Tissue Culture and Plant Regeneration of Blue Grama Grass, Bouteloua gracilis (H.B.K.) Lag. ex Steud

• Published in: In vitro cellular & developmental biology. Plant 2001-03, Vol.37 (2), p.182-189
• Main Authors: Gerardo Armando Aguado-Santacruz, Cabrera-Ponce, José Luis, Victor Olalde-Portugal, Ma. Rosario Sánchez-González, Márquez-Guzmán, Judith, Herrera-Estrella, Luis
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biotechnology; Callus; Embryos; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Grasses; In vitro propagation: entire plant regeneration from tissues and cell cultures; Methods. Procedures. Technologies; Plant cells and fungal cells; Plantlets; Plants; Protocols and Research Reports; Regeneration; Seedlings; Somatic embryogenesis; Tissue culture techniques
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-001-0032-z

As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H.B.K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid (2,4-D), Picloram (4-amino-3,5,6-trichloropicolinic acid),$N^6-benzyladenine (BA)$or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations containing$1 mg l^{-1} (4.52 \mu M) 2,4-D plus 0.5 mg l^{-1} (2.22 \mu M) BA$, and$2 mg l^{-1} (8.88 \mu M) BA plus 1 mg l^{-1} (4.14 \mu M) Picloram$with or without$40 mg l^{-1} (296.08 \mu M) adenine$. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient treatments for induction of embryogenic callus contained$2 mg l^{-1} (9.05 \mu M) 2,4-D combined$with$0.25 (1.11 \mu M)$or$0.50 mg l^{-1} (2.22 \mu M) BA$, or$1 mg l^{-1} (4.52 \mu M) 2,4-D$with$0.50 mg l^{-1} (2.22 \mu M) BA$. Regeneration was achieved in hormone-free Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus$1 mg l^{-1} (1.44 \mu M) gibberellic acid$. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from$9 for 2 mg l^{-1} (9.05 \mu M) 2,4-D to 62.2$for induction medium containing$2 mg l^{-1} (8.28 \mu M) Picloram$,$1 mg l^{-1} (4.44 \mu M) BA$and$40 mg l^{-1} (296.08 \mu M) adenine$. Regenerated plants grown in soil under greenhouse conditions reached maturity and produced seeds.