Abstract
Manipulation of plant natural product biosynthesis through genetic engineering is an attractive but technically challenging goal. Here, we demonstrate that different secondary metabolites can be produced in cultured maize cells by ectopic expression of the appropriate regulatory genes. Cell lines engineered to express the maize transcriptional activators C1 and R accumulate two cyanidin derivatives, which are similar to the predominant anthocyanin found in differentiated plant tissues. In contrast, cell lines that express P accumulate various 3-deoxy flavonoids. Unexpectedly, P-expressing cells in culture also accumulate phenylpropanoids and green fluorescent compounds that are targeted to different subcellular compartments. Two endogenous biosynthetic genes (c2 and a1, encoding chalcone synthase and flavanone/dihydroflavonol reductase, respectively) are independently activated by ectopic expression of either P or C1/R, and there is a dose-response relationship between the transcript level of P and the degree to which c2 or a1 is expressed. Our results support a simple model showing how the gene encoding P may act as a quantitative trait locus controlling insecticidal C-glycosyl flavone level in maize silks, and they suggest how p1 might confer a selective advantage against insect predation in maize.
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- Athma P., Grotewold E., Peterson T. Insertional mutagenesis of the maize P gene by intragenic transposition of Ac. Genetics. 1992 May;131(1):199–209. doi: 10.1093/genetics/131.1.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Athma P., Peterson T. Ac induces homologous recombination at the maize P locus. Genetics. 1991 May;128(1):163–173. doi: 10.1093/genetics/128.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett M. K. SNAREs and the specificity of transport vesicle targeting. Curr Opin Cell Biol. 1995 Aug;7(4):581–586. doi: 10.1016/0955-0674(95)80016-6. [DOI] [PubMed] [Google Scholar]
- Bennett M. K., Scheller R. H. A molecular description of synaptic vesicle membrane trafficking. Annu Rev Biochem. 1994;63:63–100. doi: 10.1146/annurev.bi.63.070194.000431. [DOI] [PubMed] [Google Scholar]
- Bodeau J. P., Walbot V. Regulated transcription of the maize Bronze-2 promoter in electroporated protoplasts requires the C1 and R gene products. Mol Gen Genet. 1992 Jun;233(3):379–387. doi: 10.1007/BF00265434. [DOI] [PubMed] [Google Scholar]
- Britsch L. Purification and characterization of flavone synthase I, a 2-oxoglutarate-dependent desaturase. Arch Biochem Biophys. 1990 Oct;282(1):152–160. doi: 10.1016/0003-9861(90)90099-k. [DOI] [PubMed] [Google Scholar]
- Byrne P. F., McMullen M. D., Snook M. E., Musket T. A., Theuri J. M., Widstrom N. W., Wiseman B. R., Coe E. H. Quantitative trait loci and metabolic pathways: genetic control of the concentration of maysin, a corn earworm resistance factor, in maize silks. Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8820–8825. doi: 10.1073/pnas.93.17.8820. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campbell M. M., Sederoff R. R. Variation in Lignin Content and Composition (Mechanisms of Control and Implications for the Genetic Improvement of Plants). Plant Physiol. 1996 Jan;110(1):3–13. doi: 10.1104/pp.110.1.3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chopra S., Athma P., Peterson T. Alleles of the maize P gene with distinct tissue specificities encode Myb-homologous proteins with C-terminal replacements. Plant Cell. 1996 Jul;8(7):1149–1158. doi: 10.1105/tpc.8.7.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Christensen A. H., Sharrock R. A., Quail P. H. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol Biol. 1992 Feb;18(4):675–689. doi: 10.1007/BF00020010. [DOI] [PubMed] [Google Scholar]
- Deboo G. B., Albertsen M. C., Taylor L. P. Flavanone 3-hydroxylase transcripts and flavonol accumulation are temporally coordinate in maize anthers. Plant J. 1995 May;7(5):703–713. doi: 10.1046/j.1365-313x.1995.07050703.x. [DOI] [PubMed] [Google Scholar]
- Dooner H. K., Robbins T. P., Jorgensen R. A. Genetic and developmental control of anthocyanin biosynthesis. Annu Rev Genet. 1991;25:173–199. doi: 10.1146/annurev.ge.25.120191.001133. [DOI] [PubMed] [Google Scholar]
- Eberhardt T. L., Bernards M. A., He L., Davin L. B., Wooten J. B., Lewis N. G. Lignification in cell suspension cultures of Pinus taeda. In situ characterization of a gymnosperm lignin. J Biol Chem. 1993 Oct 5;268(28):21088–21096. [PubMed] [Google Scholar]
- Forkmann G., Dangelmayr B. Genetic control of chalcone isomerase activity in flowers of Dianthus caryophyllus. Biochem Genet. 1980 Jun;18(5-6):519–527. doi: 10.1007/BF00484399. [DOI] [PubMed] [Google Scholar]
- Goff S. A., Klein T. M., Roth B. A., Fromm M. E., Cone K. C., Radicella J. P., Chandler V. L. Transactivation of anthocyanin biosynthetic genes following transfer of B regulatory genes into maize tissues. EMBO J. 1990 Aug;9(8):2517–2522. doi: 10.1002/j.1460-2075.1990.tb07431.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grotewold E., Athma P., Peterson T. A possible hot spot for Ac insertion in the maize P gene. Mol Gen Genet. 1991 Nov;230(1-2):329–331. doi: 10.1007/BF00290684. [DOI] [PubMed] [Google Scholar]
- Grotewold E., Athma P., Peterson T. Alternatively spliced products of the maize P gene encode proteins with homology to the DNA-binding domain of myb-like transcription factors. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4587–4591. doi: 10.1073/pnas.88.11.4587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grotewold E., Drummond B. J., Bowen B., Peterson T. The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell. 1994 Feb 11;76(3):543–553. doi: 10.1016/0092-8674(94)90117-1. [DOI] [PubMed] [Google Scholar]
- Grotewold E., Peterson T. Isolation and characterization of a maize gene encoding chalcone flavonone isomerase. Mol Gen Genet. 1994 Jan;242(1):1–8. doi: 10.1007/BF00277341. [DOI] [PubMed] [Google Scholar]
- Herbers K., Sonnewald U. Manipulating metabolic partitioning in transgenic plants. Trends Biotechnol. 1996 Jun;14(6):198–205. doi: 10.1016/0167-7799(96)10027-5. [DOI] [PubMed] [Google Scholar]
- Herman E. M., Lamb C. J. Arabinogalactan-rich glycoproteins are localized on the cell surface and in intravacuolar multivesicular bodies. Plant Physiol. 1992 Jan;98(1):264–272. doi: 10.1104/pp.98.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howles P. A., Sewalt VJH., Paiva N. L., Elkind Y., Bate N. J., Lamb C., Dixon R. A. Overexpression of L-Phenylalanine Ammonia-Lyase in Transgenic Tobacco Plants Reveals Control Points for Flux into Phenylpropanoid Biosynthesis. Plant Physiol. 1996 Dec;112(4):1617–1624. doi: 10.1104/pp.112.4.1617. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kacser H., Burns J. A. The control of flux. Symp Soc Exp Biol. 1973;27:65–104. [PubMed] [Google Scholar]
- Kacser H., Burns J. A. The molecular basis of dominance. Genetics. 1981 Mar-Apr;97(3-4):639–666. doi: 10.1093/genetics/97.3-4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klein M., Weissenböck G., Dufaud A., Gaillard C., Kreuz K., Martinoia E. Different energization mechanisms drive the vacuolar uptake of a flavonoid glucoside and a herbicide glucoside. J Biol Chem. 1996 Nov 22;271(47):29666–29671. doi: 10.1074/jbc.271.47.29666. [DOI] [PubMed] [Google Scholar]
- Larson R. L., Coe E. H., Jr Gene-dependent flavonoid glucosyltransferase in maize. Biochem Genet. 1977 Feb;15(1-2):153–156. doi: 10.1007/BF00484558. [DOI] [PubMed] [Google Scholar]
- Larson R., Bussard J. B., Coe E. H., Jr Gene-dependent flavonoid 3'-hydroxylation in maize. Biochem Genet. 1986 Aug;24(7-8):615–624. doi: 10.1007/BF00504338. [DOI] [PubMed] [Google Scholar]
- Levings C. S., Stuber C. W. A maize gene controlling silk browning in response to wounding. Genetics. 1971 Dec;69(4):491–498. doi: 10.1093/genetics/69.4.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lloyd A. M., Walbot V., Davis R. W. Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1. Science. 1992 Dec 11;258(5089):1773–1775. doi: 10.1126/science.1465611. [DOI] [PubMed] [Google Scholar]
- Ludwig S. R., Bowen B., Beach L., Wessler S. R. A regulatory gene as a novel visible marker for maize transformation. Science. 1990 Jan 26;247(4941):449–450. doi: 10.1126/science.247.4941.449. [DOI] [PubMed] [Google Scholar]
- Marrs K. A., Alfenito M. R., Lloyd A. M., Walbot V. A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2. Nature. 1995 Jun 1;375(6530):397–400. doi: 10.1038/375397a0. [DOI] [PubMed] [Google Scholar]
- Menssen A., Höhmann S., Martin W., Schnable P. S., Peterson P. A., Saedler H., Gierl A. The En/Spm transposable element of Zea mays contains splice sites at the termini generating a novel intron from a dSpm element in the A2 gene. EMBO J. 1990 Oct;9(10):3051–3057. doi: 10.1002/j.1460-2075.1990.tb07501.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moreno M. A., Chen J., Greenblatt I., Dellaporta S. L. Reconstitutional mutagenesis of the maize P gene by short-range Ac transpositions. Genetics. 1992 Aug;131(4):939–956. doi: 10.1093/genetics/131.4.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moyano E., Martínez-Garcia J. F., Martin C. Apparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in antirrhinum flowers. Plant Cell. 1996 Sep;8(9):1519–1532. doi: 10.1105/tpc.8.9.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nash J., Luehrsen K. R., Walbot V. Bronze-2 gene of maize: reconstruction of a wild-type allele and analysis of transcription and splicing. Plant Cell. 1990 Nov;2(11):1039–1049. doi: 10.1105/tpc.2.11.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niederberger P., Prasad R., Miozzari G., Kacser H. A strategy for increasing an in vivo flux by genetic manipulations. The tryptophan system of yeast. Biochem J. 1992 Oct 15;287(Pt 2):473–479. doi: 10.1042/bj2870473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paris N., Stanley C. M., Jones R. L., Rogers J. C. Plant cells contain two functionally distinct vacuolar compartments. Cell. 1996 May 17;85(4):563–572. doi: 10.1016/s0092-8674(00)81256-8. [DOI] [PubMed] [Google Scholar]
- Quattrocchio F., Wing J. F., Leppen HTC., Mol JNM., Koes R. E. Regulatory Genes Controlling Anthocyanin Pigmentation Are Functionally Conserved among Plant Species and Have Distinct Sets of Target Genes. Plant Cell. 1993 Nov;5(11):1497–1512. doi: 10.1105/tpc.5.11.1497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwarz-Sommer Z., Shepherd N., Tacke E., Gierl A., Rohde W., Leclercq L., Mattes M., Berndtgen R., Peterson P. A., Saedler H. Influence of transposable elements on the structure and function of the A1 gene of Zea mays. EMBO J. 1987 Feb;6(2):287–294. doi: 10.1002/j.1460-2075.1987.tb04752.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snyder B. A., Nicholson R. L. Synthesis of phytoalexins in sorghum as a site-specific response to fungal ingress. Science. 1990 Jun 29;248(4963):1637–1639. doi: 10.1126/science.248.4963.1637. [DOI] [PubMed] [Google Scholar]
- Tuerck J. A., Fromm M. E. Elements of the maize A1 promoter required for transactivation by the anthocyanin B/C1 or phlobaphene P regulatory genes. Plant Cell. 1994 Nov;6(11):1655–1663. doi: 10.1105/tpc.6.11.1655. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whetten R., Sederoff R. Lignin Biosynthesis. Plant Cell. 1995 Jul;7(7):1001–1013. doi: 10.1105/tpc.7.7.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Tunen A. J., Mur L. A., Recourt K., Gerats A. G., Mol J. N. Regulation and manipulation of flavonoid gene expression in anthers of petunia: the molecular basis of the Po mutation. Plant Cell. 1991 Jan;3(1):39–48. doi: 10.1105/tpc.3.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
