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

1565355/1565232毕氏拟除虫菊酯/拟除虫菊酯是一组由菊花近缘植物丹皮菊花合成的六种类似的天然杀虫剂。除虫菊酯对人类、家畜和畜舍动物极其安全，已被用于抗击疟疾和其他昆虫传播的疾病。它们也是可生物降解的，在环境中不会持久，防止出现对除虫菊酯具有抗药性的昆虫。目前获得除虫菊酯的方法是将植物种植到开花，收集花朵，然后从干燥的花瓣中提取化合物。由于除虫菊酯在花瓣中浓度低，总体产量低，可供销售的数量有限，价格较高。研究人员将使用来自T.cinerariifolum和其他植物的基因来重建导致六种天然除虫菊酯的生物合成途径，并在番茄中创造新的变异除虫菊酯，这种番茄具有高合成除虫菊酯早期前体的能力。因此，这项工作的目的是开发番茄作为一种制造高价值有机化合物的系统，并增加天然、安全和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基因工程、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，研究人员将参与面向年轻人和成年人的外联计划，以描述研究项目、其结果和潜在的社会效益。拟除虫菊酯类杀虫剂是菊科植物银杏花合成的天然杀虫剂。除虫菊酯是由两种单萜酸(菊花酸和除虫菊酸)中的一种和茉莉酸衍生的三种醇中的一种(吡咯酮、焦油酮和茉莉酮)组成的酯。研究人员将在番茄中重建菊花酸的生物合成途径，该途径本身并不具有菊花酸合成的基因，但会产生早期的前体二甲基烯丙基二磷酸(DMAPP)。番茄之所以被选为生产系统，是因为它的两个器官--果实和毛状体--大量产生DMAPP。这条途径将通过识别和部署灰霉和其他植物物种有效地将DMAPP转化为菊花酸的基因和酶，以及通过识别和抑制将途径中间产物转移到非生产性副产品的基因和酶来简化。该项目还将检查转基因番茄品系是否生产菊酸酯，并开发方法来改造番茄以生产天然的除虫菊酯以及用于测试可能的杀虫性能的新型菊酸酯。最后，用番茄生产的菊酸在体外用酶和非酶两种方法生产天然的除虫菊酯。这项工作将导致高价值的天然和安全杀虫剂的高产生产。它还将推进更好地了解植物新陈代谢的总体目标，从而使通过基因工程技术在植物中生产特定化合物变得更具可预测性。该奖项由CBET部门的生物技术和生化工程项目获得，由分子和细胞生物科学部的系统和合成生物学项目共同资助。

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.