Coleman Heather D, Ellis Dave D, Gilbert Margarita, Mansfield Shawn D
Department of Wood Science, University of British Columbia, 2424 Main Mall, Vancouver, BC, V6T 1Z4, Canada.
Plant Biotechnol J. 2006 Jan;4(1):87-101. doi: 10.1111/j.1467-7652.2005.00160.x.
The effects of the overexpression of sucrose synthase (SuSy) and UDP-glucose pyrophosphorylase (UGPase) on plant growth and metabolism were evaluated in tobacco (Nicotiana tabacum cv. Xanthi). T(1) transgenic plants expressing either gene under the control of a tandem repeat cauliflower mosaic virus 35S promoter (2x35S) or a xylem-localized 4CL promoter (4-coumarate:CoA ligase; 4CL) were generated, and reciprocally crossed to generate plants expressing both genes. Transcript levels, enzyme activity, growth parameters, fibre properties and carbohydrate content of stem tissue were quantified. The expression profiles of both genes confirmed the expression pattern of the promoters: 2x35S expressed more strongly in leaves, while 4CL expression was highest in stem tissue. In-depth plant characterization revealed that the single-transgene lines showed significant increases in the height growth compared with corresponding control lines. The double-transgene plants demonstrated an additive effect, proving to be even taller than the single-transgene parents. Several of these lines had associated increases in soluble sugar content. Although partitioning of storage carbohydrates into starch or cellulose was not observed, the increased height growth and increases in soluble carbohydrates suggest a role for SuSy as a marker in sink strength and lend credit to the function of UGPase in a similar role. The up-regulation of these two genes, although not increasing the percentage cellulose content, was effective in increasing the total biomass, and thus the overall cellulose yield, from a given plant.
在烟草(烟草品种Xanthi)中评估了蔗糖合酶(SuSy)和UDP-葡萄糖焦磷酸化酶(UGPase)过表达对植物生长和代谢的影响。构建了在串联重复花椰菜花叶病毒35S启动子(2x35S)或木质部定位的4CL启动子(4-香豆酸:辅酶A连接酶;4CL)控制下表达任一基因的T(1)转基因植株,并进行正反交以产生同时表达这两个基因的植株。对茎组织的转录水平、酶活性、生长参数、纤维特性和碳水化合物含量进行了定量分析。两个基因的表达谱证实了启动子的表达模式:2x35S在叶片中表达更强,而4CL在茎组织中表达最高。深入的植物特性分析表明,与相应的对照品系相比,单转基因品系的株高生长显著增加。双转基因植株表现出累加效应,比单转基因亲本更高。其中一些品系的可溶性糖含量也相应增加。虽然未观察到储存碳水化合物向淀粉或纤维素的分配,但株高生长增加和可溶性碳水化合物增加表明SuSy作为库强度标记物的作用,也证明了UGPase在类似作用中的功能。这两个基因的上调虽然没有增加纤维素含量的百分比,但有效地增加了给定植物的总生物量,从而提高了整体纤维素产量。