Targeted gene suppression by RNA interference: An efficient method for production of high-amylose potato lines

Research output: Contribution to journalArticle

Abstract

Production of high-amylose potato lines can be achieved by inhibition of two genes coding for starch branching enzymes. The use of antisense technology for gene inhibition have yielded a low frequency of high-amylose lines that mostly was correlated with high numbers of integrated T-DNA copies. To investigate whether the production of high-amylose lines could be improved, RNA interference was used for gene inhibition of the genes Sbe1 and Sbe2. Two constructs with 100 bp segments (pHAS2) or 200 bp segments (pHAS3) of both branching enzyme genes were cloned as inverted repeats controlled by a potato granule-bound starch synthase promoter. The construct pHAS3 was shown to be very efficient, yielding high-amylose quality in more than 50% of the transgenic lines. An antisense construct, included in the study as a comparator, resulted in only 3% of the transgenic lines being of high-amylose type. Noticeable was also that pHAS3 yielded low T-DNA copy inserts with an average of 83% of backbone-free transgenic lines being single copy events.

Details

Authors
  • Mariette Andersson
  • M Melander
  • P Pojmark
  • H Larsson
  • Leif Bülow
  • P Hofvander
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology

Keywords

  • gene silencing, Solanum tuberosum, plant biotechnology, transformation, plant, transgenic, RNA interference, high-amylose starch
Original languageEnglish
Pages (from-to)137-148
JournalJournal of Biotechnology
Volume123
Issue number2
Publication statusPublished - 2006
Publication categoryResearch
Peer-reviewedYes