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First published online February 27, 2003; 10.1104/pp.102.017020

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Plant Physiol, March 2003, Vol. 131, pp. 1054-1063

crinkle, a Novel Symbiotic Mutant That Affects the Infection Thread Growth and Alters the Root Hair, Trichome, and Seed Development in Lotus japonicus1

Myra L. Tansengco, Makoto Hayashi,* Masayoshi Kawaguchi, Haruko Imaizumi-Anraku, and Yoshikatsu Murooka

Osaka University, Graduate School of Engineering, Department of Biotechnology, Yamadaoka 2-1, Suita, Osaka 565-0871, Japan (M.L.T., M.H., Y.M.); Niigata University, Faculty of Science, Department of Environmental Sciences, Ninomachi 8050, Ikarashi, Niigata City, Japan (M.K.); and National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan (H.I.-A.)

To elucidate the mechanisms involved in Rhizobium-legume symbiosis, we examined a novel symbiotic mutant, crinkle (Ljsym79), from the model legume Lotus japonicus. On nitrogen-starved medium, crinkle mutants inoculated with the symbiont bacterium Mesorhizobium loti MAFF 303099 showed severe nitrogen deficiency symptoms. This mutant was characterized by the production of many bumps and small, white, uninfected nodule-like structures. Few nodules were pale-pink and irregularly shaped with nitrogen-fixing bacteroids and expressing leghemoglobin mRNA. Morphological analysis of infected roots showed that nodulation in crinkle mutants is blocked at the stage of the infection process. Confocal microscopy and histological examination of crinkle nodules revealed that infection threads were arrested upon penetrating the epidermal cells. Starch accumulation in uninfected cells and undeveloped vascular bundles were also noted in crinkle nodules. Results suggest that the Crinkle gene controls the infection process that is crucial during the early stage of nodule organogenesis. Aside from the symbiotic phenotypes, crinkle mutants also developed morphological alterations, such as crinkly or wavy trichomes, short seedpods with aborted embryos, and swollen root hairs. crinkle is therefore required for symbiotic nodule development and for other aspects of plant development.


1 This work was supported by grants from the Ministry of Education, Culture, Sports and Technology of Japan (to M.L.T.), and the Core Research for Evolutional Science and Technology (CREST) from Japan Science and Technology Corporation (to M.H. and M.K.).

* Corresponding author; e-mail hayashi{at}bio.eng.osaka-u.ac.jp; fax 81-6-6879-7418.

© 2003 American Society of Plant Biologists



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