AOBPreview published online on October 7, 2007
Annals of Botany, doi:10.1093/aob/mcm243
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Isolation and Characterization of Arbuscules from Roots of an Increased-arbuscule-forming Mutant of Lotus japonicus
1 Faculty of Bioresources, Mie University, Tsu, Mie 514-8507, Japan
2 Graduate School of Arts and Sciences, University of Tokyo, Komaba, Meguro-ku, Tokyo 153-8902, Japan
3 National Institute of Agrobiological Resources, Kannondai, Tsukuba, Ibaraki 305-8602, Japan
4 Laboratory of Soil Science, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo, Tokyo 113-8657, Japan
5 School of Earth and Geographical Sciences, M087, The University of Western Australia, Crawley, WA 6009, Australia
* For correspondence. E-mail solaiman{at}cyllene.uwa.edu.au
Received: 4 June 2007 Returned for revision: 23 July 2007 Accepted: 20 August 2007
Background and Aims: Previous methods for isolation of arbuscules from mycorrhizal roots are time-consuming, complex and expensive. Therefore, a simple, rapid and inexpensive method for the isolation of metabolically active arbuscules from plant root of an increased-arbuscule-forming mutant of Lotus japonicus (Ljsym78-2) is described.
Methods: Roots of the L. japonicus mutant plants Ljsym78-2 colonized by Glomus sp. were separated from soil, washed with water, immersed in CaSO4 before being cut into 5-mm pieces and homogenized with a Waring blender at 6000 rpm for 30 s. The arbuscules were purified by separation from plant tissues with a 50-µm nylon mesh, finally collecting on a 30-µm nylon mesh. Enzyme histochemical staining showed that the collected arbuscules had succinate dehydrogenase, alkaline phosphatase and acid phosphatase activities.
Key Results and Conclusions: The enzymic activity of the arbuscules was not affected after the isolation process. The establishment of this simple, rapid and inexpensive method for the isolation of metabolically active arbuscules will be useful to clarify the biochemical processes occurring in nutrient exchange at the arbuscular interface.
Key words: Arbuscular mycorrhiza, arbuscule isolation, Glomus sp., increased-arbuscule-forming mutant, Lotus japonicus