Abstract
Fusarium head blight (scab), primarily caused by Fusarium graminearum, is a devastating disease of wheat (Triticum aestivum L.) worldwide. Wheat sharp eyespot, mainly caused by Rhizoctonia cerealis, is one of the major diseases of wheat in China. The defensin RsAFP2, a small cyteine-rich antifungal protein from radish (Raphanus sativus), was shown to inhibit growth in vitro of agronomically important fungal pathogens, such as F. graminearum and R. cerealis. The RsAFP2 gene was transformed into Chinese wheat variety Yangmai 12 via biolistic bombardment to assess the effectiveness of the defensin in protecting wheat from the fungal pathogens in multiple locations and years. The genomic PCR and Southern blot analyses indicated that RsAFP2 was integrated into the genomes of the transgenic wheat lines and heritable. RT-PCR and Western blot proved that the RsAFP2 was expressed in these transgenic wheat lines. Disease tests showed that four RsAFP2 transgenic lines (RA1–RA4) displayed enhanced resistance to F. graminearum compared to the untransformed Yangmai 12 and the null-segregated plants. Assays on Q-RT-PCR and disease severity showed that the express level of RsAFP2 was associated with the enhanced resistance degree. Two of these transgenic lines (RA1 and RA2) also exhibited enhanced resistance to R. cerealis. These results indicated that the expression of RsAFP2 conferred increased resistance to F. graminearum and R. cerealis in transgenic wheat.
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Anand A, Zhou T, Trick HN, Gill BS, Bockus WW, Muthukrishnan S (2003) Greenhouse and field testing of transgenic wheat plants stably expressing genes for thaumatin-like protein, chitinase and glucanase against Fusarium graminearum. J Exp Bot 54:1101–1111
Bai GH, Shaner G (1994) Scab of wheat: prospects for control. Plant Dis 78:760–766
Cai SB, Ren LJ, Yan W, Wu JZ, Chen HG, Wu XY, Zhang XY (2006) Germplasm development and mapping of resistance to sharp eyespot (Rhizoctonia cerealis) in wheat. Sci Agric Sin 39:928–934 (in Chinese with English abstract)
Chen WP, Chen PD, Liu DJ, Kynast R, Friebe B, Velazhahan R, Muthukrishnan S, Gill BS (1999) Development of wheat scab symptoms is delayed in transgenic wheat plants that constitutively express a rice thaumatin-like protein gene. Theor Appl Genet 99:755–760
Chen L, Zhang ZY, Liang HX, Liu HX, Du LP, Xu HJ, Xin ZY (2008) Overexpression of TiERF1 enhances resistance to sharp eyespot in transgenic wheat. J Exp Bot 59:4195–4204
Christensen AH, Quail PH (1996) Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transg Res 5:213–218
Jha S, Chattoo BB (2010) Expression of a plant defensin in rice confers resistance to fungal phytopathogens. Trans Res 19:373–384
Koike M, Okamoto T, Tsuda S, Imai R (2002) A novel plant defensin-like gene of winter wheat is specifically induced during cold acclimation. Biochem Biophysic Res Communic 298:46–53
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CT method. Methods 25:402–408
Lu Y, Liu BY, Jing JX, Zhang ZZ (2010) Expression of RsAFP2 in Escherichia coli and its antifungal assay. J Plant Genet Resour 11:84–88 (in Chinese with English abstract)
Mackintosh CA, Lewis J, Radmer LE, Shin S, Heinen SJ, Smith LA, Wyckoff MN, Dill-Macky R, Evans CK, Kravchenko S, Baldridge GD, Zeyen RJ, Muehlbauer GJ (2007) Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight. Plant Cell Rep 26:479–488
Mauch F, Mauch-Mani B, Boller T (1988) Antifungal hydrolases in pea tissue: inhibition of fungal growth by combinations of chitinase and β-1, 3-glucanase. Plant Physiol 88:936–942
Osborn RW, De Samblanx GW, Thevissen K, Goderis I, Torrekens S, Van Leuven F, Attenborough S, Rees SB, Broekaert WF (1995) Isolation and characterisation of plant defensins from seeds of Asteraceae, Fabaceae, Hippocastanaceae and Saxifragaceae. FEBS Lett 368:257–262
Sharp PJ, Kries M, Shewry PR, Gale MD (1988) Location of β-amylase sequences in wheat and its relatives. Theor Appl Genet 75:286–290
Sharp PJ, Chao S, Desai S, Gale MD (1989) The isolation, characterization and application in the Triticeae of a set of wheat RFLP probes identifying each homoeologous chromosome arm. Theor Appl Genet 78:342–348
Shin S, Mackintosh CA, Lewis J, Heinen SJ, Radmer L, Dill-Macky R, Baldridge GD, Zeyen RJ, Muehlbauer GJ (2008) Transgenic wheat expressing a barley class II chitinase gene has enhanced resistance against Fusarium graminearum. J Exp Bot 59:2371–2378
Terras FRG, Goderis IJ, Van Leuven F, Vanderleyden J, Cammue BPA, Broekaert WF (1992) In vitro antifungal activity of a radish (Raphanus sativus L.) seed protein homologous to nonspecific lipid transfer proteins. Plant Physiol 100:1055–1058
Terras FRG, Eggermont K, Kovaleva V, Raikhel NV, Osborn RW, Kester A, Rees SB, Torrekem S, van Leuven FV, Vanderleyden J, Cammue BPA, Broekaert WF (1995) Small cysteine-rich antifungal proteins from radish: their role in host defense. Plant Cell 7:573–588
Thevissen K, Warnecke DC, Francois IEJA, Leipelt M, Heinz E, Ott C, Zähringer U, Thomma BPHJ, Ferket KKA, Cammue BPA (2004) Defensins from insects and plants interact with fungal glucosylceramides. J Biol Chem 279:3900–3905
Tuite J, Shaner G, Everson RJ (1990) Wheat scab in soft red winter wheat in Indiana in 1986 and its relation to quality measurements. Plant Dis 74:959–962
Xu HJ, Pang JL, Ye XG, Du LP, Li LC, Xin ZY, Ma YZ, Chen JP, Chen J, Cheng SH, Wu YH (2001) Study on the gene transfer of Nib8 into wheat for its resistance to the yellow mosaic virus by bombardment. Acta Agron Sin 27:684–689 (in Chinese with English abstract)
Zhou MP, Hayden MJ, Zhang ZY, Lu WZ, Ma HX (2010) Saturation and mapping of a major Fusarium head blight resistance QTL on chromosome 3BS of Sumai 3 wheat. J Appl Genet 51:19–25
Acknowledgments
The authors are grateful to Dr. Yong Zhang for help in evaluating the FHB disease reactions. This study was supported by a National “Key Sci-Tech” project (grant no. 2008ZX08002-001).
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Zhao Li and Miaoping Zhou contributed equally to this work.
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Li, Z., Zhou, M., Zhang, Z. et al. Expression of a radish defensin in transgenic wheat confers increased resistance to Fusarium graminearum and Rhizoctonia cerealis . Funct Integr Genomics 11, 63–70 (2011). https://doi.org/10.1007/s10142-011-0211-x
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DOI: https://doi.org/10.1007/s10142-011-0211-x