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Identification of Novel Putative Regulatory Genes Induced During Alfalfa Nodule Development with a Cold-Plaque Screening Procedure

May 1998 , Volume 11 , Number  5
Pages  358 - 366

Florian Frugier , 1 Adam Kondorosi , 1 , 2 and Martin Crespi 1

1Institut des Sciences Végétales, Centre National de la Recherche Scientifique, F-91198 Gif-sur-Yvette Cedex, France; and 2Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, Szeged, PO Box 521, H-6701 Szeged, Hungary

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Accepted 24 January 1998.

Until now very few plant genes with possible regulatory functions during nodule development have been isolated. We have used a modified cold-plaque screening method to identify new transcripts expressed at low levels that are induced during nodulation. Several clones were isolated and characterized by their mRNA expression patterns during nodule development and in spontaneous nodules. Sequence homology with known genes of other organisms indicated that transcripts corresponded to (i) “basic” genes probably required during the growth of the nodule organ (e.g., structural proteins), (ii) genes related to the metabolic adaptations taking place during nodule morphogenesis and function (e.g., carbonic anhydrase), and (iii) genes containing regulatory motifs and/or homologies (three clones out of the 20 identified). The latter genes encode a zinc-finger-containing protein, a putative protein kinase, and a Wilm's tumor (WT) suppressor homologue, respectively. Expression of the kinase and WT suppressor homologues was induced early in nodulation, although the latter was activated transiently. Accumulation of the Zn-finger gene transcripts was detected at a later stage of development and seems to be regulated in a complex manner. Hence, using a cold-plaque screening procedure, we could identify genes that may play regulatory roles in the signal transduction pathways activated during nodule development.

Additional keyword: nodulins.

© 1998 The American Phytopathological Society