Diploid potato (Solanum tuberosum L.) as a model crop to study transgene expression.
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.
Nadolska-Orczyk
Anna
A
Plant Transformation and Cell Engineering Department, Plant Breeding and Acclimatization Institute, Radzików, Błonie, Poland. a.orczyk@ihar.edu.pl
Pietrusinska
Aleksandra
A
Binka-Wyrwa
Agnieszka
A
Kuc
Dominik
D
Orczyk
Wacław
W
D000613:Aminobutyrates / Q000494:pharmacology; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000235:genetics; D004171:Diploidy; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005966:Glucuronidase / Q000378:metabolism; D053209:Herbicide Resistance; D007613:Kanamycin Resistance / Q000187:drug effects; D018515:Plant Leaves / Q000187:drug effects; D018547:Plant Stems / Q000187:drug effects; D030821:Plants, Genetically Modified; D012038:Regeneration / Q000187:drug effects; D011198:Solanum tuberosum / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D014170:Transformation, Genetic / Q000187:drug effects; D019076:Transgenes / Q000235:genetics*
D000613:Aminobutyrates; D004269:DNA, Bacterial; C036483:T-DNA; C003121:phosphinothricin; D005966:Glucuronidase