Diploid potato ( Solanum tuberosum L.) as a model crop to study transgene expression

Cellular & Molecular Biology Letters

Wacław Orczyk¹  Aleksandra Pietrusinska¹  Dominik Kuc¹  Agnieszka Binka-Wyrwa¹  Anna Nadolska-Orczyk¹ 

[1] Plant Transformation and Cell Engineering Department, Plant Breeding and Acclimatization Institute, Radzików, Błonie, Poland$$

关键词: Binary vectors;    Gene expression;    Genetic transformation;    Polyploid;   

DOI  :  10.2478/s11658-006-0064-9

学科分类：分子生物学，细胞生物学和基因

来源: Uniwersytet Wroclawski * Wydzial Biotechnologii / University of Wroclaw, Faculty of Biotechnology

【 摘 要 】

This paper presents a method of Agrobacterium-mediated transformation for two diploid breeding lines of potato, and gives a detailed analysis of reporter gene expression. In our lab, these lines were also used to obtain tetraploid somatic hybrids. We tested four newly prepared constructs based on the pGreen vector system containing the selection gene nptII or bar under the 35S or nos promoter. All these vectors carried gus under 35S. We also tested the pDM805 vector, with the bar and gus genes respectively under the Ubi1 and Act1 promoters, which are strong for monocots. The selection efficiency (about 17%) was highest in the stem and leaf explants after transformation with pGreen where nptII was under 35S. About half of the selected plants were confirmed via PCR and Southern blot analysis to be transgenic and, depending on the combination, 0 to 100% showed GUS expression. GUS expression was strongest in multi-copy transgenic plants where gus was under Act1. The same potato lines carrying multi-copy bar under Ubi1 were also highly resistant to the herbicide Basta. The suggestion of using Agrobacterium-mediated transformation of diploid lines of potato as a model crop is discussed herein.

【 授权许可】

Unknown