Evaluation of foliar fungal endophyte incidence in field-grown transgenicBtwhite spruce trees

Abstract

A total of 770 transgenic Bt white spruce needles were collected and plated on potato dextrose agar to determine their foliar endophyte diversity. The ribosomal internal transcribed spacer regions for 310 foliar endophytes were amplified by polymerase chain reaction (PCR) and digested using CfoI and MspI, which created 21 restriction groups. Isolates from each restriction group were sequenced and compared with reference sequences in GenBank. Eighteen sequence groups were obtained, of which five were identified at the species level. The most common endophytic fungi identified by PCR-RFLP was Lophodermium piceae (incidence of 74.5%). The second and third most common ones were Hypoxylon fragiforme (3.63%) and Lophodermium nitens (3.18%). A statistical analysis performed on the most common endophyte groups showed no statistical difference in endophyte frequency or distribution between the control white spruce needles (nontransgenic) and saplings with constructs containing the reporter gene GUS or the Bt Cry1A(b) gene and kanamycin.