Journal Article

Rapid and reproducible Agrobacterium-mediated transformation of sorghum

Fromm, M, Howe, A, Sato, S, Dweikat, I, Clemente, T

2006

Overview

A rapid and reproducible Agrobacterium-mediated transformation protocol for sorghum has been developed. The protocol uses the nptII selectable marker gene with either of the aminoglycosides geneticin or paromomycin. A screen of various A. tumefaciens strains revealed that a novel C58 nopaline chromosomal background carrying the chrysanthopine disarmed Ti plasmid pTiKPSF2, designated NTL4/Chry5, was most efficient for gene transfer to sorghum immature embryos. A NTL4/Chry5 transconjugant harboring the pPTN290 binary plasmid, which carries nptII and GUSPlus TM expression cassettes, was used in a series of stable transformation experiments with Tx430 and C2-97 sorghum genotypes and approximately 80% of these transformation experiments resulted in the recovery of at least one transgenic event. The transformation frequencies among the successful experiments ranged from 0.3 to 4.5%, with the average transformation frequency being approximately 1% for both genotypes. Over 97% of the transgenic events were successfully established in the greenhouse and were fully fertile. Co-expression of GUSPlus occurred in 89% of the transgenic T0 events. Seed set for the primary transgenic plants ranged from 145 to 1400 seed/plant. Analysis of T1 progeny demonstrated transmission of the transgenes in a simple Mendelian fashion in the majority of events.

• Publisher: Springer,Springer Nature B.V
• Subject: Agrobacterium radiobacter / Agrobacterium tumefaciens / Agrobacterium tumefaciens - genetics / beta-glucuronidase / Biological and medical sciences / Biotechnology / Blotting, Southern / Chromosome Segregation / Chromosome Segregation - genetics / Chromosomes, Plant / Chromosomes, Plant - genetics / embryo (plant) / embryology / Embryonic Development / Experiments / forage crops / Fundamental and applied biological sciences. Psychology / gene expression regulation / Genetic engineering / Genetic technics / genetic transformation / genetics / Genotype / Genotypes / grain sorghum / Methods. Procedures. Technologies / microbiology / micropropagation / neomycin phosphotransferase II / plant regeneration / Plant Tumor-Inducing Plasmids / Plant Tumor-Inducing Plasmids - genetics / Plants, Genetically Modified / reporter genes / Reproducibility of Results / Sorghum / Sorghum - embryology / Sorghum - genetics / Sorghum - microbiology / Sorghum bicolor / Time Factors / Transformation, Genetic / Transgenic animals and transgenic plants / Transgenic plants