Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants.
Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants.
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DOI：
10.34133/2021/9891082
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发表时间：
2021
影响因子：
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通讯作者：
Orzáez D
中科院分区：
文献类型：
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作者：
Mateos-Fernández R;Moreno-Giménez E;Gianoglio S;Quijano-Rubio A;Gavaldá-García J;Estellés L;Rubert A;Rambla JL;Vazquez-Vilar M;Huet E;Fernández-Del-Carmen A;Espinosa-Ruiz A;Juteršek M;Vacas S;Navarro I;Navarro-Llopis V;Primo J;Orzáez D
以植物为基础的昆虫性信息素生物生产已被提出作为一种创新策略，以提高农业害虫防治的可持续性。在这里，我们描述了转基因植物产生(Z)-11-十六烯醇（Z11-16OH）和(Z)-11-十六烯乙酸酯（Z11-16OAc）的工程，这是许多鳞翅目性信息素混合物中的两种主要挥发性成分。我们组装了编码信息素生物合成途径的多基因DNA构建体，并将其稳定地转化为烟叶植物。这些构建体含有不同构型的transitella Amyelois AtrΔ11去饱和酶基因、棉铃虫Helicoverpa fatty酰基还原酶HarFAR基因和褐杨二酰基甘油乙酰转移酶EaDAct基因。所有产生信息素的植株均呈现矮化表型，矮化程度与信息素水平相关。除一条外，所有恢复的系都产生高水平的Z11-16OH，但极低水平的Z11-16OAc，可能是EaDAct基因水平反复截断的结果。只有1株株（SxPv1.2）具有完整的信息素通路，产生中等水平的Z11-16OAc （11.8 μg -1 FW）和高水平的Z11-16OH （111.4 μg -1）。Z11-16OAc的产生伴随着矮化表型的部分恢复。利用SxPv1.2估算挥发性信息素的释放速率，Z11-16OH和Z11-16OAc的挥发性信息素释放速率分别为8.48 ng g-1 FW / d和9.44 ng g-1 FW / d。我们的研究结果表明，信息素释放是信息素生物分配策略的限制因素，并为生物技术改进建立了路线图。
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.