Particle bombardment transformation of some Turkish wheat cultivars with TaNAC69-1 and TtNAMB2 genes

Abstract

Wheat is one of the most important crop plants in the world. The production of wheat is seriously affected by biotic (pests, bacterial and fungal diseases) and abiotic (drought, salinity and freezing) stresses. NAC family proteins (NAM/ATAF1-2/CUC2) are plant specific transcription factors and related with development, defense and abiotic stress responses. TaNAC69-1 and TtNAMB2 genes are the transcription factors from this protein family. The aim of this study was to transform of some Turkish wheat cultivars with TaNAC69-1 and TtNAMB2 using particle bombardment. After isolation of TaNAC69-1 from Triticum aestivum and TtNAMB2 from Triticum turgidum, these genes were cloned into pAHC25, an overexpression vector for monocots. pAHC25 includes bar gene as a plant selectable marker gene. Plasmids including TaNAC69- 1 and TtNAMB2 were introduced into immature inflorescences based calli of Yure˘gir-89 (T. aestivum) and Kızıltan-91 (T. turgidum) cultivars and mature embryo based calli of Mirzabey-2000 (T. turgidum) cultivar via particle bombardment. The bombarded calli were regenerated previously optimized tissue culture and regeneration conditions. After transformation of immature inflorescence based callus, the regeneration rate was about 16.2% for Yure˘gir- 89 and Kızıltan-91. The rooting rate for these two cultivars was 3.1%. The regeneration rate was 15.6% for Mirzabey-2000, with a 2.7% rooting rate. Putative transgenic plants were transferred to the greenhouse for maturation. The studies on the molecular characterization of putative transgenic plants have been continued.