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Molecular dissection of pathway components unravel atisine biosynthesis in a non-toxic Aconitum species, A. heterophyllum Wall

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ABSTRACT

Aconitum heterophyllum is an important component for various herbal drug formulations due to the occurrence of non-toxic aconites including marker compound, atisine. Despite huge pharmacological potential, the reprogramming of aconites production is limited due to lack of understanding on their biosynthesis. To address this problem, we have proposed here the complete atisine biosynthetic pathway for the first time connecting glycolysis, MVA/MEP, serine biosynthesis and diterpene biosynthetic pathways. The transcript profiling revealed phosphorylated pathway as a major contributor towards serine production in addition to repertoire of genes in glycolysis (G6PI, PFK, ALD and ENO), serine biosynthesis (PGDH and PSAT) and diterpene biosynthesis (KO and KH) sharing a similar pattern of expression (2-4-folds) in roots compared to shoots vis-à-vis atisine content (0–0.37 %). Quantification of steviol and comparative analysis of shortlisted genes between roots of high (0.37 %) vs low (0.14 %) atisine content accessions further confirmed the route of atisine biosynthesis. The results showed 6-fold increase in steviol content and 3–62-fold up-regulation of all the selected genes in roots of high content accession ascertaining their association towards atisine production. Moreover, significant positive correlations were observed between selected genes suggesting their co-expression and crucial role in atisine biosynthesis. This study, thus, offers unprecedented opportunities to explore the selected candidate genes for enhanced production of atisine in cultivated plant cells.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-016-0417-7) contains supplementary material, which is available to authorized users.

No MeSH data available.


HPLC analysis of steviol quantification in A. heterophyllum accessions. a HPLC chromatogram and UV spectra of steviol standard, b HPLC chromatogram and UV spectra of high content accession, c HPLC chromatogram and UV spectra of low content accession
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Fig2: HPLC analysis of steviol quantification in A. heterophyllum accessions. a HPLC chromatogram and UV spectra of steviol standard, b HPLC chromatogram and UV spectra of high content accession, c HPLC chromatogram and UV spectra of low content accession

Mentions: Atisine was not detected in the shoots of A. heterophyllum plants (Malhotra et al. 2014). In contrast, the level of atisine in roots of high content accession was 0.37 %, which was 3-fold (p < 0.001) greater compared to that in roots of low content accession (0.14 %) (Malhotra et al. 2014). Further, this study showed that the level of steviol in roots of high content accession was 0.06 %, which was 6.0-fold (p < 0.001) greater as compared to that in roots of low content accession (0.01 %). The HPLC chromatograms and UV spectra of the steviol standard and the samples are provided in Fig. 2. It was evident from the results that steviol could be involved in the biosynthesis of atisine as latter has been found to be biosynthesized and accumulated exclusively in roots of A. heterophyllum.Fig. 2


Molecular dissection of pathway components unravel atisine biosynthesis in a non-toxic Aconitum species, A. heterophyllum Wall
HPLC analysis of steviol quantification in A. heterophyllum accessions. a HPLC chromatogram and UV spectra of steviol standard, b HPLC chromatogram and UV spectra of high content accession, c HPLC chromatogram and UV spectra of low content accession
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835424&req=5

Fig2: HPLC analysis of steviol quantification in A. heterophyllum accessions. a HPLC chromatogram and UV spectra of steviol standard, b HPLC chromatogram and UV spectra of high content accession, c HPLC chromatogram and UV spectra of low content accession
Mentions: Atisine was not detected in the shoots of A. heterophyllum plants (Malhotra et al. 2014). In contrast, the level of atisine in roots of high content accession was 0.37 %, which was 3-fold (p < 0.001) greater compared to that in roots of low content accession (0.14 %) (Malhotra et al. 2014). Further, this study showed that the level of steviol in roots of high content accession was 0.06 %, which was 6.0-fold (p < 0.001) greater as compared to that in roots of low content accession (0.01 %). The HPLC chromatograms and UV spectra of the steviol standard and the samples are provided in Fig. 2. It was evident from the results that steviol could be involved in the biosynthesis of atisine as latter has been found to be biosynthesized and accumulated exclusively in roots of A. heterophyllum.Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

Aconitum heterophyllum is an important component for various herbal drug formulations due to the occurrence of non-toxic aconites including marker compound, atisine. Despite huge pharmacological potential, the reprogramming of aconites production is limited due to lack of understanding on their biosynthesis. To address this problem, we have proposed here the complete atisine biosynthetic pathway for the first time connecting glycolysis, MVA/MEP, serine biosynthesis and diterpene biosynthetic pathways. The transcript profiling revealed phosphorylated pathway as a major contributor towards serine production in addition to repertoire of genes in glycolysis (G6PI, PFK, ALD and ENO), serine biosynthesis (PGDH and PSAT) and diterpene biosynthesis (KO and KH) sharing a similar pattern of expression (2-4-folds) in roots compared to shoots vis-&agrave;-vis atisine content (0&ndash;0.37&nbsp;%). Quantification of steviol and comparative analysis of shortlisted genes between roots of high (0.37&nbsp;%) vs low (0.14&nbsp;%) atisine content accessions further confirmed the route of atisine biosynthesis. The results showed 6-fold increase in steviol content and 3&ndash;62-fold up-regulation of all the selected genes in roots of high content accession ascertaining their association towards atisine production. Moreover, significant positive correlations were observed between selected genes suggesting their co-expression and crucial role in atisine biosynthesis. This study, thus, offers unprecedented opportunities to explore the selected candidate genes for enhanced production of atisine in cultivated plant cells.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-016-0417-7) contains supplementary material, which is available to authorized users.

No MeSH data available.