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A bacterial artificial chromosome ( BAC ) genomic approach reveals partial clustering of the furanocoumarin pathway genes in parsnip
Author(s) -
Roselli Sandro,
Olry Alexandre,
Vautrin Sonia,
Coriton Olivier,
Ritchie Dave,
Galati Gianni,
Navrot Nicolas,
Krieger Célia,
Vialart Guilhem,
Bergès Hélène,
Bourgaud Frédéric,
Hehn Alain
Publication year - 2017
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.13450
Subject(s) - bacterial artificial chromosome , biology , gene , chromosome , furanocoumarin , genetics , circular bacterial chromosome , cluster analysis , genome , botany , computer science , artificial intelligence
Summary Furanocoumarins are specialized metabolites that are involved in the defense of plants against phytophagous insects. The molecular and functional characterization of the genes involved in their biosynthetic pathway is only partially complete. Many recent reports have described gene clusters responsible for the biosynthesis of specialized metabolites in plants. To investigate possible co‐localization of the genes involved in the furanocoumarin pathway, we sequenced parsnip BAC clones spanning two different gene loci. We found that two genes previously identified in this pathway, CYP 71 AJ 3 and CYP 71 AJ 4 , were located on the same BAC , whereas a third gene, Ps PT 1 , belonged to a different BAC clone. Chromosome mapping using fluorescence in situ hybridization ( FISH ) indicated that Ps PT 1 and the CYP 71 AJ 3 ‐ CYP 71 AJ 4 clusters are located on two different chromosomes. Sequencing the BAC clone harboring Ps PT 1 led to the identification of a gene encoding an Fe( II ) α‐ketoglutarate‐dependent dioxygenase (Ps DIOX ) situated in the neighborhood of Ps PT 1 and confirmed the occurrence of a second gene cluster involved in the furanocoumarin pathway. This enzyme metabolizes p ‐coumaroyl CoA, leading exclusively to the synthesis of umbelliferone, an important intermediate compound in furanocoumarin synthesis. This work provides an insight into the genomic organization of genes from the furanocoumarin biosynthesis pathway organized in more than one gene cluster. It also confirms that the screening of a genomic library and the sequencing of BAC clones represent a valuable tool to identify genes involved in biosynthetic pathways dedicated to specialized metabolite synthesis.