Abstract
A procedure has been developed for the successful regeneration of plants from mesophyll protoplasts of Arabidopsis thaliana line WS. The protocol is an improved version of that of Damm and Willmitzer (1988).
The main changes in original procedure are as follows:
-
(1)
A mixture of Cellulase Y-C (0.5%) and Pectolyase Y-23 (0.05%) is used for the isolation of protoplasts. Use of these enzymes reduces the incubation time to 50 min.
-
(2)
α-Naphthaleneacetic acid is used as the auxin throughout cultures of protoplasts and calli.
-
(3)
Protoplasts and calli are incubated under dim white light (0.8–8 μW/cm2) during culture.
With these modifications, we were able consistently to obtain regenerated shoots from about 70% of calli that had been transferred to shoot-forming medium even though the plating efficiency was rather low (about 0.5–1.5%).
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- NAA:
-
α-naphthaleneacetic acid
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- BAP:
-
6-benzylaminopurine
- Kin:
-
kinetin
References
Akama K, Shiraishi H, Ohta S, Nakamura K, Okada K, Shimura Y (1992) Plant Cell Rep 12: 7–11
Altmann T, Damm B, Halfter U, Willmitzer L, Morris PC (1992) In: Koncz C, Chua NH, Schell J (eds.) Methods in Arabidopsis Research, Chapter 12. World Scientific Publishing, Singapore, pp. 310–330
Damm B, Willmitzer L (1988) Mol Gen Genet 213: 15–20
Estelle MA, Somerville C (1987) Mol Gen Genet 206: 200–206
Feldmann KA (1992) In: Koncz C, Chua NH, Schell J (eds.) Methods in Arabidopsis Research, Chapter 10. World Scientific Publishing, Singapore, pp. 275–289
Feldmann KA, Marks MD (1987) Mol Gen Genet 208: 1–9
Fromm M, Callis J, Taylor LP, Walbot V (1987) Methods Enzymol 153: 351–366
Gamborg OL, Miller RA, Ojima K (1968) Exp Cell Res 50: 151–158
Karesch H, Bilang R, Potrykus I (1991) Plant Cell Rep 9: 575–578
Koncz CS, Mayerhofer R, Koncz-Kalman Z, Nawrath C, Reiss B, Redai GP, Schell J (1990) EMBO J. 9: 1337–1346
Li LC, Kohlenbach HW (1982) Plant Cell Rep 1: 209–211
Lloyd AM, Barnason AR, Rogers SG, Byrne MC, Frawley RT, Horsch RB (1986) Science 234: 464–466
Masson J, Paszkowski J (1992) The Plant J 2 (5): 829–833
Marton L, Browse J (1991) Plant Cell Rep 10: 235–239
Murashige T, Skoog F (1962) Physiol Plant 15: 473–497
Siemens J, Torres M, Morgner M, Sacristan MD (1993) Plant Cell Rep 12: 569–572
Valvekens D, Van Montagu M, Van Lijsebettens M (1988) Proc Natl Acad Sci USA 85: 5536–5540
Xuan LE, Menczel L (1980) Z Pflanzenphysiol 96: 77–80
Author information
Authors and Affiliations
Additional information
Communicated by A. Komamine
Rights and permissions
About this article
Cite this article
Sando, S., Goto, N. A novel method for regenerating plants from mesophyll protoplasts of Arabidopsis thaliana line WS. Plant Cell Reports 14, 75–80 (1994). https://doi.org/10.1007/BF00233765
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00233765