Molecular characterization of mutations in white-flowered torenia plants
Tóm tắt
Torenia (Torenia fournieri Lind.) is a model plant increasingly exploited in studies in various disciplines, including plant engineering, biochemistry, physiology, and ecology. Additionally, cultivars with different flower colors have been bred and made commercially available. Flower color in torenia is mainly attributed to the accumulation of anthocyanins, but the molecular mechanisms inducing flower color mutations in torenia have not been well elucidated. In this study, we therefore attempted to identify the cause of white coloration in torenia by comparing the white-flowered cultivar Crown White (CrW) with Crown Violet (CrV), a violet-flowered variety. In an expression analysis, no flavanone 3-hydroxylase (TfF3H) transcript accumulation was detected in CrW petals. Sequence analyses revealed that a novel long terminal repeat (LTR)-type retrotransposable element, designated as TORE1 (Torenia retrotransposon 1), is inserted into the 5′-upstream region of the TfF3H gene in CrW. A transient expression assay using torenia F3H promoters with or without TORE1 insertion showed that the TORE1 insertion substantially suppressed F3H promoter activity, suggesting that this insertion is responsible for the absence of F3H transcripts in white petals. Furthermore, a transformation experiment demonstrated that the introduction of a foreign gentian F3H cDNA, GtF3H, into CrW was able to recover pink-flower pigmentation, indicating that F3H deficiency is indeed the cause of the colorless flower phenotype in CrW. Detailed sequence analysis also identified deletion mutations in flavonoid 3′-hydroxylase (TfF3′H) and flavonoid 3′,5′- hydroxylase (TfF3′5′H) genes, but these were not directly responsible for white coloration in this cultivar. Taken together, a novel retrotransposable element, TORE1, inserted into the F3H 5′-upstream region is the cause of deficient F3H transcripts in white-flowered torenia, thereby leading to reduced petal anthocyanin levels. This is the first report of a retrotransposable element involved in flower color mutation in the genus Torenia.
Tài liệu tham khảo
Grotewold E: The Science of Flavonoids. London/New York: Springer-Verlag 2006.
Dixon RA, Liu C, Jun JH: Metabolic engineering of anthocyanins and condensed tannins in plants. Curr Opin Biotechnol. 2013, 24: 329-335. 10.1016/j.copbio.2012.07.004.
Dixon RA, Pasinetti GM: Flavonoids and isoflavonoids: from plant biology to agriculture and neuroscience. Plant Physiol. 2010, 154: 453-457. 10.1104/pp.110.161430.
Petroni K, Tonelli C: Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci. 2011, 181: 219-229. 10.1016/j.plantsci.2011.05.009.
Aida R: Torenia fournieri (torenia) as a model plant for transgenic studies. Plant Biotechnol. 2008, 25: 541-545. 10.5511/plantbiotechnology.25.541.
Nishihara M, Shimoda T, Nakatsuka T, Arimura G: Frontiers of torenia research: innovative ornamental traits and study of ecological interaction networks through genetic engineering. Plant Methods. 2013, 9: 23-10.1186/1746-4811-9-23.
Aida R, Yoshida K, Kondo T, Kishimoto S, Shibata M: Copigmentation gives bluer flowers on transgenic torenia plants with the antisense dihydroflavonol-4-reductase gene. Plant Sci. 2000, 160: 49-56. 10.1016/S0168-9452(00)00364-2.
Nakamura N, Fukuchi-Mizutani M, Fukui Y, Ishiguro K, Suzuki K, Suzuki H, Okazaki K, Shibata D, Tanaka Y: Generation of pink flower varieties from blue Torenia hybrida by redirecting the flavonoid biosynthetic pathway from delphinidin to pelargonidin. Plant Biotechnol. 2010, 27: 375-383. 10.5511/plantbiotechnology.10.0610a.
Ueyama Y, Suzuk IK, Fukuchi-Mizutani M, Fukui Y, Miyazaki K, Ohkawa H, Kusumi T, Tanaka Y: Molecular and biochemical characterization of torenia flavonoid 3'- hydroxylase and flavone synthase II and modification of flower color by modulating the expression of these genes. Plant Sci. 2002, 163: 253-263. 10.1016/S0168-9452(02)00098-5.
Nagira Y, Shimamura K, Hirai S, Shimanuki M, Kodama H, Ozeki Y: Identification and characterization of genes induced for anthocyanin synthesis and chlorophyll degradation in regenerated torenia shoots using suppression subtractive hybridization, cDNA microarrays, and RNAi techniques. J Plant Res. 2006, 119: 217-230. 10.1007/s10265-006-0264-4.
Sasaki K, Aida R, Yamaguchi H, Shikata M, Niki T, Nishijima T, Ohtsubo N: Functional divergence within class B MADS-box genes TfGLO and TfDEF in Torenia fournieri Lind. Mol Genet Genomics. 2010, 284: 399-414. 10.1007/s00438-010-0574-z.
Sasaki K, Yamaguchi H, Aida R, Shikata M, Abe T, Ohtsubo N: Mutation in Torenia fournieri Lind. UFO homolog confers loss of TfLFY interaction and results in a petal to sepal transformation. Plant J. 2012, 71: 1002-1014. 10.1111/j.1365-313X.2012.05047.x.
Narumi T, Aida R, Niki T, Nishijima T, Mitsuda N, Hiratsu K, Ohme-Takagi M, Ohtsubo N: Chimeric AGAMOUS repressor induces serrated petal phenotype in Torenia fournieri similar to that induced by cytokinin application. Plant Biotechnol. 2008, 25: 45-53. 10.5511/plantbiotechnology.25.45.
Narumi T, Aida R, Koyama T, Yamaguchi H, Sasaki K, Shikata M, Nakayama M, Ohme-Takagi M, Ohtsubo N: Arabidopsis chimeric TCP3 repressor produces novel floral traits in Torenia fournieri and Chrysanthemum morifolium. Plant Biotechnol. 2011, 28: 131-140. 10.5511/plantbiotechnology.11.0121a.
Takeuchi H, Higashiyama T: A species-specific cluster of defensin-like genes encodes diffusible pollen tube attractants in Arabidopsis. PLoS Biol. 2012, 10: e1001449-10.1371/journal.pbio.1001449.
Suzuki K, Xue H, Tanaka Y, Fukui Y, Fukuchi-Mizutani M, Murakami Y, Katsumoto Y, Tsuda S, Kusumi T: Flower color modifications of Torenia hybrida by cosuppression of anthocyanin biosynthesis genes. Mol Breed. 2000, 6: 239-246. 10.1023/A:1009678514695.
Fukusaki E, Kawasaki K, Kajiyama S, An CI, Suzuki K, Tanaka Y, Kobayashi A: Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference. J Biotechnol. 2004, 111: 229-240. 10.1016/j.jbiotec.2004.02.019.
Nakamura N, Fukuchi-Mizutani M, Miyazaki K, Suzuki K, Tanaka Y: RNAi suppression of the anthocyanidin synthase gene in Torenia hybrida yields white flowers with higher frequency and better stability than antisense and sense suppression. Plant Biotechnol. 2006, 23: 13-17. 10.5511/plantbiotechnology.23.13.
Ono E, Fukuchi-Mizutani M, Nakamura N, Fukui Y, Yonekura-Sakakibara K, Yamaguchi M, Nakayama T, Tanaka T, Kusumi T, Tanaka Y: Yellow flowers generated by expression of the aurone biosynthetic pathway. Proc Natl Acad Sci U S A. 2006, 103: 11075-11080. 10.1073/pnas.0604246103.
Gerats AG, Huits H, Vrijlandt E, Marana C, Souer E, Beld M: Molecular characterization of a nonautonomous transposable element (dTph1) of petunia. Plant Cell. 1990, 2: 1121-1128.
Clegg MT, Durbin ML: Flower color variation: a model for the experimental study of evolution. Proc Natl Acad Sci U S A. 2000, 97: 7016-7023. 10.1073/pnas.97.13.7016.
Martin C, Prescott A, Mackay S, Bartlett J, Vrijlandt E: Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J. 1991, 1: 37-49. 10.1111/j.1365-313X.1991.00037.x.
Hudson A, Critchley J, Erasmus Y: The genus Antirrhinum (snapdragon): a flowering plant model for evolution and development. CSH Protocols. 2008, 2008: pdb emo100-
Hoshino A, Abe Y, Saito N, Inagaki Y, Iida S: The gene encoding flavanone 3-hydroxylase is expressed normally in the pale yellow flowers of the Japanese morning glory carrying the speckled mutation which produce neither flavonol nor anthocyanin but accumulate chalcone, aurone and flavanone. Plant Cell Physiol. 1997, 38: 970-974. 10.1093/oxfordjournals.pcp.a029260.
Iida S, Morita Y, Choi JD, Park KI, Hoshino A: Genetics and epigenetics in flower pigmentation associated with transposable elements in morning glories. Adv Biophys. 2004, 38: 141-159.
Inagaki Y, Hisatomi Y, Suzuki T, Kasahara K, Iida S: Isolation of a Suppressor-mutator/Enhancer-like transposable element, Tpn1, from Japanese morning glory bearing variegated flowers. Plant Cell. 1994, 6: 375-383.
Hoshino A, Park KI, Iida S: Identification of r mutations conferring white flowers in the Japanese morning glory (Ipomoea nil). J Plant Res. 2009, 122: 215-222. 10.1007/s10265-008-0202-8.
Matsubara K, Kodama H, Kokubun H, Watanabe H, Ando T: Two novel transposable elements in a cytochrome P450 gene govern anthocyanin biosynthesis of commercial petunias. Gene. 2005, 358: 121-126.
Itoh Y, Higeta D, Suzuki A, Yoshida H, Ozeki Y: Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation (Dianthus caryophyllus). Plant Cell Physiol. 2002, 43: 578-585. 10.1093/pcp/pcf065.
Momose M, Nakayama M, Itoh Y, Umemoto N, Toguri T, Ozeki Y: An active hAT transposable element causing bud mutation of carnation by insertion into the flavonoid 3'-hydroxylase gene. Mol Genet Genomics. 2013, 288: 175-184. 10.1007/s00438-013-0742-z.
Nishizaki Y, Matsuba Y, Okamoto E, Okamura M, Ozeki Y, Sasaki N: Structure of the acyl-glucose-dependent anthocyanin 5-O-glucosyltransferase gene in carnations and its disruption by transposable elements in some varieties. Mol Genet Genomics. 2011, 286: 383-394. 10.1007/s00438-011-0655-7.
Nakatsuka T, Nishihara M, Mishiba K, Yamamura S: Two different mutations are involved in the formation of white-flowered gentian plants. Plant Sci. 2005, 169: 949-958. 10.1016/j.plantsci.2005.06.013.
Nakatsuka T, Haruta KS, Pitaksutheepong C, Abe Y, Kakizaki Y, Yamamoto K, Shimada N, Yamamura S, Nishihara M: Identification and characterization of R2R3-MYB and bHLH transcription factors regulating anthocyanin biosynthesis in gentian flowers. Plant Cell Physiol. 2008, 49: 1818-1829. 10.1093/pcp/pcn163.
Nishihara M, Hikage T, Yamada E, Nakatsuka T: A single-base substitution suppresses flower color mutation caused by a novel miniature inverted-repeat transposable element in gentian. Mol Genet Genomics. 2011, 286: 371-382. 10.1007/s00438-011-0652-x.
Nishijima T, Morita Y, Sasaki K, Nakayama M, Yamaguchi H, Ohtsubo N, Niki T, Niki T: A torenia (Torenia fournieri Lind. ex Fourn.) Novel mutant ‘Flecked’ produces variegated flowers by insertion of a DNA transposon into an R2R3-MYB gene. J Japan Soc Hort Sci. 2013, 39: 39-50.
Xiao H, Jiang N, Schaffner E, Stockinger EJ, van der Knaap E: A retrotransposon-mediated gene duplication underlies morphological variation of tomato fruit. Science. 2008, 319: 1527-1530. 10.1126/science.1153040.
Rico-Cabanas L, Martinez-Izquierdo JA: CIRE1, a novel transcriptionally active Ty1-copia retrotransposon from Citrus sinensis. Mol Genet Genomics. 2007, 277: 365-377. 10.1007/s00438-006-0200-2.
Nakatsuka T, Yamada E, Saito M, Fujita K, Nishihara M: Heterologous expression of gentian MYB1R transcription factors suppresses anthocyanin pigmentation in tobacco flowers. Plant Cell Rep. 2013, 32: 1925-1937. 10.1007/s00299-013-1504-4.
Koes R, Verweij W, Quattrocchio F: Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci. 2005, 10: 236-242. 10.1016/j.tplants.2005.03.002.
Britsch L, Ruhnau-Brich B, Forkmann G: Molecular cloning, sequence analysis, and in vitro expression of flavanone 3β-hydroxylase from Petunia hybrida. J Biol Chem. 1992, 267: 5380-5387.
van Houwelingen A, Souer E, Spelt K, Kloos D, Mol J, Koes R: Analysis of flower pigmentation mutants generated by random transposon mutagenesis in Petunia hybrida. Plant J. 1998, 13: 39-50.
Zabala G, Vodkin LO: The wp mutation of Glycine max carries a gene-fragment-rich transposon of the CACTA superfamily. Plant Cell. 2005, 17: 2619-2632. 10.1105/tpc.105.033506.
Dedio J, Saedler H, Forkmann G: Molecular cloning of the flavanone 3β-hydroxylase gene (FHT) from carnation (Dianthus caryophyllus) and analysis of stable and unstable FHT mutants. Theor Appl Genet. 1995, 90: 611-617.
Kumar A, Bennetzen JL: Plant retrotransposons. Annu Rev Genet. 1999, 33: 479-532. 10.1146/annurev.genet.33.1.479.
Casacuberta JM, Santiago N: Plant LTR-retrotransposons and MITEs: control of transposition and impact on the evolution of plant genes and genomes. Gene. 2003, 311: 1-11.
Huang CR, Burns KH, Boeke JD: Active transposition in genomes. Annu Rev Genet. 2012, 46: 651-675. 10.1146/annurev-genet-110711-155616.
Rebollo R, Horard B, Hubert B, Vieira C: Jumping genes and epigenetics: Towards new species. Gene. 2010, 454: 1-7. 10.1016/j.gene.2010.01.003.
Oliver KR, McComb JA, Greene WK: Transposable elements: powerful contributors to angiosperm evolution and diversity. Genome Biol Evol. 2013, 5: 1886-1901. 10.1093/gbe/evt141.
Grandbastien M, Casacuberta J: Plant transposable elements. Impact on genome structure and function. Springer Berlin Heidelberg: Springer 2012.
Uchiyama T, Fujino K, Ogawa T, Wakatsuki A, Kishima Y, Mikami T, Sano Y: Stable transcription activities dependent on an orientation of Tam3 transposon insertions into Antirrhinum and yeast promoters occur only within chromatin. Plant Physiol. 2009, 151: 1557-1569. 10.1104/pp.109.142356.
McClelland M, Nelson M, Raschke E: Effect of site-specific modification on restriction endonucleases and DNA modification methyltransferases. Nucleic Acids Res. 1994, 22: 3640-3659. 10.1093/nar/22.17.3640.
Jones AL, Thomas CL, Maule AJ: De novo methylation and co-suppression induced by a cytoplasmically replicating plant RNA virus. EMBO J. 1998, 17: 6385-6393. 10.1093/emboj/17.21.6385.
Vitte C, Panaud O: Formation of solo-LTRs through unequal homologous recombination counterbalances amplifications of LTR retrotransposons in rice Oryza sativa L. Mol Biol Evol. 2003, 20: 528-540. 10.1093/molbev/msg055.
Devos KM, Brown JK, Bennetzen JL: Genome size reduction through illegitimate recombination counteracts genome expansion in Arabidopsis. Genome Res. 2002, 12: 1075-1079. 10.1101/gr.132102.
Ma J, Bennetzen JL: Rapid recent growth and divergence of rice nuclear genomes. Proc Natl Acad Sci U S A. 2004, 101: 12404-12410. 10.1073/pnas.0403715101.
Toda K, Yang D, Yamanaka N, Watanabe S, Harada K, Takahashi R: A single-base deletion in soybean flavonoid 3'-hydroxylase gene is associated with gray pubescence color. Plant Mol Biol. 2002, 50: 187-196. 10.1023/A:1016087221334.
Takahashi R, Githiri SM, Hatayama K, Dubouzet EG, Shimada N, Aoki T, Ayabe S, Iwashina T, Toda K, Matsumura H: A single-base deletion in soybean flavonol synthase gene is associated with magenta flower color. Plant Mol Biol. 2007, 63: 125-135.
Morita Y, Hoshino A, Kikuchi Y, Okuhara H, Ono E, Tanaka Y, Fukui Y, Saito N, Nitasaka E, Noguchi H, Iida S: Japanese morning glory dusky mutants displaying reddish-brown or purplish-gray flowers are deficient in a novel glycosylation enzyme for anthocyanin biosynthesis, UDP-glucose:anthocyanidin 3-O-glucoside-2''-O-glucosyltransferase, due to 4-bp insertions in the gene. Plant J. 2005, 42: 353-363. 10.1111/j.1365-313X.2005.02383.x.
Hoshino A, Morita Y, Choi JD, Saito N, Toki K, Tanaka Y, Iida S: Spontaneous mutations of the flavonoid 3'-hydroxylase gene conferring reddish flowers in the three morning glory species. Plant Cell Physiol. 2003, 44: 990-1001. 10.1093/pcp/pcg143.
Durst F, Nelson DR: Diversity and evolution of plant P450 and P450-reductases. Drug Metabol Drug Interact. 1995, 12: 189-206.
Shimoda T, Nishihara M, Ozawa R, Takabayashi J, Arimura G: The effect of genetically enriched (E)-β-ocimene and the role of floral scent in the attraction of the predatory mite Phytoseiulus persimilis to spider mite-induced volatile blends of torenia. New Phytol. 2012, 193: 1009-1021. 10.1111/j.1469-8137.2011.04018.x.
Nakatsuka T, Nishihara M, Mishiba K, Yamamura S: Temporal expression of flavonoid biosynthesis-related genes regulates flower pigmentation in gentian plants. Plant Sci. 2005, 168: 1309-1318. 10.1016/j.plantsci.2005.01.009.
Hartmann U, Sagasser M, Mehrtens F, Stracke R, Weisshaar B: 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. 2005, 57: 155-171. 10.1007/s11103-004-6910-0.
Nakatsuka T, Saito M, Yamada E, Fujita K, Kakizaki Y, Nishihara M: Isolation and characterization of GtMYBP3 and GtMYBP4, orthologues of R2R3-MYB transcription factors that regulate early flavonoid biosynthesis, in gentian flowers. J Exp Bot. 2012, 63: 6505-6517. 10.1093/jxb/ers306.
Aida R: A protocol for transformation of Torenia. Methods Mol Biol. 2012, 847: 267-274. 10.1007/978-1-61779-558-9_23.