Abstract
The endosperm plays an important role in seed formation and germination, especially in rice (Oryza sativa). We used a high-throughput sequencing technique (RNA-Seq) to reveal the molecular mechanisms involved in rice endosperm development. Three cDNA libraries were taken from rice endosperm at 3, 6 and 10 days after pollination (DAP), which resulted in the detection of 21,596, 20,910 and 19,459 expressed gens, respectively. By ERANGE, we identified 10,371 differentially expressed genes (log2Ratio ≥1, FDR ≤0.001). The results were compared against three public databases (Gene Ontology, Kyoto Encyclopedia of Genes and Genomes and MapMan) in order to annotate the gene descriptions, associate them with gene ontology terms and to assign each to pathways. A large number of genes related to ribosomes, the spliceosome and oxidative phosphorylation were found to be expressed in the early and middle stages. Plant hormone, galactose metabolism and carbon fixation related genes showed a significant increase in expression at the middle stage, whereas genes for defense against disease or response to stress as well as genes for starch/sucrose metabolism were strongly expressed during the later stages of endosperm development. Interestingly, most metabolic pathways were down-regulated between 3 and 10 DAP except for those involved in the accumulation of material, such as starch/sucrose and protein metabolism. We also identified the expression of 1,118 putative transcription factor genes in endosperm development. The RNA-Seq results provide further systematic understanding of rice endosperm development at a fine scale and a foundation for future studies.
Similar content being viewed by others
References
Balwierz PJ, Carninci P, Daub CO, Kawai J, Hayashizaki Y, Van Belle W, Beisel C, Van Nimwegen E (2009) Methods for analyzing deep sequencing expression data: constructing the human and mouse promoterome with deepCAGE data. Genome Biol 10:R79. doi:10.1186/gb-2009-10-7-r79
Bellin D, Ferrarini A, Chimento A, Kaiser O, Levenkova N, Bouffard P, Delledonne M (2009) Combining next-generation pyrosequencing with microarray for large scale expression analysis in non-model species. BMC Genomics 10:555. doi:10.1186/1471-2164-10-555
Berger F (1999) Endosperm development. Curr Opin Plant Biol 2:28–32. doi:10.1016/S1369-5266(99)80006-5
Bethke PC, Lonsdale JE, Fath A, Jones RL (1999) Hormonally regulated programmed cell death in barley aleurone cells. Plant Cell 11:1033–1046. doi:10.1105/tpc.11.6.1033
Bindea G, Mlecnik B, Hackl H, Charoentong P, Tosolini M, Kirilovsky A, Fridman WH, Pages F, Trajanoski Z, Galon J (2009) ClueGO: a Cytoscape plug-into decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics 25:1091–1093. doi:10.1093/bioinformatics/btp101
Bleeker PM, Spyropoulou EA, Diergaarde PJ, Volpin H, De Both MT, Zerbe P, Bohlmann J, Falara V, Matsuba Y, Pichersky E, Haring MA, Schuurink RC (2011) RNA-seq discovery, functional characterization, and comparison of sesquiterpene synthases from Solanum lycopersicum and Solanum habrochaites trichomes. Plant Mol Biol 77:323–336. doi:10.1007/s11103-011-9813-x
Cavel E, Pillot M, Pontier D, Lahmy S, Bies-Etheve N, Vega D, Grimanelli D, Lagrange T (2011) A plant-specific transcription factor IIB-related protein, pBRP2, is involved in endosperm growth control. PLoS ONE 6:e17216. doi:10.1371/journal.pone.0017216
Chen X, Wang Y, Li J, Jiang A, Cheng Y, Zhang W (2009) Mitochondrial proteome during salt stress-induced programmed cell death in rice. Plant Physiol Biochem 47:407–415. doi:10.1016/j.plaphy.2008.12.021
Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676. doi:10.1093/bioinformatics/bti610
Gaur VS, Singh US, Kumar A (2010) Transcriptional profiling and in silico analysis of Dof transcription factor gene family for understanding their regulation during seed development of rice Oryza sativa L. Mol Biol Rep 38:2827–2848. doi:10.1007/s11033-010-0429-z
Gilchrist DG (1998) Programmed cell death in plant disease: the purpose and promise of cellular suicide. Annu Rev Phytopathol 36:393–414. doi:10.1146/annurev.phyto.36.1.393
Girke T, Todd J, Ruuska S, White J, Benning C, Ohlrogge J (2000) Microarray analysis of developing Arabidopsis seeds. Plant Physiol 124:1570–1581. doi:10.1104/pp.124.4.1570
Goldberg RB, Paiva GD, Yadegari R (1994) Plant embryogenesis: zygote to seed. Science 266:605–614. doi:10.1126/science.266.5185.605
Hatsugai N, Kuroyanagi M, Yamada K, Meshi T, Tsuda S, Kondo M, Nishimura M, Hara-Nishimura I (2004) A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death. Science 305:855–858. doi:10.1126/stke.2452004tw287
Ishimaru T, Matsuda T, Ohsugi R, Yamagishi T (2003) Morphological development of rice caryopses located at the different positions in a panicle from early to middle stage of grain filling. Funct Plant Biol 30:1139–1149. doi:10.1071/FP03122
Jiao BB, Wang JJ, Zhu XD, Zeng LJ, Li Q, He ZH (2012) A novel protein RLS1 with NB-ARM domains is involved in chloroplast degradation during leaf senescence in rice. Mol Plant 5:205–217. doi:10.1093/mp/ssr081
Johnson AA, Kyriacou B, Callahan DL, Carruthers L, Stangoulis J, Lombi E, Tester M (2011) Constitutive overexpression of the OsNAS gene family reveals single-gene strategies for effective iron-and zinc-biofortification of rice endosperm. PLoS ONE 6:e24476. doi:10.1371/journal.pone.0024476
Kim SG, Kim SY, Park CM (2007) A membrane-associated NAC transcription factor regulates salt-responsive flowering via FLOWERING LOCUS T in Arabidopsis. Planta 226:647–654. doi:10.1007/s00425-007-0513-3
Kondou H, Ooka H, Yamada H, Satoh K, Kikuchi S, Takahara Y, Yamamoto K (2006) Microarray analysis of gene expression at initial stages of rice seed development. Breed Sci 56:235–242. doi:10.1270/jsbbs.56.235
Lan SY, Zhong FX, Yang ZM, Jin DM, Xu ZX (2004) The starchy endosperm denucleation by a process of programmed cell death during rice grain development. Acta Biol Exp Sin 37:34–44
Laudencia-Chingcuanco DL, Stamova BS, You FM, Lazo GR, Beckles DM, Anderson OD (2007) Transcriptional profiling of wheat caryopsis development using cDNA microarrays. Plant Mol Biol 63:651–668. doi:10.1007/s11103-006-9114-y
Le BH, Wagmaister JA, Kawashima T, Bui AQ, Harada JJ, Goldberg RB (2007) Using genomics to study legume seed development. Plant Physiol 144:562–574. doi:10.1104/pp.107.100362
Lee SC, Luan S (2011) ABA signal transduction at the crossroad of biotic and abiotic stress responses. Plant, Cell Environ 35:53–60. doi:10.1111/j.1365-3040.2011.02426.x
Li D, Liu H, Zhang H, Wang X, Song F (2008) OsBIRH1, a DEAD-box RNA helicase with functions in modulating defence responses against pathogen infection and oxidative stress. J Exp Bot 59:2133–2146. doi:10.1093/jxb/ern072
Li R, Zhu H, Ruan J, Qian W, Fang X, Shi Z, Li Y, Li S, Shan G, Kristiansen K, Li S, Yang H, Wang J, Wang J (2009) De novo assembly of human genomes with massively parallel short read sequencing. Genome Res 20:265–272. doi:10.1101/gr.097261.109
Li X, Gao X, Wei Y, Deng L, Ouyang Y, Chen G, Li X, Zhang Q, Wu C (2011) Rice APOPTOSIS INHIBITOR5 coupled with two DEAD-box adenosine 5’-triphosphate-dependent RNA helicases regulates tapetum degeneration. Plant Cell 23:1416–1434. doi:10.1105/tpc.110.082636
Li J, Chu H, Zhang Y, Mou T, Wu C, Zhang Q, Xu J (2012) The rice HGW gene encodes a ubiquitin-associated (UBA) domain protein that regulates heading date and grain weight. PLoS ONE 7:e34231. doi:10.1371/journal.pone.0034231
Lister R, Gregory BD, Ecker JR (2009) Next is now: new technologies for sequencing of genomes, transcriptomes, and beyond. Curr Opin Plant Biol 12:107–118. doi:10.1016/j.pbi.2008.11.004
Liu X, Guo T, Wan X, Wang H, Zhu M, Li A, Su N, Shen Y, Mao B, Zhai H, Mao L, Wan J (2010) Transcriptome analysis of grain-filling caryopses reveals involvement of multiple regulatory pathways in chalky grain formation in rice. BMC Genomics 11:730. doi:10.1186/1471-2164-11-730
Lord CE, Gunawardena AH (2011) Environmentally induced programmed cell death in leaf protoplasts of Aponogeton madagascariensis. Planta 233:407–421. doi:10.1007/s00425-010-1304-9
Luo M, Taylor JM, Spriggs A, Zhang H, Wu X, Russell S, Singh M, Koltunow A (2011) A genome-wide survey of imprinted genes in rice seeds reveals imprinting primarily occurs in the endosperm. PLoS Genet 7:e1002125. doi:10.1371/journal.pgen.1002125
Marguerat S, Bahler J (2010) RNA-seq: from technology to biology. Cell Mol Life Sci 67:569–579. doi:10.1007/s00018-009-0180-6
Martinez M, Abraham Z, Carbonero P, Diaz I (2005) Comparative phylogenetic analysis of cystatin gene families from arabidopsis, rice and barley. Mol Genet Genomics 273:423–432. doi:10.1007/s00438-005-1147-4
Mena M, Vicente-Carbajosa J, Schmidt RJ, Carbonero P (1998) An endosperm-specific DOF protein from barley, highly conserved in wheat, binds to and activates transcription from the prolamin-box of a native B-hordein promoter in barley endosperm. Plant J 16:53–62. doi:10.1046/j.1365-313x.1998.00275.x
Morrison IN, Kuo J, O’Brien TP (1975) Histochemistry and fine structure of developing wheat aleurone cells. Planta 123:105–116. doi:10.1007/BF00383859
Olsen OA, Potter RH, Kalla R (1992) Histo-differentiation and molecular biology of developing cereal endosperm. Seed Sci Res 2:117–131. doi:10.1017/S0960258500001240
Qi Y, Wang H, Zou Y, Liu C, Liu Y, Wang Y, Zhang W (2011) Over-expression of mitochondrial heat shock protein 70 suppresses programmed cell death in rice. FEBS Lett 585:231–239. doi:10.1016/j.febslet.2010.11.051
Steffens B, Sauter M (2005) Epidermal cell death in rice is regulated by ethylene, gibberellin, and abscisic acid. Plant Physiol 139:713–721. doi:10.1104/pp.105.064469
Venu R, Sreerekha M, Nobuta K, Belo A, Ning Y, An G, Meyers BC, Wang GL (2011) Deep sequencing reveals the complex and coordinated transcriptional regulation of genes related to grain quality in rice cultivars. BMC Genomics 12:190. doi:10.1186/1471-2164-12-190
Verza NC, Silve TR, Neto GC, Nogueira FT, Fisch PH, Rosa VE, Rebello MM, Vettore AL, Silva FR, Arruda P (2005) Endosperm-preferred expression of maize genes as revealed by transcriptome-wide analysis of expressed sequence tags. Plant Mol Biol 59:363–374. doi:10.1007/s11103-005-8924-7
Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63. doi:10.1038/nrg2484
Wang L, Feng Z, Wang X, Wang X, Zhang X (2010a) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26:136–138. doi:10.1093/bioinformatics/btp612
Wang Y, Zha X, Zhang S, Qian X, Dong X, Sun F, Yang J (2010b) Down-regulation of the OsPDCD5 gene induced photoperiod-sensitive male sterility in rice. Plant Sci 178:221–228. doi:10.1016/j.plantsci.2009.12.001
Wei CX, Xu ZX, Lan SY (2002) Ultrastructural features of nucleus degradation during programmed cell death of starchy endosperm cells in rice. Acta Bot Sin 44:1396–1402
Wu M, Huang J, Xu S, Ling T, Xie Y, Shen W (2011) Haem oxygenase delays programmed cell death in wheat aleurone layers by modulation of hydrogen peroxide metabolism. J Exp Bot 62:235–248. doi:10.1093/jxb/erq261
Xu SB, Li T, Deng ZY, Chong K, Xue Y, Wang T (2008) Dynamic proteomic analysis reveals a switch between central carbon metabolism and alcoholic fermentation in rice filling grains. Plant Physiol 148:908–925. doi:10.1104/pp.108.125633
Xu H, Gao Y, Wang J (2012) Transcriptomic analysis of rice (Oryza sativa) developing embryos using the RNA-Seq technique. PLoS ONE 7:e30646. doi:10.1371/journal.pone.0030646
Xue LJ, Zhang JJ, Xue HW (2012) Genome-wide analysis of the complex transcriptional networks of rice developing seeds. PLoS ONE 7:e31081. doi:10.1371/journal.pone.0031081
Ye J, Fang L, Zheng H, Zhang Y, Chen J, Zhang Z, Wang J, Li S, Li R, Bolund L, Wang J (2006) WEGO: a web tool for plotting GO annotations. Nucleic Acids Res 34:293–297. doi:10.1093/nar/gkl031
Young TE, Gallie DR (1999) Analysis of programmed cell death in wheat endosperm reveals differences in endosperm development between cereals. Plant Mol Biol 39:915–926. doi:10.1023/A:1006134027834
Young TE, Gallie DR, DeMason DA (1997) Ethylene-mediated programmed cell death during maize endosperm development of wild-type and shrunken2 Genotypes. Plant Physiol 115:737–751. doi:10.1104/pp.115.2.737
Acknowledgments
This work was supported by the State Key Basic Research and Development Plan of China (2013CB126900).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
11103_2013_9_MOESM1_ESM.xls
Supplementary material 1 (XLS 3314 kb). Supplemental Table 1 A total of 23,836 genes derived from three cDNA libraries. The GeneID, length of gene, RPKM of three stages and the expression pattern are presented in this table
11103_2013_9_MOESM2_ESM.xls
Supplementary material 2 (XLS 1246 kb). Supplemental Table 2 A table listing 10,371 differentially expressed genes (DEGs) during rice endosperm development. We use FDR < 0.001 and the absolute value of log2Ratio ≥ 1 as the threshold to judge the significance of gene expression difference. The RPKM of three stages and expression pattern for DEGs are presented in the table
11103_2013_9_MOESM3_ESM.xls
Supplementary material 3 (XLS 51 kb). Supplemental Table 3 Overview of all 123 KEGG pathways. All 10,371 DEGs are assigned to 123 KEGG pathways. The KEGG functional class, number, RPKM of three stages, expression type and expression pattern of members are presented in this table
11103_2013_9_MOESM4_ESM.xls
Supplementary material 4 (XLS 67 kb). Supplemental Table 4 Detail of all 211 branch pathways in 35 MapMan major pathways. The MapMan functional class, number, RPKM of three stages, expression type and expression pattern of members are presented in the table
11103_2013_9_MOESM5_ESM.xls
Supplementary material 5 (XLS 34 kb). Supplemental Table 5 All 55 transcription factor (TF) families found in the result. 1,118 putative TF genes are identified in all 23,836 genes. The TF family name, number, RPKM, expression pattern and expression pattern of TF family’s members are presented in the table
11103_2013_9_MOESM6_ESM.xls
Supplementary material 6 (XLS 2896 kb). Supplemental Table 6 Detail of top 100 up-regulated genes between R1 and R3. The gene ID, gene description and difference RPKM (R3-R1) are presented in the table
11103_2013_9_MOESM7_ESM.xls
Supplementary material 7 (XLS 25 kb). Supplemental Table 7 Thirteen PCD-related genes identified in the RNA-Seq result. The name of gene, gene ID, description and RPKM of three stages are presented in the table
11103_2013_9_MOESM8_ESM.doc
Supplementary material 8 (DOC 43 kb). Supplemental Table 8 Primers used in the analysis of rice endosperm development. UBQ5 is used as internal control to standardize the results
Rights and permissions
About this article
Cite this article
Gao, Y., Xu, H., Shen, Y. et al. Transcriptomic analysis of rice (Oryza sativa) endosperm using the RNA-Seq technique. Plant Mol Biol 81, 363–378 (2013). https://doi.org/10.1007/s11103-013-0009-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11103-013-0009-4