Production of Amylopectin and High- Amylose Starch in Separate Potato Genotypes

Hofvander, P. 2004. Production of amylopectin and high-amylose starch in separate potato genotypes. Doctor ́s dissertation. ISSN 1401-6249, ISBN 91-576-6775-6 Starch is one of the most important processed products from agriculture. Two main outlets can be identified; starch is either enzymatically processed for the production of sweeteners and as raw material for fermentation or channelled to various applications as dry starch. Native or chemically modified starches are utilized in food as well as non-food applications, where the specific physicochemical properties are main determinants for their respective use. Starch consists of two different molecules, amylose and amylopectin. To be able to take the full benefit of the unique properties of either component it is of interest to divide the production of amylose and amylopectin into separate plant genotypes. In the presented work, potatoes producing either amylopectin or high-amylose starch were achieved using genetic modification. For potato transformation a highly efficient protocol was developed for a herbicide selection gene instead of the commonly used nptII antibiotic selection gene. In order to achieve respective starch qualities, the expression of genes important for amylopectin or amylose synthesis was silenced. Antisense technology as well as the expression of dsRNA was investigated where the expression of dsRNA was determined to be at least ten-fold more efficient for gene silencing. An added benefit of dsRNA expression was that a higher fraction of silenced transgenic lines compared to the use antisense were associated with single copy T-DNA integrations. One amylopectin potato line was furthermore characterized regarding genetic and chemical composition. The T-DNA was found integrated as an inverted repeat with the inverted repeat region extending into potato chromosomal DNA. This transgenic locus was found to be more consistent with integration into a doublestranded chromosomal break than insertion by a mechanism nicking one strand of the locus. The high-amylose trait generally resulted in a higher tuber fresh weight yield, much elevated sugar levels and a decreased starch content. Amylose levels were obtained where very limited amounts of material recognizable as amylopectin could be found. The production of amylopectin and amylose was divided into separate genotypes but additional factors are needed to be able to produce amylose at levels comparable to starch contents of cultivated potatoes.