AJPS  Vol.8 No.8 , July 2017
Evaluation of a Brassica napus Auxin-Repressed Gene Induced by Flea Beetle Damage and Sclerotinia sclerotiorum Infection
Abstract: Biotic stresses negatively affect canola growth and production. Flea beetle damage and Sclerotinia sclerotiorum (S. sclerotiorum) infection are two of the worst biotic stresses for canola. Auxin Repressed Proteins (ARPs) responsive to several abiotic stresses have been reported. However, information about ARPs induced by Flea beetle damage and S. sclerotiorum infection, their roles in biotic stress tolerance are still lacking in canola. ESTs for an Auxin Repressed Protein 1 (BnARP1) were highly represented (expressed) in a Brassica napus subtractive library developed after leaf damage by the crucifer flea beetle (Phyllotreta cruciferae). Expression of this gene was under different developmental control in B. napus, and it was co-induced in B. napus by flea beetle feeding, S. sclerotiorum infection, drought and cold. A total of 25 BnARP genes were represented in different B. napus stress and development EST libraries and indicated larger, diversified families than known earlier. Dwarf phenotypes, primary root growth inhibition, lateral root enhancement, reduced sensitivity to 2, 4-D, and reduced PIN1 and LOX expression in transgenic Arabidopsis expression lines suggest that BnARP1 is an auxin repressor that prevents auxin transport and supports an interaction between the auxin and jasmonate signalling pathways. And the increased survival after S. sclerotiorum infection in transgenic over-expression Arabidopsis suggests that BnARP1 could play a role in S. sclerotiorum tolerance through connecting auxin and jasmonate signalling pathways.
Cite this paper: Wu, L. , Yu, M. , Holowachuk, J. , Sharpe, A. , Lydiate, D. , Hegedus, D. and Gruber, M. (2017) Evaluation of a Brassica napus Auxin-Repressed Gene Induced by Flea Beetle Damage and Sclerotinia sclerotiorum Infection. American Journal of Plant Sciences, 8, 1921-1952. doi: 10.4236/ajps.2017.88130.

[1]   Grillo, G., Stotz, H.U., Pittendrigh, B.R., Kroymann, J., Weniger, K., Fritsche, J., Bauke, A. and Mitchell-Olds, T. (2000) Induced Plant Defense Responses against Chewing Insects. Ethylene Signaling Reduces Resistance of Arabidopsis against Egyptian Cotton Worm but Not Diamondback Moth. Plant Physiology, 124, 1007-1017.

[2]   Kessler, A. and Baldwin, I.T. (2002) Plant Responses to Insect Herbivory: the Emerging Molecular Analysis. Annual Review of Plant Biology, 53, 299-328.

[3]   Walling, L.L. (2000) The Myriad Plant Responses to Herbivores. Journal of Plant Growth Regulation, 19, 195-216.

[4]   Moran, P.J. and Thompson, G.A. (2001) Molecular Responses to Aphid Feeding in Arabidopsis in Relation to Plant Defense Pathways. Plant Physiology, 125, 1074-1085.

[5]   Xu, Y., Chang, P.-F.L., Liu, D., Narasimhan, M.L., Raghothama, K.G., Hasegawa, P.M. and Bressan, R.A. (1994) Plant Defense Genes Are Synergistically Induced by Ethylene and Methyl Jasmonate. Plant Cell, 6, 1077-1085.

[6]   O’Donnell, P.J., Calvert, C., Atzorn, R., Wasternack, C., Leyser, H.M.O. and Bowles, D.J. (1996) Ethylene as a Signal Mediating the Wound Response of Tomato Plants. Science, 274, 1914-1917.

[7]   Penninckx, I.A., Thomma, B.P., Buchala, A., Métraux, J-P. and Broekaert, W.F. (1998) Concomitant Activation of Jasmonate and Ethylene Response Pathways Is Required for Induction of a Plant Defensin Gene in Arabidopsis. Plant Cell, 10, 2103-2113.

[8]   Kahl, J., Siemens, D.H., Aerts, R.J., Gäbler, R., Kühnemann, F., Preston, C.A. and Baldwin, I.T. (2000) Herbivore-Induced Ethylene Suppresses a Direct Defense but Not a Putative Indirect Defense against an Adapted Herbivore. Planta, 210, 336-342.

[9]   Guo, H. and Ecker, J.R. (2004) The Ethylene Signaling Pathway: New Insights. Current Opinion in Plant Biology, 7, 40-49.

[10]   Johnson, P.R. and Ecker, J.R. (1998) The Ethylene Gas Signal Transduction Pathway: A Molecular Perspective. Annual Review of Genetics, 32, 227-254.

[11]   Shinozaki, K., Yamaguchi-Shinozaki, K. and Seki, M. (2003) Regulatory Network of Gene Expression in the Drought and Cold Stress Responses. Current Opinion in Plant Biology, 6, 410-417.

[12]   Anderson, J.P., Badruzsaufari, E., Schenk, P.M., Manners, J.M., Desmond, O.J., Ehlert, C., Maclean, D.J., Ebert, P.R. and Kazan, K. (2004) Antagonistic Interaction between Abscisic Acid and Jasmonate-Ethylene Signaling Pathways Modulates Defense Gene Expression and Disease Resistance in Arabidopsis. Plant Cell, 16, 3460-3479.

[13]   Davies, P.J. (1995) The Plant Hormones: Their Nature, Occurrence, and Functions. In: Davies, P.J., Ed., Plant Hormones: Physiology, Biochemistry, and Molecular Biology, Kluwer Academic Publishers, Dordrecht, The Netherlands, 1-5.

[14]   Leyser, O. (2001) Auxin Signalling: The Beginning, the Middle and the End. Current Opinion in Plant Biology, 4, 382-386.

[15]   Zhao, Y. (2010) Auxin Biosynthesis and Its Role in Plant Development. Annual Review of Plant Biology, 61, 49-64.

[16]   Abel, S., Oeller, P.W. and Theologis, A. (1994) Early Auxin-Induced Genes Encode Short-Lived Nuclear Proteins. Proceedings of the National Academy of Sciences of the United States of America, 91, 326-330.

[17]   Overwoode, P.I., Ikushima, Y., Alonso, J.M., Chare, A., Chan, C., Ecker, J.R., Hughest, B., Liu, A., Onodera, C., Quach, H., Smith, A., Yu, G. and Theologis, A. (2005) Functional Genomic Analysis of the AUXIN/INDOLE-3-ACETIC ACID Gene Family Members in Arabidopsis thaliana. Plant Cell, 17, 3282-3300.

[18]   Ulmasov, T., Hagen, G. and Guilfoyle, T.J. (1999) Activation and Repression of Transcription by Auxin-Response Factors. Proceedings of the National Academy of Sciences of the United States of America, 96, 5844-5849.

[19]   Sieberer, T., Seifert, G.J., Hauser, M.T., Grisafi, P., Fink, G.R. and Luschnig, C. (2000) Post-Transcriptional Control of the Arabidopsis Auxin Efflux Carrier EIR1 requires AXR1. Current Biology, 10, 1595-1598.

[20]   Tiryaki, I. and Staswick, P.E. (2002) An Arabidopsis Mutant Defective in Jasmonate Response Is Allelic to the Auxin-Signaling Mutant axr1. Plant Physiology, 130, 887-894.

[21]   Reddy, A.S.N. and Poovaiah, B.W. (1990) Molecular Cloning and Sequencing of a cDNA for an Auxin-Repressed Messenger-RNA-Correlation between Fruit Growth and Repression of the Auxin Regulated Gene. Plant Molecular Biology, 14, 127-136.

[22]   Stafstrom, J.P., Ripley, B.D., Devitt, M.L. and Drake, B. (1998) Dormancy-Associated Gene Expression in Pea Axillary Buds. Planta, 205, 547-552.

[23]   Kebrom, T.H., Burson, B.L., and Finlayson, S.A. (2006) Phytochrome B Represses Toesinte Branched1 Expression and Induces Sorghum Axillary Bud Outgrowth in Response to Light Signals. Plant Physiology, 140, 1109-1117.

[24]   Steiner, C., Bauer, J., Amrhein, N. and Bucher, M. (2003) Two Novel Genes Are Differentially Expressed during Early Germination of the Male Gametophyte of Nicotiana tabacum. Biochimica et Biophysica Acta, 1625, 123-133.

[25]   Park, S. and Han, K.-H. (2003) An Auxin-Repressed Gene (RpARP) from Black Locust (Robinia pseudoacacia) Is Post-Transcriptionally Regulated and Negatively Associated with Shoot Elongation. Tree Physiology, 23, 815-823.

[26]   Kim, H.B., Lee, H., Oh, C.J., Lee, N.H. and An, C.S. (2007) Expression of EuNOD-ARP1 Encoding Auxin-Repressed Protein Homolog Is Upregulated by Auxin and Localized to the Fixation Zone in Root Nodules of Elaeagnus umbellata. Molecules and Cells, 23, 115-121.

[27]   Shimizu, M., Suzuki, K., Miyazawa, Y., Fujii, N. and Takahashi, H. (2006) Differential Accumulation of the mRNA of the Auxin-Repressed Gene CsGRP1 and the Auxin-Induced Peg Formation during Gravimorphogenesis of Cucumber Seedlings. Planta, 225, 13-22.

[28]   Tatematsu, K., Ward, S., Leyser, O., Kamiya, Y. and Nambara, E. (2005) Identification of cis-Elements That Regulate Gene Expression during Initiation of Axillary Bud Outgrowth in Arabidopsis. Plant Physiology, 138, 757-766.

[29]   Hwang, E.W., Kim, K.A., Park, S.C., Jeong, M.J., Byun, M.O. and Kwon, H.B. (2005) Expression Profiles of Hot Pepper (Capsicum annum) Genes under Cold Stress Condition. Journal of Biosciences, 30, 657-667.

[30]   Mantri, N.L., Ford, R., Coram, T.E. and Pang, E.C.K. (2007) Transcriptional Profiling of Chickpea Genes Differentially Regulated in Response to High-Salinity, Cold and Drought. BMC Genomics, 8, 303.

[31]   Chen, L., Ren, F., Zhong, H., Jiang, W. and Li, X. (2010) Identification and Expression Analysis of Genes in Response to High-Salinity and Drought Stresses in Brassica napus. Acta Biochim Biophys Sin, 42, 154-164.

[32]   Lee, J., Han, C.-T. and Hur, Y. (2013) Molecular Characterization of the Brassica rapa Auxin-Repressed, Superfamily Genes, BrARP1 and BrDRM1. Molecular Biology Reports, 40, 197-209.

[33]   Gruber, G., Wu, L., Links, M., Gjetvaj, B., Durkin, J., Lydiate, D. and Hegedus, D. (2012) Analysis of Expressed Sequence Tags in Brassica napus Cotyledons Damaged by Crucifer Flea Beetle Feeding. Genome, 55, 118-133.

[34]   Moreno-Hagelsieb, G. and Latimer, K. (2008) Choosing BLAST Options for Better Detection of Orthologs as Reciprocal Best Hits. Bioinformatics, 24, 319-324.

[35]   Clough, S.J. and Bent, A.F. (1998) Floral Dip: A Simplified Method for Agrobacterium-Mediated Transformation of Arabidopsis thaliana. The Plant Journal, 16, 735-743.

[36]   Klimyuk, V.I., Carroll, B.J., Thomas, C.M. and Jones, J.D.G. (1993) Alkali Treatment for Rapid Preparation of Plant Material for Reliable PCR Analysis. The Plant Journal, 3, 493-494.

[37]   Murray, M.G. and Thompson, W.F. (1980) Rapid Isolation of High Molecular Weight Plant DNA. Nucleic Acids Research, 8, 4321-4325.

[38]   Southern, E.M. (1975) Detection of Specific Sequences among DNA Fragments Separated by Gel Electrophoresis. Journal of Molecular Biology, 98, 503-517.

[39]   Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. II Edition, Cold Spring Harbour Laboratory. Press, Cold Spring Harbour, New York.

[40]   Fukaki, H., Tameda, S., Masuda, H. and Tasaka, M. (2002) Lateral Root Formation Is Blocked by a Gain-of-Function Mutation in the SOLITARY-ROOT/IAA14 Gene of Arabidopsis. The Plant Journal, 29, 153-168.

[41]   Okushima, Y., Fukaki, H., Onoda, M., Theologis, A. and Tasaka, M. (2007) ARF7 and ARF19 Regulate Lateral Root Formation via Direct Activation of LBD/ASL Genes in Arabidopsis. Plant Cell, 19, 118-130.

[42]   Palaniswamy, P., Lamb, R.J. and McVetty, P.B.E. (1992) Screening for Antixenosis Resistance to Flea Beetles, Phyllotreta cruciferae (Goeze) (Coleoptera: Chrysomelidae) in Rapeseed and Related Ccrucifers. The Canadian Entomologist, 124, 895-906.

[43]   Hallett, R.H., Ray, H., Holowachuk, J., Soroka, J.J. and Gruber, M.Y. (2005) Bioassay for Assessing Resistance of Arabidopsis thaliana L. (Heynh.) to the Adult Crucifer Flea Beetle, Phyllotreta cruciferae (Goeze) (Coleoptera: Chrysomelidae). Canadian Journal of Plant Science, 85, 225-235.

[44]   Vogel, J.T., Zarka, D.G., Buskirk, H.A.V., Fowler, S.G. and Thomashow, M.F. (2005) Roles of the CBF2 and ZAT12 Transcription Factors in Configuring the Low Temperature Transcriptome of Arabidopsis. The Plant Journal, 41, 195-211.

[45]   Pedras, M.S.C. and Ahiahonu, P.W.K. (2004) Phytotoxin Production and Phytoalexin Elicitation by the Phytopathogenic Fungus Sclerotinia sclerotiorum. Journal of Chemical Ecology, 30, 2163-2179.

[46]   SAS Institute, Inc (2001) The SAS System. Version 9.0., SAS Institute, Inc., Cary, North Carolina.

[47]   Bailey, L.C., Searls, D.B. and Overton, G.C. (1998) Analysis of EST-Driven Gene Annotation in Human Genomic Sequence. Genome Research, 8, 362-376.

[48]   Lamb, R.J. (1984) Effects of Flea Beetles, Phyllotreta spp. (Chrysomelidae: Coleoptera), on the Survival, Growth, Seed Yield and Quality of Canola, Rape and Yellow Mustard. The Canadian Entomologist, 116, 269-280.

[49]   Parkin, I., Gulden, S.M., Sharpe, A.G., Lukens, L., Trick, M., Osborn, T.C. and Lydiate, D.J. (2005) Segmental Structure of the Brassica napus Genome Based on Comparative Analysis with Arabidopsis thaliana. Genetics, 171, 765-781.

[50]   Nakazawa, M., Yabe, N., Ichikawa, T., Yamamoto, Y.Y., Yoshizumi, T., Hasunuma, K. and Matsui, M. (2001) DFL1, an Auxin-Responsive GH3 Gene Homologue, Negatively Regulates Shoot Cell Elongation and Lateral Root Formation, and Positively Regulates the Light Response of Hypocotyl Length. The Plant Journal, 25, 213-221.

[51]   Gälweiler, L., Guan, C., Müller, A., Wisman, E., Mendgen, K., Yephremov, A. and Palme, K. (1998) Regulation of Polar Auxin Transport by AtPIN1 in Arabidopsis Vascular Tissue. Science, 282, 2226-2230.

[52]   Jones, A.M., Im, K.H., Savka, M.A., Wu, M.J., DeWitt, N.G., Shillito, R. and Binns, A.N. (1998) Auxin-Dependent Cell Expansion Mediated by Overexpressed Auxin-Binding Protein 1. Science, 282, 1114-1117.

[53]   Creelman, R.A. and Mullet, J.E. (1997) Biosynthesis and Action of Jasmonates in Plants. Annual Review of Plant Physiology and Plant Molecular Biology, 48, 355-381.

[54]   Tiwari, S.B., Hagen, G. and Guilfoyle, T. (2003) The Roles of Auxin Response Factor Domains in Auxin-Responsive Transcription. Plant Cell, 15, 533-543.