A transposon, Ping, is integrated into intron 4 of the DROOPING LEAF gene of rice, weakly reducing its expression and causing a mild drooping leaf phenotype

Abstract

The YABBY gene DROOPING LEAF (DL) regulates midrib formation in the leaves and carpel specification in the flowers of rice (Oryza sativa L). We found a new dl allele (dl-5) that caused a mild phenotype in the descendants of a mutable line, IM294. In plants homozygous for this allele, midrib structures were formed but their sizes were reduced. Molecular analysis revealed that a transposon, Ping, was inserted in the fourth intron of DL. Together with mPing and Pong, Ping is a member of a transposon family that was first identified as a group of active transposable elements in rice. Our finding of the Ping insertion in the DL gene is a first indication that Ping is active in planta, and that it can be transposed and integrated in a new locus. Ping seems to be still active because it was excised from intron 4 of DL at a relatively high frequency in rice calli. Real-time PCR analysis and in situ hybridization indicated that DL transcript levels were reduced in dl-5 without alterations in the spatial expression pattern of the DL gene. The reduction of DL expression may be due to inefficient splicing of the large intron caused by Ping insertion. By comparing the expression levels of DL and leaf phenotypes in the dl mutants with different severities, we confirmed our previous hypothesis that DL promotes cell proliferation in the central region of leaf primordia, and that this cell proliferation is critical for midrib formation in the mature leaves.