Hostname: page-component-586b7cd67f-dlnhk Total loading time: 0 Render date: 2024-11-26T07:20:58.131Z Has data issue: false hasContentIssue false
  • English
  • Français

Wheat egg in vitro fusion with wheat and green bristlegrass sperm

Published online by Cambridge University Press:  20 May 2019

Dong Xiao Li ,
Hai Yan Hu ,
Zhen Gang Ru  and
Hui Qiao Tian Open the ORCID record for Hui Qiao Tian [Opens in a new window]
Dong Xiao Li
Affiliation:
Henan Institute of Science and Technology, Xinxiang 453003, China
Hai Yan Hu
Affiliation:
Henan Institute of Science and Technology, Xinxiang 453003, China Collaborative Innovation Center of Modern Biological Breeding, Henan Province
Zhen Gang Ru
Affiliation:
Henan Institute of Science and Technology, Xinxiang 453003, China Collaborative Innovation Center of Modern Biological Breeding, Henan Province
Hui Qiao Tian*
Affiliation:
School of Life Sciences, Xiamen University, Xiamen, 361005, China
*
*Address for correspondence: Hui Qiao Tian. School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China. Tel: +011 86 592 2186486. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

Isolated gametes can be used to investigate fertilization mechanisms, and probe distant hybridization between different species. Pollen grains of wheat and Setaria viridis are tricellular, containing sperm cells at anthesis. Sperm from these plants were isolated by breaking open pollen grains in a osmotic solution. Wheat ovules were digested in an enzyme solution for 20 min, and then transferred to an isolation solution without enzymes to separate egg cells from ovules. The fusion of wheat egg cells with wheat and S. viridis sperm was conducted using an electro-fusion apparatus. Under suitable osmotic pressure (10% mannitol), calcium concentration of 0.001% (CaCl2·2H2O), and a 30–35 V alternating electric field for 15 s, egg cells and sperm adhered to each other and became arranged in a line. Electroporation of the plasma membrane of egg cells and sperm using a 300–500 V direct-current electric field (45 µs amplitude pulse) caused them to fuse.

Keywords

EggGreen bristlegrassIn vitro fusionSpermWheat
Type
Research Article
Information
Zygote , Volume 27 , Issue 3 , June 2019 , pp. 126 - 130
Copyright
© Cambridge University Press 2019 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Clarke, JH, Chevre, AM, Landgren, M Glimelius, K (1999) Characterization of sexual progenies of male-sterile somatic hybrids between Brassica napus and Brassica tournefortii . Theor Appl Genet 99, 605–610.Google Scholar
Dresselhaus, T, Cords, S Lörz, H (1999) A transcript encoding translation initiation factor eIF-5A is stored in unfertilized egg cells of maize. Plant Mol Biol 39, 1063–1071.Google Scholar
Dumas, C Faure, JE (1995) Use of in vitro fertilization and zygote culture in crop improvement. Curr Opin Biotechnol 6, 183–188.Google Scholar
Evans, D, Sharp, W Flick, C (2011) Plant regeneration from cell cultures. Hort Rev 3, 214–314.Google Scholar
Faure, JE, Digonnet, C Dumas, C (1994) An in vitro system for adhesion and fusion of maize gametes. Science 263, 1598–1600.Google Scholar
Khalequzzaman, M Haq, N (2005) Isolation and in vitro fusion of egg and sperm cells in Oryza sativa . Plant Physiol Biochem 43, 69–75.Google Scholar
Kovács, M, Barnabás, B Kranz, E (1995) Electro-fused isolated wheat (Triticum aestivum L.) gametes develop into multicellular structures. Plant Cell Rep 15, 178–180.Google Scholar
Kranz, E Lörz, H (1993) In vitro fertilization with isolated, single gametes results in zygotic embryogenesis and fertile maize plants. Plant Cell 5, 739–746.Google Scholar
Kranz, E Lörz, H (1994) In vitro fertilization of maize by single egg and sperm cell protoplast fusion mediated by high calcium and high pH. Zygote 2, 125–128.Google Scholar
Kranz, E, Bautor, J Lörz, H (1991) In vitro fertilization of single isolated, gametes of maize mediated by electrofusion. Sex Plant Reprod 4, 12–16.Google Scholar
Kranz, E, Wiegen, P Lörz, H (1995) Early cytological events after induction of cell division in egg cells and zygote development following in vitro fertilization with angiosperm gametes. Plant J 8, 9–23.Google Scholar
Liu, Y, Liu, D, Zhou, A He, C (2013) The summary of plant cell fusion and its application. Bot Res 2, 99–105.Google Scholar
Skirvin, RM, McPheeters, KD Norton, D (1989) Source and frequency of somaclonal variation. Hortic Sci 29, 1232–1237.Google Scholar
Uchiumi, K, Uemura, I Okamoto, T (2007 ) Establishment of an in vitro fertilization system in rice (Oryza sativa L.) Planta 226, 581–589.Google Scholar
Vielle-Calzada, JP, Baskar, R Grossniklaus, U (2000) Delayed activation of the paternal genome during seed development. Nature 404, 91–94.Google Scholar
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, 103–115.Google Scholar
Yu, JJ, Chen, C Tian, HQ (2012) The primary attempt of in vitro fertilization of Torenia fournieri . Plant Physiol J 48, 1015–1020 (in Chinese).Google Scholar

Li et al. supplementary material

Li et al. supplementary material 2

Download Li et al. supplementary material(Video)
Video 5.5 MB