Molecular cloning and mRNA localization of tomato pollen profilin

Abstract

The actin cytoskeleton plays an important role in the growth of pollen tube. The actin-binding protein profilin could play a role in regulating the organization of the actin filaments. Using the RT-PCR technique, we isolated a cDNA clone (designated LePro 1) encoding profilin from pollen grains of tomato (Lycopersicon esculentum Mill. cv. Moneymaker). Sequence analysis of the insert shows 87% similarity to tobacco ntPro2, 78% to timothy grass profilin, 77% to Arabidopsis AthPRF4, 77% to maize ZmPro3, and 73% to birch profilin. Both quantitative PCR and RNA gel blot analyses demonstrated that LePro 1 is expressed in a tissue- or cell-type specific manner in the tomato plant. In situ hybridization of 2 microns thick anther sections using a non-radioactive labeling method reveals that LePro 1 is expressed only in pollen grains, with undetectable transcription in other parts of the anther or the other organs. Phylogenetic analysis of amino acid sequences of 18 plant profilins indicates that two distinct profilin gene classes are present in higher plants. One is pollen-specific, another is constitutive. LePro 1 belongs to the former class.