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酵母源转化酶在烟草和拟南芥植物细胞壁中的表达导致碳水化合物积累、光合作用受抑制,并强烈影响转基因烟草植物的生长和表型。

Expression of a yeast-derived invertase in the cell wall of tobacco and Arabidopsis plants leads to accumulation of carbohydrate and inhibition of photosynthesis and strongly influences growth and phenotype of transgenic tobacco plants.

作者信息

von Schaewen A, Stitt M, Schmidt R, Sonnewald U, Willmitzer L

机构信息

Institut für Genbiologische Forschung Berlin GmbH, FRG.

出版信息

EMBO J. 1990 Oct;9(10):3033-44. doi: 10.1002/j.1460-2075.1990.tb07499.x.

Abstract

Chimeric genes consisting of the coding sequence of the yeast invertase gene suc 2 and different N-terminal portions of the potato-derived vacuolar protein proteinase inhibitor II fused to the 35S CaMV promoter and the poly-A site of the octopine synthase gene were transferred into tobacco and Arabidopsis thaliana plants using Agrobacterium based systems. Regenerated transgenic plants display a 50- to 500-fold higher invertase activity compared to non-transformed control plants. This invertase is N-glycosylated and efficiently secreted from the plant cell leading to its apoplastic location. Whereas expression of the invertase does not lead to drastic changes in transgenic Arabidopsis thaliana plants, transgenic tobacco plants show dramatic changes with respect to development and phenotype. Expression of the invertase leads to stunted growth due to reduction of internodal distances, to development of bleached and/or necrotic regions in older leaves and to suppressed root formation. In mature leaves, high levels of soluble sugars and starch accumulate. These carbohydrates do not show a diurnal turnover. The accumulation of carbohydrate is accompanied by an inhibition of photosynthesis, and in tobacco, by an increase in the rate of respiration. Measurements in bleached versus green areas of the same leaf show that the bleached section contains high levels of carbohydrates and has lower photosynthesis and higher respiration than green sections. It is concluded that expression of invertase in the cell wall interrupts export and leads to an accumulation of carbohydrates and inhibition of photosynthesis.

摘要

由酵母转化酶基因suc 2的编码序列与马铃薯来源的液泡蛋白蛋白酶抑制剂II的不同N端部分组成的嵌合基因,与35S CaMV启动子和章鱼碱合酶基因的多聚A位点融合,通过基于农杆菌的系统转入烟草和拟南芥植株中。与未转化的对照植株相比,再生的转基因植株显示出高50至500倍的转化酶活性。这种转化酶进行N-糖基化修饰,并从植物细胞中有效分泌,从而定位于质外体。虽然转化酶的表达在转基因拟南芥植株中不会导致剧烈变化,但转基因烟草植株在发育和表型方面显示出显著变化。转化酶的表达导致节间距离缩短,从而使植株生长受阻,老叶出现白化和/或坏死区域,并且根的形成受到抑制。在成熟叶片中,可溶性糖和淀粉大量积累。这些碳水化合物没有昼夜周转。碳水化合物的积累伴随着光合作用的抑制,在烟草中还伴随着呼吸速率的增加。对同一片叶子的白化区域和绿色区域的测量表明,白化部分含有高水平的碳水化合物,其光合作用低于绿色部分,呼吸作用高于绿色部分。得出的结论是,细胞壁中转化酶的表达会中断输出,导致碳水化合物积累并抑制光合作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14bb/552027/056ba8e94111/emboj00237-0019-a.jpg

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