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番茄磷转运蛋白的功能分析及细胞特异性表达

Functional analysis and cell-specific expression of a phosphate transporter from tomato.

作者信息

Daram P, Brunner S, Persson B L, Amrhein N, Bucher M

机构信息

Federal Institute of Technology (ETH) Zurich, Institute of Plant Sciences, Lindau, Switzerland.

出版信息

Planta. 1998 Oct;206(2):225-33. doi: 10.1007/s004250050394.

Abstract

For a better understanding of the molecular and biochemical processes involved in orthophosphate (Pi) uptake at the root/soil interface, we cloned a Pi-transporter c DNA (LePT1) from a root air-specific cDNA library of tomato (Lycopersicon esculentum Mill.). The corresponding protein belongs to the growing family of ion transporters with twelve putative transmembrane domains. It is highly homologous to recently isolated Pi transporters from higher plants, yeast and fungi. When expressed in a Pi-uptake-deficient yeast mutant, the L. esculentum phosphate transporter 1 (LePT1) protein exhibits an apparent Km of 31 MicroM. The transporter is still active at submicromolar Pi concentrations and mediates highest Pi uptake at pH 5. The activity of LePT1 is dependent on the electrochemical membrane potential mediated by the yeast P-type H + - ATPase. Transcript levels of LePT1 in tomato seedlings are detectable in all vegetative organs under Pi-sufficient conditions, with highest concentrations in root hairs. In situ hybridization studies demonstrate cell-specific expression of LePT1 in the tomato root. The LePT1 mRNA is detectable in peripheral cell layers such as rhizodermal and root cap cells. Under Pi-deprivation condition, mRNA levels are also detectable in young stelar tissue. This work presents molecular and biochemical evidence for distinct root cells playing an important role in Pi acquisition at the root/soil interface.

摘要

为了更好地理解根/土界面正磷酸盐(Pi)吸收过程中涉及的分子和生化过程,我们从番茄(Lycopersicon esculentum Mill.)根特异cDNA文库中克隆了一个Pi转运蛋白cDNA(LePT1)。相应的蛋白质属于具有12个推定跨膜结构域的离子转运蛋白家族,该家族仍在不断增加。它与最近从高等植物、酵母和真菌中分离出的Pi转运蛋白高度同源。当在Pi吸收缺陷型酵母突变体中表达时,番茄磷酸转运蛋白1(LePT1)蛋白的表观Km值为31微摩尔。该转运蛋白在亚微摩尔Pi浓度下仍具有活性,在pH 5时介导最高的Pi吸收。LePT1的活性依赖于酵母P型H⁺-ATP酶介导的电化学膜电位。在Pi充足条件下,番茄幼苗中LePT1的转录水平在所有营养器官中均可检测到,在根毛中的浓度最高。原位杂交研究表明LePT1在番茄根中具有细胞特异性表达。LePT1 mRNA在根表皮和根冠细胞等外周细胞层中可检测到。在Pi缺乏条件下,在幼嫩的中柱组织中也可检测到mRNA水平。这项工作提供了分子和生化证据,证明不同的根细胞在根/土界面Pi获取中起重要作用。

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