082 Infection and Transformation of Rhododendron by Agrobacterium tumefaciens Strain B6
The objective of this study was to determine if selected strains of Agrobacterium could infect microshoots of Rhododendron catawbiense. Fifteen microshoot stems of R. catawbiense var. album `America', `Joe Paterno', and `Cunningham's White' were inoculated with two drops (about 25 μL) of wild type Agrobacterium tumefaciens strains C58 or B6 or with wild type A. rhizogenes strain E8/73. Five control shoots were inoculated with 1.2 mM KH2PO4 buffer. Microshoots were grown on woody plant medium (WPM) supplemented with 4.9 μM 2iP. Six weeks after inoculation galls that formed were excised from the microshoots and placed on WPM that lacked plant growth regulators but contained 300 mg·L-1 cefotaxime. In another study, these wild-type bacterial strains were genetically modified by inserting the pBINm-gfp5-ER plasmid, which contained genes coding for NPTII and green fluorescence protein (GFP), into the bacteria. These modified strains were inoculated on 15 stems of the three rhododendron cultivars and one variety. Calluses that formed were excised, placed on basal WPM with cefotaxime, and allowed to proliferate. Wild type C58 induced galls to form on `Joe Paterno', R.c. album, and `Cunningham's White' stems, whereas wild type B6 caused galls to form only on the latter two types of rhododendron. Wild-type E8/73 failed to induce gall formation on the rhododendrons. Only genetically modified B6 caused galls to form on only `Cunningham's White' microshoots (seven of 15 inoculated stems). Three of these galls fluoresced green under ultraviolet light. Physical presence of the NPTII and GFP genes in the plant genome was determined by polymerase chain reaction. This study demonstrated that R. catawbiense is susceptible to Agrobacterium infection, and this plant can be genetically transformed.