Abstract
Plants produce a wide array of natural products, many of which are likely to be useful bioactive structures. Unfortunately, these complex natural products usually occur at very low abundance and with restricted tissue distribution, thereby hindering their evaluation. Here, we report a novel approach for enhancing the accumulation of natural products based on activation tagging by Agrobacterium-mediated transformation with a T-DNA that carries cauliflower mosaic virus 35S enhancer sequences at its right border. Among approximately 5000 Arabidopsis activation-tagged lines, we found a plant that exhibited intense purple pigmentation in many vegetative organs throughout development. This upregulation of pigmentation reflected a dominant mutation that resulted in massive activation of phenylpropanoid biosynthetic genes and enhanced accumulation of lignin, hydroxycinnamic acid esters, and flavonoids, including various anthocyanins that were responsible for the purple color. These phenotypes, caused by insertion of the viral enhancer sequences adjacent to an MYB transcription factor gene, indicate that activation tagging can overcome the stringent genetic controls regulating the accumulation of specific natural products during plant development. Our findings suggest a functional genomics approach to the biotechnological evaluation of phytochemical biodiversity through the generation of massively enriched tissue sources for drug screening and for isolating underlying regulatory and biosynthetic genes.
MeSH Terms
Amino Acid Sequence
Arabidopsis/genetics,metabolism
Arabidopsis Proteins
Base Sequence
DNA Primers
DNA, Bacterial
DNA-Binding Proteins/genetics
Genes, Plant
Molecular Sequence Data
Phenylpropionates/metabolism
Plant Proteins/genetics
Proto-Oncogene Proteins c-myb
Sequence Homology, Amino Acid
Chemicals
ATR1 protein, Arabidopsis
Arabidopsis Proteins
CPC protein, Arabidopsis
DNA Primers
DNA, Bacterial
DNA-Binding Proteins
Phenylpropionates
Plant Proteins
Proto-Oncogene Proteins c-myb
T-DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Borevitz J O
Plant Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037, USA.
Xia Y
Blount J
Dixon R A
Lamb C
References (29)
29 references, click to expand
-
Developmental expression and substrate specificities of alfalfa caffeic acid 3-O-methyltransferase and caffeoyl coenzyme A 3-O-methyltransferase in relation to lignification.
Plant Physiol. 1998 Jul;117(3):761-70
PMID: 9662519
-
Dual methylation pathways in lignin biosynthesis
Plant Cell. 1998 Dec;10(12):2033-46
PMID: 9836743
-
Altering expression of cinnamic acid 4-hydroxylase in transgenic plants provides evidence for a feedback loop at the entry point into the phenylpropanoid pathway.
Plant Physiol. 2000 Jan;122(1):107-16
PMID: 10631254
-
Activation tagging in Arabidopsis.
Plant Physiol. 2000 Apr;122(4):1003-13
PMID: 10759496
-
The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein.
Plant Cell. 1999 Jul;11(7):1337-50
PMID: 10402433
-
Functional complementation of anthocyanin sequestration in the vacuole by widely divergent glutathione S-transferases.
Plant Cell. 1998 Jul;10(7):1135-49
PMID: 9668133
-
Maize R2R3 Myb genes: Sequence analysis reveals amplification in the higher plants.
Genetics. 1999 Sep;153(1):427-44
PMID: 10471724
-
Phloem-Specific Expression of Tyrosine/Dopa Decarboxylase Genes and the Biosynthesis of Isoquinoline Alkaloids in Opium Poppy.
Plant Cell. 1995 Nov;7(11):1811-1821
PMID: 12242361
-
Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and C1.
Science. 1992 Dec 11;258(5089):1773-5
PMID: 1465611
-
Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant.
Plant Cell. 1993 Dec;5(12):1795-805
PMID: 8305872
-
Engineering secondary metabolism in maize cells by ectopic expression of transcription factors.
Plant Cell. 1998 May;10(5):721-40
PMID: 9596632
-
Quantitative relationship between phenylalanine ammonia-lyase levels and phenylpropanoid accumulation in transgenic tobacco identifies a rate-determining step in natural product synthesis.
Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7608-12
PMID: 8052628
-
Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana.
Plant J. 1998 Oct;16(2):263-76
PMID: 9839469
-
A common gene regulates pigmentation pattern in diverse plant species.
Cell. 1992 Mar 6;68(5):955-64
PMID: 1547495
-
BAS1: A gene regulating brassinosteroid levels and light responsiveness in Arabidopsis.
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15316-23
PMID: 10611382
-
The an11 locus controlling flower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast, plants, and animals.
Genes Dev. 1997 Jun 1;11(11):1422-34
PMID: 9192870
-
Allele-specific interactions between ttg and gl1 during trichome development in Arabidopsis thaliana.
Genetics. 1999 Apr;151(4):1591-604
PMID: 10101180
-
More than 80R2R3-MYB regulatory genes in the genome of Arabidopsis thaliana.
Plant J. 1998 May;14(3):273-84
PMID: 9628022
-
Analysis of Arabidopsis mutants deficient in flavonoid biosynthesis.
Plant J. 1995 Nov;8(5):659-71
PMID: 8528278
-
A myb gene required for leaf trichome differentiation in Arabidopsis is expressed in stipules.
Cell. 1991 Nov 1;67(3):483-93
PMID: 1934056
-
Activation tagging of the floral inducer FT.
Science. 1999 Dec 3;286(5446):1962-5
PMID: 10583961
-
Role of the regulatory gene pl in the photocontrol of maize anthocyanin pigmentation.
Plant Cell. 1993 Dec;5(12):1807-16
PMID: 8305873
-
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal Biochem. 1976 May 7;72:248-54
PMID: 942051
-
Plant-cell cultures: fact and fantasy.
Biochem Soc Trans. 1983 Jan;11(1):23-8
PMID: 6337885
-
ORCA3, a jasmonate-responsive transcriptional regulator of plant primary and secondary metabolism.
Science. 2000 Jul 14;289(5477):295-7
PMID: 10894776
-
Development of several epidermal cell types can be specified by the same MYB-related plant transcription factor.
Development. 1998 Sep;125(17):3497-508
PMID: 9693152
-
Flavonoid and isoflavonoid distribution in developing soybean seedling tissues and in seed and root exudates.
Plant Physiol. 1991 Feb;95(2):594-603
PMID: 16668024
-
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):543-53
PMID: 8313474
-
Analysis of bHLH and MYB domain proteins: species-specific regulatory differences are caused by divergent evolution of target anthocyanin genes.
Plant J. 1998 Feb;13(4):475-88
PMID: 9680994