same chromosome number as the mother plant as 2n = 40. During acclimatization, 80% of the plantlets survived after one month of transplantation. 

scholarly journals Lippia graveolens (Lamiales: Verbenaceae) and Oryganum vulgare (Lamiales: Lamiaceae) obtained by means of the in vitro propagation technique

2021 ◽  
Author(s):  
María A. Aguilar Morales ◽  
Armandina De la Cruz Olvera ◽  
E. Archundia-Garduño ◽  
Rosy G. Cruz Monterrosa ◽  
Mayra Díaz-Ramírez ◽  
...  
Keyword(s):  
Activated Carbon ◽  
Culture Media ◽  
Callus Formation ◽  
Block Design ◽  
Leaf Explants ◽  
Basal Medium ◽  
Culture Technique ◽  
Ms Medium ◽  
Antioxidant Agents

Objective: The objective of this study was to establish the method of propagation of Oryganum vulgare and Lippia graveolens employing a plant tissue culture technique that decreased the phenolization percentages and increased the multiplication coefficients. Design/ methodology/ approach: The in vitro germination percentage was evaluated in both MS and MS medium + activated carbon. Microcuttings (small shoots) of both species were established in base medium added with different antioxidant agents to decrease the phenolization of explants; the treatments were arranged in a completely randomized block  design. For the propagation phase, a completely randomized factorial design was used, where the auxin/cytokinin phytoregulators, type of explants (axillary buds and leaves), and the species (Lippia graveolens and Oryganum vulgare)  were considered as factors. Results: maximum germination (63.3% ±12.5) was obtained on day 15 ​​in both culture media for L. graveolens and O. vulgare. The use of antioxidant agents mainly activated carbon, increased the in vitro establishment and activation of vegetative buds in both species by up to 90%. There were significant differences in the variables evaluated regarding the treatments, the explant, and the species in the multiplication phase. The combination 1.0/ 0.5 mg L-1 BA/AIB induces callus formation for both species. When used as leaf explants, callus formation was potentiated. Study Limitations / Implications: The results presented are advances from a long-term experiment. Findings/conclusions: The germination of L. graveolens seeds can be achieved in MS medium after 15 days. Microcuttings of both L. graveolens and O. vulgare were successfully established in MS basal medium enriched with 1 g L-1 charcoal that showed low oxidation percentages and induced up to 90% the production of shoots in the explants. The mixture of 1.0/0.5 mg L-1 BA/AIB induces callus formation for both species; when this medium is in contact with leaves as an explant, its formation is potentiated, achieving diameters up to 15 mm. In order to achieve the induction of shoots and roots, buds should be established in MS medium enriched with 0.5 mg L-1 IBA for both species; this mixture encreased the multiplication coefficients

scholarly journals Secondary Metabolite Production in Callus Cultures of Stevia rebaudiana Bertoni

1970 ◽  
Vol 45 (3) ◽  
pp. 243-248 ◽  
Author(s):  
B Janarthanam ◽  
M Gopalakrishnan ◽  
T Sekar
Keyword(s):  
Cultured Cells ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Dry Weight ◽  
Callus Cultures ◽  
Ms Medium ◽  
Cell Biomass ◽  
Mother Plants ◽  
Main Components

Stevia rebaudiana is and an important non-caloric sweetening herb contains diterpene glycosides need to be explored for its commercialization. The evolving commercial importance of secondary metabolites in recent years has resulted in a great demand in the Pharma industry. Callus cultures were established from nodal and leaf explants. Leaf explants showed better callus initiation than nodal explants. Maximum callus biomass was observed in MS medium supplemented with 2, 4-D 1.0 mg/l. Further screening of callus culture was carried out on Murashige and Skoog (MS) medium with different concentration and combinations of 2, 4-D, NAA, IAA, IBA, BA and KN individually and in combinations. Remarkable callus biomass of 11.6 g/l dry weight (182.3 g/l fresh weight) was observed in MS media containing 0.5 mg/l 2, 4-D, 0.5 mg/l NAA and 1.0 mg/l KN. The harvested cell biomass was subjected to extraction of active principles. In this study, cell biomass extracts were compared with extracts from leaves of mother plants of Stevia rebaudiana. HPLC analysis of these extracts showed that the main components of the active principles namely Stevioside were present in sufficiently large amounts in the undifferentiated cultured cells. Keywords: In vitro culture; Biomass; Stevia rebaudiana; Stevioside DOI: 10.3329/bjsir.v45i3.6532Bangladesh J. Sci. Ind. Res. 45(3), 243-248, 2010

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