Abstract

Although microbes are generally found as endophytes in many plant tissues, the plant shoot meristems have been considered virtually sterile. Plant tissue culture, which utilizes mostly the meristems, has nevertheless given numerous references to microbial existence in these tissues. Since the bud-derived tissue cultures of Scots pine (Pinus sylvestris L.) also become easily occupied by microbes, microbial presence was considered to be one potential cause of the low regeneration capacity of tissues from mature trees. The origin of the microbes occurring in tissue cultures of buds of Scots pine was established in this study. One of the microbes, classified as Hormonema dematioides (Ascomycota), was localized in the scale tissues of a bud. Several other microbes were detected, and were associated especially with the meristematic tissues of Scots pine buds. This group was comprised of the yeast Rhodotorula minuta, and bacteria belonging to Methylobacterium spp., the Pseudomonas fluorescens subgroup, and a Mycobacterium sp. These endophytes were located particularly in the outermost cells of meristems, in the epithelial cells of resin ducts, and in the cells of the developing stem, in the vicinity of the meristems. These endophytes were less frequently found in the vascular tissue or in the intercellular spaces of cells, which are typical locations for the previously known endophytic bacteria.

The meristem-associated endophytes were discovered to affect growth of pine tissues, and some of them produced substances that are suitable as precursors for phytohormone synthesis. Additionally, pure cultures of both bacterial and fungal endophytes showed antagonism in vitro against pathogens. When the correlation between the presence endophytes and the degeneration of bud-derived tissue cultures of Scots pine was studied, it was discovered that the endophytes grew uncontrollably once a tissue culture was initiated from the bud. A high level of chitinase production was also detected in these tissue cultures, which was considered to be indicative of a defense reaction. However, the endophytes were not found to colonize excessively in every tissue, but all bud-derived tissue cultures of Scots pine eventually degenerate. Therefore, it was concluded that the endophytes may not exclusively be considered responsible for the degeneration of the cultures.