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

Andersson, Mariette LU ; Melander, M ; Pojmark, P ; Larsson, H ; Bülow, Leif LU and Hofvander, P (2006) In Journal of Biotechnology 123(2). p.137-148
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... (More)
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. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
gene silencing, Solanum tuberosum, plant biotechnology, transformation, plant, transgenic, RNA interference, high-amylose starch
in
Journal of Biotechnology
volume
123
issue
2
pages
137 - 148
publisher
Elsevier
external identifiers
  • wos:000237881800002
  • pmid:16466822
  • scopus:33646137372
ISSN
1873-4863
DOI
10.1016/j.jbiotec.2005.11.001
language
English
LU publication?
yes
id
0fe23155-ad7b-475b-a0da-80e11d4b3c66 (old id 408339)
date added to LUP
2016-04-01 12:22:45
date last changed
2021-05-25 04:33:14
@article{0fe23155-ad7b-475b-a0da-80e11d4b3c66,
  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.},
  author       = {Andersson, Mariette and Melander, M and Pojmark, P and Larsson, H and Bülow, Leif and Hofvander, P},
  issn         = {1873-4863},
  language     = {eng},
  number       = {2},
  pages        = {137--148},
  publisher    = {Elsevier},
  series       = {Journal of Biotechnology},
  title        = {Targeted gene suppression by RNA interference: An efficient method for production of high-amylose potato lines},
  url          = {http://dx.doi.org/10.1016/j.jbiotec.2005.11.001},
  doi          = {10.1016/j.jbiotec.2005.11.001},
  volume       = {123},
  year         = {2006},
}