An easy method using glutaraldehyde-introduced fluorescence for the microscopic analysis of plant biotrophic interactions.
The method introduced in this article makes use of the glutaraldehyde-induced auto-fluorescence of proteins after cross-linking with glutaraldehyde for the analysis of cellular and sub-cellular structures. Because the interface of biotrophic interactions is rich in proteins, the method presented is particularly suitable for the analysis of such interactions; we have exemplified its usefulness by analyzing (1) the root feeding sites induced in roots from Arabidopsis thaliana by the root-knot nematode Meloidogyne incognita; (2) leaves from Cucurbita pepo infected by powdery mildew and (3) roots from Nicotiana tabacum colonized by the arbuscular mycorrhizal fungus Glomus intraradices. The use of confocal and multi-photon laser scanning microscopy allows three-dimensional reconstructions from optical sections of complex biotrophic interactions. In the case of root-knot nematode feeding sites, our method enabled us to simultaneously study the development of the plant xylem elements (using lignin auto-fluorescence), the nematode feeding site and the nematode itself.
Fester
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Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO 63132, USA.
Berg
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Taylor
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D000818:Animals; D017360:Arabidopsis / Q000469:parasitology*; D028464:Cucurbita / Q000382:microbiology*; D005453:Fluorescence; D005656:Fungal Proteins / Q000032:analysis; D005658:Fungi / Q000737:chemistry* / Q000254:growth & development; D005976:Glutaral / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D018515:Plant Leaves / Q000382:microbiology; D018517:Plant Roots / Q000382:microbiology; D011506:Proteins / Q000032:analysis; D014026:Nicotiana / Q000382:microbiology*; D014431:Tylenchoidea / Q000737:chemistry* / Q000254:growth & development
D005656:Fungal Proteins; D011506:Proteins; D005976:Glutaral