Plant Physiol. Illumina
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Characterization of LeMir, a Root-Knot Nematode-Induced Gene in Tomato with an Encoded Product Secreted from the Root1

Eric D. Brenner2, Kris N. Lambert3, Isgouhi Kaloshian4, and Valerie M. Williamson*

Department of Nematology, University of California, Davis, California 95616

A tomato gene that is induced early after infection of tomato (Lycopersicon esculentum Mill.) with root-knot nematodes (Meloidogyne javanica) encodes a protein with 54% amino acid identity to miraculin, a flavorless protein that causes sour substances to be perceived as sweet. This gene was therefore named LeMir (L. esculentum miraculin). Sequence similarity places the encoded protein in the soybean trypsin-inhibitor family (Kunitz). LeMir mRNA is found in root, hypocotyl, and flower tissues, with the highest expression in the root. Rapid induction of expression upon nematode infection is localized to root tips. In situ hybridization shows that LeMir is expressed constitutively in the root-cap and root-tip epidermis. The LeMir protein product (LeMir) was produced in the yeast Pichia pastoris for generation of antibodies. Western-blot analysis showed that LeMir expression is up-regulated by nematode infection and by wounding. LeMir is also expressed in tomato callus tissue. Immunoprint analysis revealed that LeMir is expressed throughout the seedling root, but that levels are highest at the root/shoot junction. Analysis of seedling root exudates revealed that LeMir is secreted from the root into the surrounding environment, suggesting that it may interact with soil-borne microorganisms.

1   This work was supported by grants from the U.S. Department of Agriculture-National Research Initiative Competitive Grants Program (nos. 92-37302-7635 and 94-37302-0569).
2   Present address: Department of Biology, 1009 Main Building, Washington Square, New York University, New York, NY 10003.
3   Present address: Department of Plant Biology, 311 Koshland Hall, University of California, Berkeley, CA 94720.
4   Present address: Department of Nematology, University of California, Riverside, CA 92521.
*   Corresponding author; e-mail vmwilliamson{at}ucdavis.edu; fax 1-530-752-5809.

Plant Physiol. (1998) 118: 237-247
Copyright Clearance Center:   0032-0889/98/118//11
© 1998 American Society of Plant Physiologists




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