Application of biotechnological tools to model plant: brachypodium distachyon
Soğutmaz Özdemir, Bahar (2009) Application of biotechnological tools to model plant: brachypodium distachyon. [Thesis]
Official URL: http://192.168.1.20/record=b1301613 (Table of Contents)
Brachypodium distachyon has recently emerged as a model plant species for the grass family (Poaceae) which includes cereal crops and forage grasses. Tn the present study, it was aimed to create inbred lines of Brachypodium distachyon that were collected from different geographic regions of Turkey and characterize this diverse collection morphologically and cytologically. Besides, an efficient tissue culture and a plant transformation system using microprojectile bombardment and Agrobacterium- mediated gene transfer methods for three selected genotypes were established. Phenotypic characterization of the lines and macro-/microelement analysis in Brachypodium seeds showed extensive variation and significant differences. Callus formation percentages (50% to 100%) of the mature embryos and regeneration capacities of these calli (11.7% to 52.3%) were found to be genotype dependent. Each genotype displayed different responses to tissue culture conditions. Bombardment of the calli with the GUS (fJ-glucorinidase) gene revealed an average transient transformation efficiency of 28.7% within all three genotypes. Agrobacterium tumefaciens strains (EHA105, LBA4404 and AGL1), first two conferring resistance to kanamycin (nptll gene) and the later to glufosinate ammonium based herbicides (bar gene), were used for the transformation of calli that were wounded by microprojectile bombardment with two different sizes of gold particles or cutting with a blade. The GUS expression ranged from 0.0 to 10.4 (GUS foci/callus). All parameters were found to have significant effect on transient gene expression efficiency. Our well-characterized germplasm collection (146 inbred lines and 116 of them diploid) is freely available for scientific community and the lines that were used for the first time in this research for plant tissue culture and transformation studies could aid for further genetic and genomics studies in Brachypodium and other grass species including energy grasses.
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