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The isolation and mapping of a novel hydroxycinnamoyltransferase in the globe artichoke chlorogenic acid pathway.

Comino C, Hehn A, Moglia A, Menin B, Bourgaud F, Lanteri S, Portis E - BMC Plant Biol. (2009)

Bottom Line: The leaves of globe artichoke and cultivated cardoon (Cynara cardunculus L.) have significant pharmaceutical properties, which mainly result from their high content of polyphenolic compounds such as monocaffeoylquinic and dicaffeoylquinic acid (DCQ), and a range of flavonoid compounds.A novel acyltransferase involved in the biosynthesis of CGA in globe artichoke has been isolated, characterized and mapped.This is a good basis for our effort to understand the genetic basis of phenylpropanoid (PP) biosynthesis in C. cardunculus.

View Article: PubMed Central - HTML - PubMed

Affiliation: DiVaPRA Plant Genetics and Breeding, University of Torino, Grugliasco (Torino), Italy. cinzia.comino@unito.it

ABSTRACT

Background: The leaves of globe artichoke and cultivated cardoon (Cynara cardunculus L.) have significant pharmaceutical properties, which mainly result from their high content of polyphenolic compounds such as monocaffeoylquinic and dicaffeoylquinic acid (DCQ), and a range of flavonoid compounds.

Results: Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyltransferase (HQT) encoding genes have been isolated from both globe artichoke and cultivated cardoon (GenBank accessions DQ915589 and DQ915590, respectively) using CODEHOP and PCR-RACE. A phylogenetic analysis revealed that their sequences belong to one of the major acyltransferase groups (anthranilate N-hydroxycinnamoyl/benzoyltransferase). The heterologous expression of globe artichoke HQT in E. coli showed that this enzyme can catalyze the esterification of quinic acid with caffeoyl-CoA or p-coumaroyl-CoA to generate, respectively, chlorogenic acid (CGA) and p-coumaroyl quinate. Real time PCR experiments demonstrated an increase in the expression level of HQT in UV-C treated leaves, and established a correlation between the synthesis of phenolic acids and protection against damage due to abiotic stress. The HQT gene, together with a gene encoding hydroxycinnamoyl-CoA:shikimate/quinate hydroxycinnamoyltransferase (HCT) previously isolated from globe artichoke, have been incorporated within the developing globe artichoke linkage maps.

Conclusion: A novel acyltransferase involved in the biosynthesis of CGA in globe artichoke has been isolated, characterized and mapped. This is a good basis for our effort to understand the genetic basis of phenylpropanoid (PP) biosynthesis in C. cardunculus.

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Related in: MedlinePlus

Phylogenetic analysis of acyltransferases. The tree was constructed by the neighbour-joining method with 10000 bootstrap replicates. The length of the lines indicates the relative distances between nodes. Protein sequences used for the alignment are: Dc_HCBT, anthranilate N-hydroxycinnamoyl/benzoyltransferase from Dianthus caryophyllus (CAB06427); Ib_HCBT, N-hydroxycinnamoyl/benzoyltransferase from Ipomoea batatas (BAA87043); At_HCT, shikimate/quinate hydroxycinnamoyltransferase from Arabidopsis. thaliana (ABH04595); Nt_HCT, shikimate/quinate hydroxycinnamoyltransferase from Nicotiana tabacum (CAD47830); Nt_HQT, hydroxycinnamoyl CoA quinate transferase from Nicotiana tabacum (CAE46932); Le_HQT, hydroxycinnamoyl CoA quinate transferase from Lycopersicum esculentum (CAE46933); Cca_HQT, hydroxycinnamoyl CoA quinate transferase from Coffea canephora (ABO77956); Cca_HCT, shikimate/quinate hydroxycinnamoyltransferase from Coffea canephora (ABO47805); NP_179497 and NP_200592, Arabidopsis thaliana genes encoding putative acyltransferases; Cc_(artichoke)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. scolymus (AAZ80046); Cc_(cardoon)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. altilis (AAZ80047); Cc_(artichoke)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. scolymus (ABK79689, this work) and Cc_(cardoon)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. altilis (ABK79690, this work).
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Figure 3: Phylogenetic analysis of acyltransferases. The tree was constructed by the neighbour-joining method with 10000 bootstrap replicates. The length of the lines indicates the relative distances between nodes. Protein sequences used for the alignment are: Dc_HCBT, anthranilate N-hydroxycinnamoyl/benzoyltransferase from Dianthus caryophyllus (CAB06427); Ib_HCBT, N-hydroxycinnamoyl/benzoyltransferase from Ipomoea batatas (BAA87043); At_HCT, shikimate/quinate hydroxycinnamoyltransferase from Arabidopsis. thaliana (ABH04595); Nt_HCT, shikimate/quinate hydroxycinnamoyltransferase from Nicotiana tabacum (CAD47830); Nt_HQT, hydroxycinnamoyl CoA quinate transferase from Nicotiana tabacum (CAE46932); Le_HQT, hydroxycinnamoyl CoA quinate transferase from Lycopersicum esculentum (CAE46933); Cca_HQT, hydroxycinnamoyl CoA quinate transferase from Coffea canephora (ABO77956); Cca_HCT, shikimate/quinate hydroxycinnamoyltransferase from Coffea canephora (ABO47805); NP_179497 and NP_200592, Arabidopsis thaliana genes encoding putative acyltransferases; Cc_(artichoke)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. scolymus (AAZ80046); Cc_(cardoon)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. altilis (AAZ80047); Cc_(artichoke)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. scolymus (ABK79689, this work) and Cc_(cardoon)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. altilis (ABK79690, this work).

Mentions: CODEHOP [37] was used to target conserved acyltransferases in globe artichoke (see arrows in Fig. 2), resulting in the amplification of an incomplete internal acyltransferase-like sequence, which was extended in both globe artichoke and cultivated cardoon via a RACE strategy. Full-length cDNA sequences have been deposited in Genbank (DQ915589, DQ915590). The genes are of identical length (1335 bp) and their translation product is a 444 residue peptide with a molecular mass of ~50 kDa. The best matches obtained from a local alignment search within a non-redundant protein database (blastp) were with a sweet potato HCBT (70% identity, 85% similarity), and a tobacco HQT (72% identity, 84% similarity), which belongs to a multifunctional superfamily of plant acyltransferases [38]. The sequences contain a HTLSD peptide (aa 163–168, black boxes in Fig. 2), as does the HCT isolated by Comino et al. [32], matching the highly conserved HXXXD motif characteristic for acyl transfer proteins. The DFGWG block [38,39] present in other acyltransferases of the BAHD family [40] is present from aa 391 to 395 (Fig. 2, black boxes). Phylogenetic analyses confirmed that the isolated sequence showed a high degree of similarity with other already isolated HQT sequences [25,41] and with HCTs from globe artichoke [32], coffee [41], tobacco and Arabidopsis [24] (Fig. 3).


The isolation and mapping of a novel hydroxycinnamoyltransferase in the globe artichoke chlorogenic acid pathway.

Comino C, Hehn A, Moglia A, Menin B, Bourgaud F, Lanteri S, Portis E - BMC Plant Biol. (2009)

Phylogenetic analysis of acyltransferases. The tree was constructed by the neighbour-joining method with 10000 bootstrap replicates. The length of the lines indicates the relative distances between nodes. Protein sequences used for the alignment are: Dc_HCBT, anthranilate N-hydroxycinnamoyl/benzoyltransferase from Dianthus caryophyllus (CAB06427); Ib_HCBT, N-hydroxycinnamoyl/benzoyltransferase from Ipomoea batatas (BAA87043); At_HCT, shikimate/quinate hydroxycinnamoyltransferase from Arabidopsis. thaliana (ABH04595); Nt_HCT, shikimate/quinate hydroxycinnamoyltransferase from Nicotiana tabacum (CAD47830); Nt_HQT, hydroxycinnamoyl CoA quinate transferase from Nicotiana tabacum (CAE46932); Le_HQT, hydroxycinnamoyl CoA quinate transferase from Lycopersicum esculentum (CAE46933); Cca_HQT, hydroxycinnamoyl CoA quinate transferase from Coffea canephora (ABO77956); Cca_HCT, shikimate/quinate hydroxycinnamoyltransferase from Coffea canephora (ABO47805); NP_179497 and NP_200592, Arabidopsis thaliana genes encoding putative acyltransferases; Cc_(artichoke)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. scolymus (AAZ80046); Cc_(cardoon)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. altilis (AAZ80047); Cc_(artichoke)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. scolymus (ABK79689, this work) and Cc_(cardoon)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. altilis (ABK79690, this work).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 3: Phylogenetic analysis of acyltransferases. The tree was constructed by the neighbour-joining method with 10000 bootstrap replicates. The length of the lines indicates the relative distances between nodes. Protein sequences used for the alignment are: Dc_HCBT, anthranilate N-hydroxycinnamoyl/benzoyltransferase from Dianthus caryophyllus (CAB06427); Ib_HCBT, N-hydroxycinnamoyl/benzoyltransferase from Ipomoea batatas (BAA87043); At_HCT, shikimate/quinate hydroxycinnamoyltransferase from Arabidopsis. thaliana (ABH04595); Nt_HCT, shikimate/quinate hydroxycinnamoyltransferase from Nicotiana tabacum (CAD47830); Nt_HQT, hydroxycinnamoyl CoA quinate transferase from Nicotiana tabacum (CAE46932); Le_HQT, hydroxycinnamoyl CoA quinate transferase from Lycopersicum esculentum (CAE46933); Cca_HQT, hydroxycinnamoyl CoA quinate transferase from Coffea canephora (ABO77956); Cca_HCT, shikimate/quinate hydroxycinnamoyltransferase from Coffea canephora (ABO47805); NP_179497 and NP_200592, Arabidopsis thaliana genes encoding putative acyltransferases; Cc_(artichoke)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. scolymus (AAZ80046); Cc_(cardoon)_HCT, hydroxycinnamoyl CoA quinate transferase from Cynara cardunculus var. altilis (AAZ80047); Cc_(artichoke)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. scolymus (ABK79689, this work) and Cc_(cardoon)_HQT, quinate hydroxycinnamoyltransferase from Cynara cardunculus var. altilis (ABK79690, this work).
Mentions: CODEHOP [37] was used to target conserved acyltransferases in globe artichoke (see arrows in Fig. 2), resulting in the amplification of an incomplete internal acyltransferase-like sequence, which was extended in both globe artichoke and cultivated cardoon via a RACE strategy. Full-length cDNA sequences have been deposited in Genbank (DQ915589, DQ915590). The genes are of identical length (1335 bp) and their translation product is a 444 residue peptide with a molecular mass of ~50 kDa. The best matches obtained from a local alignment search within a non-redundant protein database (blastp) were with a sweet potato HCBT (70% identity, 85% similarity), and a tobacco HQT (72% identity, 84% similarity), which belongs to a multifunctional superfamily of plant acyltransferases [38]. The sequences contain a HTLSD peptide (aa 163–168, black boxes in Fig. 2), as does the HCT isolated by Comino et al. [32], matching the highly conserved HXXXD motif characteristic for acyl transfer proteins. The DFGWG block [38,39] present in other acyltransferases of the BAHD family [40] is present from aa 391 to 395 (Fig. 2, black boxes). Phylogenetic analyses confirmed that the isolated sequence showed a high degree of similarity with other already isolated HQT sequences [25,41] and with HCTs from globe artichoke [32], coffee [41], tobacco and Arabidopsis [24] (Fig. 3).

Bottom Line: The leaves of globe artichoke and cultivated cardoon (Cynara cardunculus L.) have significant pharmaceutical properties, which mainly result from their high content of polyphenolic compounds such as monocaffeoylquinic and dicaffeoylquinic acid (DCQ), and a range of flavonoid compounds.A novel acyltransferase involved in the biosynthesis of CGA in globe artichoke has been isolated, characterized and mapped.This is a good basis for our effort to understand the genetic basis of phenylpropanoid (PP) biosynthesis in C. cardunculus.

View Article: PubMed Central - HTML - PubMed

Affiliation: DiVaPRA Plant Genetics and Breeding, University of Torino, Grugliasco (Torino), Italy. cinzia.comino@unito.it

ABSTRACT

Background: The leaves of globe artichoke and cultivated cardoon (Cynara cardunculus L.) have significant pharmaceutical properties, which mainly result from their high content of polyphenolic compounds such as monocaffeoylquinic and dicaffeoylquinic acid (DCQ), and a range of flavonoid compounds.

Results: Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyltransferase (HQT) encoding genes have been isolated from both globe artichoke and cultivated cardoon (GenBank accessions DQ915589 and DQ915590, respectively) using CODEHOP and PCR-RACE. A phylogenetic analysis revealed that their sequences belong to one of the major acyltransferase groups (anthranilate N-hydroxycinnamoyl/benzoyltransferase). The heterologous expression of globe artichoke HQT in E. coli showed that this enzyme can catalyze the esterification of quinic acid with caffeoyl-CoA or p-coumaroyl-CoA to generate, respectively, chlorogenic acid (CGA) and p-coumaroyl quinate. Real time PCR experiments demonstrated an increase in the expression level of HQT in UV-C treated leaves, and established a correlation between the synthesis of phenolic acids and protection against damage due to abiotic stress. The HQT gene, together with a gene encoding hydroxycinnamoyl-CoA:shikimate/quinate hydroxycinnamoyltransferase (HCT) previously isolated from globe artichoke, have been incorporated within the developing globe artichoke linkage maps.

Conclusion: A novel acyltransferase involved in the biosynthesis of CGA in globe artichoke has been isolated, characterized and mapped. This is a good basis for our effort to understand the genetic basis of phenylpropanoid (PP) biosynthesis in C. cardunculus.

Show MeSH
Related in: MedlinePlus