Skip to main content
SpringerLink
Log in

Somatic Embryogenesis in Common BeanPhaseolus vulgaris L.

  • Protocol
  • First Online:
Plant Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1815))

Abstract

Common bean Phaseolus vulgaris L. has been shown to be a recalcitrant plant to induce somatic embryogenesis (SE) under in vitro conditions. An alternative strategy to yield SE is based upon the use of a cytokinin (benzyladenine) coupled with osmotic stress adaptation instead of the auxin-inducing SE in common bean. Here we described the induction of proembryogenic masses (PEM) derived from the apical meristem and cotyledonary zone of zygotic embryos, from which secondary SE indirect embryogenesis emerged. Maturation of SE was achieved by using osmotic stress medium and converted to plants. Long-term recurrent SE was demonstrated by propagation of PEM at early stages of SE. This protocol is currently being applied for stable genetic transformation by means of Agrobacterium tumefaciens and biobalistics as well as basic biochemical and molecular biology research.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 279.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Food and Agricultural Organization of the United Nations (FAO) (1999) PHASEOLUS BEAN: Post-harvest Operations. Organisation: Centro Internacional de Agricultura tropical (CIAT) www.cgiar.org/ciat. Author: AL. Jones. Edited by AGSI/FAO: Danilo Mejia (Technical), Beverly Lewis (Language and style)

  2. Veltcheva M, Svetleva D, Petkova SP, Perl A (2005) In vitro regeneration and genetic transformation of common bean (Phaseolus vulgaris L.)-problems and progress. Sci Hort 107:2–10. https://doi.org/10.1016/j.scienta.2005.07.005

    Article  CAS  Google Scholar 

  3. Delgado-Sánchez P, Saucedo-Ruiz M, Guzmán-Maldonado SH et al (2006) An organogenic plant regeneration system for common bean (Phaseolus vulgaris L.). Plant Sci 170:822–827. https://doi.org/10.1016/j.plantsci.2005.11.015

    Article  CAS  Google Scholar 

  4. Arellano J, Fuentes SI, Castillo-España P, Hernández G (2009) Regeneration of different cultivars of common bean (Phaseolus vulgaris L.) via indirect organogenesis. Plant Cell Tiss Org 96:11–18. https://doi.org/10.1007/s11240-008-9454-1

    Article  CAS  Google Scholar 

  5. Gatica-Arias AM, Muñoz-Valverde J, Ramírez-Fonseca P, Valdez-Melara M (2010) In vitro plant regeneration system for common bean (Phaseolus vulgaris): effect of N6-benzylaminopurine and adenine sulphate. Electron J Biotechnol 13:a06. https://doi.org/10.4067/S0717-34582010000100006

    Article  Google Scholar 

  6. Kwapata K, Sabzikar R, Stcklen MB, Kelly JD (2010) In vitro regeneration and morphogenesis studies in common bean. Plant Cell Tissue Organ Cult 100:97–105. https://doi.org/10.1007/s11240-009-9624-9

    Article  CAS  Google Scholar 

  7. Collado R, Veitía N, Bermúdez-Caraballoso I et al (2013) Efficient in vitro plant regeneration via indirect organogenesis for different common bean cultivars. Sci Hort 153:109–116. https://doi.org/10.1016/j.scienta.2013.02.007

    Article  CAS  Google Scholar 

  8. Russell DR, Wallace KM, Bathe JH et al (1993) Stable transformation of Phaseolus vulgaris via electric-discharge mediated particle acceleration. Plant Cell Rep 12:165–169. https://doi.org/10.1007/BF00239099

    Article  PubMed  CAS  Google Scholar 

  9. Aragao FJL, Barros LMG, Brasileiro ACM et al (1996) Inheritance of foreign genes in transgenic bean (Phaseolus vulgaris L.) co-transformed via particle bombardment. Theor Appl Genet 93:142–150. https://doi.org/10.1007/BF00225739

    Article  Google Scholar 

  10. Aragao FJL, Ribeiro SG, Barros LMG et al (1998) Transgenic beans (Phaseolus vulgaris L.) engineered to express viral antisense RNAs show delayed and attenuated symptoms to bean golden mosaic geminivirus. Mol Breed 4:491–499. https://doi.org/10.1023/A:1009613607559

    Article  CAS  Google Scholar 

  11. Kim JW, Minamikawa T (1996) Transformation and regeneration of French bean plants by the particle bombardment process. Plant Sci 117:131–138. https://doi.org/10.1016/0168-9452(96)04403-2

    Article  CAS  Google Scholar 

  12. Bonfim K, Faria JC, Nogueira EOPL, Mendes EA, Aragao FJL (2007) RNAi-mediated resistance to bean golden mosaic virus in genetically engineered common bean (Phaseolus vulgaris). Mol Plant Microbe In 20:717–726. https://doi.org/10.1094/MPMI-20-6-0717

    Article  CAS  Google Scholar 

  13. Christou P (1990) Morphological description of transgenic soybean chimeras created by the delivery, integration and expression of foreign DNA using electric discharge particle acceleration. Ann Bot 66:379–386. https://doi.org/10.1093/oxfordjournals.aob.a088039

    Article  CAS  Google Scholar 

  14. Krastanova S, Perrin M, Barbier P et al (1995) Transformation of grapevine rootstocks with the coat protein gene of grapevine fanleaf nepovirus. Plant Cell Rep 13:357–360. https://doi.org/10.1007/BF00231936

    Article  Google Scholar 

  15. Xiang YZ, Ying HS, Zhi JC, Xian SZ (2008) Cell fate switch during in vitro plant organogenesis. J Integ Plant Biol 50:816–824. https://doi.org/10.1111/j.1744-7909.2008.00701.x

    Article  CAS  Google Scholar 

  16. Steward FC, Mapes MO, Smith J (1958) Growth and organized development of cultured cells. I. Growth and division of freely suspended cells. Am J Bot 45:693–703

    Article  Google Scholar 

  17. Haccius B (1978) Question of unicellular origin of non-zygotic embryos in callus cultures. Phytomorphology 28:74–81

    Google Scholar 

  18. Raghavan V (1976) Adventive embryogenesis: induction of diploid embryoids. In: Sutchliffe JF (ed) Experimental embryogenesis in vascular plants. Academic Press, London, pp 349–381

    Google Scholar 

  19. Xu Z, Zhang C, Zhang X et al (2013) Transcriptome profiling reveals auxin and cytokinin regulating somatic embryogenesis in different sister lines of cotton cultivar CCR124. J Integr Plant Biol 55:631–642. https://doi.org/10.1111/jipb.12073

    Article  PubMed  CAS  Google Scholar 

  20. Allavena A, Rossetti L (1983) Efforts in somatic embryogenesis of Phaseolus vulgaris L. Acta Hort 131:239–246. https://doi.org/10.17660/ActaHortic.1983.131.27

    Article  Google Scholar 

  21. Martins IS, Sondahl MR (1984) Early stages of somatic embryo differentiation from callus cells of bean (Phaseolus vulgaris L.) grown in liquid medium. J Plant Physiol 117:97–103. https://doi.org/10.1016/S0176-1617(84)80021-8

    Article  PubMed  CAS  Google Scholar 

  22. Saunders JW, Hosfield GL, Levi A (1987) Morphogenetic effects of 2,4-dichlorophenoxyacetic acid on pinto bean (Phaseolus vulgaris L.) leaf explants in vitro. Plant Cell Rep 6:46–49. https://doi.org/10.1007/BF00269737

    Article  PubMed  CAS  Google Scholar 

  23. Hoyos RA (1990) Somatic embryogenesis in common bean (Phaseolus vulgaris L.): influence of media and environmental factors on globular and mature embryoid formation. Department of Crop and Soil Science. Michigan State University, USA

    Google Scholar 

  24. Collado R, Garcia LR, Angenon G et al (2011) Somatic embryos formation from immature cotyledons in Phaseolus vulgaris cv. CIAP 7247. Biotechnol Veg 11:235–240

    Google Scholar 

  25. Malik KA, Saxena PK (1992) Somatic embryogenesis and shoot regeneration from intact seedlings of Phaseolus acutifolius A., P. aureus (L.) Wilczek, P. coccineus, and P. wrightii L. Plant Cell Rep 11:163–168. https://doi.org/10.1007/BF00232172

    Article  PubMed  CAS  Google Scholar 

  26. Garcia LR, Pérez J, Kosky RG et al (2010) Regeneración de plantas via embriogénesis somática en Phaseolus acutifolius A. Gray. Biotechnol Veg 10:143–149

    Google Scholar 

  27. Nafie EM, Taha HS, Mansur RM (2013) Impact of 24-epibrassinolide on callogenesis and regeneration via somatic embryogenesis in Phaseolus vulgaris L. cv Brunca. World Appl Sci J 24:188–200. https://doi.org/10.5829/idosi.wasj.2013.24.02.13191

    Article  Google Scholar 

  28. Fehér A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Plant Cell Tiss Org 74:201–228. https://doi.org/10.1023/A:1024033216561

    Article  Google Scholar 

  29. Karami O, Saidi A (2010) The molecular basis for stress-induced acquisition of somatic embryogenesis. Mol Biol Rep 37:2493–2507. https://doi.org/10.1007/s11033-009-9764-3

    Article  PubMed  CAS  Google Scholar 

  30. Kamada H, Ishikawa K, Saga H, Harada H (1993) Induction of somatic embryogenesis in carrot by osmotic stress. Plant Tiss Cult Lett 10:38–44. https://doi.org/10.5511/plantbiotechnology1984.10.38

    Article  CAS  Google Scholar 

  31. Lou H, Kako S (1995) Role of high sugar concentrations in inducing somatic embryogenesis from cucumber cotyledons. Sci Hort 64:11–20. https://doi.org/10.1016/0304-4238(95)00833-8

    Article  CAS  Google Scholar 

  32. Ikeda-Iwai M, Umehara M, Satoh S, Kamada H (2003) Stress-induced somatic embryogenesis in vegetative tissues of Arabidopsis thaliana. Plant J 34:107–114. https://doi.org/10.1046/j.1365-313X.2003.01702.x

    Article  PubMed  CAS  Google Scholar 

  33. Karami O, Deljou A, Esna-Ashari M, Ostad-Ahmadi P (2006) Effect of sucrose concentrations on somatic embryogenesis in carnation (Dianthus caryophillus L.). Sci Hort 110:340–344. https://doi.org/10.1016/j.scienta.2006.07.029

    Article  CAS  Google Scholar 

  34. Dudits D., J. Györgyey, L. Bögre y L. Bakó, (1995) Molecular biology of somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants, Kluwer Academic Publishers, Dordrecht, pp 267–308. doi: https://doi.org/10.1007/978-94-011-0485-2_8

    Chapter  Google Scholar 

  35. Werner T, Schmülling T (2009) Cytokinin action in plant development. Curr Opin Plant Biol 12:527–538. https://doi.org/10.1016/j.pbi.2009.07.002

    Article  PubMed  CAS  Google Scholar 

  36. Ha S, Vankova R, Yamaguchi-Shinozaki K et al (2012) Cytokinins: metabolism and function in plant adaptation to environmental stresses. Trends Plant Sci 17:172–179. https://doi.org/10.1016/j.tplants.2011.12.005

    Article  PubMed  CAS  Google Scholar 

  37. Cabrera-Ponce JL, Lopez L, León-Ramírez CG et al (2015) Stress induced acquisition of somatic embryogenesis in common bean Phaseolus vulgaris L. Protoplasma 252:559–570. https://doi.org/10.1007/s00709-014-0702-4

    Article  PubMed  Google Scholar 

  38. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

    Article  CAS  Google Scholar 

  39. Barraza A, Cabrera-Ponce JL, Gamboa-Becerra R et al (2015) The Phaseolus vulgaris PvTRX1h gene regulates plant hormone biosynthesis in embryogenic callus from common bean. Front Plant Sci 6:577. https://doi.org/10.3389/fpls.2015.00577

    Article  PubMed  PubMed Central  Google Scholar 

  40. Kawashima T, Goldberg RB (2009) The suspensor: not just pending the embryo. Trends Plant Sci 15:23–30. https://doi.org/10.1016/j.tplants.2009.11.002

    Article  PubMed  CAS  Google Scholar 

  41. Quiroz-Figueroa F, Rojas-Herrera R, Galaz-Avalos R, Loyola-Vargas V (2006) Embryo production through somatic embryogenesis can be used to study cell differentiation in plants. Plant Cell Tiss Org 86:285–301. https://doi.org/10.1007/s11240-006-9139-6

    Article  Google Scholar 

  42. Urao T, Yakubov B, Satoh R et al (1999) A transmembrane hybrid-type histidine kinase in Arabidopsis functions as an osmosensor. Plant Cell 11:1743–1754. https://doi.org/10.1105/tpc.11.9.1743

    Article  CAS  Google Scholar 

  43. Tran LSP, Urao T, Qin F 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. https://doi.org/10.1073/pnas.0706547105

    Article  PubMed  PubMed Central  Google Scholar 

  44. Jeon J, Kim YN, Kim S et al (2010) A subset of cytokinins two-component signaling system plays a role in cold temperature stress response in Arabidopsis. J Biol Chem 285:23371–23386

    Article  CAS  Google Scholar 

  45. Müller B, Sheen J (2008) Cytokinin and auxin interaction in root stem-cell specification during early embryogenesis. Nature 453:1094–1097. https://doi.org/10.1038/nature06943

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  46. Hwang I, Sheen J (2001) Two-component circuitry in Arabidopsis cytokinin signal transduction. Nature 413:383–389. https://doi.org/10.1038/35096500

    Article  PubMed  CAS  Google Scholar 

  47. Buechel S, Eibfried A, To JP et al (2010) Role of A-type ARABIDOPSIS RESPONSE REGULATORS in meristem maintenance and regeneration. Eur J Cell Biol 89:279–284. https://doi.org/10.1016/j.ejcb.2009.11.016

    Article  PubMed  CAS  Google Scholar 

  48. Su YH, Li YB, Bai B, Zhang XS (2015) Establishment of embryonic shoot-root axis in auxin and cytokinin response during Arabidopsis somatic embryogenesis. Front Plant Sci 5:792. https://doi.org/10.3389/fpls.2014.00792

    Article  PubMed  PubMed Central  Google Scholar 

  49. Murch SJ, KrishnaRaj S, Saxena PK (1997) Thidiazuron-induced morphogenesis of regal geranium (Pelargonium domesticum): a potential stress response. Physiol Plant 101:183–191. https://doi.org/10.1111/j.1399-3054.1997.tb01835.x

    Article  CAS  Google Scholar 

  50. Victor JMR, Murthy BNS, Murch SJ et al (1999) Role of endogenous purine metabolism in thidiazuron induced somatic embryogenesis of peanut (Arachis hypogea L.). Plant Growth Regul 28:41–47. https://doi.org/10.1023/A:1006251531319

    Article  CAS  Google Scholar 

  51. Ashihara H, Stasolla C, Loukanina N, Thorpe T (2001) Purine metabolism during white spruce somatic embryo development: salvage of adenine, adenosine and inosine. Plant Sci 160:647–657. https://doi.org/10.1016/S0168-9452(00)00441-6

    Article  PubMed  CAS  Google Scholar 

  52. Genga A, Allavena A (1991) Factors affecting morphogenesis from immature cotyledon of Phaseolus coccineus L. Plant Cell Tiss Org 27:189–196. https://doi.org/10.1007/BF00041289

    Article  CAS  Google Scholar 

  53. El Dawayati MM, Abd El Bar OH, Zaid ZE, Zein El Din AFM (2012) In vitro morpho-histological studies of newly developed embryos from abnormal malformed embryos of date palms cv. Gundila under dessication effect of polyethelyne glycol treatments. Ann Agric Sci 57:117–128. https://doi.org/10.1016/j.aoas.2012.08.005

    Article  Google Scholar 

  54. Yeung EC, Brown DCW (1982) The osmotic environment of developing embryos of Phaseolus vulgaris. Z Pflanzenphysiol 106:149–156. https://doi.org/10.1016/S0044-328X(82)80077-9

    Article  Google Scholar 

  55. Geerts P, Mergeai G, Baudoin JP (1999) Rescue of early-shaped embryos and plant regeneration of Phaseolus polyanthus Greenm. And Phaseolus vulgaris L. Biotechnol Agron Soc Environ 3:141–148

    Google Scholar 

  56. Roberts DR, Sutton BCS, Flinn BS (1990) Synchronous and high frequency germination of interior spruce somatic embryos following partial drying at high relative humidity. Can J Bot 68:1086–1090. https://doi.org/10.1139/b90-136

    Article  Google Scholar 

  57. Attree SM, Moore D, Sawhney VK, Fowke LC (1991) Enhanced maturation and desiccation tolerance of white spruce (Picea glauca (Moench) Voss) somatic embryos: effects of a non-plasmolysing water stress and abscisic acid. Ann Bot 68:519–525. https://doi.org/10.1093/oxfordjournals.aob.a088291

    Article  Google Scholar 

  58. Flinn BR, Roberts DR, Taylor IEP (1991) Evaluation of somatic embryos on interior spruce. Characterization and developmental regulation of storage proteins. Physiol Plant 82:624–632. https://doi.org/10.1111/j.1399-3054.1991.tb02956.x

    Article  CAS  Google Scholar 

Download references

Acknowledgment

This work was supported in part by SAGARPA (Grant 2003-199). We thank Miss Sandra Chavez-León for her excellent technical assistance and Dra. Rosa Maria Rangel-Cano for the critical review of the manuscript.

Author information

Authors and Affiliations

  1. Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del IPN, CP, Guanajuato, Mexico

    José Luis Cabrera-Ponce, Itzel Anayetzi González-Gómez, Claudia G. León-Ramírez, José A. Sánchez-Arreguín & Alba E. Jofre y Garfias

Authors
  1. José Luis Cabrera-Ponce
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Itzel Anayetzi González-Gómez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Claudia G. León-Ramírez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. José A. Sánchez-Arreguín
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alba E. Jofre y Garfias
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Centro de Investigación Científica de Yucatán, Unidad de Bioquímica y Biología, Molecular de Plantas, Mérida, Yucatán, Mexico

    Víctor M. Loyola-Vargas

  2. Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Irapuato, Guanajuato, Mexico

    Neftalí Ochoa-Alejo

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Cabrera-Ponce, J.L., González-Gómez, I.A., León-Ramírez, C.G., Sánchez-Arreguín, J.A., Jofre y Garfias, A.E. (2018). Somatic Embryogenesis in Common BeanPhaseolus vulgaris L.. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 1815. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8594-4_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8594-4_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8593-7

  • Online ISBN: 978-1-4939-8594-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation