Increasing Transient and Subsequent Stable Transgene Expression in Chrysanthemum Following Optimization of Particle Bombardment and Agroinfection Parameters.

Abstract

Transient and stable transgene expression was maximized in in vitro and greenhouse-derived chrysanthemum ‘Lineker’ and ‘Shuhou-no-chikara’ stem explants. In Agroinfection, Agrobacterium tumefaciens LBA4404-with either pBI121 or pKT2-or AGL0 (with pKT3) were utilized, while in particle bombardment pSKGN1 was utilized. Transformation efficiency was affected by both the gene introduction method and its experimental parameters and the origin and developmental state of the explant. There was a decrease in GUS focal points with a simultaneous increase in the number of blue staining areas for both ‘Lineker’ and ‘Shuhou-no-chikara’ in both Agroinfection (pKT2 and pKT3) and particle bombardment (pSKGN1) over time (0, 24, 48, 72 h and 1, 2 and 4 weeks). GUS transformation efficiency was 6.6, 26.89, 25.0 and 11.84% for Agroinfection, bombardment, sonication and Agrolistics, respectively for ‘Lineker’, and 0, 2.17, 0 and 7.69%, respectively for ‘Shuhou-no-chikara’, the highest level of stable GUS transgene expression occurring in thevenation.