Callus Induction and Establishment of Cell Suspension Culture of Cumin (Cuminum cyminum L.)

2020 ◽  
Vol 15 (2) ◽  
pp. 54-63
Author(s):  
S. Suthar Ram ◽  
P.N. Bhatt ◽  
D.P. Bhatt
2014 ◽  
Vol 20 ◽  
pp. 161-169
Author(s):  
MA Rahman ◽  
MA Bari

Context: Tissue culture is an important tool in plant biotechnology that allows for an increase in biomass or metabolite production by utilizing several techniques in callus or cell cultures. Ricin is a toxic protein that can be extracted from the castor bean as secondary metabolite. The ricin has been used experimentally in medicine to kill cancer cell. We developed callus and cell culture technique for the possible extraction of ricin from the cell suspension culture of Ricinus communis. Objectives: The present investigation aimed to develop the cell culture technique of R. communis L. cv. Shabje and tried to establish a protocol for cell suspension culture of castor for possible extraction of ricin from cell extract. Materials and Methods: The hypocotyl explants of R. communis L. cv. Shabje were used as experimental materials. Cultured on Murashige and Skoog medium supplemented with different concentrations and combinations of BAP, NAA, 2,4-D and IAA for callus induction. For cell culture, the media were used without agar with different concentrations and combinations of these hormones. Results: For callus induction the combination of BAP 2.0 mg/l + 0.5 mg/l NAA showed the best performance but in case of cell culture the combination of BAP 2.0 mg/l and 0.2 mg/l NAA showed the best result. Conclusion: The present investigation clearly established and demonstrated the method of obtaining cell suspension culture and important secondary metabolite ricin could be obtained from cell suspension culture of R. communis L. holding promises to explore cell culture industry for ricin production. DOI: http://dx.doi.org/10.3329/jbs.v20i0.17738 J. bio-sci.  20:  161-169, 2012


1970 ◽  
Vol 3 (2) ◽  
pp. 40-45
Author(s):  
M.F. Mohamad Bukhori ◽  
Norzulaani Khalid ◽  
Ch'ng Lou Ven

To explore the potential for embryogenic callus induction protocol through cell suspension culture forhigh frequency plant regeneration of Maspine pineapple (Ananas comosus L.), eight different culturemedia formulation were evaluated for their effects on the induction of somatic embryos from suckerexplants. Explants were cultured on MS medium supplemented with various media concentration(NAA, Dicamba and BAP, Picloram, Kinetin and NAA, 2,4-D, TDZ, and TDZ and BAP).Embryogenic callus induction percentage, color and texture of the callus were assessed after fivemonths of culture. The optimum medium for the proliferation of in vitro shoots from sucker explantswas MS medium supplemented with 3 mg/L BAP. Meanwhile, the optimum medium for the inductionof fastest and high percentage of embryogenic callus growth from in vitro leaf-based was MS mediumsupplemented with Picloram. Results of mean comparison showed that 3 mg/L Picloram were moreeffective on explants than 10 mg/L. Results of the double staining method proved that somaticembryogenesis occurred in MS supplemented with 3 mg/L Picloram. Under microscopic observations,the globular-stage of the embryos were revealed in callus cells which is relatively suitable forsuspension cells inoculums, indicating that the tested PGR were significantly effective for somaticembryogenesis formation in this species. Most embryogenic callus from sucker explants wasyellowish-mucilaginous-wet-friable. The developed protocol potentially leads to the production ofembryogenic callus from sucker explants and plant regeneration through somatic embryogenesis.


2018 ◽  
Vol 8 (5-s) ◽  
pp. 196-208
Author(s):  
JM Aswathy ◽  
Greeshma Murukan ◽  
Bosco Lawarence ◽  
K Murugan

Tribal people use the floral extract of Caesalpinia pulcherrima to cure liver, stomach and skin prone disorders in traditional Indian medicine. This study aimed to evaluate the effect of purified quercetin and its derivatives from in vitro cell suspension cultures of C. pulcherrima Sw. against SW 480, HeLa, MCF-7 and MCF 10A cell lines and its mode of action. Standard protocol was developed for callus induction using leaf explants. Cytotoxic effect was evaluated against SW 480, HeLa, MCF-7 and MCF 10A cells by MTT assay. Apoptosis was evaluated via Hoechst analysis,  flow cytometry, mitochondrial membrane potential and caspase 3 and 9 expression. 2, 4-D (2.5 mg/l), BAP (2.5 mg/l) + kin (1 mg/ml) was effective for remarkable callus induction. Further, cell suspension culture was established.  Effect of elicitors on cell suspension culture was also carried. Sucrose, ABA and salicylic acid (SA) at different concentrations influenced cell biomass and quercetin synthesis. Cells cultured on the medium fortified with 45 g/L sucrose without ABA/SA showed the highest quercetin content (16.5 mg/g). Quercetin was purified, fractionated by HPLC-DAD and was further analyzed by NMR revealed a major fraction of quercetin (3, 5, 7, 3’, 4’-pentahydroxyflavon). Insignificant cytotoxicity was noticed in SW 480, HeLa, MCF 10A when compared to MCF-7 cell lines exposed to different concentrations of purified quercetin for 24- 48 h. Similarly, the apoptosis by nuclei staining using Hoechst 33258 revealed a concentration dependent effect on MCF 7 cells only. This was further substantiated by caspase-9 and 3 induction and mitochondrial depolarization as revealed by flow cytometry. Overall, the results showed that quercetin and its derivatives induced effective apoptosis on MCF-7 cells. Quercetin isolated from the in vitro cell suspension culture of C. pulcherrima showed significant cytotoxicity and apoptotic activity towards MCF-7 cell lines as compared to other cell lines. Keywords:  Caesalpinia pulcherrima; quercetins; suspension culture; cytotoxicity; apoptotic.


2017 ◽  
Vol 6 (2) ◽  
pp. 38 ◽  
Author(s):  
Liyan Jin ◽  
Yang Yang ◽  
Wenjie Gao ◽  
Mingxue Gong ◽  
Jijia Wang ◽  
...  

Dendranthema indicum var. aromaticum is an important aroma plant in genus Dendrathema, and the establishment of callus cultures and cell suspension cultures is the basement of further protoplast fusion studies, which make it possible to breed new fragrant chrysanthemum. In this study, the effects of different plant growth regulating substances in different concentrations on callus induction were investigated with stem segments, leaves, petioles as explants. The results showed that the optimal explants were lower stem segments according to the percentage of callus formation, callus hardness, growth potential and shoot differentiation. The optimal induction mediums were MS supplemented with 1.0 mg.l-12.4 D and 0.2 mg.l-1 6-BA. The cell suspension culture system was established by using the subculture calli. The results showed that the suitable inoculum size was 2g and the suitable cell suspension culture medium was MS supplemented with 0.2 mg.l-1 6-BA and 0.5 mg.l-1 2,4-D. The time course of cell growth showed that the greatest cell fresh weight appeared on day 14 and the highest cell viability on day 3.


2020 ◽  
pp. 35-39
Author(s):  
M. Tsolmon ◽  
G Oyundari ◽  
Oyunbileg Yu ◽  
Kalaiselvi Senthil

Oxymatrine is one of the most important biologically active compounds and is present in Sophora alopecuroides L. The present investigation focuses on the development of an efficient tissue culture method to induce callus and cell suspension culture of S. alopecuroides by studying the effect of jasmonic acid and nitric oxide on cell suspension culture. Callus induction efficiency is high in axenic leaf explants grown in MS medium supplemented with 1.0 mg/L Kinetin (Kin), 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The cell suspension culture was developed using the same callus induction medium without agar. The maximum cell number and dry weight of suspension culture were obtained by the 9th day of incubation. The synthesis of oxymatrine is higher in jasmonic acid and nitric oxide (200 µMJA and 50 µMNO) combination (11.91 µg/g) when compared to the non-elicited control (8.3 µg/g) of callus.


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