Molecular and General Genetics MGG

, Volume 215, Issue 3, pp 501–506 | Cite as

Construction of rice cybrid plants

  • Hiromori Akagi
  • Masahiro Sakamoto
  • Tomoko Negishi
  • Tatsuhito Fujimura


The mitochondrial genomes of rice cells were transferred to a fertile rice variety (N8) from a cytoplasmic male sterile variety (CMS) by asymmetric protoplast fusion based on metabolic complementation. Protoplasts derived from CMS were X-irradiated (125 krad) and electrofused with protoplasts which had been treated with iodoacetamide. Metabolic complementation, presumably between nuclear and cytoplasmic compartments, enabled fused protoplasts to form colonies at high efficiency. Restriction digest analysis of mitochondrial DNA (mtDNA) indicated that hybrid cells carried mtDNA derived from both parents. Of the plants regenerated from hybrid calli, 68% carried a diploid chromosome set (2n=24) and the rest of them carried 48 chromosomes. All of them expressed the aryl acylamidase I deficient phenotype encoded by the recessive allele of the fertile N8 parent. These results indicate that the novel somatic hybrid plants regenerated were cybrids, deriving their nucleus from the iodoacetamide treated parent and their mitochondria from both parents.

Key words

Oryza Protoplast fusion Cybrids Mitochondrial DNA Cytoplasmic male sterile 


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  1. Barsby TL, Chuong PV, Yarrow SA, Wu S-C, Coumans M, Kemble RJ, Powell AD, Beversdorf WD, Pauls KP (1987) The combination of Polima cms and cytoplasmic triazine resistance in Brassica napus. Theor Appl Genet 73:809–814Google Scholar
  2. Clark E, Schnabelrauch L, Hanson MR, Sink KC (1986) Differential fate of plastid and mitochondrial genomes in Petunia somatic hybrids. Theor Appl Genet 72:748–755Google Scholar
  3. Dudits D, Fejer O, Hadlaczly G, Koncz C, Lazar GB, Horvath G (1980) Intergeneric gene transfer mediated by plant protoplast fusion. Mol Gen Genet 179:283–288Google Scholar
  4. Fujimura T, Sakurai M, Akagi H, Negishi T, Hirose A (1985) Regeneration of rice plants from protoplasts. Plant Tissue Culture Lett 2:74–75Google Scholar
  5. Hanson MR, Rothenberg M, Boshore ML, Nivison HY (1985) Organelle segregation and recombination following protoplasts fusion: Analysis of sterile cytoplasms. In: Zaithlin M, Day P, Hollaender A (eds) Biotechnology in Plant Science Relevance to Agriculture in the Eighties. Academic Press, New York, pp 129–144Google Scholar
  6. Hoagland RE, Graf G (1972) Enzymatic hydrolysis of herbicides in plants. Weed Sci 20:303–305Google Scholar
  7. Ichikawa H, Tanno-Suenaga L, Imamura J (1987) Selection of Daucus cybrids based on metabolic complementation between X-irradiated D. capillifolius and iodoacetamide-treated D. carota by somatic cell fusion. Theor Appl Genet 74:746–752Google Scholar
  8. Kadowaki K, Ishige T, Suzuki S, Harada K, Shynjyou C (1986) Differences in characteristics of mitochondrial DNA between normal and male sterile cytoplasms of japonica rice. Jap J Breed 36:333–339Google Scholar
  9. Kemble RJ, Barsby TL, Wong RSL, Shepard JF (1986) Mitochondrial DNA rearrangements in somatic hybrids of Solanum tuberosum and Solanum brevidens. Theor Appl Genet 72:787–793Google Scholar
  10. Leaver CJ, Gray MW (1982) Mitochondrial genome organization and expression in higher plants. Annu Rev Plant Physiol 33:373–402Google Scholar
  11. Matsunaka S (1974) Genetic background of specific herbicide tolerance in higher plants. In: Plant Growth Substances 1973, Hirokawa Publishing Co, Tokyo, pp 1182–1186Google Scholar
  12. Matthews BF, Widholm JM (1985) Organellar DNA compositions and isozyme expression in an interspecific somatic hybrid of Daucus. Mol Gen Genet 198:371–376Google Scholar
  13. Miganouna H, Virmani SS, Briquet M (1987) Mitochondrial DNA modifications associated with cytoplasmic male sterility in rice. Theor Appl Genet 74:666–669Google Scholar
  14. Miyakawa I, Aoi H, Sando N, Kuroiwa T (1984) Fluorescence microscopic studies of mitochondrial nucleoids during meiosis and sporulation in the yeast Saccharomyces cerevisiae. J Cell Sci 66:21–38Google Scholar
  15. Nchls R (1978) The use of metabolic inhibitors for the selection of fusion products in higher plant protoplasts. Mol Gen Genet 166:117–118Google Scholar
  16. Nishibayashi S, Kaeriyama J (1986) Structural stability of chromosomes in rice (Oryza sativa L.) plants regenerated from somatic tissue culture. Plant Tissue Culture Lett 3:31–34Google Scholar
  17. Ozias-Akins P, Pring DR, Vasil IK (1987) Rearrangements in the mitochondrial genome of somatic hybrid cells lines of Pennisetum americanum (L.) K. Schum. + Panicum maximum Jacq. Theor Appl Genet 74:15–20Google Scholar
  18. Pelletier G, Primard C, Uedel F, Chetrit P, Remy R, Rouselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Gen Genet 191:244–250Google Scholar
  19. Sidorov VA, Menczel L, Nagy F, Maliga P (1981) Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts. Planta 152:341–345Google Scholar
  20. Sidorov VA, Zubko MK, Kuchko AA, Komarnitsky IK, Gleba YY (1987) Somatic hybridization in potato: Use of γ-irradiated protoplasts of Solanum pinnatisectum in genetic reconstruction. Theor Appl Genet 74:364–368Google Scholar
  21. Shikanai T, Yang Z-Q, Yamada Y (1987) Properties of circular plasmid-like DNA B1 from mitochondria of cytoplasmic malesterile rice. Plant Cell Physiol 28:1243–1251Google Scholar
  22. Tabaeizadeh Z, Pring DR, Vasil IK (1987) Analysis of mitochondrial DNA from somatic hybrid cell lines of Sacchrum officinarum (sugar cane) and Pennisetum americanum (pearl millet). Plant Mol Biol 8:509–513Google Scholar
  23. Terada R, Kyouzuka J, Nishyibayashi S, Shimamoto K (1987) Plantlet regeneration from somatic hybrids of rice (Oryza sativa L.) and barnyard grass (Echinochloa oryzicola Vasing). Mol Gen Genet 210:39–43Google Scholar
  24. Umbeck PF, Gengenback BG (1983) Reservation of male-sterile T-cytoplasm maize to male fertility in tissue culture. Crop Sci 23:584–588Google Scholar
  25. Yamaguchi H, Kakiuchi H (1983) Electrophoretic analysis of mitochondrial DNA from normal and male sterile cytoplasms in rice. Jap J Genet 58:607–611Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Hiromori Akagi
    • 1
  • Masahiro Sakamoto
    • 1
  • Tomoko Negishi
    • 1
  • Tatsuhito Fujimura
    • 1
  1. 1.Plant Biotechnology DepartmentMitsui Toatsu ChemicalsChigasaki City, Kanagawa Pref.Japan

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