Skip to main navigation menu Skip to main content Skip to site footer

Research Articles

Early Access

Differentially expressed gene profiles of soybean seeds with contrasting seed coat colour

DOI
https://doi.org/10.14719/pst.12065
Submitted
30 September 2025
Published
27-04-2026

Abstract

Soybeans are a vital source of plant-based protein and are widely utilised for various agricultural and industrial purposes. One notable characteristic of soybeans is the diversity in their seed coat colour, which ranges from yellow, black, brown and green to bicoloured variants. This study aimed to identify key genes associated with seed quality by analysing transcriptome profiles of differentially expressed genes in seeds, naked seeds and seed coats of black and white soybean genotypes. The analysis revealed that several upregulated genes are involved in hormone signalling pathways and metabolic processes, such as lysine, starch, sucrose, protein and galactose metabolism. These genes also participate in biosynthetic pathways for ethylene, lipids, brassinosteroids, lignin and sulfur-containing amino acids. Such molecular activities are likely linked to the enhanced seed quality observed in black-coated cultivars, which exhibit greater longevity, improved resistance to ageing, moisture and physical stress. Furthermore, the identification of key transcription factors provided deeper insight into the regulatory mechanisms underlying these traits. This research offers a comprehensive understanding of the genomic and metabolic pathways that influence seed quality in soybeans and lays a foundation for future gene-silencing studies to further explore the biological significance of black seed coat characteristics.

 

References

  1. 1. Crane PR, Friis EM, Pedersen KR. The origin and early diversification of angiosperms. Nature. 1995;374:27–33. https://doi.org/10.1038/374027a0
  2. 2. Soltis P, Soltis D. The origin and diversification of angiosperms. Am J Bot. 2004;91:1614–26. https://doi.org/10.3732/ajb.91.10.1614
  3. 3. Radchuk V, Borisjuk L. Physical, metabolic and developmental functions of the seed coat. Front Plant Sci. 2014;5:510. https://doi.org/10.3389/fpls.2014.00510
  4. 4. Souza FH, Marcos Filho J. The seed coat as a modulator of seed environment relationships in Fabaceae. Braz J Bot. 2001;24:365–75. https://doi.org/10.1590/s0100-84042001000400002
  5. 5. Qiu HM, Chen L, Hou YL, Wang XF, Chen J, Ma XP, et al. Research progress on the genetic regulatory mechanism of seed colour in soybean (Glycine max). Acta Agron Sin. 2021;47(12):2299–313. https://doi.org/10.3724/SP.J.1006.2021.14022
  6. 6. Esau K. Anatomy of seed plants. 2nd ed. New York: John Wiley; 1977.
  7. 7. Miller SS, Bowman LA, Gijzen M, Miki BLA. Early development of the seed coat of soybean. Ann Bot. 1999;84:297–304. https://doi.org/10.1006/anbo.1999.0915
  8. 8. Wang YN, Qi GX, Zhao HK, Yuan CP, Liu XD, Li YQ, et al. Genetic diversity of soybean landraces with different seed coat colours. Mol Plant Breed. 2020;18:1–18. https://doi.org/10.13271/j.mpb.019.007984
  9. 9. McKee GW, Peiffer RA, Mohsenin NN. Seedcoat structure in Coronilla varia and its relations to hard seed. Agron J. 1977;69:53–8.
  10. 10. Eckardt NA. Tissue specific siRNAs that silence CHS genes in soybean. Plant Cell. 2009;21(10):2983–4. https://doi.org/10.1105/tpc.109.072421
  11. 11. Lepiniec L, Debeaujon I, Routaboul JM, Baudry A, Pourcel L, Nesi N, et al. Genetics and biochemistry of seed flavonoids. Annu Rev Plant Biol. 2006;57:405–30. https://doi.org/10.1146/annurev.arplant.57.032905.105252
  12. 12. Smykal P, Vanessa V, Blair MW, Aleš S, Richard DT. The role of the testa during development and in establishment of dormancy of the legume seed. Front Plant Sci. 2014;5:351. https://doi.org/10.3389/fpls.2014.00351
  13. 13. Pourcel L, Routaboul JM, Kerhoas L, Caboche M, Lepiniec L, Debeaujon I. TRANSPARENT TESTA10 encodes a laccase-like enzyme involved in oxidative polymerisation of flavonoids in Arabidopsis seed coat. Plant Cell. 2005;17:2966–80. https://doi.org/10.1105/tpc.105.035154
  14. 14. Rajjou L, Debeaujon I. Seed longevity: survival and maintenance of high germination ability of dry seeds. C R Biol. 2008;331(10):796–805.
  15. 15. Sun W, Meng X, Liang L, Jiang W, Huang Y, He J, et al. Molecular and biochemical analysis of chalcone synthase from freesia hybrid in flavonoid biosynthetic pathway. PLoS One. 2015;10(3):e0119054. https://doi.org/10.1371/journal.pone.0119054
  16. 16. Zabala G, Vodkin L. Cloning of pleiotropic T locus in soybean and two recessive alleles that differentially affect structure and expression of encoded flavonoid 3′ hydroxylase. Genetics. 2003;163:295–309. https://doi.org/10.1093/genetics/163.1.295
  17. 17. Song J, Guo Y, Yu LJ, Qiu LJ. Progress in genes related to seed coat colour in soybean. Yi Chuan. 2012;34(6):687–94. https://doi.org/10.3724/sp.j.1005.2012.00687
  18. 18. Yoshikura K, Hamaguchi Y. Anthocyanins of black soybean. Eiyo To Shokuryo. 1969;22(6):367–70. https://doi.org/10.4327/jsnfs1949.22.367
  19. 19. Winkel-Shirley B. Flavonoid biosynthesis: a colourful model for genetics, biochemistry, cell biology and biotechnology. Plant Physiol. 2001;126(2):485–93. https://doi.org/10.1104/pp.126.2.485
  20. 20. Kovinich N, Saleem A, Rintoul TL, Brown DCW, Arnason JT, Miki B. Colouring genetically modified soybean grains with anthocyanins by suppression of the proanthocyanidin genes ANR1 and ANR2. Transgenic Res. 2012;21(4):757–71. https://doi.org/10.1007/s11248-011-9566-y
  21. 21. Wang H, Murphy PA. Isoflavone content in commercial soybean foods. J Agric Food Chem. 1994;42:1666–73. https://doi.org/10.1021/jf00044a016
  22. 22. Dhaubhadel S, Gijzen M, Moy P, Farhangkhoee M. Transcriptome analysis reveals a critical role of CHS7 and CHS8 genes for isoflavone synthesis in soybean seeds. Plant Physiol. 2007;143:326–38. https://doi.org/10.1104/pp.106.086306
  23. 23. Ralston L, Subramanian S, Matsuno M, Yu O. Partial reconstruction of flavonoid and isoflavone biosynthesis in yeast using soybean type I and type II chalcone isomerases. Plant Physiol. 2005;137:1375–88. https://doi.org/10.1104/pp.104.054502
  24. 24. Akashi T, Aoki T, Ayabe S. Cloning and functional expression of a cytochrome P450 cDNA encoding 2-hydroxyisoflavanone synthase involved in biosynthesis of the isoflavonoid skeleton in licorice. Plant Physiol. 1999;121(3):821–8. https://doi.org/10.1104/pp.121.3.821
  25. 25. Jung W, Yu O, Lau SC, O’Keefe DP, Odell J, Fader G. Identification and expression of isoflavone synthase, the key enzyme for biosynthesis of isoflavones in legumes. Nat Biotechnol. 2000;18:208–12. https://doi.org/10.1038/72671
  26. 26. Kuchlan MK, Kuchlan P, Onkar M, Ramesh A, Husain SM. Influence of seed coat compactness around cotyledons, protein and mineral composition on mechanical strength of soybean (Glycine max (L.) Merrill) seed coat. Legume Res. 2018;41:246–52.
  27. 27. Song QX, Liu YF, Hu XY, Zhang WK, Ma B, Chen SY, et al. Identification of miRNAs and their target genes in developing soybean seeds by deep sequencing. BMC Plant Biol. 2011;11:5. https://doi.org/10.1186/1471-2229-11-5
  28. 28. Dhaubhadel S, McGarvey BD, Williams R, Gijzen M. Isoflavonoid biosynthesis and accumulation in developing soybean seeds. Plant Mol Biol. 2003;53:733–43. https://doi.org/10.1023/B:PLAN.0000023666.30358.ae
  29. 29. Palmer RG, Pfeiffer TW, Buss GR, Kilen TC. Qualitative genetics. In: Boerma HR, Specht JE, editors. Soybeans: improvement, production and uses. 3rd ed. Madison (WI): ASA, CSSA, SSSA; 2004. p. 137–214.
  30. 30. Yang X, Yan J, Shah T, Warburton ML, Li Q, Li L, et al. Genetic analysis and characterisation of a new maize association mapping panel for quantitative trait loci dissection. Theor Appl Genet. 2010;121(3):417–31. https://doi.org/10.1007/s00122-010-1320-y
  31. 31. Song J, Liu Z, Hong H, Ma Y, Tian L, Li X, et al. Identification and validation of loci governing seed coat color by combining association mapping and bulk segregation analysis in soybean. PLoS One. 2016;11:e0159064. https://doi.org/10.1371/journal.pone.0159064
  32. 32. Bernard RL, Weiss MG. Qualitative genetics. In: Caldwell BE, editor. Soybean: improvement, production and uses. Madison (WI): American Society of Agronomy; 1973. p. 117–54.
  33. 33. Nagamatsu A, Masuta C, Senda M, Matsuura H, Kasai A, Hong JS, et al. Functional analysis of soybean genes involved in flavonoid biosynthesis by virus induced gene silencing. Plant Biotechnol J. 2007;5:778–90. https://doi.org/10.1111/j.1467-7652.2007.00288.x
  34. 34. Toda K, Yang D, Yamanaka N, Watanabe S, Harada K, Takahashi R. A single base deletion in soybean flavonoid 3′ hydroxylase gene is associated with grey pubescence color. Plant Mol Biol. 2002;50:187–96. https://doi.org/10.1023/A:1016087221334
  35. 35. Clough SJ, Tuteja JH, Li M, Marek LF, Shoemaker RC, Vodkin LO. Features of a 103 kb gene rich region in soybean include an inverted perfect repeat cluster of CHS genes comprising the I locus. Genome. 2004;47(5):819–31. https://doi.org/10.1139/g04-049
  36. 36. Senda M. Patterning of virus infected Glycine max seed coat is associated with suppression of endogenous silencing of chalcone synthase genes. Plant Cell. 2004;16(4):807–18. https://doi.org/10.1105/tpc.019885
  37. 37. Tuteja JH. Tissue-specific gene silencing mediated by a naturally occurring chalcone synthase gene cluster in Glycine max. Plant Cell. 2004;16(4):819–35. https://doi.org/10.1105/tpc.021352.
  38. 38. Kasai A, Kasai K, Yumoto S, Senda M. Structural features of GmIRCHS, candidate of the I gene inhibiting seed coat pigmentation in soybean: implications for inducing endogenous RNA silencing of chalcone synthase genes. Plant Mol Biol. 2007;64(4):467–79. https://doi.org/10.1007/s11103-007-9169-4.
  39. 39. Tuteja JH, Zabala G, Varala K, Hudson M, Vodkin LO. Endogenous, tissue-specific short interfering RNAs silence the chalcone synthase gene family in Glycine max seed coats. Plant Cell. 2009;21:3063–77.
  40. 40. Senda M, Kasai A, Yumoto S, Akada S, Ishikawa R, Harada T, et al. Sequence divergence at chalcone synthase gene in pigmented seed coat soybean mutants of the inhibitor locus. Genes Genet Syst. 2002;77(5):341–50. https://doi.org/10.1266/ggs.77.341.
  41. 41. Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016;32(19):3047–8. https://doi.org/10.1093/bioinformatics/btw354
  42. 42. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29(1):15–21. https://doi.org/10.1093/bioinformatics/bts635
  43. 43. Liao Y, Smyth GK, Shi W. featureCounts: an efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 2014;30(7):923–30. https://doi.org/10.1093/bioinformatics/btt656
  44. 44. Blighe K, Rana S, Lewis M. Enhanced Volcano: publication-ready volcano plots with enhanced colouring and labeling. R package. 2019;Version 1.0.
  45. 45. Kassambara A, Kassambara MA. ggpubr: ‘ggplot2’ based publication-ready plots. R package. 2020;Version 0.1.
  46. 46. Singer WM, Zhang B, Mian MAR, Huang H. Soybean amino acids in health, genetics and evaluation. In: Soybean: human consumption and animal feed. 2019.
  47. 47. Aguirre M, Kiegle E, Leo G, Ezquer I. Carbohydrate reserves and seed development: an overview. Plant Reprod. 2018;31(3):263–90. https://doi.org/10.1007/s00497-018-0336-3
  48. 48. Tayade R, Kulkarni KP, Jo H, Song JT, Lee JD. Insight into the prospects for the improvement of seed starch in legumes: a review. Front Plant Sci. 2019;10:1213. https://doi.org/10.3389/fpls.2019.01213
  49. 49. Krishnan HB, Jez JM. The promise and limits for enhancing sulfur containing amino acid content of soybean seed. Plant Sci. 2018;272:14–21. https://doi.org/10.1016/j.plantsci.2018.03.030
  50. 50. Ma Y, Ma W, Hu D, Zhang X, Yuan W, He X, et al. QTL mapping for protein and sulfur-containing amino acid contents using a high-density bin map in soybean (Glycine max L. Merr.). J Agric Food Chem. 2019;67:12313–21. https://doi.org/10.1021/acs.jafc.9b04497
  51. 51. Ding Y, Zhou X, Zuo L, Wang H, Yu D. Identification and functional characterization of the sulfate transporter gene GmSULTR1;2b in soybean. BMC Genomics. 2016;17:1–19. https://doi.org/10.1186/s12864-016-2705-3
  52. 52. Bai Z, Qi T, Liu Y, Wu Z, Ma L, Liu W, et al. Alteration of S-adenosylhomocysteine levels affects lignin biosynthesis in switchgrass. Plant Biotechnol J. 2018;16:2016–26. https://doi.org/10.1111/pbi.12935
  53. 53. Malle S, Eskandari M, Morrison M, Belzile F. Genome wide association identifies several QTLs controlling cysteine and methionine content in soybean seed including some promising candidate genes. Sci Rep. 2020;10:1–14. https://doi.org/10.1038/s41598-020-78907-w
  54. 54. Zhang X, Wang Y, Yan Y, Peng H, Long Y, Zhang Y, et al. Transcriptome sequencing analysis of maize embryonic callus during early redifferentiation. BMC Genomics. 2019;20:1–22. https://doi.org/10.1186/s12864-019-5506-7
  55. 55. Miyakawa T, Hatano KI, Miyauchi Y, Suwa YI, Sawano Y, Tanokura M. A secreted protein with plant specific cysteine rich motif functions as a mannose binding lectin that exhibits antifungal activity. Plant Physiol. 2014;166:766–78. https://doi.org/10.1104/pp.114.242636
  56. 56. Belkhadir Y, Yang L, Hetzel J, Dangl JL, Chory J. The growth–defense pivot: crisis management in plants mediated by LRR-RK surface receptors. Trends Biochem Sci. 2014;39:447–56. https://doi.org/10.1016/j.tibs.2014.06.006
  57. 57. Ishibashi Y, Koda Y, Zheng SH, Yuasa T, Iwaya-Inoue M. Regulation of soybean seed germination through ethylene production in response to reactive oxygen species. Ann Bot. 2013;111:95–102. https://doi.org/10.1093/aob/mcs240
  58. 58. Ciabotti S, Silva ACBB, Juhasz ACP, Mendonça CD, Tavano OL, Mandarino JMG, et al. Chemical composition, protein profile and isoflavones content in soybean genotypes with different seed coat colors. Int Food Res J. 2016.
  59. 59. Chandra S, Talukdar A, Taak Y, Yadav RR, Saini M, Sipani NS. Seed longevity studies in wild type, cultivated and inter specific recombinant inbred lines (RILs) of soybean (Glycine max (L.) Merr.). Genet Resour Crop Evol. 2022;69:399–409. https://doi.org/10.1007/s10722-021-01240-2
  60. 60. Ohnishi T, Godza B, Watanabe B, Fujioka S, Hategan L, Ide K, et al. CYP90A1/CPD, a brassinosteroid biosynthetic cytochrome P450 of Arabidopsis, catalyzes C-3 oxidation. J Biol Chem. 2012;287:31551–60. https://doi.org/10.1074/jbc.M112.392720
  61. 61. Ahammed GJ, Gantait S, Mitra M, Yang Y, Li X. Role of ethylene crosstalk in seed germination and early seedling development: a review. Plant Physiol Biochem. 2020;151:124–31. https://doi.org/10.1016/j.plaphy.2020.03.016
  62. 62. Yin W, Dong N, Niu M, Zhang X, Li L, Liu J, et al. Brassinosteroid regulated plant growth and development and gene expression in soybean. Crop J. 2019;7:411–8. https://doi.org/10.1016/j.cj.2018.10.003
  63. 63. Chen H, Kim HU, Weng H, Browse J. Malonyl-CoA synthetase, encoded by ACYL ACTIVATING ENZYME13, is essential for growth and development of Arabidopsis. Plant Cell. 2011;23:2247–62. https://doi.org/10.1105/tpc.111.086140
  64. 64. Griebel T, Zeier J. A role for β-sitosterol to stigmasterol conversion in plant pathogen interactions. Plant J. 2010;63:254–68. https://doi.org/10.1111/j.1365-313X.2010.04235.x
  65. 65. Huth C, Mertz-Henning LM, Lopes SJ, Tabaldi LA, Rossato LV, Krzyzanowski FC, et al. Susceptibility to weathering damage and oxidative stress on soybean seeds with different lignin contents in the seed coat. J Seed Sci. 2016;38:296–304. https://doi.org/10.1590/2317-1545v38n4162115
  66. 66. Bellaloui N, Mengistu A, Fisher DK, Abel CA. Soybean seed composition constituents as affected by drought and Phomopsis in Phomopsis susceptible and resistant genotypes. J Crop Improv. 2012;26:428–53. https://doi.org/10.1080/15427528.2011.651774
  67. 67. Kuchlan MK, Dadlani M, Samuel DVK. Seed coat properties and longevity of soybean seeds. J New Seeds. 2010;11:239–49. https://doi.org/10.1080/1522886X.2010.497960
  68. 68. Ariyoshi Y, Itoyama H, Nakagawa ACS, Ario N, Kondo Y, Tomita Y, et al. Regulation of brassinosteroid on pod growth through cell hypertrophy in soybean (Glycine max (L.) Merr.). Plant Growth Regul. 2016;80(3):391–5. https://doi.org/10.1007/s10725-016-0176-9
  69. 69. Cabianca A, Müller L, Pawlowski K, Dahlin P. Changes in the plant β-sitosterol/stigmasterol ratio caused by the plant parasitic nematode Meloidogyne incognita. Plants. 2021;10:292. https://doi.org/10.3390/plants10020292
  70. 70. Wang M, Xu X, Zhang X, Sun S, Wu C, Hou W, et al. Functional analysis of GmCPDs and investigation of their roles in flowering. PLoS One. 2015;10:e0118476. https://doi.org/10.1371/journal.pone.0118476
  71. 71. Ertekin M, Kirdar E. Effects of seed coat colour on seed characteristics of honeylocust (Gleditsia triacanthos). Afr J Agric Res. 2010;5(17):2434–8.

Downloads

Download data is not yet available.