Summary
An endosperm derived tissue culture of maize (Zea mays L.) variety A636 has been characterised for its ability to synthesize zein protein and respond to a zein gene regulatory element. Western analysis with zein specific antibodies revealed the distinct presence of zein proteins of the 15, 19 and 21 kDa classes in this tissue, in contrast to an embryo-derived Black Mexican Sweet variety tissue culture which exhibited no zein proteins. Transient transformation studies with a cauliflower mosaic virus 35S promoter and luciferase reporter gene construct demonstrate that protoplasts from this tissue culture, but not from the embryo-derived culture, respond positively to the potential enhancer which is located approximately 300 base pairs upstream of the coding region in most zein genes of maize, thus establishing the usefulness of this culture for studies of tissue specific gene regulation.
Similar content being viewed by others
Abbreviations
- MS:
-
Murashige and Skoog
- CaMV:
-
cauliflower mosaic virus
- BMS:
-
Black Mexican Sweet
- 2,4-D:
-
2,4-dichlorophenoxy — acetic acid
- DAP:
-
days after pollination
- SDS:
-
sodium dodecyl sulfate
- PAGE:
-
polyacrylamide gel electrophoresis
- MES:
-
2-morpholinoethane-sulfonic acid
- Hepes:
-
N-2-hydroxyethylpiper-azine-N′-2-ethansulfonic acid
- Tris:
-
Tris-(hydroxymethyl)-aminomethane
- EDTA:
-
ethyl-enediaminetetra-acetic acid disodium salt
- PEG:
-
polyethylene glycol
References
DeLuca M, McElroy WD (1984) Two kinetically distinguishable ATP sites in firefly luciferase. Biochem Biophys Res Comm 123:764–770
Kriz AL, Boston RS, Larkins BA (1987) Structural and transcriptional analysis of DNA sequences flanking genes that encode 19 kilodalton zeins. Mol Gen Genet 207:90–98
Laemli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
La Rue CD (1949) Cultures of the endosperm of maize. Amer J Bot 36:798
Lyznik LA, Tsai CY (1989) Protein synthesis in endosperm cell cultures of maize (Zea mays L). Plant Science 63:105–114
Maier U-G, Brown, JWS, Toloczyki, C and Feix G. (1987) Binding of a nuclear factor to a consensus sequence in the 5′ flanking region of zein genes from maize. EMBO J 6:17–22
Maier U-G, Brown JWS, Schmitz LM, Schwall M, Dietrich G, Feix G (1988) Mapping of tissue-dependent and independent protein binding sites to the 5′ upstream region of a zein gene. Mol Gen Genet 212:241–245.
Maniatis T, Fritsch EF, Sambrook, J (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26:101–106
Ow DW, Jacobs JD, Howell SH (1987) Functional regions of the cauliflower mosaic virus 35S RNA promoter determined by use of the firefly luciferase gene as a reporter of promoter activity. Proc Natl Acad Sci USA 84:4870–4874
Quayle TJA, Brown JWS, Feix G (1989) Analysis of distal flanking regions of maize 19 kDa zein genes. Gene 80:249–258
Schwall M, Feix G (1988) Zein promoter activity in transiently transformed protoplasts from maize. Plant Science 56:161–166
Shannon JC, Bately JW (1973) Inbred and hybrid effects on establishment of in vitro cultures of Zea mays L. endosperm. Crop Science 13:491–493
Shannon JC, Liu J-W (1979) A simplified medium for the growth of maize (Zea mays) endosperm tissue in suspension culture. Physiol Plant 40:285–291
Shimamoto K, Ackermann M, Dierks-Ventling C (1983) Expression of zein in long term endosperm cultures of maize. Plant Physiol 73:915–920
Wandelt C, Feix G (1989) Sequence of a 21 kd zein gene from maize containing an inframe stop codon. Nucleic Acids Research 17:2354
Wienand U, Feix G (1978) Electrophoretic fractionation and translation in vitro of poly(rA)-containing RNA from maize endosperm: evidence for two mRNAs coding for zein protein. Eur J Biochem 92:605–611
Author information
Authors and Affiliations
Additional information
Communicated by H. Lörz
Rights and permissions
About this article
Cite this article
Quayle, T.J.A., Hetz, W. & Feix, G. Characterization of a maize endosperm culture expressing zein genes and its use in transient transformation assays. Plant Cell Reports 9, 544–548 (1991). https://doi.org/10.1007/BF00232328
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00232328