Colonization strategy of the endophytic plant growth-promoting strains of Pseudomonas fluorescens and Klebsiella oxytoca on the seeds, seedlings and roots of the epiphytic orchid, Dendrobium nobile Lindl.

AIMS: Orchids form strong mycorrhizal associations, but their interactions with bacteria are poorly understood. We aimed to investigate the distribution of plant growth promoting rhizobacteria (PGPR) at different stages of orchid development and to study if there is any selective specificity in choosing PGPR partners. METHODS AND RESULTS: Colonization patterns of gfp-tagged Pseudomonas fluorescens and Klebsiella oxytoca were studied on roots, seeds, and seedlings of Dendrobium nobile. Endophytic rhizobacteria rapidly colonized velamen and core parenchyma entering through exodermis and the passage cells, whereas at the early stages, they stayed restricted to the surface and the outer layers of the protocorms and rhizoids. The highest amounts of auxin (indole-3-acetic acid) were produced by K. oxytoca and P. fluorescens in the nitrogen-limiting and NO3 -containing media respectively. Bacterization of D. nobile seeds resulted in promotion of their in vitro germination. The plant showed no selective specificity to the tested strains. Klebsiella oxytoca demonstrated more intense colonization activity and more efficient growth promoting impact under tryptophan supplementation, while P. fluorescens revealed its growth-promoting capacity without tryptophan. CONCLUSIONS: Both strategies are regarded as complementary, improving adaptive potentials of the orchid when different microbial populations colonize the plant. SIGNIFICANCE AND IMPACT OF THE STUDY: This study enlarges our knowledge on orchid-microbial interactions, and provides new features on application of the nonorchid PGPR in orchid seed germination and conservation.

FEATURES OF EMBRYONIC DEVELOPMENT OF DIENIA OPHRYDIS (ORCHIDACEAE).

On example of Dienia ophrydis (J. Köenig) Seidenf (Orchidaceae), we have described a new type of embryogenesis of orchids ­ Dienia-type, which is differs from Liparis-type learned earlier in the tribe Malaxideae. Embryogenesis of Dienia-type is characterized by 1) the development of a single-celled suspensor formed by cb-derivative, 2) linear arrangement of germ cells in the tetrad stage, 3) the special structure of the embryo in the stages of tetrads and octants (l, lR, m, ci, cb), and 4) the absence of ci and cb cell division. The convergent similarity of embryogenesis of Dienia- and Caryophyllaceae-types is proposed. A number of specific for D. ophrydis structures of embryo sac and embryo, including «petassum¼, «fitting¼ and «suspensor coat¼ are described for the first time. Petassum represents remains of the cell walls of pollen tube, and perhaps of filamentous apparatus of synergids, plugging the micropyle side of the fertilized embryo sac. The only cell of suspensor has a specific appendix («fitting¼), that connects it with the embryo itself. There is «suspensor coat¼ which surrounds the only suspensor cell, including «fitting¼, but does not extend to the basal cells of the embryo itself.

[Localization of associative cyanobacteria on the roots of epiphytic orchids]. / Osobennosti lokalizatsii assotsiativnykh tsianobakterii na korniakh épifitnykh orkhidei.

This work is the first study of the localization of phototrophic microorganisms in the rhizoplane and velamen of epiphytic orchids, namely on the aerial and substrate roots of Acampe papillosa and Dendrobium moschatum and on the aerial roots of the Phalaenopsis amabilis and Dendrobium phalaenopsis. The composition of the bacterial community on the plant roots depended on the conditions of plant growth. Under conditions simulating climate of moist tropical forests, the aerial roots proved to be populated with phototrophic microorganisms among which cyanobacteria predominated. Interlaced fungal hyphae and filamentous cyanobacteria formed a sheath on the surface of aerial roots. The nitrogen-fixing capacity of the sheath of aerial roots was studied by the example of P. amabilis.

[Associative cyanobacteria isolated from the roots of epiphytic orchids]. / Assotsiativnye cyanobakterii, vydelennye s kornei épifitnykh orkhidei.

Associative cyanobacteria were isolated from the rhizoplane and velamen of the aerial roots of the epiphytic orchids Acampe papillosa, Phalaenopsis amabilis, and Dendrobium moschatum and from the substrate roots of Acampe papillosa and Dendrobium moschatum. Cyanobacteria were isolated on complete and nitrogen-free variants of BG-11 medium. On all media and in all samples, cyanobacteria of the genus Nostoc predominated. Nostoc, Anabaena, and Calothrix were isolated from the surface of the A. papillosa aerial roots, whereas the isolates from the substrate roots were Nostoc, Oscillatoria, and representatives of the LPP-group (Lyngbia, Phormidium, and Plectonema, incapable of nitrogen fixation). On the D. moschatum substrate roots, Nostoc and LPP-group representatives were also found, as well as Fischerella. On the aerial roots of P. amabilis and D. phalaenopsis grown in a greenhouse simulating the climate of moist tropical forest, cyanobacteria were represented by Nostoc, LPP-group, and Scytonema in the D. phalaenopsis and by Nostoc, Scytonema, Calothrix, Spirulina, Oscillatoria, and the LPP-group in P. amabilis. For D. moschatum, the spectra of cyanobacteria populating the substrate root zhizophane and the substrate (pine bark) were compared. In the parenchyma of the aerial roots of P. amabilis, fungal hyphae and/or their half-degraded remains were detected, which testifies to the presence of mycorrhizal fungi this plant. This phenomenon is attributed to the presence of a sheath formed by cyanobacteria and serving as a substrate for fungi.

[Microbiota of the Orchid rhizoplane]. / Mikrobiota poverkhnosti kornei Orkhidnykh.

Six bacterial strains isolated from the underground roots of the terrestrial orchid Calanthe vestita var. rubrooculata were found to belong to the genera Arthrobacter, Bacillus, Mycobacterium, and Pseudomonas. Strains isolated from the aerial roots of the epiphytic orchid Dendrobium moschatum were classified into the genera Bacillus, Curtobacterium, Flavobacterium, Nocardia, Pseudomonas, Rhodococcus, and Xanthomonas. The rhizoplane of the terrestrial orchid was also populated by cyanobacteria of the genera Nostoc and Oscillatoria, whereas that of the epiphytic orchid was populated by one genus, Nostoc. In orchids occupying different econiches the spectra of the bacterial genera revealed differed. The microbial complex of the terrestrial orchid rhizoplane differed from that of the surrounding soil.