Transferring TrxS Gene Into Beer Barley By Particle Bombardment And Inheritance Analysis Of The Progenies

In this paper, the tested materials were 4 beer barley cultivars including Stirling, Franklin, Gairdner, Harrington from Australia. The effect of Cu²⁺ concentrations, different acceptors and mannitol treatments on callus cultures and transformation frequency of barley immature embryo were studied. The two plasmids with TrxS gene and Bar gene were co-transformed by particle bombardment. The regenerated plantlets were obtained and the transformed plants progeny were also analyzed. The results showed that (1) 5.0¹0.0#M Cu²⁺ not only inhibited immature embryo germination directly, but also significantly improved the embryogenic callus development. (2)the immature embryos pre-cultured for 15 days have a higher transformation frequency than that of pre-cultured 3⁴ days and immature embryos. (3) 0.5M mannitol treatment from 6h before to 18h after bombardment could increase the transformation frequency. (4) 9 regenerated plantlets were obtained by optimum plant regeneration culture and PPT screening, which all were positive in Bialaphos resistance and 7 plants were positive individuals by the semi-nested PCR assay. (5) according to the semi-nested PCR assay and PCR-Southern blotting, the inheritance patterns of T₁ plants (LSY) showed about 3:1 segregation ratio, in accordance with Mendelian rule of dominant gene with one locus. However, the segregation ratio of T₂ plants (JY6) was about 1:1 and 8:1. It is possible that there was chimaera in T₁ plants.