Leggewie Georg, Kolbe Anna, Lemoine Rémi, Roessner Ute, Lytovchenko Anna, Zuther Ellen, Kehr Julia, Frommer Wolf B, Riesmeier Jörg W, Willmitzer Lothar, Fernie Alisdair R
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476, Golm, Germany.
Planta. 2003 May;217(1):158-67. doi: 10.1007/s00425-003-0975-x. Epub 2003 Feb 12.
The aim of this work was to examine the consequences of the heterologous expression of a spinach ( Spinacia oleracea L.) sucrose transporter ( SoSUT1) in potato ( Solanum tuberosum L.). Many studies have indicated that reduction of the expression of this class of sucrose transporter has deleterious effects on plant growth and development; however, until now the possibility of improving plant performance by enhancing the expression of this sucrose transporter has not been reported. With this intention we constructed a chimeric construct in which SoSUT1 was cloned in-frame with the myc epitope. We confirmed that this construct, SoSUT1m, was able to mediate sucrose transport by expression in the yeast strain SUSY7. SoSUT1m was expressed in wild-type potato in the sense orientation under the control of the cauliflower mosaic virus 35S promoter to evaluate the effect of an increased constitutive expression of a class-I sucrose transporter. We confirmed that these plants displayed expression of SoSUT1 at both the transcript and protein level and that microsomal fragments isolated from selected lines had an increased sucrose uptake capacity. Analysis of metabolism of these lines indicated that the leaves were characterised by a reduced sucrose level yet exhibited little change in photosynthetic rate. Furthermore, despite the observed increase in sugar (and reduction in amino acid) levels within the tubers, there was little change in either starch content or tuber yield in the transformants. In summary, the genetic manipulation described in this paper resulted in a shift in carbon partitioning in both leaves and tubers and an increased sucrose uptake rate in plasma-membrane vesicles isolated from these lines, but had little impact on tuber metabolism or morphology.
本研究旨在探究菠菜(Spinacia oleracea L.)蔗糖转运蛋白(SoSUT1)在马铃薯(Solanum tuberosum L.)中的异源表达所产生的影响。许多研究表明,这类蔗糖转运蛋白表达的降低会对植物生长发育产生有害影响;然而,迄今为止,尚未有通过增强该蔗糖转运蛋白的表达来改善植物性能的报道。出于这个目的,我们构建了一个嵌合构建体,其中SoSUT1与myc表位框内克隆。我们证实,这个构建体SoSUT1m能够通过在酵母菌株SUSY7中的表达来介导蔗糖转运。SoSUT1m在花椰菜花叶病毒35S启动子的控制下,以正义方向在野生型马铃薯中表达,以评估I类蔗糖转运蛋白组成型表达增加的效果。我们证实,这些植物在转录本和蛋白质水平上均显示出SoSUT1的表达,并且从选定株系中分离出的微粒体片段具有增加的蔗糖摄取能力。对这些株系的代谢分析表明,叶片的特征是蔗糖水平降低,但光合速率变化不大。此外,尽管观察到块茎内糖水平升高(氨基酸水平降低),但转化体的淀粉含量和块茎产量均变化不大。总之,本文所述的基因操作导致了叶片和块茎中碳分配的改变,以及从这些株系中分离出的质膜囊泡中蔗糖摄取速率的增加,但对块茎代谢或形态影响不大。