First published online June 7, 2007; 10.1104/pp.107.096727
Plant Physiology 144:2000-2008 (2007)
© 2007 American Society of Plant Biologists
PLANTS INTERACTING WITH OTHER ORGANISMS
Involvement of a Soybean ATP-Binding Cassette-Type Transporter in the Secretion of Genistein, a Signal Flavonoid in Legume-Rhizobium Symbiosis1
Akifumi Sugiyama,
Nobukazu Shitan and
Kazufumi Yazaki*
Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Gokasho, Uji 611–0011, Japan
Legume plants have an ability to fix atmospheric nitrogen into nutrients via symbiosis with soil microbes. As the initial event of the symbiosis, legume plants secrete flavonoids into the rhizosphere to attract rhizobia. Secretion of flavonoids is indispensable for the establishment of symbiotic nitrogen fixation, but almost nothing is known about the membrane transport mechanism of flavonoid secretion from legume root cells. In this study, we performed biochemical analyses to characterize the transport mechanism of flavonoid secretion using soybean (Glycine max) in which genistein is a signal flavonoid. Plasma membrane vesicles prepared from soybean roots showed clear transport activity of genistein in an ATP-dependent manner. This transport activity was inhibited by sodium orthovanadate, a typical inhibitor of ATP-binding cassette (ABC) transporters, but was hardly affected by various ionophores, such as gramicidin D, nigericin, or valinomycin, suggesting involvement of an ABC transporter in the secretion of flavonoids from soybean roots. The Km and Vmax values of this transport were calculated to be 158 µM and 322 pmol mg protein–1 min–1, respectively. Competition experiments using various flavonoids of both aglycone and glucoside varieties suggested that this ABC-type transporter recognizes genistein and daidzein, another signaling compound in soybean root exudates, as well as other isoflavonoid aglycones as its substrates. Transport activity was constitutive regardless of the availability of nitrogen nutrition. This is, to our knowledge, the first biochemical characterization of the membrane transport of flavonoid secretion from roots.
1 This work was supported by a Grant-in-Aid for Scientific Research (grant nos. 17051018 and 17027016 to K.Y.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by a research fellowship from the Japan Society for the Promotion of Science for Young Scientists (grant no. 183051 to A.S.).
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Kazufumi Yazaki (yazaki{at}rish.kyoto-u.ac.jp).
www.plantphysiol.org/cgi/doi/10.1104/pp.107.096727
* Corresponding author; e-mail yazaki{at}rish.kyoto-u.ac.jp; fax 81–774–38–3623.
Received January 29, 2007;
accepted June 1, 2007;
published June 7, 2007.
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