Uporaba dvo-fotonskega konfokalnega mikroskopa za raziskovanje encimske aktivnosti ektomikoriznih gliv in analiza slik s programom ImageJ / Use of two-confocal mycroscopy for research of enzymatic activity of ectomycorrhizal fungi and image analysis with

Authors

  • Ines Štraus Gozdarski inštitut Slovenije, Večna pot 2, 1000 Ljubljana, Slovenija
  • Marko Kreft Biotehniška fakulteta, Oddelek za biologijo, & Inštitut za patofiziologijo, Medicinska fakulteta, Univerza v Ljubljani, Večna pot 111, 1000 Ljubljana, Slovenia; Celica Biomedical, Tehnološki park 24, 1000 Ljubljana, Slovenia
  • Hojka Kraigher Gozdarski inštitut Slovenije, Večna pot 2, 1000 Ljubljana, Slovenija

DOI:

https://doi.org/10.3986/fbg0013

Abstract

Simbioza med ektomikoriznimi glivami in gozdnimi drevesi prispeva k rasti dreves in delovanju ekosistema. Ektomikorizne glive so razvile visoko diferencirane strukture, ki omogočajo različne načine privzemanja hranilnih snovi in transport le teh do korenin. Aktivnost fosfataz v mikoriznih glivah omogoča dostopnost fosfatnih spojin v tleh, do katerih rastline sicer nimajo dostopa. V raziskavi smo analizirali aktivnost na površino hif vezane fosfataze (SBP) mikoriznih gliv pri sadikah bukve (Fagus sylvatica L.) in predstavili metodo za zajemanje in analizo slik fosfatazne aktivnosti. Uporabili smo dvofotonski konfokalni laserski mikroskop LSM 7 MP, ki omogoča zajemanje slik od površine mikoriznega plašča skozi mikorizni plašč vse do površine koreninskih celic. Koreninske vršičke smo obarvali s komercialnim barvilom ELF® 97. Slike smo analizirali s prosto dostopnim programom ImageJ. V članku smo pod- robno opisali postopek analize in predstavili prednosti in slabosti izbrane metode.

Ključne besede: na površino vezana fosfataza, eksploracijski tipi, fosfat, ELF® 97, bukev


The ectomycorrhizal symbiosis of forest trees contributes to the function of ecosystem and tree growth. The symbiotic organs of ectomycorrhiza (ECM) are highly differentiated structures differing in nutrient uptake, transfer capacities and ability to promote tree growth. In the present study surface-bound phosphatase activity (SBP) was analyzed on ectomycorrhiza of beech seedlings (Fagus sylvatica L.). The method for images acquisition and their analysis was discussed. Two-photon confocal laser scanning microscope LSM 7 MP was used to study spatial distribution of SBP enzymatic activity in various thicknesses of ECM mantle in intact (non-sectioned) ECM root tips. ELF® 97 was used for staining phosphatase enzymes. Images were analyzed using the freely available program ImageJ. In this article, we describe in detail the analysis procedure and the advantages and disadvantages of the method.

Key words: surface-bound phosphatase, exploration types, phosphate, ELF® 97, beech 


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Published

2017-04-27

Issue

Vol. 57 No. 2 (2016)

Section

Articles

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