The availability of complete genome sequence of soybean has allowed research community to design the 66 K Affymetrix Soybean Array GeneChip for genome-wide expression profiling of soybean. In this study, we carried out microarray analysis of leaf tissues of soybean plants, which were subjected to drought stress from late vegetative V6 and from full bloom reproductive R2 stages. Our data analyses showed that out of 46093 soybean genes, which were predicted with high confidence among approximately 66000 putative genes, 41059 genes could be assigned with a known function. Using the criteria of a ratio change > = 2 and a q-value<0.05, we identified 1458 and 1818 upregulated and 1582 and 1688 downregulated genes in drought-stressed V6 and R2 leaves, respectively. These datasets were classified into 19 most abundant biological categories with similar proportions. There were only 612 and 463 genes that were overlapped among the upregulated and downregulated genes, respectively, in both stages, suggesting that both conserved and unconserved pathways might be involved in regulation of drought response in different stages of plant development. A comparative expression analysis using our datasets and that of drought stressed Arabidopsis leaves revealed the existence of both conserved and species-specific mechanisms that regulate drought responses. Many upregulated genes encode either regulatory proteins, such as transcription factors, including those with high homology to Arabidopsis DREB, NAC, AREB and ZAT/STZ transcription factors, kinases and two-component system members, or functional proteins, e.g. late embryogenesis-abundant proteins, glycosyltransferases, glycoside hydrolases, defensins and glyoxalase I family proteins. A detailed analysis of the GmNAC family and the hormone-related gene category showed that expression of many GmNAC and hormone-related genes was altered by drought in V6 and/or R2 leaves. Additionally, the downregulation of many photosynthesis-related genes, which contribute to growth retardation under drought stress, may serve as an adaptive mechanism for plant survival. This study has identified excellent drought-responsive candidate genes for in-depth characterization and future development of improved drought-tolerant transgenic soybeans.
References
[1]
Manavalan LP, Guttikonda SK, Tran LS, Nguyen HT (2009) Physiological and molecular approaches to improve drought resistance in soybean. Plant Cell Physiol 50: 1260–1276.
[2]
Tran LS, Mochida K (2010) Functional genomics of soybean for improvement of productivity in adverse conditions. Funct Integr Genomics 10: 447–462.
[3]
Thao NP, Tran LS (2011) Potentials toward genetic engineering of drought-tolerant soybean. Crit Rev Biotechnol doi:10.3109/07388551.2011.643463.
[4]
Sulieman S, Tran LS (2012) Asparagine: an amide of particular distinction in the regulation of symbiotic nitrogen fixation of legumes. Crit Rev Biotechnol.
[5]
Tran LS, Urao T, Qin F, Maruyama K, Kakimoto T, et al. (2007) Functional analysis of AHK1/ATHK1 and cytokinin receptor histidine kinases in response to abscisic acid, drought, and salt stress in Arabidopsis. Proc Natl Acad Sci U S A 104: 20623–20628.
[6]
Yamaguchi-Shinozaki K, Shinozaki K (2006) Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol 57: 781–803.
[7]
Valliyodan B, Nguyen HT (2006) Understanding regulatory networks and engineering for enhanced drought tolerance in plants. Curr Opin Plant Biol 9: 189–195.
[8]
Yang S, Vanderbeld B, Wan J, Huang Y (2010) Narrowing down the targets: towards successful genetic engineering of drought-tolerant crops. Mol Plant 3: 469–490.
[9]
Hadiarto T, Tran LS (2011) Progress studies of drought-responsive genes in rice. Plant Cell Rep 30: 297–310.
[10]
Ma Y, Qin F, Tran LS (2012) Contribution of Genomics to Gene Discovery in Plant Abiotic Stress Responses. Molecular Plant doi: 10.1093/mp/sss085.
[11]
Mochida K, Yoshida T, Sakurai T, Yamaguchi-Shinozaki K, Shinozaki K, et al. (2010) Genome-wide analysis of two-component systems and prediction of stress-responsive two-component system members in soybean. DNA Res 17: 303–324.
[12]
Matsui A, Ishida J, Morosawa T, Mochizuki Y, Kaminuma E, et al. (2008) Arabidopsis transcriptome analysis under drought, cold, high-salinity and ABA treatment conditions using a tiling array. Plant Cell Physiol 49: 1135–1149.
[13]
Zhou J, Wang X, Jiao Y, Qin Y, Liu X, et al. (2007) Global genome expression analysis of rice in response to drought and high-salinity stresses in shoot, flag leaf, and panicle. Plant Mol Biol 63: 591–608.
[14]
Degenkolbe T, Do PT, Zuther E, Repsilber D, Walther D, et al. (2009) Expression profiling of rice cultivars differing in their tolerance to long-term drought stress. Plant Mol Biol 69: 133–153.
[15]
Schmutz J, Cannon SB, Schlueter J, Ma J, Mitros T, et al. (2010) Genome sequence of the palaeopolyploid soybean. Nature 463: 178–183.
[16]
Le DT, Nishiyama R, Watanabe Y, Vankova R, Tanaka M, et al. (2012) Identification and Expression Analysis of Cytokinin Metabolic Genes in Soybean under Normal and Drought Conditions in Relation to Cytokinin Levels. PLoS One 7: e42411.
[17]
Le DT, Nishiyama R, Watanabe Y, Mochida K, Yamaguchi-Shinozaki K, et al. (2011) Genome-wide expression profiling of soybean two-component system genes in soybean root and shoot tissues under dehydration stress. DNA Res 18: 17–29.
[18]
Thimm O, Blasing O, Gibon Y, Nagel A, Meyer S, et al. (2004) MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J 37: 914–939.
[19]
Tran LS, Quach TN, Guttikonda SK, Aldrich DL, Kumar R, et al. (2009) Molecular characterization of stress-inducible GmNAC genes in soybean. Mol Genet Genomics 281: 647–664.
[20]
Park SY, Yu JW, Park JS, Li J, Yoo SC, et al. (2007) The senescence-induced staygreen protein regulates chlorophyll degradation. Plant Cell 19: 1649–1664.
[21]
Xu F, Meng T, Li P, Yu Y, Cui Y, et al. (2011) A soybean dual-specificity kinase, GmSARK, and its Arabidopsis homolog, AtSARK, regulate leaf senescence through synergistic actions of auxin and ethylene. Plant Physiol 157: 2131–2153.
[22]
Harb A, Krishnan A, Ambavaram MM, Pereira A (2010) Molecular and physiological analysis of drought stress in Arabidopsis reveals early responses leading to acclimation in plant growth. Plant Physiol 154: 1254–1271.
[23]
Tran LS, Nakashima K, Sakuma Y, Simpson SD, Fujita Y, et al. (2004) Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter. Plant Cell 16: 2481–2498.
[24]
Fujita Y, Fujita M, Satoh R, Maruyama K, Parvez MM, et al. (2005) AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis. Plant Cell 17: 3470–3488.
[25]
Wu Y, Deng Z, Lai J, Zhang Y, Yang C, et al. (2009) Dual function of Arabidopsis ATAF1 in abiotic and biotic stress responses. Cell Research 19: 1279–1290.
[26]
Mochida K, Yoshida T, Sakurai T, Yamaguchi-Shinozaki K, Shinozaki K, et al. (2009) In silico analysis of transcription factor repertoire and prediction of stress responsive transcription factors in soybean. DNA Res 16: 353–369.
[27]
Mochida K, Yoshida T, Sakurai T, Yamaguchi-Shinozaki K, Shinozaki K, et al. (2010) LegumeTFDB: an integrative database of Glycine max, Lotus japonicus and Medicago truncatula transcription factors. Bioinformatics 26: 290–291.
[28]
Tran LS, Nishiyama R, Yamaguchi-Shinozaki K, Shinozaki K (2010) Potential utilization of NAC transcription factors to enhance abiotic stress tolerance in plants by biotechnological approach. GM Crops 1: 32–39.
[29]
Jogaiah S, Ramsandra Govind S, Tran LS (2012) System biology-based approaches towards understanding drought tolerance in food crops. Crit Rev Biotechnol doi: 10.3109/07388551.2012.659174.
[30]
Umezawa T, Nakashima K, Miyakawa T, Kuromori T, Tanokura M, et al. (2010) Molecular basis of the core regulatory network in ABA responses: sensing, signaling and transport. Plant Cell Physiol 51: 1821–1839.
[31]
Takahashi F, Mizoguchi T, Yoshida R, Ichimura K, Shinozaki K (2011) Calmodulin-dependent activation of MAP kinase for ROS homeostasis in Arabidopsis. Mol Cell 41: 649–660.
[32]
Ma S, Gong Q, Bohnert HJ (2006) Dissecting salt stress pathways. J Exp Bot 57: 1097–1107.
[33]
Zeller G, Henz SR, Widmer CK, Sachsenberg T, Ratsch G, et al. (2009) Stress-induced changes in the Arabidopsis thaliana transcriptome analyzed using whole-genome tiling arrays. Plant J 58: 1068–1082.
[34]
Krishnaswamy SS, Srivastava S, Mohammadi M, Rahman MH, Deyholos MK, et al. (2008) Transcriptional profiling of pea ABR17 mediated changes in gene expression in Arabidopsis thaliana. BMC Plant Biol 8: 91.
Puranik S, Sahu PP, Srivastava PS, Prasad M (2012) NAC proteins: regulation and role in stress tolerance. Trends Plant Sci.
[37]
Tran LS, Nakashima K, Sakuma Y, Osakabe Y, Qin F, et al. (2007) Co-expression of the stress-inducible zinc finger homeodomain ZFHD1 and NAC transcription factors enhances expression of the ERD1 gene in Arabidopsis. Plant J 49: 46–63.
[38]
Nakashima K, Tran LS, Van Nguyen D, Fujita M, Maruyama K, et al. (2007) Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice. Plant J 51: 617–630.
[39]
Hu H, Dai M, Yao J, Xiao B, Li X, et al. (2006) Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci U S A 103: 12987–12992.
[40]
Redillas MC, Jeong JS, Kim YS, Jung H, Bang SW, et al.. (2012) The overexpression of OsNAC9 alters the root architecture of rice plants enhancing drought resistance and grain yield under field conditions. Plant Biotechnol J.
[41]
Le DT, Nishiyama R, Watanabe Y, Mochida K, Yamaguchi-Shinozaki K, et al. (2011) Genome-wide survey and expression analysis of the plant-specific NAC transcription factor family in soybean during development and dehydration stress. DNA Res 18: 263–276.
[42]
Divi UK, Krishna P (2009) Brassinosteroid: a biotechnological target for enhancing crop yield and stress tolerance. N Biotechnol 26: 131–136.
[43]
Ha S, Vankova R, Yamaguchi-Shinozaki K, Shinozaki K, Tran LS (2012) Cytokinins: metabolism and function in plant adaptation to environmental stresses. Trends Plant Sci 17: 172–179.
[44]
Wang S, Bai Y, Shen C, Wu Y, Zhang S, et al. (2010) Auxin-related gene families in abiotic stress response in Sorghum bicolor. Funct Integr Genomics 10: 533–546.
[45]
Nishiyama R, Watanabe Y, Fujita Y, Le DT, Kojima M, et al. (2011) Analysis of cytokinin mutants and regulation of cytokinin metabolic genes reveals important regulatory roles of cytokinins in drought, salt and abscisic acid responses, and abscisic acid biosynthesis. Plant Cell 23: 2169–2183.
[46]
Nishiyama R, Le DT, Watanabe Y, Matsui A, Tanaka M, et al. (2012) Transcriptome analyses of a salt-tolerant cytokinin-deficient mutant reveal differential regulation of salt stress response by cytokinin deficiency. PLoS One 7: e32124.
[47]
Peleg Z, Blumwald E (2010) Hormone balance and abiotic stress tolerance in crop plants. Curr Opin Plant Biol 14: 290–295.
[48]
Choudhary SP, Kanwar M, Bhardwaj R, Yu JQ, Tran LS (2012) Chromium stress mitigation by polyamine-brassinosteroid application involves phytohormonal and physiological strategies in Raphanus sativus L. PLoS One. 7: e33210.
[49]
Choudhary SP, Oral HV, Bhardwaj R, Yu JQ, Tran LS (2012) Interaction of brassinosteroids and polyamines enhances copper stress tolerance in raphanus sativus. J Exp Bot 63: 5659–5675.
[50]
Choudhary SP, Yu JQ, Yamaguchi-Shinozaki K, Shinozaki K, Tran LS (2012) Benefits of brassinosteroid crosstalk. Trends Plant Sci 17: 594–605.
[51]
Wang ZY (2012) Brassinosteroids modulate plant immunity at multiple levels. Proc Natl Acad Sci U S A 109: 7–8.
[52]
Choi J, Choi D, Lee S, Ryu CM, Hwang I (2011) Cytokinins and plant immunity: old foes or new friends? Trends Plant Sci 16: 388–394.
[53]
Qin F, Kodaira KS, Maruyama K, Mizoi J, Tran LS, et al. (2011) SPINDLY, a negative regulator of gibberellic acid signaling, is involved in the plant abiotic stress response. Plant Physiol 157: 1900–1913.