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

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.

scholarly journals Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Ms Medium ◽  
Regenerated Plants ◽  
Male Flowers

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

scholarly journals Comparison of Purified Anthocyanin Isolated From In Vitro Cell Suspension Culture of Begonia malabarica, Begonia rex-cultorum ‘Baby Rainbow’ and its Antioxidant Activity

2017 ◽  
Vol 9 (08) ◽  
Author(s):  
Aswathy J. M. ◽  
Murugan K.
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Hydroxyl Radicals ◽  
Cell Suspension Culture ◽  
Callus Induction ◽  
Radical Scavenging ◽  
Leaf Explants ◽  
Reducing Power ◽  
Ms Medium

Anthocyanins are the most common flavonoid molecules of vegetables and fruits, especially berries. Human consumption of anthocyanins represents the highest among the flavonoids. Epidemiological studies have suggested that the consumption of anthocyanins lowers the risk of life style disorders like cardiovascular disease, diabetes, arthritis and cancer. Begonia malabarica Lam. of Begoniaceae, is used traditionally as anti-hypoglycemic, antimicrobial, wound healing and in the treatment of anemia. Begonia rex-cultorum ‘Baby rainbow’ an ornamental species was also substituted. Experimentation of in vitro cell suspension culture, isolation, purification of anthocyanin and its antioxidant potential are targeted in the present study. Explants such as leaves and nodes were cultured on MS medium with various phytohormones for callus induction. Leaf explants of Begonia cultured on MS medium fortified with 2, 4-D and BAP showed significant callus induction and also in terms of fresh and dry weights. Significant reddish coloured callus was achieved in cultures initiated from nodal explants in MS medium supplemented with 2, 4-D.Cell suspension cultures were also established in liquid MS medium. After 14 days of culture, cell suspension was obtained with optimal biomass accumulation. Subsequently, bioactive anthocyanin was isolated, purified and fractionated from B.malabarica andB. rex-cultorum‘Baby rainbow’ using amberlite column chromatography and LC-MS/MS analysis. The major anthocyanins eluted from Begonia speciesat 4.7-5.4 min. Tandem MS of the m/z 655.3 peak was identified as anthocyanidin Malvidin-3 –diglucoside as the major compound. The other peaks identified were (584.3) Malvidin or Peonidin, (459.2) Delphinidin + Glucose, (403.2) may be Cynanidin, (287.1) Cyanindin Aglycone and other m/z 242.3, 195.1 and 144.1 were sugar derivatives or fragments. Purified anthocyanin exhibited remarkable inhibition of linoleic acid oxidation and also a concentration dependent free-radical scavenging activities were noticed against DPPH•, hydroxyl radicals and superoxide anions. The degradation of deoxyribose by hydroxyl radicals was also inhibited via iron ion chelators, rather than by directly scavenging the radicals. The results are comparable with reducing power activity of ascorbate and catechin.

Callus and cell suspension culture of Chelidonium majus L

2014 ◽  
Vol 11 (2) ◽  
pp. 150-154
Author(s):  
S Otgonpurev ◽  
Kh Altantsetseg ◽  
N Tsevegsuren
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Basal Medium ◽  
Ms Medium ◽  
Chelidonium Majus ◽  
Root Explants ◽  
Root Callus ◽  
Murashige Skoog

Chelidonium majus L has long history as a being useful for the treatment of many diseases in Asian and European countries. Aim of this study is to cultivate callus and cell suspension culture in vitro using plant phytohormones. The proliferative capacity was tested on shoot and root explants, cultivated on Murashige-Skoog basal medium testing two auxins: 2,4-diclorphenoxiacetic acid (2.4D) and napheteline acetic acid in combination with cytokinine: kinnetine (K). Calluses were developed on MS medium with 0.5 mg/l Kin, 0.5 mg/l IAA from root explants, as well when added with 0.5 mg/l NAA and 0.5 mg/l Kin from shoot explants. More biomass of cell suspension culture of shoot and root callus was accumulated on MS medium added with 0.1mg/l Kin. DOI: http://dx.doi.org/10.5564/mjas.v11i2.238 Mongolian Journal of Agricultural Sciences Vol.11(2) 2013 pp.150-154

scholarly journals The effects of genotypes and media composition on callogenesis, regeneration and cell suspension culture of chamomile (Matricaria chamomilla L.)

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11464
Author(s):  
Aqeel Ahmad ◽  
Muhammad Tahir ul Qamar ◽  
Almeera Shoukat ◽  
Mehtab Muhammad Aslam ◽  
Mohsin Tariq ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
In Vitro Regeneration ◽  
Sustainable Production ◽  
Conservation Strategy ◽  
Direct Regeneration ◽  
Ms Medium ◽  
Indirect Regeneration

Background Chamomile is an important herb being used widely for medicinal purposes. Its multitherapeutic, cosmetic, and nutritional values have been established through years of traditional and scientific use and research. Increased use of medicinal plants necessitates rational use as well as sustainable production of such genetic resources. Plant in vitro micro-propagation poses unique opportunities for sustainable production of medicinal herbs, their regrowth and conservation. The present study aimed to investigate the effects of different explants, plant growth regulators (PGRs) combinations and media type on callogenesis, in vitro regeneration and cell suspension of six chamomile genotypes to enhance its sustainable production. Methods The shoot, lateral sprout, and leaf derived explants of six chamomile genotypes including Isfahan, Shiraz, Kazeron, Goral, Sharokashari and Presso were used for direct and indirect regeneration. For indirect regeneration various doses of NAA and kinetin were used to induce calli which were cultured on MS media containing PGRs for direct and indirect regeneration. Later, cell suspension was established and morphological characterization of CrO3 stained cells was carried out using microscopy. Results and Discussion Our findings revealed that the highest callus percentage and callus volume were observed from lateral sprouts and shoots of genotype Isfahan on MS medium containing 1 mg/L NAA and 1 mg/L kinetin. The in vitro regeneration was found to be genotype dependent while 77% and 77.5% was the highest percentage for indirect and direct regeneration, respectively. Additionally, the maximum shoot number (two shoots/explant) and shoot length (2.22 cm) were also observed in Isfahan genotype. Cell suspension culture showed the highest fresh weight (18.59 g) and dry weight (1.707 g) with 0.75 g inoculum of the callus derived from lateral sprouts cultured on MS medium. Microscopy of CrO3 stained cells was carried on each 3rd day for 27 days that revealed larger and spongier cells in the early days as compared to final days when the cell number was greater but cell size was smaller. Conclusion The callogenesis, organogenesis, and cell suspension culture of chamomile may be genotype dependent. Hence, optimization of media ingredients and culture conditions is of utmost importance for devising tissue culture based conservation strategy of any chamomile genotype and secondary metabolite production.

scholarly journals 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

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.

Somatic Embryogenesis and Plant Regeneration of Wiregrass (Aristida stricta) and Creeping Bluestem (Schizachyrium scoparium var. stoloniferum)

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 479a-479
Author(s):  
Wusi Chen ◽  
Jeffrey G. Norcini ◽  
Robert S. Kalmbacher ◽  
James H. Aldrich
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Growth Regulators ◽  
Cell Suspension Culture ◽  
Embryogenic Callus ◽  
Basal Medium ◽  
Suspension Cultures ◽  
Embryoid Formation ◽  
Semi Solid

Initiation of callus and induction of embryogenesis were achieved from both wiregrass and creeping bluestem. MS basal medium containing coconut milk, sucrose, and 2,4-D were used to initiate callus from young inflorescence of wiregrass and creeping bluestem. The presence of 2,4-D was found to be essential for the induction and early development of embryoids, possibly up to the globular stage. In the case of bluestem, initiation of embryogenic callus required the presence of a low concentration of BA; using only 2,4-D resulted in more non-embryogenic callus. More globular embryos were formed when embryogenic cultures grew rapidly without subculturing, or after being transferred to a hormone-free or a reduced 2,4-D medium. Plant regeneration was carried on a hormone-free MS medium. Initiation of cell suspension and induction of embryoid formation of wiregrass were achieved. However, maintaining cell suspensions seems to have some problems. A majority of the cells were thick-walled, elongated, and non-dividing. No embryos were formed in suspension cultures planted onto solid media. Reinitiation of cell suspension culture of wiregrass is in progress. Initiation of creeping bluestem cell suspension culture was carried out in MS basal medium containing coconut milk, sucrose, and 2,4-D. The maintenance of the cell suspension cultures and induction of embryoid formation were tested under different combinations and concentrations of growth regulators. Suspension cultures were selected and planted onto semi-solid MS basal medium with or without growth regulators. Somatic embryoids formed from suspension culture 3 to 4 weeks after being planted on semi-solid medium. Germination and plant regeneration of somatic embryoid of creeping bluestem are in progress.

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