Characterization of the Mt4 gene from Medicago truncatula.

Previously we identified Mt4, a phosphate starvation inducible cDNA from Medicago truncatula which is down-regulated in roots in response to phosphate fertilization as well as colonization by arbuscular mycorrhizal (AM) fungi (AM). Here we present further studies of Mt4. Expression was highly sensitive to exogenous applications of phosphate fertilizer; transcripts were abundant in roots fertilized with nutrient solution lacking phosphate, reduced when fertilized with 0.02 or 0.1 mM phosphate and undetectable when fertilized with 1 or 5 mM phosphate. A time course experiment, to study the expression of Mt4 following colonization by AM fungi, revealed that Mt4 transcripts increased in uncolonized roots during the first three weeks of growth and then plateaued, while transcript levels in roots colonized with the AM fungus, Glomas versiforme, increased transiently and then decreased. Although the Mt4 gene is expressed exclusively in roots, transcripts were also detected in M. truncatula cell suspension cultures following phosphate starvation. A genomic clone containing the Mt4 gene and 1133 bp of the 5' flanking sequence was identified from a M. truncatula genomic library. The promoter region contains a conserved cis-element found in the promoters of phosphate starvation inducible genes of yeast and tomato. As Mt4 is the first cDNA reported to show independent regulation by both phosphate and mycorrhizal fungi, the genomic clone may provide a starting point from which to analyze the signal transduction pathways involved in these two processes.