Transgenic sorghum plants via microprojectile bombardment

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 23; pp. 11212 - 11216
• Main Authors: Casas, A.M, Kononowicz, A.K, Zehr, U.B, Tomes, D.T, Axtell, J.D, Butler, L.G, Bressan, R.A, Hasegawa, P.M
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.12.1993; National Acad Sciences; National Academy of Sciences
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; Biological and medical sciences; Biotechnology; Callus; Cells, Cultured; Corn; CULTIVO DE TEJIDOS; CULTURE DE TISSU; Deoxyribonucleic acid; DNA; DNA - administration & dosage; Edible Grain - genetics; Embryos; Fundamental and applied biological sciences. Psychology; Gene Transfer Techniques; Genes; Genes, Plant; Genetic engineering; Genetic technics; GENOTIPOS; GENOTYPE; GLICOSIDASAS; GLYCOSIDASE; HERBICIDAS; HERBICIDE; Immatures; In Vitro Techniques; Methods. Procedures. Technologies; Plant cells; Plant reproduction; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; Plants; Plants, Genetically Modified; Plasmids; RESISTANCE AUX PRODUITS CHIMIQUES; RESISTENCIA QUIMICA; Sorghum; SORGHUM BICOLOR; TRANSFERASAS; TRANSFERASE; TRANSFERENCIA DE GENES; TRANSFERT DE GENE; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; Transgenic animals and transgenic plants; Transgenic plants; VARIEDADES; VARIETE; Monocotyledones; Sorghum bicolor; Microprojectile bombardment; Gramineae; Selection; Angiospermae; Spermatophyta; Transgenic plant; Method; Optimization; Genetic transfer
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.23.11212

Transgenic sorghum plants have been obtained after microprojectile bombardment of immature zygotic embryos of a drought-resistant sorghum cultivar, P898012. DNA delivery parameters were optimized based on transient expression of R and C1 maize anthocyanin regulatory elements in scutellar cells. The protocol for obtaining transgenic plants consists of the delivery of the bar gene to immature zygotic embryos and the imposition of bialaphos selection pressure at various stages during culture, from induction of somatic embryogenesis to rooting of regenerated plantlets. One in about every 350 embryos produced embryogenic tissues that survived bialaphos treatment; six transformed callus lines were obtained from three of the eight sorghum cultivars used in this research. Transgenic (T0) plants were obtained from cultivar P898012 (two independent transformation events). The presence of the bar and uidA genes in the T0 plants was confirmed by Southern blot analysis of genomic DNA. Phosphinothricin acetyltransferase activity was detected in extracts of the T0 plants. These plants were resistant to local application of the herbicide Ignite/Basta, and the resistance was inherited in T1 plants as a single dominant locus