Skip to main content
Log in

Update on transparent testa mutants from Arabidopsis thaliana: characterisation of new alleles from an isogenic collection

Planta Aims and scope Submit manuscript

Abstract

Main conclusion

We present a comprehensive overview on flavonoid-related phenotypes of A. thaliana tt and tds mutants, provide tools for their characterisation, increase the number of available alleles and demonstrate that tds3 is allelic to tt12 and tds5 to aha10.

Flavonoid biosynthesis is one of the best-studied secondary metabolite pathways in plants. In the model system Arabidopsis thaliana it leads to the synthesis of three phenolic compound classes: flavonol glycosides, anthocyanins and proanthocyanidins (PAs). PAs appear brown in their oxidised polymeric forms, and most A. thaliana mutants impaired in flavonoid accumulation were identified through screens for lack of this seed coat pigmentation. These mutants are referred to as transparent testa (tt) or tannin-deficient seed (tds). More than 20 mutants of these types have been published, probably representing most of the genes relevant for PA accumulation in A. thaliana. However, data about the genes involved in PA deposition or oxidation are still rather scarce. Also, for some of the known mutants it is unclear if they represent additional loci or if they are allelic to known genes. For the present study, we have performed a systematic phenotypic characterisation of almost all available tt and tds mutants and built a collection of mutants in the genetic background of the accession Columbia to minimise effects arising from ecotype variation. We have identified a novel tt6 allele from a forward genetic screen and demonstrated that tds3 is allelic to tt12 and tds5 to aha10.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

AGI:

Arabidopsis Genome Initiative

Col:

Columbia

DMACA:

p-dimethylaminocinnamaldehyde

DPBA:

Diphenylboric acid 2-aminoethyl ester

EGL3:

ENHANCER OF GLABRA 3

MBW:

MYB-bHLH-WD40 repeat protein

NFZ:

Norflurazon

PA(s):

Proanthocyanidin(s)

PAP1:

Production of Anthocyanin Pigment 1

PFG:

Production of Flavonol Glycosides

tds :

tannin-deficient seed

TLC:

Thin-layer chromatography

tt :

transparent testa

Ws:

Wassilewskija

References

  • Abrahams S, Tanner GJ, Larkin PJ, Ashton AR (2002) Identification and biochemical characterization of mutants in the proanthocyanidin pathway in Arabidopsis. Plant Physiol 130:561–576

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Abrahams S, Lee E, Walker AR, Tanner GJ, Larkin PJ, Ashton AR (2003) The Arabidopsis TDS4 gene encodes leucoanthocyanidin dioxygenase (LDOX) and is essential for proanthocyanidin synthesis and vacuole development. Plant J 35:624–636

    Article  PubMed  CAS  Google Scholar 

  • Albert S, Delseny M, Devic M (1997) BANYULS, a novel negative regulator of flavonoid biosynthesis in the Arabidopsis seed coat. Plant J 11:289–299

    Article  PubMed  CAS  Google Scholar 

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prednis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schmid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  PubMed  Google Scholar 

  • Appelhagen I, Huep G, Lu GH, Strompen G, Weisshaar B, Sagasser M (2010) Weird fingers: functional analysis of WIP domain proteins. FEBS Letters 584:3116–3122

  • Appelhagen I, Jahns O, Bartelniewoehner L, Sagasser M, Weisshaar B, Stracke R (2011a) Leucoanthocyanidin dioxygenase in Arabidopsis thaliana: characterization of mutant alleles and regulation by MYB-BHLH-TTG1 transcription factor complexes. Gene 484:61–68

    Article  PubMed  CAS  Google Scholar 

  • Appelhagen I, Lu GH, Huep G, Schmelzer E, Weisshaar B, Sagasser M (2011b) TRANSPARENT TESTA1 interacts with R2R3-MYB factors and affects early and late steps of flavonoid biosynthesis in the endothelium of Arabidopsis thaliana seeds. Plant J 67:406–419

    Article  PubMed  CAS  Google Scholar 

  • Baudry A, Heim MA, Dubreucq B, Caboche M, Weisshaar B, Lepiniec L (2004) TT2, TT8, and TTG1 synergistically specify the expression of BANYULS and proanthocyanidin biosynthesis in Arabidopsis thaliana. Plant J 39:366–380

    Article  PubMed  CAS  Google Scholar 

  • Baxter IR, Young JC, Armstrong G, Foster N, Bogenschutz N, Cordova T, Peer WA, Hazen SP, Murphy AS, Harper JF (2005) A plasma membrane H+-ATPase is required for the formation of proanthocyanidins in the seed coat endothelium of Arabidopsis thaliana. Proc Natl Acad Sci USA 102:2649–2654

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Bharti A, Khurana J (2003) Molecular characterization of transparent testa (tt) mutants of Arabidopsis thaliana (ecotype Estland) impaired in flavonoid biosynthetic pathway. Plant Sci 165:1321–1332

    Article  CAS  Google Scholar 

  • Bolle C, Huep G, Kleinbolting N, Haberer G, Mayer K, Leister D, Weisshaar B (2013) GABI-DUPLO: a collection of double mutants to overcome genetic redundancy in Arabidopsis thaliana. Plant J 75:157–171

    Article  PubMed  CAS  Google Scholar 

  • Bowerman PA, Ramirez MV, Price MB, Helm RF, Winkel BS (2012) Analysis of T-DNA alleles of flavonoid biosynthesis genes in Arabidopsis ecotype Columbia. BMC Res Notes 5:485

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Brillouet JM, Romieu C, Schoefs B, Solymosi K, Cheynier V, Fulcrand H, Verdeil JL, Conéjéro G (2013) The tannosome is an organelle forming condensed tannins in the chlorophyllous organs of Tracheophyta. Ann Bot (Lond) 112:1003–1014

    Article  CAS  Google Scholar 

  • Buer CS, Sukumar P, Muday GK (2006) Ethylene modulates flavonoid accumulation and gravitropic responses in roots of Arabidopsis. Plant Physiol 140:1384–1396

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Bürger D (1971) Die morphologischen Mutanten des Göttinger Arabidopsis-Sortiments, einschließlich der Mutanten mit abweichender Samenfarbe. Arabidopsis Inf Serv 8:36–42

    Google Scholar 

  • Debeaujon I, Leon-Kloosterziel K, Koornneef M (2000) Influence of the testa on seed dormancy, germination, and longevity in Arabidopsis. Plant Physiol 122:403–413

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Debeaujon I, Peeters AJM, Léon-Kloosterziel KM, Koornneef M (2001) The TRANSPARENT TESTA12 gene of Arabidopsis encodes a multidrug secondary transporter-like protein required for flavonoid sequestration in vacuoles of the seed coat endothelium. Plant Cell 13:853–872

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Debeaujon I, Nesi N, Perez P, Devic M, Grandjean O, Caboche M, Lepiniec L (2003) Proanthocyanidin-accumulating cells in Arabidopsis testa: regulation of differentiation and role in seed development. Plant Cell 15:2514–2531

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • DeBolt S, Scheible WR, Schrick K, Auer M, Beisson F, Bischoff V, Bouvier-Navé P, Carroll A, Hematy K, Li Y, Milne J, Nair M, Schaller H, Zemla M, Somerville C (2009) Mutations in UDP-glucose:sterol glucosyltransferase in Arabidopsis cause transparent testa phenotype and suberization defect in seeds. Plant Physiol 151:78–87

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Devic M, Guilleminot J, Debeaujon I, Bechtold N, Bensaude E, Koornneef M, Pelletier G, Delseny M (1999) The BANYULS gene encodes a DFR-like protein and is a marker of early seed coat development. Plant J 19:387–398

    Article  PubMed  CAS  Google Scholar 

  • Feinbaum RL, Ausubel FM (1988) Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol 8:1985–1992

    PubMed  CAS  PubMed Central  Google Scholar 

  • Focks N, Sagasser M, Weisshaar B, Benning C (1999) Characterization of tt15, a novel transparent testa mutant of Arabidopsis thaliana (L.) Heynh. Planta 208:352–357

    Article  PubMed  CAS  Google Scholar 

  • Francisco RM, Regalado A, Ageorges A, Burla BJ, Bassin B, Eisenach C, Zarrouk O, Vialet S, Marlin T, Chaves MM, Martinoia E, Nagy R (2013) ABCC1, an ATP binding cassette protein from grape berry, transports anthocyanidin 3-O-glucosides. Plant Cell 25:1840–1854

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Garcia D, Fitz Gerald JN, Berger F (2005) Maternal control of integument cell elongation and zygotic control of endosperm growth are coordinated to determine seed size in Arabidopsis. Plant Cell 17:52–60

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827

    Article  PubMed  CAS  Google Scholar 

  • Gonzalez A, Mendenhall J, Huo Y, Lloyd A (2009) TTG1 complex MYBs, MYB5 and TT2, control outer seed coat differentiation. Developmental Biology 325:412–421

  • Gonzalez A, Mendenhall J, Huo Y, Lloyd A (2009) TTG1 complex MYBs, MYB5 and TT2, control outer seed coat differentiation. Developmental Biology 325: 412-421 Ishida T, Hattori S, Sano R, Inoue K, Shirano Y, Hayashi H, Shibata D, Sato S, Kato T, Tabata S, Okada K, Wada T (2007) Arabidopsis TRANSPARENT TESTA GLABRA2 is directly regulated by R2R3 MYB transcription factors and is involved in regulation of GLABRA2 transcription in epidermal differentiation. Plant Cell 19:2531–2543

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Jez JM, Noel JP (2002) Reaction mechanism of chalcone isomerase. pH dependence, diffusion control, and product binding differences. J Biol Chem 277:1361–1369

    Article  PubMed  CAS  Google Scholar 

  • Jez JM, Bowman ME, Dixon RA, Noel JP (2000) Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerase. Nat Struct Biol 7:786–791

    Article  PubMed  CAS  Google Scholar 

  • Johnson CS, Kolevski B, Smyth DR (2002) TRANSPARENT TESTA GLABRA2, a trichome and seed coat development gene of Arabidopsis, encodes a WRKY transcription factor. Plant Cell 14:1359–1375

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Kitamura S, Shikazono N, Tanaka A (2004) TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis. Plant J 37:104–114

    Article  PubMed  CAS  Google Scholar 

  • Kitamura S, Matsuda F, Tohge T, Yonekura-Sakakibara K, Yamazaki M, Saito K, Narumi I (2010) Metabolic profiling and cytological analysis of proanthocyanidins in immature seeds of Arabidopsis thaliana flavonoid accumulation mutants. Plant J 62:549–559

    Article  PubMed  CAS  Google Scholar 

  • Klein M, Burla B, Martinoia E (2006) The multidrug resistance-associated protein (MRP/ABCC) subfamily of ATP-binding cassette transporters in plants. FEBS Lett 580:1112–1122

    Article  PubMed  CAS  Google Scholar 

  • Kleinboelting N, Huep G, Kloetgen A, Viehoever P, Weisshaar B (2012) GABI-Kat SimpleSearch: new features of the Arabidopsis thaliana T-DNA mutant database. Nucleic Acids Res 40:D1211–D1215

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Koornneef M (1981) The complex syndrome of ttg mutants. Arabidopsis Inf Serv 18:45–51

    Google Scholar 

  • Koornneef M (1990) Mutations affecting the testa colour in Arabidopsis. Arabidopsis Inf Serv 27:1–4

    Google Scholar 

  • Koornneef M, Luiten W, de Vlaming P, Schram AW (1982) A gene controlling flavonoid 3′-hydroxylation in Arabidopsis. Arabidopsis Inf Serv 19:113–115

    Google Scholar 

  • Lepiniec L, Debeaujon I, Routaboul JM, Baudry A, Pourcel L, Nesi N, Caboche M (2006) Genetics and biochemistry of seed flavonoids. Annu Rev Plant Biol 57:405–430

    Article  PubMed  CAS  Google Scholar 

  • Lukacin R, Britsch L (1997) Identification of strictly conserved histidine and arginine residues as part of the active site in Petunia hybrida flavanone 3β-hydroxylase. Eur J Biochem 249:748–757

    Article  PubMed  CAS  Google Scholar 

  • Martens S, Preuß A, Matern U (2010) Multifunctional flavonoid dioxygenases: flavonol and anthocyanin biosynthesis in Arabidopsis thaliana L. Phytochemistry 71:1040–1049

    Article  PubMed  CAS  Google Scholar 

  • Mueller LA, Goodman CD, Silady RA, Walbot V (2000) AN9, a petunia glutathione S-transferase required for anthocyanin sequestration, is a flavonoid-binding protein. Plant Physiol 123:1561–1570

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Nesi N, Debeaujon I, Jond C, Stewart AJ, Jenkins GI, Caboche M, Lepiniec L (2002) The TRANSPARENT TESTA16 locus encodes the ARABIDOPSIS BSISTER MADS domain protein and is required for proper development and pigmentation of the seed coat. Plant Cell 14:2463–2479

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Owens DK, Crosby KC, Runac J, Howard BA, Winkel BS (2008) Biochemical and genetic characterization of Arabidopsis flavanone 3β-hydroxylase. Plant Physiol Biochem 46:833–843

    Article  PubMed  CAS  Google Scholar 

  • Pang Y, Peel G, Sharma S, Tang Y, Dixon R (2008) A transcript profiling approach reveals an epicatechin-specific glucosyltransferase expressed in the seed coat of Medicago truncatula. Proc Natl Acad Sci USA 105:14210–14215

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Pang Y, Cheng X, Huhman DV, Ma J, Peel GJ, Yonekura-Sakakibara K, Saito K, Shen G, Sumner LW, Tang Y, Wen J, Yun J, Dixon RA (2013) Medicago glucosyltransferase UGT72L1: potential roles in proanthocyanidin biosynthesis. Planta 238:139–154

    Article  PubMed  CAS  Google Scholar 

  • Peer WA, Brown DE, Tague BW, Muday GK, Taiz L, Murphy AS (2001) Flavonoid accumulation patterns of transparent testa mutants of Arabidopsis. Plant Physiol 126:536–548

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Pourcel L, Routaboul JM, Kerhoas L, Caboche M, Lepiniec L, Debeaujon I (2005) TRANSPARENT TESTA10 encodes a laccase-like enzyme involved in oxidative polymerization of flavonoids in Arabidopsis seed coat. Plant Cell 17:2966–2980

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Pourcel L, Routaboul JM, Cheynier V, Lepiniec L, Debeaujon I (2007) Flavonoid oxidation in plants: from biochemical properties to physiological functions. Trends Plant Sci 12:29–36

    Article  PubMed  CAS  Google Scholar 

  • Pourcel L, Irani NG, Lu Y, Riedl K, Schwartz S, Grotewold E (2010) The formation of anthocyanic vacuolar inclusions in Arabidopsis thaliana and implications for the sequestration of anthocyanin pigments. Mol Plant 3:78–90

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Rosso MG, Li Y, Strizhov N, Reiss B, Dekker K, Weisshaar B (2003) An Arabidopsis thaliana T-DNA mutagenised population (GABI-Kat) for flanking sequence tag based reverse genetics. Plant Mol Biol 53:247–259

    Article  PubMed  CAS  Google Scholar 

  • Routaboul JM, Kerhoas L, Debeaujon I, Pourcel L, Caboche M, Einhorn J, Lepiniec L (2006) Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana. Planta 224:96–107

    Article  PubMed  CAS  Google Scholar 

  • Routaboul JM, Dubos C, Beck G, Marquis C, Bidzinski P, Loudet O, Lepiniec L (2012) Metabolite profiling and quantitative genetics of natural variation for flavonoids in Arabidopsis. J Exp Bot 63:3749–3764

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Sagasser M, Lu G-H, Hahlbrock K, Weisshaar B (2002) A. thaliana TRANSPARENT TESTA 1 is involved in seed coat development and defines the WIP subfamily of plant zinc finger proteins. Genes Dev 16:138–149

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Saito K, Yonekura-Sakakibara K, Nakabayashi R, Higashi Y, Yamazaki M, Tohge T, Fernie AR (2013) The flavonoid biosynthetic pathway in Arabidopsis: structural and genetic diversity. Plant Physiol Biochem 72:21–34

    Article  PubMed  CAS  Google Scholar 

  • Sarkar SK, Howarth RE (1976) Specificity of the vanillin test for flavanols. J Agric Food Chem 24:317–320

    Article  PubMed  CAS  Google Scholar 

  • Schoenbohm C, Martens S, Eder C, Forkmann G, Weisshaar B (2000) Identification of the Arabidopsis thaliana flavonoid 3′-hydroxylase gene and functional expression of the encoded P450 enzyme. Biol Chem 381:749–753

    Article  PubMed  CAS  Google Scholar 

  • Sheahan JJ, Rechnitz GA (1992) Flavonoid-specific staining of Arabidopsis thaliana. BioTechniques 13:880–883

    PubMed  CAS  Google Scholar 

  • Sheahan JJ, Rechnitz GA (1993) Differential visualization of transparent testa mutants in Arabidopsis thaliana. Anal Chem 65:961–963

    Article  CAS  Google Scholar 

  • Sheahan JJ, Cheong H, Rechnitz GA (1998) The colorless flavonoids of Arabidopsis thaliana (Brassicaceae). I. A model system to study the orthodihydroxy structure. Am J Bot 85:467–475

    Article  PubMed  CAS  Google Scholar 

  • Shirley BW, Hanley S, Goodman HM (1992) Effects of ionizing radiation on a plant genome: analysis of two Arabidopsis transparent testa mutations. Plant Cell 4:333–347

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Shirley BW, Kubasek WL, Storz G, Bruggemann E, Koornneef M, Ausubel FM, Goodman HM (1995) Analysis of Arabidopsis mutants deficient in flavonoid biosynthesis. Plant J 8:659–671

    Article  PubMed  CAS  Google Scholar 

  • Stracke R, Ishihara H, Huep G, Barsch A, Mehrtens F, Niehaus K, Weisshaar B (2007) Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. Plant J 50:660–677

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Stracke R, De Vos RCH, Bartelniewoehner L, Ishihara H, Sagasser M, Martens S, Weisshaar B (2009) Metabolomic and genetic analyses of flavonol synthesis in Arabidopsis thaliana support the in vivo involvement of leucoanthocyanidin dioxygenase. Planta 229:427–445

    Article  PubMed  CAS  Google Scholar 

  • Stracke R, Jahns O, Keck M, Tohge T, Niehaus K, Fernie AR, Weisshaar B (2010) Analysis of PRODUCTION OF FLAVONOL GLYCOSIDES-dependent flavonol glycoside accumulation in Arabidopsis thaliana plants reveals MYB11-, MYB12- and MYB111-independent flavonol glycoside accumulation. New Phytol 188:985–1000

    Article  PubMed  CAS  Google Scholar 

  • Sun Y, Li H, Huang JR (2012) Arabidopsis TT19 functions as a carrier to transport anthocyanin from the cytosol to tonoplasts. Mol Plant 5:387–400

    Article  PubMed  CAS  Google Scholar 

  • Teng S, Keurentjes J, Bentsink L, Koornneef M, Smeekens S (2005) Sucrose-specific induction of anthocyanin biosynthesis in Arabidopsis requires the MYB75/PAP1 gene. Plant Physiol 139:1840–1852

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Verweij W, Spelt C, Di Sansebastiano GP, Vermeer J, Reale L, Ferranti F, Koes R, Quattrocchio F (2008) An H+ P-ATPase on the tonoplast determines vacuolar pH and flower colour. Nat Cell Biol 10:1456–1462

    Article  PubMed  CAS  Google Scholar 

  • Viotti C, Kruger F, Krebs M, Neubert C, Fink F, Lupanga U, Scheuring D, Boutte Y, Frescatada-Rosa M, Wolfenstetter S, Sauer N, Hillmer S, Grebe M, Schumacher K (2013) The endoplasmic reticulum is the main membrane source for biogenesis of the lytic vacuole in Arabidopsis. Plant Cell 25:3434–3449

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Walker AR, Davison PA, Bolognesi-Winfield AC, James CM, Srinivasan N, Blundell TL, Esch JJ, Marks MD, Gray JC (1999) The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein. Plant Cell 11:1337–1350

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Wangwattana B, Koyama K, Nishiyama Y, Kitayama M, Yamazaki M, Saito K (2008) Characterization of PAP1-upregulated glutathione S-transferase genes in Arabidopsis thaliana. Plant Biotechnol 25:191–196

    Article  CAS  Google Scholar 

  • Weisshaar B, Jenkins GI (1998) Phenylpropanoid biosynthesis and its regulation. Curr Opin Plant Biol 1:251–257

    Article  PubMed  CAS  Google Scholar 

  • Xie DY, Sharma SB, Paiva NL, Ferreira D, Dixon RA (2003) Role of anthocyanidin reductase, encoded by BANYULS in plant flavonoid biosynthesis. Science 299:396–399

    Article  PubMed  CAS  Google Scholar 

  • Xu W, Grain D, Bobet S, Le Gourrierec J, Thévenin J, Kelemen Z, Lepiniec L, Dubos C (2013a) Complexity and robustness of the flavonoid transcriptional regulatory network revealed by comprehensive analyses of MYB-bHLH-WDR complexes and their targets in Arabidopsis seed. New Phytol 202:132–144

    Article  PubMed  Google Scholar 

  • Xu W, Grain D, Le Gourrierec J, Harscoet E, Berger A, Jauvion V, Scagnelli A, Berger N, Bidzinski P, Kelemen Z, Salsac F, Baudry A, Routaboul JM, Lepiniec L, Dubos C (2013b) Regulation of flavonoid biosynthesis involves an unexpected complex transcriptional regulation of TT8 expression, in Arabidopsis. New Phytol 198:59–70

    Article  PubMed  CAS  Google Scholar 

  • Zhao J, Dixon RA (2009) MATE transporters facilitate vacuolar uptake of epicatechin 3′-O-glucoside for proanthocyanidin biosynthesis in Medicago truncatula and Arabidopsis. Plant Cell 21:2323–2340

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Zhao J, Pang Y, Dixon RA (2010) The mysteries of proanthocyanidin transport and polymerization. Plant Physiol 153:437–443

    Article  PubMed  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgments

We are grateful to Nadja Parusel for help with tt6-5 mutant analysis and Andrea Voigt for excellent technical assistance. We thank Sharon Abrahams and Anthony Ashton (CSIRO-Plant Industry, Canberra, Australia) for seeds of the tds mutant lines, NASC for providing additional seed stocks and the reviewers for their suggestions to improve the manuscript. This project was funded in part by the German Federal Ministry of Education and Research (BMBF) in the context of the German plant genomics program GABI (Förderkennzeichen 0313855).

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Bernd Weisshaar.

Additional information

Special topic: Anthocyanins. Guest editor: Stefan Martens.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Table S1 Oligonucleotides used for genotyping (DOCX 15 kb)

425_2014_2088_MOESM2_ESM.tiff

Fig. S1 Genomic structure and phenotypes of different tt3 alleles. a Gene model of tt3 alleles characterised in this study. Symbols and colours as in Fig. 4. b Left column, vanillin/HCl-treated seeds; right column, DMACA-stained mature seed as in Fig. 3 (TIFF 9696 kb)

425_2014_2088_MOESM3_ESM.tiff

Fig. S2 Genomic structure and phenotypes of different tt2 alleles. a Gene model of tt2 alleles characterised in this study. Symbols and colours as in Fig. 4. b TT2 transcript levels in tt2-5 relative to the expression in Col-0 determined by qRT-PCR. c Vanillin/HCl-treated seeds of the indicated developmental stages. d DMACA-stained mature seed as in Fig. 3. e Mature seeds as in Fig. 2 (TIFF 6376 kb)

425_2014_2088_MOESM4_ESM.tiff

Fig. S3 Genomic structure and phenotypes of different tt6 alleles. Upper panel, gene model of tt6 alleles characterised in this study. Symbols and colours as in Fig. 4. Lower panel, mature seeds as in Fig. 2 (TIFF 3306 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Appelhagen, I., Thiedig, K., Nordholt, N. et al. Update on transparent testa mutants from Arabidopsis thaliana: characterisation of new alleles from an isogenic collection. Planta 240, 955–970 (2014). https://doi.org/10.1007/s00425-014-2088-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-014-2088-0

Keywords

Navigation