Abstract
The surface of plants is covered with a cuticular wax, which contains a mixture of very-long-chain fatty acid derivatives. This wax layer provides a hydrophobic barrier which reduces non-stomatal water loss and prevents pathogen attack. The biosynthesis pathway of cuticular wax in Arabidopsis is well studied; however, little is known about the synthesis of cuticular wax in Brassica rapa. Genetic analyses indicated that the waxy characteristic is controlled by a single dominant gene. In the present study, preliminary mapping results from an F2 population consisting of 308 recessive individuals showed that the BrWax1 (Brassica Wax) gene is located on linkage group A01. We developed a set of new markers closely linked to the target gene, and used another population of 1,020 recessive F2 individuals to fine-map the BrWax1 gene to a genomic DNA fragment of approximately 86.4 kb. Fifteen genes were identified in this target region. Based on gene annotation, the Bra013809 gene was the candidate for the BrWax1 gene. Quantitative real-time PCR analysis and expression pattern of the two parents showed that the expression level of Bra013809 was much higher in the waxy phenotype than in the glossy phenotype. This result should provide not only important information for functional studies of the BrWax1 gene, but also the starting point for studying the pathway of cuticular wax biosynthesis in Brassica rapa.
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References
Aarts MGM, Keijzer CJ, Stiekema WJ et al (1995) Molecular characterization of the CER1 gene of Arabidopsis involved in epicuticular wax biosynthesis and pollen fertility. Plant Cell 7:2115–2127
Baker EA (1982) In: Cutler DJ, Alvin KL, Price CE (eds) The plant cuticle. Academic Press, London, pp 139–165
Bernard A, Joubès J (2013) Arabidopsis cuticular waxes: advances in synthesis, export and regulation. Progr Lipid Res 52:110–129
Bourdenx B, Bernard A, Domergue F, Pascal S et al (2011) Overexpression of Arabidopsis ECERIFERUM1 promotes wax very-long-chain alkane biosynthesis and influences plant response to biotic and abiotic stresses. Plant Physiol 156:29–45
Broun P, Poindexter P, Osborne E et al (2004) WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis. Proc Natl Acad Sci USA 10:4706–4711
Burow GB, Franks CD, Acosta-Martinez V, Xin ZG (2009) Molecular mapping and characterization of BLMC, a locus for profuse wax (bloom) and enhanced cuticular features of Sorghum (Sorghum bicolor (L.) Moench.). Theor Appl Genet 118:423–431
Chen XB, Goodwin M, Boroff VL et al (2003) Cloning and characterization of the WAX2 gene of Arabidopsis involved in cuticle membrane and wax production. Plant Cell 15:1170–1185
D’Auria JC (2006) Acyltransferases in plants: a good time to be BAHD. Plant Biol 9:331–340
Fiebig A, Mayfield JA, Miley NL, Chau S, Fischer RL, Preuss D (2000) Alterations in CER6, a gene identical to CUT1, differentially affect long-chain lipid content on the surface of pollen and stems. Plant Cell 12:2001–2008
Hannoufa A, Negruk V, Eisner G et al (1996) The CER3 gene of Arabidopsis thaliana is expressed in leaves, stems, roots, flowers and apical meristems. Plant J 10:459–467
Hansen JD, Pyee J, Xia Y et al (1997) The glossy1 locus of maize and an epidermis- specific cDNA from Kleinia odora define a class of receptor-like proteins required for the normal accumulation of cuticular waxes. Plant Physiol 113:1091–1100
Haslam TM, Mañas-Fernández A, Zhao LF, Kunst L (2012) Arabidopsis ECERIFERUM2 is a component of the fatty acid elongation machinery required for fatty acid extension to exceptional lengths. Plant Physiol 160:1164–1174
Holmes MG, Keiller DR (2002) Effects of pubescence and waxes on the reflectance of leaves in the ultraviolet and photosynthetic wavebands: a comparison of a range of species. Plant, Cell Environ 25:85–93
Hooker TS, Millar AA, Kunst L (2002) Significance of the expression of the CER6 condensing enzyme for cuticular wax production in Arabidopsis. Plant Physiol 129:1568–1580
Jenks MA, Eigenbrode SD, Lemieux B (2002) Cuticular waxes of Arabidopsis. In: Somerville CR, Meyerowitz EM (eds) The Arabidopsis book. American Society of Plant Biologists, Rockville, MD
Jung KH, Han MJ, Lee DY, Lee YS, Schreiber L, Franke R, Faust A, Yephremov A, Saedler H, Kim YW, HwangI AG (2006) Wax-deficient anther1 is involved in cuticle and wax production in rice anther walls and is required for pollen development. Plant Cell 18:3015–3032
Kannangara R, Branigan C, Liu Y et al (2007) The transcription factor WIN1/SHN1 regulates cutin biosynthesis in Arabidopsis thaliana. Plant Cell 19:1278–1294
Koornneef M, Hanhart CJ, Thiel F (1989) A genetic and phenotypic description of eceriferum (cer) mutants in Arabidopsis thaliana. J Hered 80:118–122
Kunst L, Samuels AL (2003) Biosynthesis and secretion of plant cuticular wax. Prog Lipid Res 42:51–80
Kunst L, Samuels L (2009) Plant cuticles shine: advances in wax biosynthesis and export. Curr Opin Plant Biol 12:721–727
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-[Delta][Delta] CT method. Methods 25(4):402–408
Mariani M, Wolters-Arts M (2000) Complex waxes. Plant Cell 12:1795–1798
Markstädter C, Federle W, Jetter R, Riederer M, Hölldobler B (2000) Chemical composition of the slippery epicuticular wax blooms on Macaranga (Euphorbiac- eae) ant-plants. Chemoecology 10:33–40
Martin JT, Juniper BE (1970) The cuticles of plants. Edward Arnold, London
Millar AA, Clemens S, Zachgo S, Giblin EM, Taylor DC, Kunst L (1999) CUT1, an Arabidopsis gene required for cuticular wax biosynthesis and pollen fertility, encodes a very-long-chain fatty acid condensing enzyme. Plant Cell 11:825–838
Moose S, Sisco P (1996) Glossy15, an APETALA2-like gene from maize that regulates leaf epidermal cell identity. Genes Dev 10:3018–3027
Müller C (2006) Plant-Insect interactions on cuticular surfaces. In: Riederer M, Müller C (eds) Biology of the plant cuticle. Blackwell, Oxford, pp 398–417
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4326
Negruk V, Yang P, Subramanian M et al (1996) Molecular cloning and characterization of the CER2 gene of Arabidopsis thaliana. Plant J 9:137–145
Oliveira AFM, Meirelles ST, Salatino A (2003) Epicuticular waxes from caatinga and cerrado species and their efficiency against water loss. Anais de Academia Brasileira de Ciencias 75:431–439
Post-Beittenmiller D (1998) The cloned Eceriferum genes of Arabidopsis and the corresponding Glossy genes in maize. Plant Physiol Biochem 36:157–166
Pruitt RE, Vielle-Calzada JP, Ploense SE et al (2000) FIDDLEHEAD, a gene required to suppress epidermal cell interactions in Arabidopsis, encodes a putative lipid biosynthetic enzyme. Proc Natl Acad Sci USA 97:1311–1316
Qin BX, Tang D, Huang J, Li M, Wu XR, Lu LL, Wang KJ, Yu HX, Chen JM, Gu MH, Cheng ZK (2011) Rice OsGL1-1 is involved in leaf cuticular wax and cuticle membrane. Mol Plant 4(6):985–995
Riederer M, Schreiber L (2001) Protecting against water loss: analysis of the barrier properties of plant cuticles. J Exp Bot 52:2023–2032
Ristic Z, Jenks MA (2002) Leaf cuticle and water loss in maize lines differing in dehydration avoidance. J Plant Physiol 159:645–651
Rowland O, Lee R, Franke R, Schreiber L, Kunst L (2007) The CER3 wax biosynthetic gene from Arabidopsis thaliana is allelic to WAX2/YRE/FLP1. FEBS Lett 581:3538–3544
Samuels L, Kunst L, Jetter R (2008) Sealing plant surfaces: cuticular wax formation by epidermal cells. Annu Rev Plant Biol 59:683–707
Sieber P, Schorderet M, Ryser U, Buchala A, Kolattukudy P, Metraux J-P, Nawrath C (2000) Transgenic Arabidopsis plants expressing a fungal cutinase show alterations in the structure and properties of the cuticle and postgenital organ fusions. Plant Cell 12:721–737
Tacke E, Korfhage C, Michel D et al (1995) Transposon tagging of the maize Glossy2 locus with the transposable element En/Spm. Plant J 8:907–917
Todd J, Post BD, Jaworski JG (1999) KCS1 encodes a fatty acid elongase 3-ketoacyl-CoA synthase affecting wax biosynthesis in Arabidopsis thaliana. Plant J 17:119–130
Wang P, Liu CY, Wang DX, Liang CH, Zhao KH, Fan JJ (2013) Isolation of resistance gene analogs from grapevine resistant to downy mildew. Sci Hort 150:326–333
Xia Y, Nikolau BJ, Schnable PS (1996) Cloning and characterization of CER2, an Arabidopsis gene that affects cuticular wax accumulation. Plant Cell 8:1291–1304
Xia Y, Nikolau BJ, Schnable PS (1997) Developmental and hormonal regulation of the Arabidopsis CER2 gene that codes for a nuclear-localized protein required for the normal accumulation of cuticular waxes. Plant Physiol 115:925–937
Xu X, Dietrich CR, Delledonne M et al (1997) Sequence analysis of the cloned glossy8 gene of maize suggests that it may code for a [beta]-ketoacyl reductase required for the biosynthesis of cuticular waxes. Plant Physiol 115:501–510
Xu X, Dietrich CR, Lessire R, Nikolau BJ, Schnable PS (2002) The endoplasmic reticulum-associated maize GL8 protein is a component of the acyl-coenzyme A elongase involved in the production of cuticular waxes. Plant Physiol 128:924–934
Zhang JY, Broeckling CD, Blancaflor EB et al (2005) Over-expression of WXP1, a putative Medicago truncatula AP2 domain-containing transcription factor gene, increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa (Medicago sativa). Plant J 42:689–707
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This work was supported by grants from the National Natural Science Foundation of China (Nos. 31272157 and 31201625).
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Zhiyong Liu and Hui Feng are co-first authors.
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Zhang, X., Liu, Z., Wang, P. et al. Fine mapping of BrWax1, a gene controlling cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis). Mol Breeding 32, 867–874 (2013). https://doi.org/10.1007/s11032-013-9914-0
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DOI: https://doi.org/10.1007/s11032-013-9914-0