Isolation and characterization of arbuscules from roots of an increased-arbuscule-forming mutant of Lotus japonicus.
OBJECTIVE
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 CaSO(4) 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-mum nylon mesh, finally collecting on a 30-mum nylon mesh. Enzyme histochemical staining showed that the collected arbuscules had succinate dehydrogenase, alkaline phosphatase and acid phosphatase activities.


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.
Senoo
Keishi
K
Faculty of Bioresources, Mie University, Tsu, Mie 514-8507, Japan.
Solaiman
Zakaria
Z
Tanaka
Satoki
S
Kawaguchi
Masayoshi
M
Imaizumi-Anraku
Haruko
H
Akao
Shoichiro
S
Tanaka
Akiyoshi
A
Obata
Hitoshi
H
D000135:Acid Phosphatase / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000070116:Lotus / Q000235:genetics / Q000382:microbiology*; D009154:Mutation; D038821:Mycorrhizae / Q000201:enzymology / Q000254:growth & development / Q000302:isolation & purification*; D018517:Plant Roots / Q000235:genetics / Q000382:microbiology*; D013385:Succinate Dehydrogenase / Q000378:metabolism
D013385:Succinate Dehydrogenase; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase