The Purple Cauliflower Arises from Activation of a MYB Transcription Factor

Oxford University Press (OUP) - Tập 154 Số 3 - Trang 1470-1480 - 2010
Li-Wei Chiu1, Xiangjun Zhou1, Sarah Burke1, Xianli Wu1, Ronald L. Prior1, Li Li1
1Department of Plant Breeding and Genetics (L.-W.C., X.Z., S.B., L.L.) and Robert W. Holley Center for Agriculture and Health, United States Department of Agriculture-Agricultural Research Service (L.L.), Cornell University, Ithaca, New York 14853; United States Department of Agriculture-Agricultural Research Service, Arkansas Children’s Nutrition Center, Little Rock, Arkansas 72202 (X.W., R.L.P.)

Tóm tắt

AbstractAnthocyanins are responsible for the color of many flowers, fruits, and vegetables. An interesting and unique Purple (Pr) gene mutation in cauliflower (Brassica oleracea var botrytis) confers an abnormal pattern of anthocyanin accumulation, giving the striking mutant phenotype of intense purple color in curds and a few other tissues. To unravel the nature of the Pr mutation in cauliflower, we isolated the Pr gene via a combination of candidate gene analysis and fine mapping. Pr encoded a R2R3 MYB transcription factor that exhibited tissue-specific expression, consistent with an abnormal anthocyanin accumulation pattern in the mutant. Transgenic Arabidopsis (Arabidopsis thaliana) and cauliflower plants expressing the Pr-D allele recapitulated the mutant phenotype, confirming the isolation of the Pr gene. Up-regulation of Pr specifically activated a basic helix-loop-helix transcription factor and a subset of anthocyanin structural genes encoding flavonoid 3’-hydroxylase, dihydroflavonol 4-reductase, and leucoanthocyanidin dioxygenase to confer ectopic accumulation of pigments in the purple cauliflower. Our results indicate that the genetic variation including a Harbinger DNA transposon insertion in the upstream regulatory region of the Pr-D allele is responsible for the up-regulation of the Pr gene in inducing phenotypic change in the plant. The successful isolation of Pr provides important information on the regulatory control of anthocyanin biosynthesis in Brassica vegetables, and offers a genetic resource for development of new varieties with enhanced health-promoting properties and visual appeal.

Từ khóa


Tài liệu tham khảo

Allan, 2008, MYB transcription factors that colour our fruit, Trends Plant Sci, 13, 99, 10.1016/j.tplants.2007.11.012

Baudry, 2006, TT8 controls its own expression in a feedback regulation involving TTG1 and homologous MYB and bHLH factors, allowing a strong and cell-specific accumulation of flavonoids in Arabidopsis thaliana, Plant J, 46, 768, 10.1111/j.1365-313X.2006.02733.x

Blazquez, 2007, Quantitative GUS activity assay of plant extracts, Cold Spring Harb Protoc, 2007

Borevitz, 2000, Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis, Plant Cell, 12, 2383, 10.1105/tpc.12.12.2383

Broun, 2005, Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis, Curr Opin Plant Biol, 8, 272, 10.1016/j.pbi.2005.03.006

Burr, 1996, The maize repressor-like gene intensifier1 shares homology with the r1/b1 multigene family of transcription factors and exhibits missplicing, Plant Cell, 8, 1249

Chandler, 1989, Two regulatory genes of the maize anthocyanin pathway are homologous: isolation of B utilizing R genomic sequences, Plant Cell, 1, 1175

Clough, 1998, Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana, Plant J, 16, 735, 10.1046/j.1365-313x.1998.00343.x

Cone, 1993, Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant, Plant Cell, 5, 1795

de Vetten, 1997, The an11 locus controlling flower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast, plants, and animals, Genes Dev, 11, 1422, 10.1101/gad.11.11.1422

Dixon, 2005, Proanthocyanidins—a final frontier in flavonoid research?, New Phytol, 165, 9, 10.1111/j.1469-8137.2004.01217.x

Dubos, 2008, MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana, Plant J, 55, 940, 10.1111/j.1365-313X.2008.03564.x

Espley, 2009, Multiple repeats of a promoter segment causes transcription factor autoregulation in red apples, Plant Cell, 21, 168, 10.1105/tpc.108.059329

Espley, 2007, Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10, Plant J, 49, 414, 10.1111/j.1365-313X.2006.02964.x

Gao, 1994, Quantitation and distribution of simple and acylated anthocyanins and other phenolics in blueberries, J Food Sci, 59, 1057, 10.1111/j.1365-2621.1994.tb08189.x

Goff, 1992, Functional analysis of the transcriptional activator encoded by the maize B gene: evidence for a direct functional interaction between two classes of regulatory proteins, Genes Dev, 6, 864, 10.1101/gad.6.5.864

Gonzalez, 2008, Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings, Plant J, 53, 814, 10.1111/j.1365-313X.2007.03373.x

Grotewold, 2006, The genetics and biochemistry of floral pigments, Annu Rev Plant Biol, 57, 761, 10.1146/annurev.arplant.57.032905.105248

Grotewold, 1994, The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset, Cell, 76, 543, 10.1016/0092-8674(94)90117-1

Grotewold, 2000, Identification of the residues in the Myb domain of maize C1 that specify the interaction with the bHLH cofactor R, Proc Natl Acad Sci USA, 97, 13579, 10.1073/pnas.250379897

Hartmann, 2005, Differential combinatorial interactions of cis-acting elements recognized by R2R3-MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes, Plant Mol Biol, 57, 155, 10.1007/s11103-004-6910-0

Koes, 2005, Flavonoids: a colorful model for the regulation and evolution of biochemical pathways, Trends Plant Sci, 10, 236, 10.1016/j.tplants.2005.03.002

Li, 2003, Molecular mapping of Or, a gene inducing beta-carotene accumulation in cauliflower (Brassica oleracea L. var. botrytis), Genome, 46, 588, 10.1139/g03-043

Li, 2001, A novel gene mutation that confers abnormal patterns of beta-carotene accumulation in cauliflower (Brassica oleracea var. botrytis), Plant J, 26, 59, 10.1046/j.1365-313x.2001.01008.x

Lu, 2006, The cauliflower Or gene encodes a DnaJ cysteine-rich domain-containing protein that mediates high levels of beta-carotene accumulation, Plant Cell, 18, 3594, 10.1105/tpc.106.046417

Ludwig, 1990, Maize R gene family: tissue-specific helix-loop-helix proteins, Cell, 62, 849, 10.1016/0092-8674(90)90259-H

Lyi, 2007, Biochemical and molecular characterization of the homocysteine S-methyltransferase from broccoli (Brassica oleracea var. italica), Phytochemistry, 68, 1112, 10.1016/j.phytochem.2007.02.007

Mathews, 2003, Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport, Plant Cell, 15, 1689, 10.1105/tpc.012963

Matsui, 2008, AtMYBL2, a protein with a single MYB domain, acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis, Plant J, 55, 954, 10.1111/j.1365-313X.2008.03565.x

Mazza, 1993, Anthocyanins in Fruits, Vegetables, and Grains

Naito, 2009, Unexpected consequences of a sudden and massive transposon amplification on rice gene expression, Nature, 461, 1130, 10.1038/nature08479

Nesi, 2000, The TT8 gene encodes a basic helix-loop-helix domain protein required for expression of DFR and BAN genes in Arabidopsis siliques, Plant Cell, 12, 1863, 10.1105/tpc.12.10.1863

Park, 2007, A bHLH regulatory gene in the common morning glory, Ipomoea purpurea, controls anthocyanin biosynthesis in flowers, proanthocyanidin and phytomelanin pigmentation in seeds, and seed trichome formation, Plant J, 49, 641, 10.1111/j.1365-313X.2006.02988.x

Payne, 2000, GL3 encodes a bHLH protein that regulates trichome development in Arabidopsis through interaction with GL1 and TTG1, Genetics, 156, 1349, 10.1093/genetics/156.3.1349

Paz-Ares, 1987, The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators, EMBO J, 6, 3553, 10.1002/j.1460-2075.1987.tb02684.x

Quattrocchio, 1999, Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color, Plant Cell, 11, 1433, 10.1105/tpc.11.8.1433

Quattrocchio, 1993, Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes, Plant Cell, 5, 1497, 10.2307/3869734

Rajagopalan, 2006, A diverse and evolutionarily fluid set of microRNAs in Arabidopsis thaliana, Genes Dev, 20, 3407, 10.1101/gad.1476406

Schwinn, 2006, A small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum, Plant Cell, 18, 831, 10.1105/tpc.105.039255

Shirley, 1992, Effects of ionizing radiation on a plant genome: analysis of two Arabidopsis transparent testa mutations, Plant Cell, 4, 333

Spelt, 2000, anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes, Plant Cell, 12, 1619, 10.1105/tpc.12.9.1619

Stracke, 2001, The R2R3-MYB gene family in Arabidopsis thaliana, Curr Opin Plant Biol, 4, 447, 10.1016/S1369-5266(00)00199-0

Sweeney, 2006, Caught red-handed: Rc encodes a basic helix-loop-helix protein conditioning red pericarp in rice, Plant Cell, 18, 283, 10.1105/tpc.105.038430

Tanaka, 2008, Biosynthesis of plant pigments: anthocyanins, betalains and carotenoids, Plant J, 54, 733, 10.1111/j.1365-313X.2008.03447.x

Toledo-Ortiz, 2003, The Arabidopsis basic/helix-loop-helix transcription factor family, Plant Cell, 15, 1749, 10.1105/tpc.013839

Udvardi, 2008, Eleven golden rules of quantitative RT-PCR, Plant Cell, 20, 1736, 10.1105/tpc.108.061143

Walker, 1999, The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein, Plant Cell, 11, 1337, 10.1105/tpc.11.7.1337

Walker, 2007, White grapes arose through the mutation of two similar and adjacent regulatory genes, Plant J, 49, 772, 10.1111/j.1365-313X.2006.02997.x

Wu, 2005, Systematic identification and characterization of anthocyanins by HPLC-ESI-MS/MS in common foods in the United States: fruits and berries, J Agric Food Chem, 53, 2589, 10.1021/jf048068b

Wu, 2005, Identification and characterization of anthocyanins by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry in common foods in the United States: vegetables, nuts, and grains, J Agric Food Chem, 53, 3101, 10.1021/jf0478861

Yuan, 2009, Transcriptional regulation of anthocyanin biosynthesis in red cabbage, Planta, 230, 1141, 10.1007/s00425-009-1013-4

Zhang, 2003, A network of redundant bHLH proteins functions in all TTG1-dependent pathways of Arabidopsis, Development, 130, 4859, 10.1242/dev.00681

Zhou, 2009, Involvement of a broccoli COQ5 methyltransferase in the production of volatile selenium compounds, Plant Physiol, 151, 528, 10.1104/pp.109.142521