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Immature Embryo Rescue and Culture

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Plant Embryo Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 710))

Abstract

Embryo culture techniques have many significant applications in plant breeding, as well as basic studies in physiology and biochemistry. Immature embryo rescue and culture is a particularly attractive technique for recovering plants from sexual crosses where the majority of embryos cannot survive in vivo or become dormant for long periods of time. Overcoming embryo inviability is the most common reason for the application of embryo rescue techniques. Recently, fruit breeding programs have greatly increased the interest in exploiting interploid hybridization to combine desirable genetic traits of complementary parents at the triploid level for the purpose of developing improved seedless fruits. However, the success of this approach has only been reported in limited number of species due to various crossing barriers and embryo abortion at very early stages. Thus, immature embryo rescue provides an alternative means to recover triploid hybrids, which usually fail to completely develop in vivo. This chapter will provide a brief discussion of the utilization of interploid crosses between a monoembryonic diploid female with an allotetraploid male in a citrus cultivar improvement program, featuring a clear and comprehensive illustration of successful protocols for immature embryo rescue and culture. The protocols will cover the complete process from embryo excision to recovered plant in the greenhouse and can easily be adapted to other plant commodities. Factors affecting the success and failure of immature embryo rescue to recover triploid progeny from interploid crosses will be discussed.

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References

  1. Bauer MJ (2006) The interploidy hybridization barrier in Zea mays L. University of Missouri-Columbia, Ph.D. dissertation

    Google Scholar 

  2. Van Huylenbroeck J, Leus L, Van Bockstaele E (2005) Interploidy crosses in Roses: use of triploids. Acta Hort 690:109–112

    Google Scholar 

  3. Jansky S (2006) Overcoming hybridization barriers in potato. Plant Breed 125:1–12

    Article  Google Scholar 

  4. Yan G, Ferguson AR, McNeilage MA, Murray BG (1997) Numerically unreduced (2n) gametes and sexual polyploidization in Actinidia. Euphytica 96:267–272

    Article  Google Scholar 

  5. Viloria Z, Grosser JW, Bracho B (2005) Immature embryo rescue, culture and seedling development of acid citrus fruit derived from interploid hybridization. Plant Cell Tiss Org Cult 82:159–167

    Article  CAS  Google Scholar 

  6. Beuselinck PR, Steiner JJ, Rim YW (2003) Morphological comparison of progeny derived from 4x-2x and 4x-4x hybridization of Lotus glaber Mill. and L. corniculatus L. Crop Sci 43:1741–1746

    Article  Google Scholar 

  7. Jaskani MJ, Khan IA, Khan MM (2005) Fruit set, seed development and embryo germination in interploid crosses of citrus. Sci Hort 107:51–57

    Article  Google Scholar 

  8. Fatta Del Bosco S, Siragusa M, Abbate L, Lucretti S, Tusa N (2007) Production and characterization of new triploid seedless progenies for mandarin improvement. Sci Hort 114:258–262

    Article  Google Scholar 

  9. Usman M, Ramzan M, Fatima B, Jaskani MJ, Khan MM (2002) Citrus germplasm enhancement by interploid hybridization 1. Reciprocal crosses of Kinnow and Succari. Int J Agri Bio 4:208–210

    Google Scholar 

  10. Raza H, Khan MM, Khan AA (2003) Seedlessness in citrus. Int J Agri Biol 5:388–391

    Google Scholar 

  11. Jaskani MJ, Khan IA, Khan MM, Abbas H (2007) Frequency of triploids in different interploidal crosses of citrus. Pak J Bot 39:1517–1522

    Google Scholar 

  12. Fatima B, Usman M, Ramzan M, Khan MM, Khan IA (2002) Interploid hybridization of Kinnow and Sweet Lime. Pak J Agri Sci 39:132–134

    Google Scholar 

  13. Reforgiato Recupero G, Russo G, Recupero S (2005) New promising citrus triploid hybrids selected from crosses between monoembryonic diploid female and tetraploid male parents. HortSci 40:516–520

    Google Scholar 

  14. Park SM, Wakana A, Hiramatsu M, Uresino K (2002) A tetraploid hybrid plant from 4x x 2x crosses in Vitis and its origin. Euphytica 126:345–353

    Article  CAS  Google Scholar 

  15. Wakana A, Sarikhani H, Hanada N, Fukudome I, Kajiwara K, Yasukochi K, Hiramatsu M, Sakai K (2007) Characteristics of seedless berries of triploid hybrid grapes (Vitis vinifera Complex) derived from eighteen crosses. J Fac Agr Kyushu Univ 52:337–344

    CAS  Google Scholar 

  16. Wakana A, Hiramatsu M, Park SM, Hanada N, Fufudome I, Xuan B (2002) Degree of abortion and germination rates in triploid seeds from crosses between diploid and tetraploid grapes (Vitis vinifera L. and V. complex). J Fac Agri Kyushu Univ 46:281–294

    Google Scholar 

  17. Wakana A, Fukudome I, Hanada N, Hiramatsu M, Sakai K, Kajiwara K (2008) ‘Bea-kei’, a new triploid seedless grape cultivar derived from a ‘Muscat Bailey A’ x ‘Kyoho’ cross. J Fac Agri Kyushu Univ 53:423–427

    Google Scholar 

  18. Wakana A, Park SM, Hiramatsu M, Hanada N, Fukudome I, Yasukochi K (2005) Characteristics of seedless berries of triploid hybrid grapes (Vitis complex) from reciprocal crosses between diploid ‘Muscat Bailey A’ and tetraploid ‘Red Pearl’. J Fac Agri Kyushu Univ 50:49–59

    Google Scholar 

  19. Yang D, Li W, Li S, Yang X, Wu J, Cao Z (2007) In vitro embryo rescue culture of F1 progenies from crosses between diploid and tetraploid grape varieties. Plant Growth Regul 51:63–71

    Article  CAS  Google Scholar 

  20. Heo JY, Park KS, Yun HK, Park SM (2007) Degree of abortion and germination percentage in seeds derived from interploid crosses between diploid and tetraploid grape cultivars. Hort Env Biotech 48:115–121

    Google Scholar 

  21. Pillay M, Ogundiwin E, Tenkouano A, Dolezel J (2006) Ploidy and genome composition of Musa germplasm at the International Institute of Tropical Agriculture (IITA). Afri J Biotech 5:1224–1232

    CAS  Google Scholar 

  22. Vuylsteke D (2000) Breeding bananas and plantains: from intractability to feasibility. Acta Hort 540:149–156

    Google Scholar 

  23. Tomekpe K, Jenny C, Escalant JV (2004) A review of conventional improvement strategies for Musa. InfoMusa 13:2–6

    Google Scholar 

  24. Jenny C, Tomekpe K, Bakry F, Escalant JV (2003) Conventional breeding of bananas. In: Jacome L, Lepoivre P, Martin D, Ortiz R, Romero R, Escalant JV (eds) Mycosphaerella leaf spot diseases of bananas: present status and outlook. The International Network for the Improvement of Banana and Plantain, Montpellier, France, pp 199–208

    Google Scholar 

  25. Olsen RT, Ranney TG (2006) Fertility and inheritance of variegated and purple foliage across a polyploidy series in Hypericum androsaemum L. J Am Soc Hort Sci 131:725–730

    Google Scholar 

  26. Olsen R (2006) Polyploid breeding in Hypericum Androsaemum L. HortSci 41:952

    Google Scholar 

  27. Jacob Y, Mastrantuono S, Ferrero F (2007) Interploid crosses in Anemone coronaria. Acta Hort 743:55–57

    Google Scholar 

  28. Kaori S, Yukio O, Michikazu H (2006) Intrasubgeneric and interploid cross compatibility in evergreen and deciduous azaleas. J Fac Agri Kyushu Univ 51:73–81

    Google Scholar 

  29. Sakai K, Ozaki Y, Ureshino K, Miyajima I, Wakana A, Okubo H (2008) Interploid crossing overcomes plastome-nuclear genome incompatibility in intersubgeneric hybridization between evergreen and deciduous azaleas. Sci Hort 115:268–274

    Article  CAS  Google Scholar 

  30. Hirsch AM, Testolin R, Brown S, Chat J, Fortune D, Bureau JM, De Nay D (2001) Embryo rescue from interspecific crosses in the genus Actinidia (Kiwifruit). Plant Cell Rep 20:508–516

    Article  CAS  Google Scholar 

  31. Van Tuyl JM (1997) Interspecific hybridization of flower bulbs: a review. Acta Hort 430:465–476

    Google Scholar 

  32. Tel-Zur N, Abbo S, Mizrahi Y (2005) Cytogenetics of semi-fertile triploid and aneuploid intergeneric vine cacti hybrid. J Hered 96:124–131

    Article  PubMed  CAS  Google Scholar 

  33. Datson PM, Murray BG, Hammett KRW (2006) Pollination systems, hybridization barriers and meiotic chromosome behavior in Nemesia hybrids. Euphytica 151:173–185

    Article  Google Scholar 

  34. Williams EG, Maheswaran G, Hutchinson JF (1987) Embryo and ovule culture in crop improvement. Plant Breed Rev 5:181–236

    Google Scholar 

  35. Sokolov VA (2006) Imprinting in plants. Russ J Genet 42:1043–1052

    Article  CAS  Google Scholar 

  36. Johnston SA, den Nijs TPM, Peloquin SJ, Hanneman RE (1980) The significance of genic balance to endosperm development in interspecific crosses. Theor Appl Genet 57:5–9

    Google Scholar 

  37. Johnston SA, Hanneman RE (1995) The genetics of triploid formation and its relationship to endosperm balance number in potato. Genome 38:60–67

    Article  PubMed  CAS  Google Scholar 

  38. Burton TL, Husband BC (2000) Fitness differences among diploids, tetraploids, and their triploid progeny in Chamerion angustifolium: mechanism of inviability and implications for polyploidy evolution. Evolution 54:1182–1191

    PubMed  CAS  Google Scholar 

  39. Navarro L, Olivares-Fuster O, Juarez J, Aleza P, Pina JA, Ballester-Olmos JF (2004) Application of biotechnology to citrus improvement in Spain. Acta Hort 632:221–234

    CAS  Google Scholar 

  40. Bridgen MP (1994) A review of plant embryo culture. HortSci 29:1243–1246

    Google Scholar 

  41. Sharma DR, Kaur R, Kumar K (1996) Embryo rescue in plants – a review. Euphytica 89:325–337

    Google Scholar 

  42. Monnier M (1990) Culture of zygotic embryos of higher plants. In: Pollard JW, Walker JM (eds) Methods in molecular biology, vol 6, Plant cell and tissue culture. Humana Press, Totowa, pp 129–139

    Google Scholar 

  43. Pierik RLM (1987) In vitro culture of higher plants. Kluwer Academic, Dordrecht, pp 139–148

    Book  Google Scholar 

  44. Sharma HC (1999) Embryo rescue following wide crosses. In: Hall RD (ed) Methods in molecular biology, vol 111, Plant cell culture protocols. Humana Press, Totowa, pp 293–307

    Google Scholar 

  45. Yao JL, Cohen D (1996) Production of triploid Zantedeschia hybrids using embryo rescue. New Zealand J Crop Hort Sci 24:297–301

    Article  Google Scholar 

  46. Ramming DW (1990) The use of embryo culture in fruit breeding. HortSci 25:393–398

    Google Scholar 

  47. Olsen RT, Ranney TG, Viloria Z (2006) Reproductive behavior of induced allotetraploid x Chitalpa and in vitro embryo culture of polyploidy progeny. J Am Soc Hort Sci 131:716–724

    Google Scholar 

  48. Chi HS (2002) The efficiencies of various embryo rescue methods in interspecific crosses of Lilium. Bot Bull Acad Sin 43:139–150

    Google Scholar 

  49. Raghavan V (2003) One hundred years of zygotic embryo culture investigations. In Vitro Cell Dev Biol Plant 39:437–442

    Article  Google Scholar 

  50. Collins GB, Grosser JW (1984) Culture of embryos. In: Vasil IK (ed) Cell Culture and Somatic Cell Genetics of Plants. Academic press, Orlando, Florida, pp 241–257

    Google Scholar 

  51. Grosser JW, Gmitter FG Jr, Fleming GH, Chandler JL (2000) Application of biotechnology to citrus cultivar improvement at the Citrus Research and Education Center. Acta Hort 535:213–220

    Google Scholar 

  52. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–495

    Article  CAS  Google Scholar 

  53. Murashige T, Tucker DPH (1969) Growth factors requirements of citrus tissue culture. Proc First Int Citrus Symp 3:1155–1161

    CAS  Google Scholar 

  54. Niedz RP, Hyndman SE, Wynn ET, Bausher MG (2002) Normalizing sweet orange [C. sinensis (L.) Osbeck) somatic embryogenesis with semi-permeable membranes. In Vitro Cell Dev Biol Plant 38:552–557

    Article  CAS  Google Scholar 

  55. Grosser JW, Gmitter FG Jr (1990) Protoplast fusion and citrus improvement. Plant Breed Rev 8:339–374

    Google Scholar 

  56. Deng XX, Grosser JW, Gmitter FG Jr (1992) Intergeneric somatic hybrid plants from protoplast fusion of Fortunella crassifolia cultivar Meiwa with Citrus sinensis cultivar Valencia. Sci Hort 49:55–62

    Article  Google Scholar 

  57. Kane ME (2000) Culture indexing for bacterial and fungal contaminants. In: Trigiano RN, Gray DJ (eds) Plant tissue culture concepts and laboratory exercise. CRC Press, Boca Raton, pp 427–431

    Google Scholar 

  58. Kunisaki JT (1974) Tissue culture of tropical ornamental plants. HortSci 12:141–142

    Google Scholar 

  59. Hartmann HT, Kester DE, Davies FT (1990) Plant propagation – principles and practices, 5th edn. Prentice Hall, New Jersey, p 199

    Google Scholar 

  60. Muthan SB, Rathore TS, Rai VR (2006) Factors influencing in vivo and in vitro micrografting of sandalwood (Santalum album L.): an endangered tree species. J For Res 11:147–151

    Article  Google Scholar 

  61. Onay A, Pirinc V, Adiyaman F, Isikalan C, Tilkat E, Basaran D (2003) In vivo and in vitro micrografting of pistachio, Pistacia vera L. cv. “Siirt”. Turk J Biol 27:95–100

    Google Scholar 

  62. Raharjo SHT, Litz RE (2005) Micrografting and ex vitro grafting for somatic embryo rescue and plant recovery in avocado (Persea americana). Plant Cell Tiss Org Cult 82:1–9

    Article  Google Scholar 

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Authors and Affiliations

  1. IFAS, Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA

    Xiuli Shen, Fred G. Gmitter Jr. & Jude W. Grosser

Authors
  1. Xiuli Shen
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  2. Fred G. Gmitter Jr.
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  3. Jude W. Grosser
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Corresponding author

Correspondence to Xiuli Shen .

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Editors and Affiliations

  1. Dept. Biological Sciences, University of Calgary, University Dr. NW., 2500, Calgary, T2N 1N4, Alberta, Canada

    Trevor A. Thorpe

  2. , Department of Biological Sciences, University of Calgary, University Dr. NW 2500, Calgary, T2N 1N4, Alberta, Canada

    Edward C. Yeung

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Shen, X., Gmitter, F.G., Grosser, J.W. (2011). Immature Embryo Rescue and Culture. In: Thorpe, T., Yeung, E. (eds) Plant Embryo Culture. Methods in Molecular Biology, vol 710. Humana Press. https://doi.org/10.1007/978-1-61737-988-8_7

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  • DOI: https://doi.org/10.1007/978-1-61737-988-8_7

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61737-987-1

  • Online ISBN: 978-1-61737-988-8

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