Plant Cell, Tissue and Organ Culture

, Volume 84, Issue 1, pp 80–89 | Cite as

Interspecific hybrids between Medicago sativa L. and annual Medicago containing Alfafa weevil resistance

  • Yuko Mizukami
  • Mitsuru kato
  • Tadashi Takamizo
  • Michio Kanbe
  • Susumu Inami
  • Kazumi Hattori
Article
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Abstract

Non-embryogenic protoplasts of Medicago rugosa and M. scutellata were electro-fused with iodoacetic acid-treated protoplasts of M. sativa (alfalfa). Putative somatic hybrid callus were obtained and some plants regenerated from both combinations. Hybridity of regenerants was confirmed by morphology, molecular means and cytological observations. Parental specific bands were recognized in somatic hybrids by Southern analysis. The somatic hybrids were perennial and their morphology was similar to M. sativa. Cytological observations were carried out on the somatic hybrids, their vegetative clones and self-pollinated offspring. Original somatic hybrids were aneuploids (2n=31–59), but during vegetative proliferation, their chromosome numbers reduced to 32. Those clones of hybrids formed seeds from M. sativa (+) M. rugosa by self-crossing. Chromosomal rearrangements within the parental genomes were observed in vegetative clones of hybrids and their S1 offspring by Genomic in situ Hybridization (GISH). Some of S1 offspring from M. sativa (+) M. rugosa showed better spring growth than parental M. sativa and tend to be tolerant to Alfalfa weevil. It was considered that these traits were introduced from the genome transferring M.␣rugosa chromosome to M. sativa. The cell fusion may still have a potential in transferring alien chromosomes in order to increase the genetic variation for crop breeding.

Keywords

alfalfa weevil resistance cell fusion chromosome rearrangement genomic in situ hybridization Medicago somatic hybrid 

Abbreviations

CTAB

cetyl trimethyl ammonium bromide

DAPI

4,6-diamino-2-phenylindole dihydro- chloride

2,4-D

2,4-dichlorophenoxi-acetic acid

GISH

Genomic in situ Hybridization

IOA

iodoac- etamide

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Notes

Acknowledgements

Authors thanked Mr. K. Fukaya for his help to this work. This research was supported in part by Ministry of Agriculture, Forestry and Fisheries, Japan.

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Copyright information

© Springer 2005

Authors and Affiliations

  • Yuko Mizukami
    • 1
    • 3
  • Mitsuru kato
    • 1
  • Tadashi Takamizo
    • 2
  • Michio Kanbe
    • 1
  • Susumu Inami
    • 1
  • Kazumi Hattori
    • 3
  1. 1.Crop InstituteAichi Agricultural research CenterNagakute AichiJapan
  2. 2.Department of Forage Crop BreedingNational Institute of Livestock and Grassland ScienceNishinasunoJapan
  3. 3.Department of Biological Mechanisms and Function, Graduate School of Bioagricultural SciencesNagoya UniversityNagoyaJapan

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