Genetically engineered hairy root cultures of Hyoscyamus senecionis and H. muticus: ploidy as a promising parameter in the metabolic engineering of tropane alkaloids

Esmaeil Dehghan, Darwin W. Reed, Patrick S. Covello, Zeinab Hasanpour, Javier Palazon, Kirsi-Marja Oksman-Caldentey, Farajollah Shahriari Ahmadi

Research output: Contribution to journalArticleScientificpeer-review

4 Citations (Scopus)

Abstract

Key message: Tetraploidy improves overexpression ofh6hand scopolamine production ofH. muticus,while inH. senecionis, pmtoverexpression and elicitation can be used as effective methods for increasing tropane alkaloids. Abstract: The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.
Original languageEnglish
Pages (from-to)1615-1626
Number of pages12
JournalPlant Cell Reports
Volume36
Issue number10
DOIs
Publication statusPublished - 1 Oct 2017
MoE publication typeA1 Journal article-refereed

Fingerprint

Hyoscyamus
Hyoscyamus muticus
tropane alkaloids
metabolic engineering
ploidy
scopolamine
tetraploidy
clones
atropine
polyploidy
tropinone
genetic stability
methyl jasmonate
transgenes
flow cytometry
biochemical pathways
alkaloids
diploidy
genetically modified organisms
biosynthesis

Keywords

  • hairy roots
  • metabolic engineering
  • methyl jasmonate
  • tetraploidy
  • tropane alkaloids

Cite this

Dehghan, Esmaeil ; Reed, Darwin W. ; Covello, Patrick S. ; Hasanpour, Zeinab ; Palazon, Javier ; Oksman-Caldentey, Kirsi-Marja ; Ahmadi, Farajollah Shahriari. / Genetically engineered hairy root cultures of Hyoscyamus senecionis and H. muticus: ploidy as a promising parameter in the metabolic engineering of tropane alkaloids. In: Plant Cell Reports. 2017 ; Vol. 36, No. 10. pp. 1615-1626.
@article{1f325288007e4ed9a8961a67dd74a7d1,
title = "Genetically engineered hairy root cultures of Hyoscyamus senecionis and H. muticus: ploidy as a promising parameter in the metabolic engineering of tropane alkaloids",
abstract = "Key message: Tetraploidy improves overexpression ofh6hand scopolamine production ofH. muticus,while inH. senecionis, pmtoverexpression and elicitation can be used as effective methods for increasing tropane alkaloids. Abstract: The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.",
keywords = "hairy roots, metabolic engineering, methyl jasmonate, tetraploidy, tropane alkaloids",
author = "Esmaeil Dehghan and Reed, {Darwin W.} and Covello, {Patrick S.} and Zeinab Hasanpour and Javier Palazon and Kirsi-Marja Oksman-Caldentey and Ahmadi, {Farajollah Shahriari}",
year = "2017",
month = "10",
day = "1",
doi = "10.1007/s00299-017-2178-0",
language = "English",
volume = "36",
pages = "1615--1626",
journal = "Plant Cell Reports",
issn = "0721-7714",
publisher = "Springer",
number = "10",

}

Genetically engineered hairy root cultures of Hyoscyamus senecionis and H. muticus: ploidy as a promising parameter in the metabolic engineering of tropane alkaloids. / Dehghan, Esmaeil; Reed, Darwin W.; Covello, Patrick S.; Hasanpour, Zeinab; Palazon, Javier; Oksman-Caldentey, Kirsi-Marja; Ahmadi, Farajollah Shahriari.

In: Plant Cell Reports, Vol. 36, No. 10, 01.10.2017, p. 1615-1626.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Genetically engineered hairy root cultures of Hyoscyamus senecionis and H. muticus: ploidy as a promising parameter in the metabolic engineering of tropane alkaloids

AU - Dehghan, Esmaeil

AU - Reed, Darwin W.

AU - Covello, Patrick S.

AU - Hasanpour, Zeinab

AU - Palazon, Javier

AU - Oksman-Caldentey, Kirsi-Marja

AU - Ahmadi, Farajollah Shahriari

PY - 2017/10/1

Y1 - 2017/10/1

N2 - Key message: Tetraploidy improves overexpression ofh6hand scopolamine production ofH. muticus,while inH. senecionis, pmtoverexpression and elicitation can be used as effective methods for increasing tropane alkaloids. Abstract: The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.

AB - Key message: Tetraploidy improves overexpression ofh6hand scopolamine production ofH. muticus,while inH. senecionis, pmtoverexpression and elicitation can be used as effective methods for increasing tropane alkaloids. Abstract: The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.

KW - hairy roots

KW - metabolic engineering

KW - methyl jasmonate

KW - tetraploidy

KW - tropane alkaloids

UR - http://www.scopus.com/inward/record.url?scp=85023756179&partnerID=8YFLogxK

U2 - 10.1007/s00299-017-2178-0

DO - 10.1007/s00299-017-2178-0

M3 - Article

VL - 36

SP - 1615

EP - 1626

JO - Plant Cell Reports

JF - Plant Cell Reports

SN - 0721-7714

IS - 10

ER -