Kurusu Takamitsu, Sakurai Yasuhiro, Miyao Akio, Hirochika Hirohiko, Kuchitsu Kazuyuki
Department of Applied Biological Science, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510 Japan.
Plant Cell Physiol. 2004 Jun;45(6):693-702. doi: 10.1093/pcp/pch082.
Cytosolic free Ca2+ serves as an important second messenger participating in signal transduction of various environmental stresses. However, molecular bases for the plasma membrane Ca2+ influx and its regulation remain largely unknown. We here identified a gene (OsTPC1) encoding a putative voltage-gated Ca2+ channel from rice, ubiquitously expressed in mature leaves, shoots and roots as well as in cultured cells. OsTPC1 rescued the Ca2+ uptake activity and growth rate of a yeast mutant cch1. To elucidate its physiological roles, we generated transgenic rice plants and cultured cells overexpressing OsTPC1 mRNA. Furthermore, a retrotransposon (Tos17) insertional knockout mutant of OsTPC1 was isolated. OsTPC1-overexpressing cells showed hypersensitivity to excess Ca2+ but higher growth rate under Ca2+ limitation, while growth of the OsTPC1-knockout cultured cells was less sensitive to extracellular free Ca2+ concentration, suggesting that OsTPC1 has Ca2+ transport activity across the plasma membrane. OsTPC1-overexpressing plants showed reduced growth and abnormal greening of roots. Growth of Ostpc1 seedlings was comparable to the control on agar plates, while significantly reduced in adult plants. These results suggest that OsTPC1 functions as a Ca2+ -permeable channel involved in the regulation of growth and development.
胞质游离Ca2+作为一种重要的第二信使,参与各种环境胁迫的信号转导。然而,质膜Ca2+内流及其调控的分子基础仍 largely未知。我们在此鉴定了一个来自水稻的编码假定电压门控Ca2+通道的基因(OsTPC1),它在成熟叶片、茎、根以及培养细胞中普遍表达。OsTPC1挽救了酵母突变体cch1的Ca2+摄取活性和生长速率。为了阐明其生理作用,我们构建了过表达OsTPC1 mRNA的转基因水稻植株和培养细胞。此外,分离出了OsTPC1的反转录转座子(Tos17)插入缺失突变体。过表达OsTPC1的细胞对过量Ca2+表现出超敏性,但在Ca2+限制条件下生长速率更高,而OsTPC1敲除的培养细胞的生长对细胞外游离Ca2+浓度不太敏感,这表明OsTPC1具有跨质膜的Ca2+转运活性。过表达OsTPC1的植株生长减缓,根部出现异常绿化。Ostpc1幼苗在琼脂平板上的生长与对照相当,但在成年植株中显著降低。这些结果表明OsTPC1作为一种Ca2+通透通道参与生长和发育的调控。