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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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Linkage groups (LG) 5 and 9 in globe artichoke maps. LG5 (a) and LG9 (b) of the globe artichoke varietal types 'Romanesco C3' (female parent, yellow LGs on the left) and 'Spinoso di Palermo' (male parent, blue LGs on the right). LGs with HCT and HQT genes are reported in gray boxes, intercross markers are shown in bold and are connected by a solid line. The LGs previously reported by Lanteri et al. [36] are presented to one side, and changed marker orders are indicated by dotted lines. Asterisks indicate markers showing significant levels of segregation distortion (*: 0.1 > P ≥ 0.05, **: 0.05 > P ≥ 0.01).
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Figure 7: Linkage groups (LG) 5 and 9 in globe artichoke maps. LG5 (a) and LG9 (b) of the globe artichoke varietal types 'Romanesco C3' (female parent, yellow LGs on the left) and 'Spinoso di Palermo' (male parent, blue LGs on the right). LGs with HCT and HQT genes are reported in gray boxes, intercross markers are shown in bold and are connected by a solid line. The LGs previously reported by Lanteri et al. [36] are presented to one side, and changed marker orders are indicated by dotted lines. Asterisks indicate markers showing significant levels of segregation distortion (*: 0.1 > P ≥ 0.05, **: 0.05 > P ≥ 0.01).

Mentions: Two single nucleotide polymorphisms (SNPs, HQTsnp359 and HCTsnp97) were identified (Fig. 5) in the HQT and HCT parental sequences. Both parents of the mapping population were heterozygous for marker HQT-snp359 (parental genotypes ab × ab), that segregated in the ratio 1:2:1 (aa:ab:bb) in the F1 individuals, with no evidence of any segregation distortion (Table 1, Fig. 6). This allowed the HQT gene to be placed on linkage group (LG) 5 in both the female and male maps (Fig. 7a). A further 14 markers were assigned to the female LG5: four microsatellites (CELMS-24, -36, -44 and CMAL-24), three S-SAPs (cyre5 markers) and 7 AFLPs, covering 62.1 cM and a mean inter-marker distance of 4.4 cM. More than 70% of intervals are < 4 cM in genetic length, with four gaps of > 6 cM. In addition to the HQT locus, the male LG5 consists of 15 markers: two SSRs (CELMS-24 and CMAL-24) one S-SAP, one M-AFLP (polyGA marker) and 11 AFLPs, spanning 69.5 cM and a mean inter-marker distance of 4.4 cM (range 1.6–7.7). Seven markers (including HQT-snp359) were shared between the parents, allowing the alignment of their LG5. The HQT locus maps close to AFLP markers e38/m47-01 and e47/m49-06 in the female map, and to the M-AFLP marker polyGA/e33-02 and the microsatellite CELMS-24 in the male map (Fig. 7a).


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)

Linkage groups (LG) 5 and 9 in globe artichoke maps. LG5 (a) and LG9 (b) of the globe artichoke varietal types 'Romanesco C3' (female parent, yellow LGs on the left) and 'Spinoso di Palermo' (male parent, blue LGs on the right). LGs with HCT and HQT genes are reported in gray boxes, intercross markers are shown in bold and are connected by a solid line. The LGs previously reported by Lanteri et al. [36] are presented to one side, and changed marker orders are indicated by dotted lines. Asterisks indicate markers showing significant levels of segregation distortion (*: 0.1 > P ≥ 0.05, **: 0.05 > P ≥ 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2664813&req=5

Figure 7: Linkage groups (LG) 5 and 9 in globe artichoke maps. LG5 (a) and LG9 (b) of the globe artichoke varietal types 'Romanesco C3' (female parent, yellow LGs on the left) and 'Spinoso di Palermo' (male parent, blue LGs on the right). LGs with HCT and HQT genes are reported in gray boxes, intercross markers are shown in bold and are connected by a solid line. The LGs previously reported by Lanteri et al. [36] are presented to one side, and changed marker orders are indicated by dotted lines. Asterisks indicate markers showing significant levels of segregation distortion (*: 0.1 > P ≥ 0.05, **: 0.05 > P ≥ 0.01).
Mentions: Two single nucleotide polymorphisms (SNPs, HQTsnp359 and HCTsnp97) were identified (Fig. 5) in the HQT and HCT parental sequences. Both parents of the mapping population were heterozygous for marker HQT-snp359 (parental genotypes ab × ab), that segregated in the ratio 1:2:1 (aa:ab:bb) in the F1 individuals, with no evidence of any segregation distortion (Table 1, Fig. 6). This allowed the HQT gene to be placed on linkage group (LG) 5 in both the female and male maps (Fig. 7a). A further 14 markers were assigned to the female LG5: four microsatellites (CELMS-24, -36, -44 and CMAL-24), three S-SAPs (cyre5 markers) and 7 AFLPs, covering 62.1 cM and a mean inter-marker distance of 4.4 cM. More than 70% of intervals are < 4 cM in genetic length, with four gaps of > 6 cM. In addition to the HQT locus, the male LG5 consists of 15 markers: two SSRs (CELMS-24 and CMAL-24) one S-SAP, one M-AFLP (polyGA marker) and 11 AFLPs, spanning 69.5 cM and a mean inter-marker distance of 4.4 cM (range 1.6–7.7). Seven markers (including HQT-snp359) were shared between the parents, allowing the alignment of their LG5. The HQT locus maps close to AFLP markers e38/m47-01 and e47/m49-06 in the female map, and to the M-AFLP marker polyGA/e33-02 and the microsatellite CELMS-24 in the male map (Fig. 7a).

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