Intergeneric somatic hybrids of rice [Oryza sativa L. (+) Porteresia coarctata (Roxb.) Tateoka]

• Published in: Theoretical and applied genetics Vol. 99; no. 3/4; pp. 570 - 577
• Main Authors: Jelodar, N.B, Balckhall, N.W, Hartman, T.P.V, Brar, D.S, Khush, G, Davey, M.R, Cocking, E.C, Power, J.B
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.08.1999; Berlin Springer Nature B.V
• Subjects: allohexaploidy; Biological and medical sciences; Classical genetics, quantitative genetics, hybrids; culture media; cytogenetic analysis; electrofusion; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; genome in situ hybridization; genomics; hexaploidy; hybrids; intergeneric hybridization; kinetin; Lolium multiflorum; naphthaleneacetic acid; nucleic acid hybridization; Oryza sativa; plant morphology; Poaceae; Porteresia coarctata; protoplast culture; protoplast fusion; protoplasts; Pteridophyta, spermatophyta; random amplified polymorphic DNA technique; regenerative ability; Rice; Seeds; shoots; somatic hybridization; sucrose; Vegetals; United Kingdom > UK; Philippines; Monocotyledones; Salt resistance; Genetic marker; Intergeneric hybridization; Random amplified polymorphic DNA marker; Cereal crop; Oryza sativa; Gramineae; Morphology; Angiospermae; Cytogenetics; Somatic hybrid; Spermatophyta; Karyotype
• ISSN: 0040-5752; 1432-2242
• DOI: 10.1007/s001220051270

Somatic hybrid plants were obtained following the electrofusion of rice (Oryza sativa L. cv 'Taipei 309', 2n = 2x = 24) cell suspension-derived protoplasts with non-dividing leaf protoplasts of Porteresia coarctata (2n = 4x = 48), a saline-tolerant wild species. Fusion-treated protoplasts were plated on the surface of cellulose nitrate filter membranes, overlaying Lolium multiflorum nurse cells. The nurse cells were embedded in KPR medium containing 0.5 mg l(-1) 2,4-dichlorophenoxyacetic acid and semi-solidified with SeaPlaque agarose. Putative somatic hybrid cell colonies were selected on the basis of their growth, whereby faster growing colonies were transferred preferentially to MS-based medium with 2.0 mg l(-1) kinetin, 0.5 mg l(-1) alpha-naphthaleneacetic acid, 30 g l(-1) sucrose and 4.0 g l(-1) SeaKem agarose to induce shoot regeneration. One hundred and nineteen regenerated plants were micropropagated clonally on MS-based medium containing 2.0 mg l(-1) 6-benzylaminopurine, 50 g l(-1) sucrose and 4.0 g l(-1) SeaKem agarose, prior to DNA extraction of plant samples. Putative somatic hybrids were initially identified by RAPD analysis, and 8 plant lines were selected for further investigation by flow cytometric ploidy determination and cytology. Plants of one line had an allohexaploid chromosome complement (2n = 6x = 72) and, following examination of its vegetative clones by GISH, were confirmed as somatic hybrids containing full chromosome complements of both O. sativa and P. coarctata.