scholarly journals ISOLATION AND CULTURE OF Celosia cristata L. CELL SUSPENSION PROTOPLASTS

2003 ◽  
Vol 9 (1) ◽  
pp. 1-6
Author(s):  
Retno Mastuti ◽  
Hiroshi Miyake ◽  
Takeshi Taniguchi ◽  
Yoji Takeoka
Keyword(s):  
Plant Regeneration ◽  
Cell Suspension ◽  
Protoplast Culture ◽  
Somatic Hybridization ◽  
Culture Media ◽  
Developmental Competence ◽  
Gelling Agent ◽  
Ms Medium ◽  
L Cell ◽  
Celosia Cristata

Developmental competence of Celosia cristata L. cell suspension-derived protoplasts was investigated. The protoplasts were isolated from 3- to 9-d old cultures in enzyme solution containing 2 percent (w/v) Cellulase YC and 0.5 percent (w/v) Macerozyme R-10 which was dissolved in washing solution (0.4 M mannitol and 10 mM CaCl2) at pH 5.6 for 3 hours. The highest number of viable protoplasts was released from 5-d old culture of a homogenous cell suspension. Subsequently, three kinds of protoplast culture media were simultaneously examined with four kinds of concentration of gelling agent. Culturing the protoplasts on KM8p medium solidified with 1.2 percent agarose significantly enhanced plating efficiency as well as microcolony formation. Afterwards, the microcalli actively proliferated into friable watery callus when they were subcultured on MS medium supplemented with 0.3 mg/l 2,4-D and 1.0 mg/l kinetin. Although the plant regeneration from the protoplasts-derived calli has not yet been obtained, the reproducible developmental step from protoplasts to callus in this study may facilitate the establishment of somatic hybridization using C. cristata as one parent.

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.

In vitro culture of pointed gourd (Trichosanthes dioica Roxb.)

1970 ◽  
Vol 35 (1) ◽  
pp. 135-142 ◽  
Author(s):  
MA Malek ◽  
D Khanam ◽  
M Khatun ◽  
MH Molla ◽  
MA Mannan
Keyword(s):  
Plant Regeneration ◽  
In Vitro Culture ◽  
Culture Media ◽  
Culture Conditions ◽  
Root Induction ◽  
Ms Medium ◽  
Trichosanthes Dioica ◽  
Pointed Gourd ◽  
Regenerated Plantlets

An experiment was conducted to study the in vitro culture of pointed gourd. Cotyledon rescued from physiologically matured seeds (PMS) and immatured seeds (IMS) of pointed gourd were used as explants. Cotyledon excised from PMS responded very well in all culture conditions. Plant regenerated from cotyledon of PMS ranged from 38 to 96% in different hormonal formulations of culture media. Highest percentage of shoot regeneration was observed in MS + 1.0 mg/l BAP and lowest in MS + 2.5 mg/l BAP. No plant regeneration was observed in cotyledon from immatured seeds. The highest percentage of root induction (99%) was achieved in half MS medium supplemented with 0.5 mg/l NAA. The regenerated plantlets were successfully established in earthen pot. Keywords: Cotyledon; in vitro; pointed gourd. DOI: 10.3329/bjar.v35i1.5874Bangladesh J. Agril. Res. 35(1) : 135-142, March 2010

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 Plant regeneration from protoplasts of Gentiana straminea Maxim

2016 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Guomin Shi ◽  
Lina Yang ◽  
Tao He
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Protoplast Culture ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Embryogenic Calli ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Solid Liquid

AbstractA protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

scholarly journals 884 PB 302 PLANT REGENERATION FROM OVARY TISSUES OF EASTER LILY

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 560e-560
Author(s):  
Jodie L. Ramsay ◽  
Donald S. Galitz ◽  
Chiwon W. Lee
Keyword(s):  
Plant Regeneration ◽  
Culture Media ◽  
Callus Formation ◽  
Lilium Longiflorum ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Flower Buds ◽  
Soil Medium ◽  
Shoot Differentiation ◽  
Easter Lily

Influences of culture media, sucrose, and growth regulator concentrations on plant regeneration from Easter lily (Lilium longiflorum L.) were investigated. Ovary tissues excised from unopened flower buds (3-10 cm long) were cultured on either B-5 medium or MS medium containing 2, 5, or 10% sucrose, 0.8% agar or Phytagel, and varying concentrations of 2,4-D, kinetin, naphthaleneacetic acid (NAA) and benzyladenine (BA). Callus formation from explants was more prolific on MS medium than on B-5 medium and when cultures were initially placed in the dark for 20 days. Cultures grew best when the medium contained 5% sucrose. Shoot differentiation from callus was maximum when MS medium contained 1 mg/liter 2,4-D and 2 mg/liter BA. Roots developed when shoots were placed on the same medium with 1 mg/liter 2,4-D, 0.1 mg/liter NAA and 0.1 mg/liter kinetin. Rooted plants were successfully transferred into soil medium in a greenhouse.

scholarly journals Efficient plant regeneration from embryogenic cell suspension culture of two deepwater rice (Oryza sativa L.) Varieties

2013 ◽  
pp. 91-103
Author(s):  
L Khaleda ◽  
M Al-Forkan
Keyword(s):  
Plant Regeneration ◽  
Cell Suspension ◽  
Culture Media ◽  
Oryza Sativa L ◽  
Cell Suspension Cultures ◽  
Cell Suspensions ◽  
Deepwater Rice ◽  
Embryogenic Calli ◽  
Rice Varieties ◽  
Embryogenic Cell

Cell suspension cultures were initiated from 28-30 d old scutellar-derived embryogenic calli of two deepwater rice varieties HAJA-1 and HAJA-8 and maintained on R2 liquid medium containing 2 mg l-1 2,4-D. The formation of fine embryogenic cell suspensions and subsequent plant regeneration were dependent on plant genotype and culture media. Variety HAJA-8 was found less viable than the other variety HAJA-1. In general, it was observed that variety HAJA-1 showed better performance compared to variety HAJA-8 to R2 medium for the initiation of embryogenic cell lines and the rate of the cell growth was higher in this variety. Plant regeneration was not obtained from cell suspensions of both varieties when cells were cultured on regeneration media semi-solidified with 0.4% (w/v) agarose. Plant regeneration from cell suspensions of the both varieties were obtained when the agrose concentration of the regeneration media was increased from 0.4 to 1% (w/v). The highest plant regeneration frequencies were obtained from cell suspensions of the varieties HAJA-1 and HAJA-8 (48%) and (42%), respectively, pre-treated with water for 5 h and cultured on MS medium supplemented with 2 mg l-1 BAP+1 mg l-1 Proline. Washing cell suspensions with water for a prolonged period of time (15 h and more) inhibited plant regeneration. DOI: http://dx.doi.org/10.3329/cujbs.v5i1.13374 The Chittagong Univ. J. B. Sci.,Vol. 5(1 &2):91-103, 2010

scholarly journals Somatic embryogenesis and plant regeneration from cell suspension cultures of Gentiana kurroo Royle.

2018 ◽  
Vol 7 (5) ◽  
pp. 2239
Author(s):  
Shiwani Kaushal ◽  
Arushdeep Sidana
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Cell Suspension ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Western Himalayas ◽  
Ms Medium ◽  
Gentiana Kurroo

Gentiana kurroo Royle, native to North- Western Himalayas, is one of the critically endangered perennial herbs, which is being overexploited due to its multiple medicinal uses. The goal of this work was to develop a protocol for somatic embryogenesis and efficient plant regeneration through cell suspension culture. Selected ranges of plant growth regulators were experimented for somatic embryogenesis. Light micrographs of the established suspension cell cultures of somatic embryos at different developmental stages were also observed under light microscope. Somatic embryogenesis was induced from the leaf explants and the maximum induction frequency (97.2%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l each of NAA, BAP and TDZ. Cell suspension cultures were established in MS liquid medium containing IAA (0.5 mg/l) + BAP (1.0 mg/l) with 89-93% viable cells. Transfer of cotyledonary somatic embryos to the MS agar medium containing 2, 4-D (0.4 mg/l) and KN (1.5 mg/l) resulted in somatic embryogenesis at high frequencies (80%) with an average of 28 ± 0.2 embryos/ callus piece. After transfer onto the half strength MS medium without growth regulators approximately 80-90% somatic embryos developed into complete plantlets. The protocol described here could be used for somatic hybridization, genetic transformation, isolation of protoplasts and large-scale propagation of G. kurroo.

scholarly journals Callus induction and plant regeneration by Brazilian triticale and wheat genotypes

1997 ◽  
Vol 20 (1) ◽  
pp. 41-44 ◽  
Author(s):  
A.L.C. Dornelles ◽  
F.I.F. Carvalho ◽  
L.C. Federizzi ◽  
C.L. Handel ◽  
F. Bered ◽  
...  
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Culture Media ◽  
Rio Grande ◽  
Immature Embryos ◽  
Ms Medium ◽  
Rio Grande Do Sul ◽  
Wheat Genotypes ◽  
The One

In order to determine the in vitro behavior of Brazilian triticale, 16 triticale genotypes, and three wheat genotypes used as checks, were sown in June 1994. The explants used were immature embryos. In addition to the genotype tests, two culture media for callus induction were also evaluated, i.e., MS (Murashige and Skoog, Physiol. Plant. 15: 473-497, 1962) medium containing 2.0 mg 2,4D/l, and MS medium containing 4.0 mg 2,4D/l. The plant regeneration protocol used was the one employed at the Laboratório de Cultura de Tecidos, Departamento de Plantas de Lavoura, Universidade Federal do Rio Grande do Sul, for wheat. Differences in plant regeneration were observed both among triticale and wheat genotypes, with triticale usually showing better regeneration than wheat. No differences were observed between the callus induction media.

Isolation, Culture and Plant Regeneration From Protoplasts of Nicotiana debneyi

1980 ◽  
Vol 7 (6) ◽  
pp. 635 ◽  
Author(s):  
WR Scowcroft ◽  
PJ Larkin
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Indole Acetic Acid ◽  
High Efficiency ◽  
Protoplast Culture ◽  
Callus Cultures ◽  
Dna Uptake ◽  
Mesophyll Protoplasts ◽  
Nicotiana Debneyi ◽  
Protoplast Division

Mesophyll protoplasts of two genetically distinct genotypes of N. debneyi were cultured with sustained division following a plating efficiency in excess of 50%. Fully fertile mature plants were regenerated from callus cultures derived from protoplasts. Shoots were induced in medium containing 1 mg/l 6-benzylaminopurine and 0.5 mg/I indole acetic acid. The repeatably high efficiency of protoplast culture was used to evaluate the quantitative effects of two drugs, kanamycin and trimethoprim, which effectively inhibited colony formation at concentrations of 100 and 50 �g/ml, respectively. An enhancer of DNA uptake, poly-L-ornithine, had virtually no effect on sustained protoplast division at a concentration of 7.5 �g/ml or less.

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