Nishimura Taisuke, Yokota Etsuo, Wada Takuji, Shimmen Teruo, Okada Kiyotaka
Department of Botany, Graduate School of Science, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, 606-8502 Japan.
Plant Cell Physiol. 2003 Nov;44(11):1131-40. doi: 10.1093/pcp/pcg158.
Eight functional actin genes are present in Arabidopsis: The functional characterization of these genes in loss-of-function mutants is difficult, because highly conserved isovariants are generally expressed in the same tissue. We isolated a novel semi-dominant mutant allele (act2-2D) of an actin gene, ACT2, with a missense mutation which causes an amino acid substitution at the surface of the ACT2 protein. ACT2 promoter::ACT2-2D transgenic plants showed the same phenotype as act2-2D, indicating that act2-2D is a dominant-negative mutant. act2-2D exhibited defects in the initiation and elongation of root hairs, the elongation of root epidermal cells, and growth in aerial portions. Specifically, radial cell expansion was reduced and occasional cell death occurred in trichoblasts but not in atrichoblasts of the root epidermis. In contrast, cell division patterns in the root meristem were not affected. act2-3, a loss-of-function ACT2 mutant, did not develop most of these morphological abnormalities. Actin filament (F-actin) bundles in root epidermal cells of act2-2D were shorter than in the wild type and in the loss-of-function mutant. We conclude that defective F-actin polymerization caused the aberrant cell morphology in a dominant-negative manner, and that ACT2 functions in cell elongation and root hair formation.
由于高度保守的同工型变体通常在相同组织中表达,因此在功能缺失突变体中对这些基因进行功能表征很困难。我们分离出了肌动蛋白基因ACT2的一个新的半显性突变等位基因(act2-2D),该突变导致错义突变,使ACT2蛋白表面发生氨基酸替换。ACT2启动子::ACT2-2D转基因植物表现出与act2-2D相同的表型,表明act2-2D是一个显性负突变体。act2-2D在根毛的起始和伸长、根表皮细胞的伸长以及地上部分的生长方面表现出缺陷。具体而言,根表皮毛细胞的径向细胞扩展减少,偶尔会发生细胞死亡,但根表皮非毛细胞中则不会。相比之下,根分生组织中的细胞分裂模式未受影响。act2-3是一个ACT2功能缺失突变体,并未出现大多数这些形态异常。act2-2D根表皮细胞中的肌动蛋白丝(F-肌动蛋白)束比野生型和功能缺失突变体中的短。我们得出结论,有缺陷的F-肌动蛋白聚合以显性负方式导致异常细胞形态,并且ACT2在细胞伸长和根毛形成中发挥作用。