Production of Intergeneric Hybrid Calli from C3 and C4 Species of Amaranthaceae through Protoplast Fusion

Japanese Journal of Crop Science

Yoji TAKEOKA¹  Retno MASTUTI¹  Takeshi TANIGUCHI¹  Hiroshi MIYAKE¹ 

关键词: Amaranthus tricolor L.;    アイソザイム;    Celosia cristata L.;    ケイトウ;    C4 species;    雑種カルス;    C3 species;    C3植物;    Electrofusion;    C4植物;    Hybrid calli;    電気融合;    Isozyme;    ハゲイトウ;    Protoplast;    プロトプラスト;   

DOI  :  10.1626/jcs.66.456

学科分类：农业科学（综合）

来源: Nihon Sakumotsu Gakkai / Crop Science Society of Japan

【 摘 要 】

Intergeneric somatic hybrid calli were obtained through electrofusion between protoplasts of Celosia cristata L. (C3 species) cell suspension and Amaranthus tricolor L. (C4 species) cotyledon. The highest binary fusion frequency, 4∼5%, was attained when the protoplasts were suspended in fusion solution containing 0.4 M sorbitol and 2.0 mM CaCl2, aligned by an AC field (200 V/cm, 0.5 MHz) for 30 s and fused by a single DC square pulse at 1.6 kV/cm for 40 μs. Fusion products were identified by characteristics of both parents in the hybrid cells : they possessed green chloroplasts or pink plastids from cotyledonary protoplasts as well as a cytoplasm-rich colorless sector derived from cell suspension protoplasts. The occurrence of two types of calli, compact and friable, obtained after culturing on proliferation medium lead to the isolation of putative hybrid calli without any artificial selection because C. cristata protoplasts regenerated only friable calli and A. tricolor protoplasts did not produce calli. A total of 14 lines of putative hybrid compact calli were obtained by screening on the basis of callus morphology. An isozyme analysis of acid phosphatase confirmed the hybridity of 14 lines of compact calli. Callus morphology was also studied by scanning electron microscopy.

【 授权许可】

Unknown