Biosynthesis of the Sex Pheromone Component (E,Z)-7,9-Dodecadienyl Acetate in the European Grapevine Moth, Lobesia botrana, Involving ∆11 Desaturation and an Elusive ∆7 Desaturase

Abstract

The European grapevine moth, Lobesia botrana, uses (E,Z)-7,9-dodecadienyl acetate as its major sex pheromone component. Through in vivo labeling experiments we demonstrated that the doubly unsaturated pheromone component is produced by ∆11 desaturation of tetradecanoic acid, followed by chain shortening of (Z)-11-tetradecenoic acid to (Z)-9-dodecenoic acid, and subsequently introduction of the second double bond by an unknown ∆7 desaturase, before final reduction and acetylation. By sequencing and analyzing the transcriptome of female pheromone glands of L. botrana, we obtained 41 candidate genes that may be involved in sex pheromone production, including the genes encoding 17 fatty acyl desaturases, 13 fatty acyl reductases, 1 fatty acid synthase, 3 acyl-CoA oxidases, 1 acetyl-CoA carboxylase, 4 fatty acid transport proteins and 2 acyl-CoA binding proteins. A functional assay of desaturase and acyl-CoA oxidase gene candidates in yeast and insect cell (Sf9) heterologous expression systems revealed that Lbo_PPTQ encodes a ∆11 desaturase producing (Z)-11-tetradecenoic acid from tetradecanoic acid. Further, Lbo_31670 and Lbo_49602 encode two acyl-CoA oxidases that may produce (Z)-9-dodecenoic acid by chain shortening (Z)-11-tetradecenoic acid. The gene encoding the enzyme introducing the E7 double bond into (Z)-9-dodecenoic acid remains elusive even though we assayed 17 candidate desaturases in the two heterologous systems.

Bioproduction of (Z,E)‐9,12‐tetradecadienyl acetate (ZETA), the major pheromone component of Plodia, Ephestia, and Spodoptera species in yeast

“…We used FAMEs as supplemented precursors because they are more soluble in the medium than free fatty acids. 34 , 35 , 36 Yeasts were cultured at 30 °C and 300 rpm. After 48 h, yeast cells were harvested by centrifugation at 4000 g and the medium was discarded.…”

Biotechnological production of the European corn borer sex pheromone in the yeast Yarrowia lipolytica

“…These were the desaturase Lbo_PPTQ from the grapevine moth Lobesia botrana, [28] Onu11 from the European corn borer O. nubilalis, [26] EpoE11 from the light brown apple moth Epiphyas postvittana, [29] CroZ11 from the oblique banded leafroller moth Choristoneura rosaceana, [30] CpaE11 from the spotted fireworm moth Choristoneura parallela, [31] Hzead11 from the corn earworm H. zea, [32] and Msex-tad11 from the tobacco hornworm Manduca sexta. [33] The resulting strains were cultivated, their lipids methanolized into FAMEs, and analyzed on GC-MS. Lbo_PPTQ expression resulted in the highest content of Z11-14:acid.…”

Evolution of the Sex Pheromone Communication System in Ostrinia Moths

“…Genes encoding other enzymes crucial for the sex pheromone biosynthesis, such as chain-shortening enzymes and acetyltransferases, of a vast majority of moths have not been functional characterized yet, except for a recent report of two acyl-CoA oxidase genes involved in chain-shortening in the European grapevine moth, Lobesia botrana [63]. Further studies are needed to identify genes encoding chain-shortening enzymes and acetyltransferases in Ostrinia moths and verify whether any differences in these genes contribute to the variation of pheromone signals.…”