Abstract
The gibberellic acid-stimulated arabidopsis (GASA) peptide family widely existed in plant species and plays various functions in regulating biological processes. Based on the transcriptome and the genome of the rubber tree, 16 genes encoding the GASA proteins were identified in Hevea brasiliensis. The phylogenetic relationship of the 16 GASA proteins of H. brasiliensis was analyzed by comparing with that of Arabidopsis thaliana and Oryza sativa. Transcriptional assay showed that the GASA genes were regulated by the fungal pathogens Colletotrichum gloeosporioides. Transient expression in Nicotiana benthamiana revealed that the GASA genes were involved in generating reactive oxygen species, indicating their roles in plant innate immunity. In addition, the quantitative real-time PCR assays suggested that the GASA genes were regulated by plant hormone in complex ways. Taken together, the present study provides the comprehensive analysis and reveals putative roles of GASA genes in fungal pathogen resistance in H. brasiliensis.
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References
Almasia NI, Bazzini AA, Hopp HE, Vazquez-Rovere C (2008) Overexpression of snakin-1 gene enhances resistance to Rhizoctoniasolani and Erwinia carotova in transgenic potato plants. Mol Plant Pathol 9:329–338. https://doi.org/10.1111/j.1364-3703.2008.00469.x
Alonso-Ramírez A, Rodríguez D, Reyes D, Jiménez JA, Nicolás G, López-Climent M, Gómez-Cadenas A, Nicolás C (2009) Evidence for a role of gibberellins in salicylic acid-modulated early plant responses to abiotic stress in Arabidopsis seeds. Plant Physiol 150:1335–1344. https://doi.org/10.1104/pp.109.139352
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. https://doi.org/10.1146/annurev.arplant.55.031903.141701
Aubert D, Chevillard MC, Dorne AM, Arlaud G, Herzog M (1998) Expression patterns of GASA genes in Arabidopsis thaliana: the GASA4 gene is up-regulated by gibberellins in meristematic regions. Plant Mol Biol 36:871–883. https://doi.org/10.1023/A:1005938624418
Ben-Nissan G, Lee JY, Borohov A, Weiss D (2004) GIP, a Petunia hybrida GA-induced cysteine-rich protein: a possible role in shoot elongation and transition to flowering. Plant J 37:229–238. https://doi.org/10.1046/j.1365-313X.2003.01950.x
Berrocal-Lobo M, Segura A, Moreno M, López G, García-Olmedo F, Molina A (2002) Snakin-2, an antimicrobial peptide from potato whose gene is locally induced by wounding and responds to pathogen infection. Plant Physiol 128:951–961. https://doi.org/10.1104/pp.010685
Bolwell GP, Bindschedler LV, Blee KA, Butt VS, Davies DR, Gardner SL, Gerrish C, Minibayeva F (2002) The apoplastic oxidative burst in response to biotic stress in plants: a three-component system. J Exp Bot 53:1367–1376. https://doi.org/10.1093/jexbot/53.372.1367
Brown P, Baxter L, Hickman R, Beynon J, Moore JD, Ott S (2013) MEME-LaB: motif analysis in clusters. Bioinformatics 29:1696–1697. https://doi.org/10.1093/bioinformatics/btt248
Cai Z, Li G, Lin C, Shi T, Zhai L, Chen Y, Huang G (2013) Identifying pathogenicity genes in the rubber tree anthracnose fungus Colletotrichum gloeosporioides through random insertional mutagenesis. Microbiol Res 168:340–350. https://doi.org/10.1016/j.micres.2013.01.005
Cutler SR, Ehrhardt DW, Griffitts JS, Somerville CR (2000) Random GFP:: cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency. Proc Natl Acad Sci U S A 97:3718–3723. https://doi.org/10.1073/pnas.97.7.3718
Fang Y, Mei H, Zhou B, Xiao X, Yang M, Huang Y, Long X, Hu S, Tang C (2016) De novo transcriptome analysis reveals distinct defense mechanisms by young and mature leaves of Hevea brasiliensis (Para rubber tree). Sci Rep 6(33151). https://doi.org/10.1038/srep33151
Finn RD, Bateman A, Clements J, Coggill P, Eberhardt RY, Eddy SR, Heger A, Hetherington K, Holm L, Mistry J, Sonnhammer ELL, Tate J, Punta M (2014) The Pfam protein families database. Nucleic Acids Res 42:D222–D230. https://doi.org/10.1093/nar/gkt1223
Furukawa T, Sakaguchi N, Shimada H (2006) Two OsGASR genes, rice GAST homologue genes that are abundant in proliferating tissues, show different expression patterns in developing panicles. Genes Genet Syst 81:171–180. https://doi.org/10.1266/ggs.81.171
Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z, Mauceli E, Hacohen N, Gnirke A, Rhind N, di Palma F, Birren BW, Nusbaum C, Lindblad-Toh K, Friedman N, Regev A (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–652. https://doi.org/10.1038/nbt.1883
Kawahara Y, de la Bastide M, Hamilton JP, Kanamori H, McCombie WR, Ouyang S et al (2013) Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data. Rice 6:4. https://doi.org/10.1186/1939-8433-6-4
Ko CB, Woo YM, Lee DJ, Lee MC, Kim CS (2007) Enhanced tolerance to heat stress in transgenic plants expressing the GASA4 gene. Plant Physiol Bioch 45:722–728. https://doi.org/10.1016/j.plaphy.2007.07.010
Kotilainen M, Helariutta Y, Mehto M, Pöllänen E, Albert VA, Elomaa P, Teeri TH (1999) GEG participates in the regulation of cell and organ shape during corolla and carpel development in Gerbera hybrida. Plant Cell 11:1093–1104. https://doi.org/10.1105/tpc.11.6.1093
Kunkel BN, Brooks DM (2002) Cross talk between signaling pathways in pathogen defense. Curr Opin Plant Biol 5:325–331. https://doi.org/10.1016/S1369-5266(02)00275-3
de la Fuente JI, Amaya I, Castillejo C, Sánchez-Sevilla JF, Quesada MA, Botella MA, Valpuesta V (2006) The strawberry gene FaGAST affects plant growth through inhibition of cell elongation. J Exp Bot 57:2401–2411. https://doi.org/10.1093/jxb/erj213
Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593. https://doi.org/10.1016/0092-8674(94)90544-4
Li H, Qin Y, Xiao X, Tang C (2011) Screening of valid reference genes for real-time RT-PCR data normalization in Hevea brasiliensis and expression validation of a sucrose transporter gene HbSUT3. Plant Sci 181:132–139. https://doi.org/10.1016/j.plantsci.2011.04.014
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Marchler-Bauer A, Derbyshire MK, Gonzales NR, Lu S, Chitsaz F, Geer LY, Geer RC, He J, Gwadz M, Hurwitz DI, Lanczycki CJ, Lu F, Marchler GH, Song JS, Thanki N, Wang Z, Yamashita RA, Zhang D, Zheng C, Bryant SH (2015) CDD: NCBI’s conserved domain database. Nucleic Acids Res 43:D222–D226. https://doi.org/10.1093/nar/gku1221
Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat Methods 5:621–628. https://doi.org/10.1038/nmeth.1226
Moyano-Cañete E, Bellido ML, García-Caparrós N, Medina-Puche L, Amil-Ruiz F, González-Reyes JA, Caballero JL, Muñoz-Blanco J, Blanco-Portales R (2013) FaGAST2, a strawberry ripening-related gene, acts together with FaGAST1 to determine cell size of the fruit receptacle. Plant Cell Physiol 54:218–236. https://doi.org/10.1093/pcp/pcs167
Roxrud I, Lid SE, Fletcher JC, Schmidt ED, Opsahl-Sorteberg HG (2007) GASA4, one of the14-member Arabidopsis GASA family of small polypeptides, regulates flowering and seed development. Plant Cell Physiol 48:471–483. https://doi.org/10.1093/pcp/pcm016
Rubinovich L, Weiss D (2010) The Arabidopsis cysteine-rich protein GASA4 promotes GA responses and exhibits redox activity in bacteria and in planta. Plant J 64:1018–1027. https://doi.org/10.1111/j.1365-313X.2010.04390.x
Segura A, Moreno M, Madueño F, Molina A, García-Olmedo F (1999) Snakin-1, a peptide from potato that is active against plant pathogens. Mol Plant Microbe Interact 12:16–23. https://doi.org/10.1094/MPMI.1999.12.1.16
Shi L, Gast RT, Gopalraj M, Olszewski NE (1992) Characterization of a shoot-specific, GA3- and ABA-regulated gene from tomato. Plant J 2:153–159. https://doi.org/10.1111/j.1365-313X.1992.00153.x
Silverstein KA, Moskal WA Jr, Wu HC, Underwood BA, Graham MA, Town CD, VandenBosch KA (2007) Small cysteine-rich peptides resembling antimicrobial peptides have been under-predicted in plants. Plant J 51:262–280. https://doi.org/10.1111/j.1365-313X.2007.03136.x
Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1:2019–2025. https://doi.org/10.1038/nprot.2006.286
Tang C, Yang M, Fang Y, Luo Y, Gao S, Xiao X, An Z, Zhou B, Zhang B, Tan X, Yeang HY, Qin Y, Yang J, Lin Q, Mei H, Montoro P, Long X, Qi J, Hua Y, He Z, Sun M, Li W, Zeng X, Cheng H, Liu Y, Yang J, Tian W, Zhuang N, Zeng R, Li D, He P, Li Z, Zou Z, Li S, Li C, Wang J, Wei D, Lai CQ, Luo W, Yu J, Hu S, Huang H (2016) The rubber tree genome reveals new insights into rubber production and species adaptation. Nat Plants 2:16073. https://doi.org/10.1038/NPLANTS.2016.73
The UniProt Consortium (2015) UniProt: a hub for protein information. Nucleic Acids Res 43:D204–D212. https://doi.org/10.1093/nar/gku989
Wang L, Wang Z, Joo SH, Kim SK, Xue Z, Xu Z, Wang Z, Chong K (2009) OsGSR1 is involved in crosstalk between gibberellins and brassinosteroids in rice. Plant J 57:498–510. https://doi.org/10.1111/j.1365-313X.2008.03707.x
Wigoda N, Ben-Nissan G, Granot D, Schwartz A, Weiss D (2006) The gibberellin-induced, cysteine-rich protein GIP2 from Petunia hybrida exhibits in planta antioxidant activity. Plant J 48:796–805. https://doi.org/10.1111/j.1365-313X.2006.02917.x
Zhang S, Wang X (2016) One new kind of phytohormonal signaling integrator: up-and-coming GASA family genes. Plant Signal Behav 12:e1226453. https://doi.org/10.1080/15592324.2016.1226453
Zhang S, Yang C, Peng J, Sun S, Wang X (2009) GASA5, a regulator of flowering time and stem growth in Arabidopsis thaliana. Plant Mol Biol 69:745–759. https://doi.org/10.1007/s11103-009-9452-7
Zheng X, Wang Q, Luo H (2017) Cloning and expression analysis of an ARF guanine nucleotide exchange factors named as HbGNOM in Hevea brasiliensis (in Chinese). Nat Sci J Hainan Univ 3:246–252. https://doi.org/10.15886/j.cnki.hdxbzkb.2017.0039
Funding
This study was supported by the National Natural Science Foundation of China (no. 31160151), the National Natural Science Foundation of China (no. 31571967), and the Natural Science Foundation of Hainan Province (no. 20163052).
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Communicated by W. Ratnam
Data archiving statement
All 16 of the HbGASA genes were acquired from the GenBank by BLAST search, and the accessions of the genes and the proteins are listed in Table 1.
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An, B., Wang, Q., Zhang, X. et al. Comprehensive transcriptional and functional analyses of HbGASA genes reveal their roles in fungal pathogen resistance in Hevea brasiliensis. Tree Genetics & Genomes 14, 41 (2018). https://doi.org/10.1007/s11295-018-1256-y
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DOI: https://doi.org/10.1007/s11295-018-1256-y