Research on orchid mycorrhizas is giardia life cycle time dynamically developing branch of mycorrhizal research. An increasing depth of knowledge is being acquired on the life cycles of individual orchid species and on the taxonomic status of their fungal partners, and also on the specificity of the plant—fungus relationship.
For certain rare species, however, there are still gaps in this basic knowledge, as in the case of the fen orchid Liparis loeselii and the bog giardia life cycle time Hammarbya paludosa. Even fewer data are available on the habitat requirements of the potential orchid-symbiotic fungi.
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The main aims of the present work were thus as follows: 1. Estimation of the natural germination ratio of the fen orchid by means of in situ germination. Detection of the symbiotic fungi of the fen orchid and bog orchid giardia life cycle time from protocorms germinated in situ. Detection of fungal groups potentially capable of forming orchid mycorrhizas in aqueous habitats in Hungary, and the investigation of their habitat requirements.
The methods used to achieve these aims will be detailed in the following section.
The seeds of the fen orchid were germinated under both in situ and ex situ conditions. For the in situ studies, the seeds were placed in two habitats currently harbouring populations of the species Pákozd, Dunaharasztiin a habitat where populations were detected from to Kistómalomin an uncertain habitat, where no orchids of this species have been detected since Vaja and in the habitat of the bog orchid population.
As regards the time between seed placement and retrieval, two types of experiment were carried out.
In all the habitats listed, jobb gyógymód a férgek ellen were placed in spring and retrieved in autumn. In the case of Lake Velence Pákozd seeds were also placed in autumn, followed by retrieval in early summer and autumn the following year and two years later in autumn. For the in situ seed placements the seeds were placed in a piece of cheese-cloth mesh size 85— µm folded giardia life cycle time two Rasmussen and Whigham The ex situ germination of fen orchid was carried out in the Botanical Garden of Eötvös Loránd University, on a piece of peat originating from Lake Velence.
Nine orchid species were used to investigate the fungi forming orchid mycorrhizas in aqueous habitats.
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The fen orchid and bog orchid species that were germinated in situ specialise in extremely wet habitats floating mat fens and bogswhile the remainder of the orchid taxa included in the experiment have a wider range of habitats.
Three of the orchids were examined both on floating mat fens and on terrestrial habitats: the lax-flowered marsh orchid Orchis laxiflora ssp.
The other four species were only found on terrestrial habitats: the fragrant orchid Gymnadenia conopseathe Crimean orchid Ophrys oestriferathe early spider orchid O. The habitats investigated were classified on the basis of their wetness floating mat fen, terrestrial fen, swamp, steppe and by using the vegetation as an indication of the soil water content WB index; Borhidi Fungal strains were isolated from the protocorms and roots of the orchids using the root segment technique Bernard and the peloton extraction technique.
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The isolated fungal strains were examined and grouped using a molecular taxonomic method. For some groups of 4 fungi, the sequences of the ribosomal ITS region can be used for identification even at the species level Frøslev et al.
Phylogenetic analysis was also carried out with the MEGA 4 program package. New results presented in the thesis: Results and discussion — The mycorrhizal fungal partner of the bog orchid Hammarbya paludosa was successfully isolated and identified from protocorms germinated in situ. Judging from its ITS sequence, the symbiont belongs to the Tulasnella genus of fungi.
The orchid seeds germinated both on the reed-sedge peat of the two floating mat fen habitats of the fen orchid in Pákozd Lake Velence and Dunaharaszti Soroksár branch of the Danubeand on the peat moss of the Bábtava habitat of the bog orchid.
This provided extremely important information on giardia life cycle time vital phase in the life cycle of the species that had not previously been investigated Wheeler et al. Further studies will be needed to decide whether the more frequent occurrence of the fungus resulted in the settlement of the orchids, or whether the development of successful symbiosis resulted in the multiplication of the fungus in the vicinity of the orchids.
This is in agreement with the findings of Bonnardeaux et al. The apparent fungal partner species specificity of the fen orchid, a floating mat fen specialist in Hungary, could be due to the fact that the number of potential fungal partners declines in extremely wet habitats, so the possibility that other fungal species could be satisfactory symbionts for fen orchid cannot be excluded.
It can thus be concluded that there are micro climatic reasons for the fact that the fen orchid is only found on floating mat fens in Hungary.
No field knowledge was previously available on germination, the first phase in the life cycle of fen orchids. The in situ germination method elaborated for orchids was successfully employed to observe the germination processes taking place in the natural habitats of fen orchids and to determine the magnitude of germination.
Studies on the mycorrhizal partners of rare orchids raise nature protection problems, since conventional fungus isolation techniques destroy the orchid plants. By contrast, the in situ examination of seed germination does not endanger the natural orchid population. The application of this method allowed the mycorrhizal fungus partners of these rare orchids to be identified at the only location in Hungary where a few tens of bog orchids are to be found and at two fen orchid habitats.
Aqueous habitats are rapidly being lost not only in Hungary, but throughout Europe.