Plant Growth Regulation

, Volume 69, Issue 2, pp 167–176 | Cite as

Abscisic acid is required for somatic embryo initiation through mediating spatial auxin response in Arabidopsis

  • Ying Hua Su
  • Yu Xiao Su
  • Ying Gao Liu
  • Xian Sheng Zhang
Original paper


Abscisic acid (ABA) regulates many aspects of plant development, including somatic embryo (SE) initiation. However, mechanisms of ABA functions on SE initiation have remained to be investigated. In this study, we examined the endogenous ABA contents of calli in Arabidopsis during the SE inductive process. We further found that the capacity for SE initiation was strongly impaired by treatment of fluridone, a potent inhibitor of ABA biosynthesis, as well as by mutation of ABA biosynthetic gene ABA2, suggesting that ABA is required for SE initiation. Furthermore, treatment of fluridone inhibited local auxin biosynthesis and auxin polar transport in the embryonic calli, resulting in the disturbance of auxin response pattern and the decreased regeneration frequency of SEs. However, application of exogenous ABA in the medium almost recovered patterns of auxin response and SE initiation. Thus, the results suggest that ABA functions on SE initiation through mediating both auxin biosynthesis and polar transport for establishment of auxin response pattern in callus. Our study provides new information for understanding mechanisms of SE initiation.


Somatic embryo initiation ABA Auxin biosynthesis Auxin polar transport Auxin response 



This research was supported by grants from the National Natural Science Foundation (NNSF) of China (90917015, 31000652, and 31170272). No conflict of interest declared.


  1. Agusti J, Herold S, Schwarz M, Sanchez P, Ljung K, Dun EA, Brewer PB, Beveridge CA, Sieberer T, Sehr EM, Greb T (2011) Strigolactone signaling is required for auxin-dependent stimulation of secondary growth in plants. Proc Natl Acad Sci USA 108:20242–20247PubMedCrossRefGoogle Scholar
  2. Ammirato PV (1977) Hormonal control of somatic embryo development from cultured cells of caraway. Plant Physiol 59:579–586PubMedCrossRefGoogle Scholar
  3. Belin C, Megies C, Hauserová E, Lopez-Molina L (2009) Abscisic acid represses growth of the Arabidopsis embryonic axis after germination by enhancing auxin signaling. Plant Cell 21:2253–2268PubMedCrossRefGoogle Scholar
  4. Birnbaum KD, Alvarado AS (2008) Slicing across kingdoms: regeneration in plants and animals. Cell 132:697–710PubMedCrossRefGoogle Scholar
  5. Braybrook SA, Harada JJ (2008) LECs go crazy in embryo development. Trends Plant Sci 13:624–630PubMedCrossRefGoogle Scholar
  6. Cheng Y, Dai X, Zhao Y (2007) Auxin synthesized by the YUCCA flavin monooxygenases is essential for embryogenesis and leaf formation in Arabidopsis. Plant Cell 19:2430–2439PubMedCrossRefGoogle Scholar
  7. Fedina IS, Tsonev TD, Guleva EI (1994) ABA as modulator of the response of Pisum sativum to salt stress. J Plant Physiol 143:245–249CrossRefGoogle Scholar
  8. Finkelstein RR, Gampala SS, Rock CD (2002) Abscisic acid signaling in seeds and seedlings. Plant Cell 14:S15–S45PubMedGoogle Scholar
  9. Friml J, Vieten A, Sauer M, Weijers D, Schwarz H, Hamann T, Offringa R, Jürgens G (2003) Efflux-dependent auxin gradients establish the apical-basal axis of Arabidopsis. Nature 426:147–153PubMedCrossRefGoogle Scholar
  10. González-Guzmán M, Apostolova N, Bellés JM, Barrero JM, Piqueras P, Ponce MR, Micol JL, Serrano R, Rodríguez PL (2002) The short-chain alcohol dehydrogenase ABA2 catalyzes the conversion of xanthoxin to abscisic aldehyde. Plant Cell 14:1833–1846PubMedCrossRefGoogle Scholar
  11. Gutmann M, von Aderkas P, Label P, Lelu MA (1996) Effects of abscisic acid on somatic embryo maturation of hybrid larch. J Exp Bot 47:1905–1917CrossRefGoogle Scholar
  12. Heisler MG, Ohno C, Das P, Sieber P, Reddy GV, Long JA, Meyerowitz EM (2005) Patterns of auxin transport and gene expression during primordium development revealed by live imaging of the Arabidopsis inflorescence meristem. Curr Biol 15:1899–1911PubMedCrossRefGoogle Scholar
  13. Ikeda-Iwai M, Satoh S, Kamada H (2002) Establishment of a reproducible tissue culture system for the induction of Arabidopsis somatic embryos. J Exp Bot 53:1575–1580PubMedCrossRefGoogle Scholar
  14. Jenik PD, Gillmor CS, Lukowitz W (2007) Embryonic patterning in Arabidopsis thaliana. Annu Rev Cell Dev Biol 23:207–236PubMedCrossRefGoogle Scholar
  15. Jimnez VM, Guevara E, Herrera J, Bangerth F (2005) Evolution of endogenous hormone concentration in embryogenic cultures of carrot during early expression of somatic embryogenesis. Plant Cell Rep 23:567–572CrossRefGoogle Scholar
  16. Karami O, Saidi A (2010) The molecular basis for stress-induced acquisition of somatic embryogenesis. Mol Biol Rep 37:2493–2507PubMedCrossRefGoogle Scholar
  17. Karami O, Aghavaisi B, Pour AM (2009) Molecular aspects of somatic-to-embryogenic transition in plants. J Chem Biol 2:177–190PubMedCrossRefGoogle Scholar
  18. Kikuchi A, Sanuki N, Higashi K, Koshiba T, Kamada H (2006) Abscisic acid and stress treatment are essential for the acquisition of embryogenic competence by carrot somatic cells. Planta 223:637–645PubMedCrossRefGoogle Scholar
  19. Koornneef M, Karssen CM (1994) Seed dormancy and germination. In: Meyerowitz EM, Somerville CR (eds) Arabidopsis. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 313–334Google Scholar
  20. Liu CM, Xu ZH, Chua NH (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5:621–630PubMedGoogle Scholar
  21. Liu Y, Ye N, Liu R, Chen M, Zhang J (2010) H2O2 mediates the regulation of ABA catabolism and GA biosynthesis in Arabidopsis seed dormancy and germination. J Exp Bot 61:2979–2990PubMedCrossRefGoogle Scholar
  22. Matusova R, Rani K, Verstappen FWA, Franssen MCR, Beale MH, Bouwmeester HJ (2005) The strigolactone germination stimulants of the plant-parasitic Striga and Orobanche spp. are derived from the carotenoid pathway. Plant Physiol 139:920–934PubMedCrossRefGoogle Scholar
  23. Meinke DW (1991) Perspectives on genetic analysis of plant embryogenesis. Plant Cell 3:857–866PubMedGoogle Scholar
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  25. Nakabayashi K, Okamoto M, Koshiba T, Kamiya Y, Nambara E (2005) Genome-wide profiling of stored mRNA in Arabidopsis thaliana seed germination: epigenetic and genetic regulation of transcription in seed. Plant J 41:697–709PubMedCrossRefGoogle Scholar
  26. Nambara E, Marion-Poll A (2003) ABA action and interactions in seeds. Trends Plant Sci 8:213–217PubMedCrossRefGoogle Scholar
  27. Nambara E, Marion-Poll A (2005) Abscisic acid biosynthesis and metabolism. Annu Rev Plant Biol 56:165–185PubMedCrossRefGoogle Scholar
  28. Nishiwaki M, Fujino K, Koda Y, Masuda K, Kikuta Y (2000) Somatic embryogenesis induced by the simple application of abscisic acid to carrot (Daucus carota L.) seedlings in culture. Planta 211:756–759PubMedCrossRefGoogle Scholar
  29. Penfield S, Li Y, Gilday AD, Graham S, Graham IA (2006) Arabidopsis ABA INSENSITIVE4 regulates lipid mobilization in the embryo and reveals repression of seed germination by the endosperm. Plant Cell 18:1887–1899PubMedCrossRefGoogle Scholar
  30. Popova L (1996) Effect of fluridone on plant development, leaf anatomy and plastid ultrastructure of barley plants. Bulg J Plant Physiol 22:3–12Google Scholar
  31. Raz V, Bergervoet JHW, Koornneef M (2001) Sequential steps for developmental arrest in Arabidopsis seeds. Development 128:243–252PubMedGoogle Scholar
  32. Reinert J (1958) Morphogenese und ihre kontrolle an gewebekulturen aus carotten. Naturwissenschaften 45:344–345CrossRefGoogle Scholar
  33. Rock CD, Quatrano RS (1995) The role of hormones during seed development. In: Davies PJ (ed) Plant hormones: physiology, biochemistry and molecular biology, 2nd edn. Kluwer, Dordrecht, pp 671–697Google Scholar
  34. Ruyter-Spira C, Kohlen W, Charnikhova T, van Zeijl A, van Bezouwen L, de Ruijter N, Cardoso C, Lopez-Raez JA, Matusova R, Bours R, Verstappen F, Bouwmeester H (2011) Physiological effects of the synthetic strigolactone analog GR24 on root system architecture in Arabidopsis: another belowground role for strigolactones? Plant Physiol 155:721–734PubMedCrossRefGoogle Scholar
  35. Saab IN, Sharp RE, Pritchard J (1992) Effect of inhibition of abscisic acid accumulation on spatial distribution of elongation in the primary root and mesocotyl of maize at low water potentials. Plant Physiol 99:26–33PubMedCrossRefGoogle Scholar
  36. Schwartz SH, Qin X, Zeevaart JAD (2003) Elucidation of the indirect pathway of abscisic acid biosynthesis by mutants, genes, and enzymes. Plant Physiol 131:1591–1601PubMedCrossRefGoogle Scholar
  37. Seo M, Peeters AJM, Koiwai H, Oritani T, Marion-Poll A, Zeevaart JAD, Koornneef M, Kamiya Y, Koshiba T (2000) The Arabidopsis aldehyde oxidase 3 (AAO3) gene product catalyzes the final step in abscisic acid biosynthesis in leaves. Proc Natl Acad Sci USA 97:12908–12913PubMedCrossRefGoogle Scholar
  38. Shiota H, Satoh R, Watabe K, Harada H, Kamada H (1998) C-ABI3, the carrot homologue of the Arabidopsis ABI3, is expressed during both zygotic and somatic embryogenesis and functions in the regulation of embryo-specific ABA-inducible genes. Plant Cell Physiol 39:1184–1193PubMedCrossRefGoogle Scholar
  39. Shkolnik-Inbar D, Bar-Zvi D (2010) ABI4 mediates abscisic acid and cytokinin inhibition of lateral root formation by reducing polar auxin transport in Arabidopsis. Plant Cell 22:3560–3573PubMedCrossRefGoogle Scholar
  40. Sieburth LE, Meyerowitz EM (1997) Molecular dissection of the AGAMOUS control region shows that cis elements for spatial regulation are located intragenically. Plant Cell 9:355–365PubMedGoogle Scholar
  41. Stepanova AN, Yun J, Robles LM, Novak O, He WR, Guo HW, Ljung K, Alonso JM (2011) The Arabidopsis YUCCA1 flavin monooxygenase functions in the indole-3-pyruvic acid branch of auxin biosynthesis. Plant Cell 23:3961–3973PubMedCrossRefGoogle Scholar
  42. Steward FC, Mapes MO, Smith J (1958) Growth and organized development of cultured cells. II. Growth and division of freely suspended cells. Am J Bot 45:693–703CrossRefGoogle Scholar
  43. Su YH, Zhao XY, Liu YB, Zhang CL, O′Neill SD, Zhang XS (2009) Auxin-induced WUS expression is essential for embryonic stem cell renewal during somatic embryogenesis in Arabidopsis. Plant J 59:448–460PubMedCrossRefGoogle Scholar
  44. Suzuki M, McCarty DR (2008) Functional symmetry of the B3 network controlling seed development. Curr Opin Plant Biol 11:548–553PubMedCrossRefGoogle Scholar
  45. Thompson AJ, Jackson AC, Symonds RC, Mulholland BJ, Dadswell AR, Blake PS, Burbidge A, Taylor IB (2000) Ectopic expression of a tomato 9-cis-epoxycarotenoid dioxygenase gene causes over-production of abscisic acid. Plant J 23:363–374PubMedCrossRefGoogle Scholar
  46. Tian L, Brown DCW (2000) Improvement of soybean somatic embryo development and maturation by abscisic acid treatment. Can J Plant Sci 80:271–276CrossRefGoogle Scholar
  47. Wang L, Hua D, He J, Duan Y, Chen Z, Hong X, Gong Z (2011) Auxin Response Factor2 (ARF2) and its regulated homeodomain gene HB33 mediate abscisic acid response in Arabidopsis. PLoS Genet 7:e1002172. doi: 10.1371/journal.pgen.1002172 PubMedCrossRefGoogle Scholar
  48. Zhao YD (2010) Auxin biosynthesis and its role in plant development. Annu Rev Plant Bio 61:49–64CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Ying Hua Su
    • 1
  • Yu Xiao Su
    • 1
  • Ying Gao Liu
    • 1
  • Xian Sheng Zhang
    • 1
  1. 1.State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Life SciencesShandong Agricultural UniversityTaianChina

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