Leaf callus induction and suspension culture establishment in lychee (Litchi chinensis Sonn.) cv. Huaizhi

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

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Potential Embryogenic Callus Induction Protocol Through Cell Suspension Culture For High Frequency Plant Regeneration Of Maspine Pineapple (Ananas comosus L.)

Borneo Journal of Resource Science and Technology ◽

10.33736/bjrst.245.2013 ◽

1970 ◽

Vol 3 (2) ◽

pp. 40-45

Author(s):

M.F. Mohamad Bukhori ◽

Norzulaani Khalid ◽

Ch'ng Lou Ven

Keyword(s):

Plant Regeneration ◽

Suspension Culture ◽

Cell Suspension ◽

Cell Suspension Culture ◽

Callus Induction ◽

Embryogenic Callus ◽

Ananas Comosus ◽

Ms Medium ◽

Embryogenic Callus Induction

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.

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Optimization of germination, callus induction, and cell suspension culture of African locust beans Parkia biglobosa (Jacq.) Benth

Journal of Genetic Engineering and Biotechnology ◽

10.1016/j.jgeb.2017.10.012 ◽

2018 ◽

Vol 16 (1) ◽

pp. 191-201 ◽

Cited By ~ 2

Author(s):

Mohamed S. Abbas ◽

Hattem M. El-Shabrawi ◽

Amira Sh. Soliman ◽

Mai A. Selim

Keyword(s):

Suspension Culture ◽

Cell Suspension ◽

Cell Suspension Culture ◽

Callus Induction ◽

Parkia Biglobosa

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DIFFERENTIAL EFFECT OF PURIFIED QUERCETIN AND ITS DERIVATIVES FROM IN VITRO CELL SUSPENSION CULTURES OF CAESALPINIA PULCHERRIMA SW. AGAINST SELECTED CANCER CELL LINES AND ITS MODE OF ACTION

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v8i5-s.1950 ◽

2018 ◽

Vol 8 (5-s) ◽

pp. 196-208

Author(s):

JM Aswathy ◽

Greeshma Murukan ◽

Bosco Lawarence ◽

K Murugan

Keyword(s):

Flow Cytometry ◽

Suspension Culture ◽

Cell Suspension ◽

Cell Lines ◽

Cell Suspension Culture ◽

Callus Induction ◽

Mode Of Action ◽

Cell Suspension Cultures ◽

Mcf 7

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.

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