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Plant and Cell Physiology Advance Access published online on June 13, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcj069
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Plant and Cell Physiology 2006 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received April 17, 2006
Accepted May 25, 2006

Rapid Paper

Knockdown of An Arbuscular Mycorrhiza-Inducible Phosphate Transporter Gene of Lotus japonicus Suppresses Mutualistic Symbiosis

Daisuke Maeda 1, Kanae Ashida 2, Keita Iguchi 3, Svetlana A. Chechetka 4, Ayaka Hijikata 4, Yasuhiro Okusako 4, Yuichi Deguchi 5, Katsura Izui 6, and Shingo Hata 4 *

1 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan; Present address: Takii Plant Breeding and Experiment Station, 1360 Hari, Konan, Shiga, 520-3231 Japan
2 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan; Present address: National Agricultural Research Center for Western Region, Fukuyama, Hiroshima, 721-8514 Japan
3 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan; Present address: Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan
4 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan
5 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan; Present address: Friedrich Miescher Institute, Maulbeerstrasse 66, Basel, CH-4058 Switzerland
6 Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan; Present address: Department of Biotechnological Science, Kinki University, Kinokawa, Wakayama, 649-6493 Japan

* To whom correspondence should be addressed.
Shingo Hata, E-mail: shing{at}kais.kyoto-u.ac.jp


  Abstract

cDNA for a major arbuscular mycorrhiza (AM)-inducible phosphate (Pi) transporter of Lotus japonicus, LjPT3, was isolated from Glomus mosseae-colonized roots. The LjPT3 transcript was expressed in arbuscule-containing cells of the inner cortex. The transport activity of the gene product was confirmed by the complementation of a yeast mutant that lacks high-affinity Pi transporters. In contrast to most AM-inducible Pi transporters thus far reported, LjPT3 has an amino acid sequence that has much in common with those of other members of the Pht1 family of plant Pi transporters, like StPT3 of potato [Rausch et al. (2001) Nature 414: 462-470]. To better understand the physiological role of this AM-inducible Pi transporter, knockdown transformants of the gene were prepared through hairy root transformation and RNA-interference. Under Pi-limiting conditions, the transformants showed a reduction of Pi uptake via AM and growth retardation. The transformants also exhibited a decrease in G. mosseae arbuscules. Additionally, when Mesorhizobium loti was inoculated into the knockdown transformants in combination with G. mosseae, necrotic root nodules were observed. Based on these findings, we consider that the genetically engineered host plants had monitored insufficient Pi uptake via AM or low expression of LjPT3, excluding the existing fungi and rhizobia and/or preventing further development of the fungal and nodule structures.

Keywords: Arbuscule; Host sanction; Lotus japonicus; Phosphate uptake; RNAi; Root nodule.
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