Tegeder M, Wang X D, Frommer W B, Offler C E, Patrick J W
Department of Biological Sciences, University of Newcastle, NSW, Australia.
Plant J. 1999 Apr;18(2):151-61. doi: 10.1046/j.1365-313x.1999.00439.x.
The anatomy of developing pea seeds is characterized by transfer cells present in both coats and cotyledons at the maternal/filial interface. To determine the nature and cellular localization of sucrose transporters in pea seeds, a full-length clone of a sucrose/H+ symporter (PsSUT1) was isolated from a cotyledon cDNA library. Northern blot analyses of different organs showed that PsSUT1 is expressed in non-seed tissues, including sucrose sinks and sources. Within developing seeds, transcripts of PsSUT1 and PsAHA1 genes were detected in all tissues, while transcripts of a sucrose binding protein (GmSBP) were confined to cotyledon epidermal transfer cells. Signal intensities of PsSUT1 and PsAHA1 transcripts and protein products were most pronounced in the thin-walled parenchyma cells of seed coats and epidermal transfer cells of cotyledons. For cotyledons, the highest transporter densities were localized to those portions of plasma membranes lining the wall ingrowth regions of epidermal transfer cells. Responses of [14C]sucrose influx to metabolic inhibitors indicated that proton-coupled sucrose transport was operative in both seed coats and cotyledons. Cotyledon epidermal transfer cells were shown to support the highest sucrose flux. Maximal transport activity was found to account for the sucrose flux differences between seed tissues. Intercellular movement of the symplasmic tracer, 5-(6)-carboxyfluorescein (CF), demonstrated that symplasmic pathways interconnect the vascular tissues to thin-walled parenchyma transfer cells of seed coats and, for cotyledons, epidermal transfer cells to storage parenchyma cells.
发育中的豌豆种子的解剖结构特点是在母本/子代界面的种皮和子叶中都存在转移细胞。为了确定豌豆种子中蔗糖转运蛋白的性质和细胞定位,从子叶cDNA文库中分离出蔗糖/H⁺同向转运体(PsSUT1)的全长克隆。对不同器官的Northern印迹分析表明,PsSUT1在非种子组织中表达,包括蔗糖库和源。在发育中的种子内,PsSUT1和PsAHA1基因的转录本在所有组织中都能检测到,而蔗糖结合蛋白(GmSBP)的转录本则局限于子叶表皮转移细胞。PsSUT1和PsAHA1转录本及蛋白产物的信号强度在种皮的薄壁实质细胞和子叶的表皮转移细胞中最为明显。对于子叶,最高的转运体密度定位于表皮转移细胞壁内生长区域的质膜部分。[¹⁴C]蔗糖流入对代谢抑制剂的反应表明,质子偶联的蔗糖转运在种皮和子叶中都起作用。子叶表皮转移细胞被证明支持最高的蔗糖通量。发现最大转运活性解释了种子组织之间的蔗糖通量差异。共质体示踪剂5-(6)-羧基荧光素(CF)的细胞间移动表明,共质体途径将维管组织与种皮的薄壁实质转移细胞连接起来,对于子叶来说,将表皮转移细胞与贮藏实质细胞连接起来。