Advertisement

In Vitro Pollination and Fertilization

  • Sant Saran Bhojwani
  • Prem Kumar Dantu
Chapter

Abstract

In angiosperms, the female gamete (egg) is formed and remains fixed at the micropylar end of the embryo sac deeply embedded in the sporophytic tissues of the ovule, which is enclosed in the ovary well removed from the stigma. The male gametes (sperms) are enclosed in the pollen grain. To effect fertilization, the pollen germinate on the stigma to form a pollen tube that transports the two non-motile sperms to the embryo sac and delivers them in the vicinity of the egg. Whereas one of the sperms fertilizes the egg (syngamy), the other fertilizes the central cell (triple fusion). The fertilized egg (zygote) develops into an embryo, the progenitor of the next generation, and the fertilized central cell forms the endosperm tissue, the main source of nutrition for the developing and germinating embryo. Thus, in the angiosperms the gametes, the process of double fertilization, zygote, early stages of embryo and endosperm development are not readily accessible to study the cellular and molecular aspects of fertilization and embryogenesis. Therefore, for almost 100 years since the discovery of double fertilization in angiosperms, by Nawaschin (1898), not much progress could be made in this area. Whatever little information is known is based mainly on mutant analysis in Arabidopsis. Sexual incompatibility is a serious handicap in developing desirable hybrids. In this the pollen fails to germinate on the stigma or the pollen tube gets arrested or bursts before reaching the ovary. Two in vitro techniques developed to overcome this problem are in vivo pollination (IVP) and In vitro fertilization (IVF). IVF, developed in nineties involved isolation of male and female gametes and their in vitro fusion (fertilization) culture of the in vitro zygote to regenerate full plants. Complete IVF technique has been developed in only two plants, namely maize and rice. This technique is proving to be an invaluable aid to directly observe and analyse fertilization and post-fertilization process in flowering plants which is not possible under in vivo conditions.

Keywords

Pollen Tube Central Cell Sperm Cell Pollen Tube Growth Female Gamete 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Suggested Further Reading

  1. Hoshino Y, Scholten S, Von Wiegen P, Lörz H, Kranz E (2004) Fertilization-induced changes in the microtubular architecture of the maize egg cell and zygote—an immunocytochemical approach adapted to single cells. Sex Plant Reprod 17:89–95CrossRefGoogle Scholar
  2. Kanta K, Rangaswamy NS, Maheshwari P (1962) Test-tube fertilization in a flowering plant. Nature 194:1214–1217CrossRefGoogle Scholar
  3. Kovács M, Barnabás B, Kranz E (1995) Electrofused isolated wheat (Triticum aestivum L.) gametes develop into multicellular structures. Plant Cell Rep 15:178–180CrossRefGoogle Scholar
  4. Kranz E (1999) In vitro fertilization with isolated single gametes. In: Hall RD (ed) Methods in molecular biology vol. 111. Plant Cell Culture protocols. Humana Press, New YorkGoogle Scholar
  5. Kranz E (2001) In vitro fertilization. In: Bhojwani SS. Soh WY (eds) Current trends in the embryology of angiosperms. Kluwer Academic Publishers, DordrechtGoogle Scholar
  6. Kranz E, Kumlehn J (1999) Angiosperm fertilization, embryo and endosperm development in vitro. Plant Sci 142:183–197CrossRefGoogle Scholar
  7. Kranz E, Scholten S (2008) In vitro fertilization: analysis of early post-fertilization development using cytological and molecular techniques. Sex Plant Reprod 21:67–77CrossRefGoogle Scholar
  8. Kranz E, Hoshino Y, Okamoto T (2008) In vitro fertilization with isolated higher plant gametes. In: Suárez MF, Bozhkov PV (eds) Methods in molecular biology vol. 427. Plant Embryogenesis. Humana Press, New YorkGoogle Scholar
  9. Márton ML, Dresselhaus T (2008) A comparison of early molecular fertilization mechanisms in animals and flowering plants. Sex Plant Reprod 21:37–52CrossRefGoogle Scholar
  10. Okamoto T, Kranz E (2005) In vitro fertilization—a tool to dissect cell specification from a higher plant zygote. Curr Sci 89:1861–1869Google Scholar
  11. Uchiumi T, Komatsu S, Koshiba T, Okamoto T (2006) Isolation of gametes and central cells from Oryza sativa L. Sex Plant Reprod 19:37–45CrossRefGoogle Scholar
  12. Uchiumi T, Uemura I, Okamoto T (2007) Establishment of an in vitro fertilization system in rice (Oryza sativa L.). Planta 226:581–589PubMedCrossRefGoogle Scholar
  13. Wang YY, Kuang A, Russell SD, Tian HQ (2006) In vitro fertilization as a tool for investigating sexual reproduction of angiosperms. Sex Plant Reprod 19:103–115CrossRefGoogle Scholar

Copyright information

© Springer India 2013

Authors and Affiliations

  1. 1.Department of BotanyDayalbagh Educational Institute (Deemed University)AgraIndia

Personalised recommendations