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BMC Plant Biology

, 14:280

Genetics and crop biotechnology


BackgroundScopoletin and its glucoside scopolin are important secondary metabolites synthesized in plants as a defense mechanism against various environmental stresses. They belong to coumarins, a class of phytochemicals with significant biological activities that is widely used in medical application and cosmetics industry. Although numerous studies showed that a variety of coumarins occurs naturally in several plant species, the details of coumarins biosynthesis and its regulation is not well understood. It was shown previously that coumarins predominantly scopolin and scopoletin occur in Arabidopsis thaliana Arabidopsis roots, but until now nothing is known about natural variation of their accumulation in this model plant. Therefore, the genetic architecture of coumarins biosynthesis in Arabidopsis has not been studied before.

ResultsHere, the variation in scopolin and scopoletin content was assessed by comparing seven Arabidopsis accessions. Subsequently, a quantitative trait locus QTL mapping was performed with an Advanced Intercross Recombinant Inbred Lines AI-RILs mapping population EstC Est-1 × Col. In order to reveal the genetic basis of both scopolin and scopoletin biosynthesis, two sets of methanol extracts were made from Arabidopsis roots and one set was additionally subjected to enzymatic hydrolysis prior to quantification done by high-performance liquid chromatography HPLC. We identified one QTL for scopolin and five QTLs for scopoletin accumulation. The identified QTLs explained 13.86% and 37.60% of the observed phenotypic variation in scopolin and scopoletin content, respectively. In silico analysis of genes located in the associated QTL intervals identified a number of possible candidate genes involved in coumarins biosynthesis.

ConclusionsTogether, our results demonstrate for the first time that Arabidopsis is an excellent model for studying the genetic and molecular basis of natural variation in coumarins biosynthesis in plants. It additionally provides a basis for fine mapping and cloning of the genes involved in scopolin and scopoletin biosynthesis. Importantly, we have identified new loci for this biosynthetic process.

KeywordsCoumarins Natural variation Plant-environment interaction Scopoletin Scopolin Secondary metabolism QTL mapping Abbreviations2OGD2-oxoglutarate-dependent dioxygenase

4CL14-coumarate:CoA ligase 1

4CL24-coumarate:CoA ligase 2

4CL34-coumarate:CoA ligase 3

4CL54-coumarate:CoA ligase 5

ACOS5Acyl-CoA synthetase 5

AI-RILsAdvanced intercross recombinant inbred lines

C3Hp-coumaroyl 3-hydroxylase

CCoAOMT1Caffeoyl coenzyme A dependent O-methyltransferase 1

CCoAOMT7Caffeoyl coenzyme A dependent O-methyltransferase 7

CYPCytochrome P450 superfamily of monooxygenases

F6-H1Feruloyl-CoA 6-hydroxylase 1

F6-H2Feruloyl-CoA 6-hydroxylase 2

GC-MSGas Chromatography-Mass Spectrometry

HCTShikimate O-hydroxycinnamoyltransferase

HPLCHigh-performance liquid chromatography

LODLogarithm of odds

MSMurashige and Skoog medium

MQMMultiple QTL mapping

MYBSuperfamily of transcription factors

NASCNottingham Arabidopsis stock centre

OMT1Caffeate O-methyltransferase 1

TSM1Tapetum-specific O- methyltransferase

PVEPhenotypic variance explained

SIMSimple interval mapping

QTLQuantitative trait loci

WRKYSuperfamily of transcription factors

Electronic supplementary materialThe online version of this article doi:10.1186-s12870-014-0280-9 contains supplementary material, which is available to authorized users.

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Autor: Joanna Siwinska - Leszek Kadzinski - Rafal Banasiuk - Anna Gwizdek-Wisniewska - Alexandre Olry - Bogdan Banecki - Ewa Lojko


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