Transgenic plants of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. ex Steud., from microprojectile bombardment of highly chlorophyllous embryogenic cells

For the first time, the production of transgenic plants of the forage grass blue grama, Bouteloua gracilis [H.B.K.] Lag. ex Steud., is reported. Transgenic plants containing a gus Colon, two colons nptll fusion driven by a double CaMV35S promoter were obtained by microprojectile bombardment of the h...

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Bibliographic Details
Published in:Theoretical and applied genetics 2002-04, Vol.104 (5), p.763-771
Main Authors: Aguado Santacruz, GA, Rascon Cruz, Q, Cabrera Ponce, JL, Martinez Hernandez, A, Olalde Portugal, V, Herrera Estrella, L
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Chlorophyll
Cloning
Fundamental and applied biological sciences. Psychology
Genetic aspects
Genetic engineering
Genetic technics
Grasses
Methods. Procedures. Technologies
Physiological aspects
Plant genetics
Plasmids
Quantitative genetics
Transgenic animals and transgenic plants
Transgenic plants
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Summary:For the first time, the production of transgenic plants of the forage grass blue grama, Bouteloua gracilis [H.B.K.] Lag. ex Steud., is reported. Transgenic plants containing a gus Colon, two colons nptll fusion driven by a double CaMV35S promoter were obtained by microprojectile bombardment of the highly chlorophyllous embryogenic cell line 'TIANSJ98'. Transformed B. gracilis cell lines resisted a lethal concentration of 160 mg/l of kanamycin for at least 8 months. Chlorophyll development and growth rate were used as useful criteria for discriminating transformed from non-transformed clones. Stable integration of the transgene in the blue grama genome was demonstrated by PCR and Southern-hybridization analysis. Expression of the NPTll protein in transgenic plants grown under greenhouse conditions was confirmed indirectly by spraying kanamycin (150-250 mg/l) on plant foliage, and directly by ELISA immunological tests. Control plants sprayed with kanamycin showed foliar necrosis and reduced growth (tillering) compared to plants containing the transgene. NPTll was found in transgenic plants in levels ranging between 12.6 and 29.6 ng/mg FW of cells, as determined by ELISA reactions.
ISSN:0040-5752
1432-2242
DOI:10.1007/s00122-001-0803-2