Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants

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.

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Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants

10.1101/2021.03.31.437903 ◽

2021 ◽

Author(s):

Ruben Mateos-Fernandez ◽

Elena Moreno Gimenez ◽

Silvia Gianoglio ◽

Alfredo Quijano-Rubio ◽

Jose Gavalda-Garcia ◽

...

Keyword(s):

Nicotiana Benthamiana ◽

Sex Pheromones ◽

Limiting Factor ◽

Volatile Components ◽

Partial Recovery ◽

Amyelois Transitella ◽

Innovative Strategy ◽

Euonymus Alatus ◽

Pheromone Release ◽

Insect Sex Pheromones

Plant-based bio-production 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-hexadecen-1-ol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multiple multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them in Nicotiana benthamiana plants. The constructs comprised the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera farnesyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, whose severity 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 harbouring an intact pheromone pathway and producing moderate levels of Z11-16OAc (11.8 μg g-1 FW), next to 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 bio-dispenser strategies and establish a roadmap for biotechnological improvements.

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Development of a chimeric odour blend for attracting gravid malaria vectors

Malaria Journal ◽

10.1186/s12936-021-03797-w ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Betelehem Wondwosen ◽

Mengistu Dawit ◽

Yared Debebe ◽

Habte Tekie ◽

Sharon R. Hill ◽

...

Keyword(s):

Vector Control ◽

Binary Logistic Regression ◽

Ground Level ◽

Malaria Vectors ◽

Volatile Components ◽

Innovative Strategy ◽

Breeding Sites ◽

Logistic Regression Models ◽

Plant Feeding ◽

Global Vector

Abstract Background Odour-based tools targeting gravid malaria vectors may complement existing intervention strategies. Anopheles arabiensis are attracted to, and stimulated to oviposit by, natural and synthetic odours of wild and domesticated grasses associated with mosquito breeding sites. While such synthetic odour lures may be used for vector control, these may have limited efficacy when placed in direct competition with the natural source. In this study, workflows developed for plant-feeding pests was used to design and evaluate a chimeric odour blend based on shared attractive compounds found in domesticated grass odours. Methods Variants of a synthetic odour blend, composed of shared bioactive compounds previously identified in domesticated grasses, was evaluated sequentially in a two-choice olfactometer to identify a ratio-optimized attractive blend for malaria vectors. During this process, blends with ratios that were significantly more attractive than the previously identified synthetic rice blend were compared to determine which was most attractive in the two-choice olfactometer. To determine whether all volatile components of the most attractive blend were necessary for maximal attraction, subtractive assays were then conducted, in which individual components were removed for the most attractive blend, to define the final composition of the chimeric blend. Binary logistic regression models were used to determine significance in all two-choice assays. The chimeric blend was then assessed under field conditions in malaria endemic villages in Ethiopia, to assess the effect of dose, trap type, and placement relative to ground level. Field data were analyzed both descriptively and using a Welch-corrected t-test. Results A ratio-optimized chimeric blend was identified that significantly attracted gravid An. arabiensis under laboratory conditions. In the field, trap captures of An. arabiensis and Anopheles pharoensis were dependent on the presence of the lure, trap type (CDC, BG Sentinel and Suna traps), placement relevant to ground level, with low release rates generally luring more mosquitoes. Conclusions The workflow designed for the development of chimeric lures provides an innovative strategy to target odour-mediated behaviours. The chimeric lure identified here can be used in existing trapping systems, and be customized to increase sustainability, in line with goals of the Global Vector Control Response Group.

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Microanalytical Methodology Relating to the Identification of Insect Sex Pheromones and Related Behavior-Control Chemicals

Journal of Chromatographic Science ◽

10.1093/chromsci/13.7.314 ◽

1975 ◽

Vol 13 (7) ◽

pp. 314-321 ◽

Cited By ~ 18

Author(s):

M. Beroza

Keyword(s):

Sex Pheromones ◽

Behavior Control ◽

Insect Sex Pheromones ◽

Insect Sex

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Development of methods for application of pheromonic materials for monitoring and managing the number of apple phytophages

Horticulture and viticulture ◽

10.31676/0235-2591-2021-2-47-53 ◽

2021 ◽

pp. 47-53

Author(s):

I. S. Agasieva ◽

V. Ya. Ismailov ◽

A. S. Nastasiy ◽

M. V. Nefedova

Keyword(s):

Species Composition ◽

Sex Pheromones ◽

Apple Tree ◽

Plant Protection ◽

Central Zone ◽

Apple Orchard ◽

Species Specificity ◽

Insect Sex Pheromones ◽

Natural Dynamics ◽

And Control

Synthetic sex pheromones have found wide application in plant protection as a mean of early detection of pests that allows observation of the phenology of insects to optimise protective measures. Insect sex pheromones can be applied for different purposes in protecting crops such as monitoring, determining the species composition and combating harmful species (by using sex pheromones for disorientation). The present work studies the species composition, dynamics of the number of basic pests of an apple tree, synchronisation of the seasonal and circadian activity of phytophages of an apple tree in the central zone of the Krasnodar Territory. The results of the field assessment of the male complex disorientation method of apple and eastern moths are presented. It was shown that the species-specificity of sex pheromones in the apple orchard depends on the faunistic diversity of Lepidoptera species with similar pheromone systems that develop at a given point in space and time. It was revealed that the behaviour of this complex changes during the season and over years, depending on the climate and natural dynamics of insect populations. The quantitative ratio and species-specificity of pheromones will probably be different in ecosystems with various species composition and different geographic zones. A novelty of this research is the division of the studied Lepidoptera phytophage species into three groups according to a decrease in the absolute species-specificity in the forest biotope in comparison with the garden one. The most widespread and coinciding in terms of summer synchronicity are apple (Cydia pomonella L.), plum (Grapholitha funebrana Tr.), eastern (Grapholitha molesta Tr.) and pomegranate moth (Euzophera bigella Zell.). The disorientation method, applied using a complex system of dispensers with apple and eastern moth pheromones, showed that installing 500 dispensers/ha allowed 99.3 % efficiency to be achieved. Fruit damage amounted to 1.2 % and 2.7 % in the experimental and control groups, respectively. The duration of the disorienting effect of the pheromone formulations lasted for over 4 months.

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