Journal Article

Dynamics of Periarbuscular Membranes Visualized with a Fluorescent Phosphate Transporter in Arbuscular Mycorrhizal Roots of Rice

Yoshihiro Kobae and Shingo Hata

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 51, issue 3, pages 341-353
Published in print March 2010 | ISSN: 0032-0781
Published online January 2010 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcq013
Dynamics of Periarbuscular Membranes Visualized with a Fluorescent Phosphate Transporter in Arbuscular Mycorrhizal Roots of Rice

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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In arbuscular mycorrhizal (AM) symbiosis, host plants supply photosynthates to AM fungi and, in return, they receive inorganic nutrients such as phosphate from finely branched fungal arbuscules. Plant cortical cells envelope arbuscules with periarbuscular membranes that are continuous with the plant plasma membranes. We prepared transgenic rice (Oryza sativa) plants that express a fusion of green fluorescent protein with rice AM-inducible phosphate transporter, OsPT11–GFP, and grew them with AM fungi. The fluorescence of the fusion transporter was observed in the arbuscule branch domain, where active nutrient exchange seems to occur. In contrast, a signal was not detected around intracellular hyphal coils on colonization by either Glomus mosseae or Gigaspora rosea, making the difference between Arum- and Paris-type mycorrhizae ambiguous. We also invented a simple device involving glass-bottomed Petri dishes for in planta observation of fluorescent proteins in living AM roots with an inverted fluorescence microscope. The plant bodies remain completely intact, avoiding any stressful procedure such as cutting, staining, etc. Since rice roots exhibit a very low level of autofluorescence, the device enabled clear time-lapse imaging to analyze the formation, function and degeneration of arbuscules. In cortical cells, arbuscules seemed to be functional for only 2–3 d. Suddenly, the arbuscular branches became fragile and they shrank. At this stage, however, the periarbuscular membranes appeared intact. Then, the fluorescence of the transporter disappeared within only 2.5–5.5 h. The collapse of arbuscules occurred in the subsequent several days. Thus, our device has a great advantage for investigation of dynamic features of AM symbiosis.

Keywords: Arbuscule; In planta imaging; Mycorrhizal symbiosis; Oryza sativa; Phosphate transporter

Journal Article.  7034 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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