Obiadalla-Ali Hazem, Fernie Alisdair R, Lytovchenko Anna, Kossmann Jens, Lloyd James R
Willmitzer Department, Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Golm, Germany.
Planta. 2004 Jul;219(3):533-40. doi: 10.1007/s00425-004-1257-y. Epub 2004 Apr 2.
A potato (Solanum tuberosum L.) cDNA coding for the chloroplastic isoform of fructose 1,6-bisphosphatase (cp-FBPase) was utilized to repress its activity in tomatoes (Lycopersicon esculentum Mill.) using antisense techniques. The patatin B33 promoter was used to ensure fruit specificity of the antisense effect. Transgenic plants were isolated in which fructose 1,6-bisphosphatase activity was reduced by more than 50% of the control in green fruits. Immunoblots indicated that the plastidial isoform was almost completely eliminated in the most strongly inhibited lines. Fruits of the transgenic plants were analyzed for levels of metabolites during fruit development. Glucose and fructose concentrations were increased in green fruits in the transgenic lines, but unchanged at later stages of development. The sucrose concentration was low, and was not significantly altered in the transgenic lines. There was net degradation of starch over the developmental period, but the starch content was not decreased. In green fruit the levels of hexose phosphates were unchanged, whilst the level of 3-phosphoglyceric acid was significantly increased in one line. Most importantly the deduced ratio of hexose phosphate to 3-phosphoglyceric acid decreased, consistent with an in vivo inhibition of fructose 1,6-bisphosphatase activity. One consequence of this reduction of in vivo activity of cp-FBPase was that the average weight of fully ripe fruits was significantly decreased by up to 20% in all transgenic lines in comparison with the control.
利用编码果糖1,6 - 二磷酸酶叶绿体同工型(cp - FBPase)的马铃薯(Solanum tuberosum L.)cDNA,采用反义技术抑制其在番茄(Lycopersicon esculentum Mill.)中的活性。使用马铃薯块茎蛋白B33启动子确保反义效应的果实特异性。分离出转基因植株,其绿色果实中果糖1,6 - 二磷酸酶活性比对照降低了50%以上。免疫印迹表明,在抑制最强的株系中,质体同工型几乎完全消失。对转基因植株果实发育过程中的代谢物水平进行了分析。转基因株系绿色果实中的葡萄糖和果糖浓度升高,但在发育后期未发生变化。蔗糖浓度较低,转基因株系中无显著改变。在整个发育时期淀粉有净降解,但淀粉含量未降低。绿色果实中磷酸己糖水平未变,而在一个株系中3 - 磷酸甘油酸水平显著升高。最重要的是,推测的磷酸己糖与3 - 磷酸甘油酸的比值降低,这与体内果糖1,6 - 二磷酸酶活性受到抑制一致。cp - FBPase体内活性降低的一个结果是,与对照相比,所有转基因株系中完全成熟果实的平均重量显著降低了多达20%。