Callus formation from mesophyll protoplasts of Pisum sativum

1973 ◽  
Vol 51 (11) ◽  
pp. 2105-2106 ◽  
Author(s):  
F. Constabel ◽  
J. W. Kirkpatrick ◽  
O. L. Gamborg

Protoplasts isolated from mesophyll of Pisum sativum L. cv. Century, and cultured in 0.2-ml droplets of B5 medium with 1.0 mg/liter (2,4-dichlorophenoxy)acetic acid (2,4-D) or 1-naphthaleneacetic acid (NAA) and 2.0 mg/liter kinetin regenerated cell walls within 2–3 days. The resulting cells began to divide and form calli after 19 days of culture. Protoplast survival depended on keeping the leaf material in the dark for at least 30 h before use.

Planta ◽  
1975 ◽  
Vol 124 (2) ◽  
pp. 177-189 ◽  
Author(s):  
Henry Wilkins ◽  
Malcolm B. Wilkins

HortScience ◽  
1990 ◽  
Vol 25 (5) ◽  
pp. 569-571 ◽  
Author(s):  
A. Raymond Miller ◽  
Craig K. Chandler

A protocol was developed for excising and culturing cotyledon explants from mature achenes of strawberry (Fragaria × ananassa Duch.). Cotyledon explants formed callus with multiple shoot buds on agar-solidified Murashige and Skoog media containing several combinations of hormones (1 μm 2,4-D; 10 μm 2,4-D; 1 μm BA + 1 μm 2,4-D; 1 μm BA + 10 μm 2,4-D; 5 μm BA; 5 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μ m 2,4-D; 5 μ m BA + 5 μm NAA; 5 μ m BA + 15 μ m NAA). After three subcultures, only tissues maintained on the medium containing 5 μm BA + 5 μm NAA continued to form shoots. Tissues transferred to other media eventually died (1 μm 2,4-D; 1 μ m BA + 10 μ m 2,4-D; 5 μ m BA; 5 μ m BA + 1 μ m 2,4-D), became unorganized (1 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μm 2,4-D; 5 μm BA + 15 μm NAA), or formed roots (10 μm 2,4-D). Whole plantlets were produced by transferring callus with buds to medium lacking hormones. The rapid regeneration of clonal plantlets from cotyledon explants may be useful for reducing variability in future developmental studies. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); (2,4-dichlorophenoxy) acetic acid (2,4-D); and 1-naphthaleneacetic acid (NAA).


1997 ◽  
Vol 75 (3) ◽  
pp. 492-500 ◽  
Author(s):  
Delphine Popiers ◽  
Frédéric Flandre ◽  
Brigitte S. Sangwan-Norreel

In vitro regeneration of pea (Pisum sativum L.), a regeneration recalcitrant legume, was optimised using thidiazuron. Buds were initiated from the meristems of the cotyledonary nodes of embryo axes, isolated from mature seeds, and subcultured on Murashige and Skoog medium supplemented with 13.3 μM 6-benzylaminopurine, 16.1 μM α-naphthaleneacetic acid, and 0.2 μM 2,3,5-triiodobenzoic acid. Proliferation of buds was preceded by the formation of white nodular-like protrusions. These structures were cut transversally in fine slices and subcultured on the same medium or in presence of thidiazuron that produces a second wave of secondary budding. The best results (90–110 buds per expiant) were obtained with 10 μM thidiazuron. The capacity of regeneration was genotype independent and reproducible. Buds elongated on the initial medium, then formed roots in presence of 5.37 μM α-naphthaleneacetic acid. and developed into viable plants. Key words: Pisum sativum L., regeneration, meristems, embryo axes, thidiazuron.


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