scholarly journals Tissue Culture Regeneration of a Medicinal Plant from Mexico: Piper auritum Kunth.

HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 207-209 ◽  
Author(s):  
Fabiola Domínguez ◽  
Xavier Lozoya ◽  
James Simon
Keyword(s):  
Leaf Tissue ◽  
Adventitious Shoot ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Whole Plant ◽  
Half Strength ◽  
Piper Auritum ◽  
Growing Conditions

An efficient whole plant regeneration method from callus cultures of Piper auritum was achieved through organogenesis derived from leaf tissue. Proliferating callus and shoot cultures derived from leaf tissue explants placed on Murashige and Skoog (MS) medium supplemented with 2.0 mg·L–1 2, 4-dichlorophenoxyacetic acid (2,4-D) plus 1.5 mg·L–1 kinetin. Optimum combination of hormones (mg·L–1) for shoot induction was 0.5 2,4-D: 1.5 mg·L–1 kinetin (by volume), that resulted in a high rooting rate (49.6 shoots per explant). All of the plants elongated when using a medium consisting of 0.1 mg·L–1 2,4-D plus 1 mg·L–1 kinetin. Elongated shoots were successfully rooted (100%) on half-strength MS medium supplemented with 2.0 mg·L–1 indole-3-acetic acid. All plantlets survived to the growing conditions of a greenhouse. This study demonstrates that leaf tissue of P. auritum is competent for adventitious shoot regeneration and establishes an efficient and useful protocol for the multiplication and conservation of P. autirum for further investigation of its medicinally active constituents.

EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION VIA SOMATIC EMBRYOGENESIS FROM IMMATURE LEAF-DERIVED PROTOPLASTS OF GROUNDNUT (ARACHIS HYPOGAEA L.)

1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Arachis Hypogaea ◽  
Callus Cultures ◽  
Alternative Methods ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
High Yields ◽  
2,4 Dichlorophenoxyacetic Acid

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.

scholarly journals Production of transgenic bean callus via genetic transformation by DNA-coated tungsten particles

2003 ◽  
Vol 9 (3-4) ◽  
Author(s):  
E. Eissa Ahmed
Keyword(s):  
Gene Expression ◽  
Genetic Transformation ◽  
Transient Gene Expression ◽  
Callus Cultures ◽  
Ms Medium ◽  
Mannitol Dehydrogenase ◽  
Dehydrogenase Gene ◽  
Half Strength ◽  
Media Data ◽  
Callus Lines

Callus cultures were induced from hypocotyl of young bean seedlings. The B5 medium completed with 1 mg/1 KIN and 2mg/1 2,4-D proved the best. Callus developed and maintenaned on B5 medium supplemented with 1mg/1 kinetin and 2mg/I 2,4-D. The B5 medium supplemented with 1mg/1 KIN and 2mg/1 2,4-D induced much more callus than half strength MS medium supplemented with 0.5 or 0.75mg/1 BA and 0.1 mg/1 NAA. The results demonstrate that GeneboosterTM is convenient method to obtain transient gene expression in callus of bean. The results have shown that the bean callus shot by GeneboosterTM can be transformed to get (kanamycin-resistant and stress mannitol­tolerant) calli. The presence of mannitol-dehydrogenase gene (mt/) was verified by PCR, showing the integration of mt/ gene carried by two plasmids. Co-transformed calli were selected after bombardment on kanamycin, mannitol and (kanamycin+mannitop-containing media. Data of molecular analysis (PCR) confirmed the insertion of mtl gene in the genome of mannitol-tolerant callus lines.

scholarly journals ENHANCED REGENERATION OF SHOOTS FROM PEACH CELLS INFECTED WITH A SHOOTY MUTANT STRAIN OF AGROBACTERIUM.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1150d-1150
Author(s):  
A. Smigocki ◽  
F. Hammerschlag
Keyword(s):  
Prunus Persica ◽  
Northern Analysis ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Gene Transcripts ◽  
Half Strength ◽  
Low Levels ◽  
2,4 Dichlorophenoxyacetic Acid

Immature `Redhaven' peach (Prunus persica L. Batsch) embryos were infected with Agrobacterium tumefaciens strain tms328::Tn5 carrying the functional cytokinin gene. Shoots were regenerated from callus grown on MS medium without added phytohormones and subsequently rooted on half-strength MS medium with 2.8 -naphthaleneacetic acid. These plants exhibited an increased frequency of branching in vitro. Low levels of cytokinin gene transcripts were detected in these cells by Northern analysis, and using an ELISA assay, the cytokinins zeatin and zeatinriboside were determined to be on the average 30-fold higher. From these results, the expression of the cytokinin gene appears to promote growth of cells in the absence of phytohormones thus serving as a marker for transformation and a promoter of morphogenesis without a 2,4-dichlorophenoxyacetic acid inductive step.

Reduction of fertile regenerants from protoplasts of Triticum tauschii (Coss.) Schmal.

2000 ◽  
Vol 48 (4) ◽  
pp. 501
Author(s):  
Shoukat Afshar-Sterle ◽  
James F. Kollmorgen ◽  
Geoffrey B. Fincher
Keyword(s):  
Cell Division ◽  
Somatic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Cell Aggregates ◽  
Ms Medium ◽  
Triticum Tauschii ◽  
Cell Divisions ◽  
Aegilops Squarrosa ◽  
Half Strength ◽  
2,4 Dichlorophenoxyacetic Acid

Four suspension cell lines generated from two accessions of Triticum tauschii (Coss.) Schmal. (Aegilops squarrosa, 2n = 2x = 14, DD genome) were used to develop an efficient protocol for producing fertile regenerants from protoplasts. Protoplasts were isolated from each cell line by incubating fine cell aggregates (<500 µm in diameter) in a solution containing 3% Cellulase ‘Onozuka’ RS, 0.5% Macerozyme R10 and 0.2% Pectolyase Y23. Cell division occurred when the protoplasts were cultured at a density of 1.0–1.5 x 106 protoplasts mL–1 in half-strength MS medium supplemented with 1.1 mg L–1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.6 M glucose and 1.2% agarose. The first cell divisions were observed after 5–7 days. Cell colonies were observed after 14 days and these grew quickly into large clumps when transferred to half-strength MS medium supplemented with 2.2 mg L-1 2,4-D, 30 g L–1 sucrose and solidified with 0.25% Phytagel. The colonies produced somatic embryos within 21–28 days of transfer to this medium. Somatic embryos were transferred to hormone-free MS medium for regeneration into plantlets. Although many regenerants produced shrivelled seeds, 9 of 16 were fertile and produced normal seeds.

scholarly journals Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl

2010 ◽  
Vol 53 (3) ◽  
pp. 679-686 ◽  
Author(s):  
Claudia Simões ◽  
Norma Albarello ◽  
Cátia Henriques Callado ◽  
Tatiana Carvalho de Castro ◽  
Elisabeth Mansur
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Formation ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Fold Reduction ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Callus Formation

This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved following transfer to growth regulator-free MS medium solidified with 2 g.L-1 phytagel. An acclimatization rate of 53% was found three months after transfer to ex vitro conditions and the recovered plants presented a normal phenotypic aspect.

scholarly journals Plant regeneration from immature embryo culture of Trifolium michelianum savi. - histological observations on adventitious shoot induction

2014 ◽  
Vol 65 (3-4) ◽  
pp. 261-266 ◽  
Author(s):  
Robert Konieczny
Keyword(s):  
Plant Regeneration ◽  
Embryo Culture ◽  
Callus Formation ◽  
Immature Embryo ◽  
Adventitious Shoot ◽  
Meristematic Tissue ◽  
Whole Plant ◽  
Half Strength ◽  
Immature Embryo Culture ◽  
Trifolium Michelianum

Whole plant regeneration via organogenesis in immature embryo culture of <em>Trifolium michelianum</em> Savi. was obtained. The shoots were induced directly from the hypocotyl or indirectly through callus on EC6 medium supplemented with 0.5 and 5 mg/l BAP, respectively. Sporadically the callus formation was preceded by shoot regeneration. Both shoots obtained directly and via callus regenerated into plants when subcultured on half-strength Murashige and Skoog (MS) medium, free of growth regulators. Histological observations revealed that the shoots were induced directly from the ring of meristematic tissue which was formed at the periphery of the hypocotyl or indirectly from the superficial cells of the callus. Irrespective of the mode of regeneration the shoots were of multicellular origin.

scholarly journals In Vitro Propagation of Miscanthus sinensis

HortScience ◽  
1990 ◽  
Vol 25 (10) ◽  
pp. 1291-1293 ◽  
Author(s):  
N.J. Gawel ◽  
C.D. Robacker ◽  
W.L. Corley
Keyword(s):  
In Vitro Propagation ◽  
Miscanthus Sinensis ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Callus Initiation ◽  
Half Strength ◽  
Modified Ms Medium ◽  
D 20 ◽  
2,4 Dichlorophenoxyacetic Acid

Immature inflorescences of Miscanthus sinensis Andress. `Gracillimus', `Variegatus', and `Zebrinus' were cultured on modified MS medium with 9.0 μm 2,4-D, 20 g sucrose/liter, 2.0 g Gelrite/liter, and 0.75 g MgCl2/liter. Organogenesis was observed 8 to 12 weeks after callus initiation. Shoots were rooted on half-strength MS medium without growth regulators. After rooting, tillers were initiated. When transferred to soil, plants matured to flowering quickly and retained their variegation patterns. Propagation through in vitro tillering is suggested. Chemical name used: 2,4-dichlorophenoxyacetic acid (2,4-D).

scholarly journals Assessment of somaclonal variation in indirect morphogenesis-derived plants of Arracacia xanthorrhiza

2019 ◽  
Vol 54 ◽  
Author(s):  
Jan Vitamvas ◽  
Iva Viehmannova ◽  
Petra Hlasna Cepkova ◽  
Hana Mrhalova ◽  
Katerina Eliasova
Keyword(s):  
Somaclonal Variation ◽  
Simple Sequence Repeat ◽  
Inter Simple Sequence Repeat ◽  
Issr Markers ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Half Strength ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Simple Sequence ◽  
Issr Primers

Abstract: The objective of this work was to induce and detect somaclonal variation in arracacha (Arracacia xanthorrhiza) plants regenerated via indirect morphogenesis, in order to evaluate the potential of this technique to produce new genotypes for breeding purposes of this crop. Calli were induced from petiole segments on Murashige & Skoog (MS) medium supplied with 0.1 mg L-1 2,4-dichlorophenoxyacetic acid. The regeneration of plants via indirect morphogenesis was carried out on half-strength MS medium without plant growth regulators. Fifteen randomly chosen plants were subjected to flow cytometry and “inter-simple sequence repeat” (ISSR) analysis. Ploidy level remained stable in all tested regenerants (2n=4x=44), with no changes in the genome. Eighteen ISSR primers produced a total of 1,584 fragments in all samples. Two ISSR primers produced four polymorphic fragments in 26.7% of the tested samples. Somaclonal variation in arracacha is a result of plant regeneration via indirect morphogenesis and can be detected by ISSR markers.

scholarly journals A Laboratory Exercise to Demonstrate Direct and Indirect Shoot Organogenesis using Internodes of Myriophyllum aquaticum

1994 ◽  
Vol 4 (3) ◽  
pp. 317-320 ◽  
Author(s):  
Michael E. Kane ◽  
Nancy L. Philman ◽  
Matthew A. Jenks
Keyword(s):  
Shoot Organogenesis ◽  
Callus Formation ◽  
Adventitious Shoot ◽  
Naphthaleneacetic Acid ◽  
Shoot Cultures ◽  
Myriophyllum Aquaticum ◽  
Laboratory Exercise ◽  
Half Strength ◽  
Organogenesis In Vitro

Only a few plants are suitable for reliably demonstrating rapid direct and indirect shoot organogenesis in vitro. A laboratory exercise has been developed using internodes of Myriophyllum aquaticum, an amphibious water garden plant. Stock shoot cultures are established and maintained in vitro from nodal explants cultured on agar-solidified medium consisting of half-strength Murashige & Skoog salts (MS) and 30 g·liter-1 sucrose. Students use these cultures as the source of internode explants. Explants are cultured on agar-solidified full-strength MS with 30 g·liter-1 sucrose, 100 mg·liter-1 myo-inositol, and 0.4 mg·liter-1 thiamine·HCL and factorial combinations of 0 to 10 μM 2iP and 0 to 1.0 μM NAA. Adventitious shoot development occurs directly from the explant epidermis within 4 days and is promoted in media supplemented with 2iP alone. Cytokinin-supplemented media amended with NAA induce organogenetic callus formation, but reduce 2iP promotion of direct shoot organogenesis. After 4 weeks, shoot organogenesis on the various media is quantified and can be analyzed statistically. Chemical names used: N-(3-methyl-2-butenyl)-1H-purin-6-amine (2iP); α-naphthaleneacetic acid (NAA).

scholarly journals In Vitro Regeneration and Mass Propagation of Ruta graveolens L.—A Multipurpose Shrub

HortScience ◽  
2005 ◽  
Vol 40 (5) ◽  
pp. 1478-1480 ◽  
Author(s):  
Mohd Faisal ◽  
Naseem Ahmad ◽  
Mohammad Anis
Keyword(s):  
High Frequency ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ruta Graveolens ◽  
Mass Propagation ◽  
Regeneration Frequency ◽  
Whole Plant ◽  
Half Strength

A protocol for rapid in vitro propagation of Ruta graveolens L. through high-frequency shoot induction from nodal explants was established. Proliferation of shoots from nodal segments was achieved on Murashige and Skoog medium supplemented with various concentrations of BA, Kin, IAA, and NAA, either singly or in various combinations. The highest shoot regeneration frequency (98.5%) and the highest number of shoots per explant (40.2 ± 2.8) was obtained on MS medium supplemented with 10 μm BA and 2.5 μm NAA. In vitro regenerated shoots rooted best on half-strength MS medium containing 0.5 μm IBA. Rooted shoots, following acclimatization in the greenhouse, were successfully transferred to field conditions, and 90% of plants survived. The efficient in vitro regeneration of the whole plant can be used as a fast and reliable method to transform R. graveolens genetically for its active principles.

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