Abstract
Pheromones are environmentally friendly alternatives to traditional pesticides for pest control. They are widely applied for insect monitoring, mating disruption and mass trapping. Nicotiana benthamiana and N. tabacum are potential green biomass production platforms of moth sex pheromones. Using these two Nicotiana species as plant factories, we expressed biosynthetic genes of plant and insect origin in leaf tissue. Moth sex pheromone precursors (E)-11-tetradecenoic acid, (Z)-11-tetradecenoic acid and (Z)-11-hexadecenoic acid were produced by introducing the acyl-ACP thioesterases CpuFatB1 from Cuphea pulcherrima or CpaFatB2 from C. palustris and the fatty acyl desaturases Ave∆11 from Argyrotaenia velutinana, CpaE11 from Choristoneura parallela or Atr∆11 from Amyelois transitella, under the control of CaMV-35S promoter. Among the Nicotiana spp. transformants, the best line produced (Z)-11-hexadecenoic acid at 17.6% of total fatty acids in leaves, during flowering stage, corresponding to 335 µg of (Z)-11-hexadecenoic acid per gram of fresh leaf. The (Z)-11-hexadecenoic acid production lines from N. benthamiana were selected for further propagation to obtain homozygous lines. In the N. benthamiana T2 generation, the production quantity of (Z)-11-hexadecenoic acid was stable. Our study demonstrates the feasibility of stable transformation of N. benthamiana for production of moth pheromone precursors in vegetative tissue.
Highlights
The insect order Lepidoptera contains more than 160,000 described moth and butterfly species (Nieukerken et al 2011)
The successful production of several insect sex pheromone precursors in N. tabacum and N. benthamiana by stable transformation was demonstrated in this study
All of the transformed plastid thioesterases and fatty acyl desaturases were functionally active in producing pheromone precursor in both Nicotiana species
Summary
The insect order Lepidoptera contains more than 160,000 described moth and butterfly species (Nieukerken et al 2011). Species of moths are among the most damaging pests of food and fiber crops. Due to the numerous problems associated with use of conventional pesticides (Brittain and Potts 2011; Bull 1982), synthetic pheromones have emerged as an alternative for insect control (Reddy and Guerrero 2000). Tons of synthetic pheromones are produced for application in pest management (Weatherston and Stewart 2002). Use of hazardous chemicals and generation of by-products during conventional chemical synthesis of pheromones (Mori 2007, 2010) may cause pollution problems. High costs for synthesis limit the use of pheromones in many crops
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