Summary
High yields of viable pea protoplasts were produced from suspension cultured cells and the conditions for the optimum transient expression of the chloramphenicol acetyltransferase (CAT) gene fused to the CaMV 35S promoter after electroporation were investigated. Conditions for elicitor induction of a member of the phenylalanine ammonia-lyase (PAL) gene family in pea was also investigated using a chimeric gene carrying 480 bp of the putative promoter region of gPAL1 connected to bacterial cat gene and nos terminator. CAT activity was considerably induced by the treatment with fungal elicitor (>100 μg/ml glucose equivalent) isolated from Mycosphaerella pinodes, a pea pathogen.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CAT:
-
chloramphenicol acetyltransferase
- PAL:
-
phenylalanine ammonia-lyase
- CM:
-
acetylated chloramphenicol
- GSH:
-
reduced glutathione
- NOS:
-
nopaline synthase
- ES:
-
electroporation solution
- CaMV:
-
cauliflower mosaic virus
- GUS:
-
β-glucuronidase
- CHS:
-
chalcone synthase
- 2,4-D:
-
2, 4-dichlorophenoxyacetic acid
References
Alton N, Vaphek D. (1979) Nature 282: 864–869.
Arnold VS, Eriksson T (1977) Physiol Plant 39: 257–260.
Bevan M, Shufflebottom D, Edwards K, Jefferson R, Schuch W (1989) EMBO J 8: 1899–1906.
Choudhary DA, Kessmann H, Lamb JC, Dixon AR (1990) Plant Cell Reports 9: 42–46.
Constabel F, Kirkpatrick WJ, Gamborg OL (1973) Can J Bot 51: 2105–2107.
Dangle JL, Hauffe KD, Lipphardt S, Hahlbrock K, Scheel D (1987) EMBO J 6: 2551–2556.
Dron M, Clouse SD, Dixon RA, Lawton HA, Lamb CJ (1988) Proc Natl Acad Sci USA 85: 6738–6742.
Gamborg OL, Miller RA, Ojima K (1868) Exp cell Res 50: 151.
Gorman CM, Moffat L, Howard B (1982) Mol Cell Biol 2: 1044–1051
Hahlbrock K, Scheel D (1989) Ann Rev Plant Physiol Plant Mol Biol 40: 347–369.
Hobbs AS, Jackson AJ, Baliski SD, Delong OMC, Mahon DJ (1990) Plant Cell Reports 9: 17–20.
Hofmann GA (1987) In Electroporation and Electrofusion in Cell Biology (Eds. Jordon C, Neumahn E, Sowess A) Plenum Publishing Co pp 1–26.
Lamb CJ, Lawton MA, Dron M, Dixon RA (1989) Cell 56: 215–224.
Lehminger-Mertens R, Jacobsen H (1989) In vitro Cellular Develop Biol 25: 571–574.
Lois R, Dietrich A, Hahlbrock K, Schulz W (1989) EMBO J 8:1641–1648.
Murashige T, Skoog F (1962) Physiol Plant 15: 473.
Oku H, Shiraishi T, Ouchi S (1977) Naturwissenschaften 64: 643.
Oku H, Shiraishi T, Ouchi S (1987) In Molecular Determinants of Plant Diseases. (eds. Nishimura S, Asada T, Doke N) Japan Sci Soc Press Tokyo/Springer-Verlag, Berlin. pp. 145–156.
Otsuki Y (1990) Culture System of Rice Protoplast: A Laboratory Manual Nohrinsuisangizyutsuzyouhoukyoukai Tokyo
Puonti-Kaerlas J, Eriksson T (1988) Plant Cell Reports 7: 242–245.
Schulze-Lefert P, Dangl JL, Becker-Andre M, Hahlbrock K, Schulz W (1989) EMBO J 8: 651–656.
Shiraishi T, Oku H, Yamashita M, Ouchi S (1978) Ann Phytopath Soc Japan 44: 659–665.
Takebe I, Otsuki Y, Aoki S (1968) Plant Cell Physiol 9:115–124.
Yamada T, Hashimoto H, Shiraishi T, Oku H (1989) Mol Plant Micr Intr 2: 256–261.
Yoshioka H, Shiraishi T, Yamada T, Ichinose Y, Oku, H. (1990) Plant Cell Physiol 31: 1139–1146.
Author information
Authors and Affiliations
Additional information
Communicated by E. D. Earle
Present address: Research Institute, Takasago Perfumery Inc, 5-31-36, Kamata, Minato, Tokyo, 144, Japan Toso Inc, 4560 Oaza-Tomita, Shin-nanyo, Yamaguchi, 746, Japan Central Laboratory of Green Complex, Kasetsert University, Kamphaensaen, Nakohn Pathom, Thailand
Rights and permissions
About this article
Cite this article
Hashimoto, T., Yamada, T., Tada, A. et al. Transient expression in electroporated pea protoplasts: Elicitor responsiveness of a phenylalanine ammonia-lyase promoter. Plant Cell Reports 11, 183–187 (1992). https://doi.org/10.1007/BF00232529
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00232529