Production of transgenic wheat through particle bombardment of scutellar tissues: Frequency is influenced by culture duration

Abstract

Summary A transformation system for hexaploid wheat was developed by using scutellar tissues of cultured immature embryos via particle bombardment. The scutellar tissues of immature embryos were cultured for 1 to 9 d prior to bombardment with two plasmids, pBI221 or p Act1 -F, containing the s-glucuronidase (GUS) gene and pARK22 containing the bialaphos resistant gene ( bar ). Transient expression of the reporter GUS gene was not influenced by culture duration prior to transformation. On the other hand, transgenic wheat plants of both spring and winter cultivars were obtained from immature embryos bombarded after culture for more than 5 d, but not from those receiving less than 5 d. The integration and inheritance of the bar gene as a selectable marker gene were confirmed by the combination of PCR amplification and Southern analysis of T 0 and T 1 plants. Some of the bialaphos resistant plants showed the expression of the GUS gene co-transformed with the bar gene.