Collaborative Research: Production of known and novel, safe, and biodegradable pyrethrin-type insecticides in tomato

1565355/1565232 Pichersky/Last除虫菊酯是一组六种类似的天然杀虫剂，由植物Tanacetum cinerariifolium的花合成，Tanacetum cinerariifolium是菊花的亲戚。除虫菊酯对人类、家畜和牲畜都非常安全，已被用于对抗疟疾和其他虫媒疾病。它们也是可生物降解的，在环境中是非持久性的，可以防止出现对除虫菊酯有抗药性的昆虫。目前，通过种植植物直到开花，收集花朵并从干燥的花瓣中提取化合物来获得除虫菊酯。由于花瓣中除虫菊酯的浓度低，总的除虫菊酯产量低，可供销售的量有限，并且价格高。 研究人员将利用T.在番茄和其它植物中重建导致六种天然除虫菊素的生物合成途径，以及在番茄中产生新的具有高合成除虫菊素早期前体能力的变异除虫菊素。 因此，这项工作的目的是开发番茄作为一个系统，使高价值的有机化合物一般，并增加天然，安全，可生物降解的杀虫剂的可用范围。此外，这项研究将为本科生和研究生以及博士后研究人员提供遗传工程，植物生物化学，分析化学和分子生物学的跨学科培训机会。此外，研究人员还将参与面向年轻人和成年人的推广计划，以描述研究项目，其结果和潜在的社会效益。 除虫菊是一种天然杀虫剂，由菊科植物菊醋藤（Tanacetum cinerariifolium）的花合成。除虫菊酯是由两种单萜酸（菊酸和除虫菊酸）之一和茉莉酸衍生的三种醇（除虫菊酮、cinerolone和茉莉酮）之一组成的酯。研究人员将在番茄中重建胆酸的生物合成途径，番茄本身没有指定胆酸合成的基因，但会产生早期前体二甲基烯丙基二磷酸（DMAPP）。选择番茄作为生产系统是因为它的两个器官，果实和毛状体，大量生产DMAPP。该途径将通过识别和部署T.本发明的目的是通过鉴定和抑制将途径中间体转化为非生产性副产物的基因和酶，来研究有效地将DMAPP转化为胆酸的西洋参和其他植物物种。该项目还将检查工程番茄品系是否产生蝶啶酸酯，并开发工程番茄的方法，以生产天然除虫菊酯酯以及新型蝶啶酸酯，用于测试可能的杀虫特性。最后，将在体外用番茄产生的胆酸酶促和非酶促产生天然除虫菊酯。这项工作将导致高产量生产高价值的天然和安全的农药。该奖项由CBET生物技术和生物化学工程项目组颁发，由分子和细胞生物科学部门的系统和合成生物学项目组共同资助。

1565355/1565232 Pichersky/Last Pyrethrins are a group of six similar natural insecticides synthesized by the flowers of the plant Tanacetum cinerariifolium, a relative of chrysanthemum. Pyrethrins are extremely safe to humans and to domestic and barn animals, and have been used to fight malaria and other insect-borne diseases. They are also biodegradable and non-persistent in the environment, preventing the appearance of pyrethrin-resistant insects. Pyrethrins are currently obtained by growing the plants until they bloom, collecting the flowers, and extracting the compounds from dry petals. Because the concentration of pyrethrins in the petals is low, overall pyrethrin yield is low, the amount available for sale is limited, and the price is high. The researchers will employ genes from T. cinerariifolium and other plants to reconstruct the biosynthetic pathway leading to the six natural pyrethrins, as well as to create new variant pyrethrins, in tomato that have a high capacity for synthesizing an early precursor of pyrethrins. Thus, this work aims to develop tomato as a system for making high-value organic compounds in general, and increase the available range of natural, safe, and biodegradable insecticides. In addition, this research will provide interdisciplinary training opportunities in genetic engineering, plant biochemistry, analytical chemistry, and molecular biology to undergraduate and graduate students as well as post-doctoral researchers. Furthermore, the researchers will engage in outreach programs to young people and adults to describe the research project, its results, and potential societal benefits. Pyrethrins are natural insecticides synthesized by the flowers of the plant Tanacetum cinerariifolium (Asteraceae). A pyrethrin is an ester made up of one of two monoterpenoid acids (chrysanthemic and pyrethric acids) and one of three alcohols derived from jasmonic acid (pyrethrolone, cinerolone, and jasmolone). The researchers will recreate the biosynthetic pathway to chrysanthemic acid in tomato, which by itself does not have the genes specifying the synthesis of chrysanthemic acid but makes the early precursor dimethylallyl diphosphate (DMAPP). Tomato was chosen as a production system because two of its organs, fruits and trichomes, make DMAPP in large amounts. The pathway will be streamlined by identifying and deploying genes and enzymes from T. cinerariifolium and other plant species that efficiently convert DMAPP to chrysanthemic acid, and by identifying and inhibiting genes and enzymes that divert pathway intermediates to non-productive side-products. The project will also examine if the engineered tomato lines produce esters of chrysanthemic acid, and develop approaches to engineer tomato to make natural pyrethrin esters as well as novel esters of chrysanthemic acid for testing possible insecticidal properties. Finally, natural pyrethrins will be produced enzymatically and non-enzymatically in vitro with tomato-produced chrysanthemic acid. This work will result in the high-yield production of highly valuable natural and safe pesticides. It will also advance the general goal of better understanding of plant metabolism so that the production of specific compounds in plants by genetic engineering techniques can become more predictable.This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biosciences.