Wan Pin-Jun, Fu Kai-Yun, Lü Feng-Gong, Wang Xin-Xin, Guo Wen-Chao, Li Guo-Qing
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou, China.
Pest Manag Sci. 2015 Oct;71(10):1387-96. doi: 10.1002/ps.3941. Epub 2014 Dec 5.
Leptinotarsa decemlineata is an able disperser by flight. Novel control strategies must be explored to control the damage and inhibit the dispersal efficiently. Proline is a major energy substrate during flight. Δ-Pyrroline-5-carboxylate dehydrogenase (P5CDh) catalyses the second step of proline degradation for the production of ATP.
A full-length Ldp5cdh cDNA was cloned. Ldp5cdh was ubiquitously expressed in the eggs, the first through fourth larval instars, wandering larvae, pupae and adults. In the adults, Ldp5cdh mRNA was widely distributed in thorax muscles, midgut, foregut, hindgut, Malpighian tubules, ventral ganglion, fat body and epidermis, with the expression levels from the highest to the lowest. Two double-stranded RNAs (dsRNAs) (dsLdp5cdh1 and dsLdp5cdh2) targeting Ldp5cdh were constructed and bacterially expressed. Ingestion of dsLdp5cdh1 and dsLdp5cdh2 successfully silenced Ldp5cdh, significantly increased the contents of proline, arginine and alanine, but strongly decreased the contents of asparate, asparagine, glutamate and glutamine in the haemolymph. Moreover, knocking down Ldp5cdh significantly reduced ATP content, decreased flight speed, shortened flight distance and increased adult mortality.
It seems that identified Ldp5cdh encodes a functional P5CDh enzyme, and Ldp5cdh may serve as a potential target for dsRNA-based pesticide for controlling the damage and dispersal of L. decemlineata adults. © 2014 Society of Chemical Industry.
马铃薯甲虫是一种善于飞行扩散的昆虫。必须探索新的控制策略，以有效控制其危害并抑制其扩散。脯氨酸是飞行过程中的主要能量底物。Δ-吡咯啉-5-羧酸脱氢酶（P5CDh）催化脯氨酸降解的第二步以产生ATP。
克隆了全长的Ldp5cdh cDNA。Ldp5cdh在卵、一至四龄幼虫、预蛹、蛹和成虫中均有广泛表达。在成虫中，Ldp5cdh mRNA广泛分布于胸肌、中肠、前肠、后肠、马氏管、腹神经节、脂肪体和表皮，表达水平由高到低。构建并在细菌中表达了两种靶向Ldp5cdh的双链RNA（dsRNAs）（dsLdp5cdh1和dsLdp5cdh2）。摄入dsLdp5cdh1和dsLdp5cdh2成功沉默了Ldp5cdh，显著增加了血淋巴中脯氨酸、精氨酸和丙氨酸的含量，但强烈降低了天冬氨酸、天冬酰胺、谷氨酸和谷氨酰胺的含量。此外，敲低Ldp5cdh显著降低了ATP含量，降低了飞行速度，缩短了飞行距离，并增加了成虫死亡率。
鉴定出的Ldp5cdh似乎编码一种功能性的P5CDh酶，并且Ldp5cdh可能作为基于dsRNA的杀虫剂的潜在靶标，用于控制马铃薯甲虫成虫的危害和扩散。©2014化学工业协会。
