Research Article | Open Access

Volume 2021 |Article ID 9891082 | https://doi.org/10.34133/2021/9891082

Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants

Rubén Mateos-FernándeziD ,¹ Elena Moreno-Giménez,¹ Silvia GianoglioiD ,¹ Alfredo Quijano-RubioiD ,¹ Jose Gavaldá-GarcíaiD ,¹ Lucía EstellésiD ,¹ Alba Rubert,¹ José Luis Rambla,^1,2 Marta Vazquez-VilariD ,¹ Estefanía Huet,¹ Asunción Fernández-del-Carmen,¹ Ana Espinosa-Ruiz,¹ Mojca Juteršek,^3,4 Sandra Vacas,⁵ Ismael Navarro,⁶ Vicente Navarro-Llopis,⁵ Jaime PrimoiD ,⁵ and Diego Orzáez iD ¹

¹Institute for Plant Molecular and Cell Biology (IBMCP), Consejo Superior de Investigaciones Científicas (CSIC) - Universidad Politécnica de Valencia (UPV), Valencia, Spain
²Jaume I University, Castellon de la Plana, Spain
³Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia
⁴Jožef Stefan International Postgraduate School, Ljubljana, Slovenia
⁵Centro de Ecología Química Agrícola, Instituto Agroforestal del Mediterráneo, Universitat Politècnica de València, Valencia, Spain
⁶Ecología Y Protección Agrícola SL, Valencia, Spain

Abstract

Plant-based bioproduction of insect sex pheromones has been proposed as an innovative strategy to increase the sustainability of pest control in agriculture. Here, we describe the engineering of transgenic plants producing (Z)-11-hexadecenol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them into Nicotiana benthamiana plants. The constructs contained the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera fatty acyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, the severity of which correlated with pheromone levels. All but one of the recovered lines produced high levels of Z11-16OH, but very low levels of Z11-16OAc, probably as a result of recurrent truncations at the level of the EaDAct gene. Only one plant line (SxPv1.2) was recovered that harboured an intact pheromone pathway and which produced moderate levels of Z11-16OAc (11.8 μg g-1 FW) and high levels of Z11-16OH (111.4 μg g-1). Z11-16OAc production was accompanied in SxPv1.2 by a partial recovery of the dwarf phenotype. SxPv1.2 was used to estimate the rates of volatile pheromone release, which resulted in 8.48 ng g-1 FW per day for Z11-16OH and 9.44 ng g-1 FW per day for Z11-16OAc. Our results suggest that pheromone release acts as a limiting factor in pheromone biodispenser strategies and establish a roadmap for biotechnological improvements.