Yu Xiudao, Gowda Siddarame, Killiny Nabil
Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred, FL, USA.
School of Agricultural Engineering/Henan Collaborative Innovation Center of Water Security for Water Source Region of Mid-line of South-to-North Diversion Project, Nanyang Normal University, Nanyang, Henan, China.
Pest Manag Sci. 2017 Sep;73(9):1846-1853. doi: 10.1002/ps.4549. Epub 2017 Mar 14.
Asian citrus psyllid, Diaphorina citri Kuwayama, is the most important economic pest of citrus because it transmits Candidatus Liberibacter asiaticus (CLas), the causal agent of huanglongbing (HLB). Silencing genes by RNA interference (RNAi) is a promising approach for controlling D. citri. RNAi-based insect management strategies depend on the selection of suitable target genes.
The muscle protein 20 gene DcMP20 was characterized from D. citri in an effort to impair proper muscle development through RNAi. Phylogenetic analysis showed that DcMP20 was more closely related to MP20 from Drosophila compared with its counterpart from other insect species. Developmental expression analysis revealed that transcription of DcMP20 was development dependent and reached a maximum level in the last instar (fourth-fifth) of the nymphal stage. The extent of RNAi in D. citri was dose dependent, with dsRNA-DcMP20 at 75 ng µL being sufficient to knock down endogenous DcMP20 expression, which resulted in significant mortality and reduced body weight that positively correlated with the silencing of DcMP20. No effect was found when dsRNA-GFP or water was used, indicating the specific effect of dsRNA-DcMP20.
Our results suggest that dsRNA can be delivered to D. citri through soaking, and DcMP20 is an effective RNAi target to be used in the management of D. citri. © 2017 Society of Chemical Industry.
亚洲柑橘木虱(Diaphorina citri Kuwayama)是柑橘最重要的经济害虫,因为它传播柑橘黄龙病(HLB)的病原体——亚洲韧皮杆菌(Candidatus Liberibacter asiaticus,CLas)。通过RNA干扰(RNAi)使基因沉默是控制亚洲柑橘木虱的一种有前景的方法。基于RNAi的昆虫管理策略依赖于合适靶基因的选择。
对亚洲柑橘木虱的肌肉蛋白20基因DcMP20进行了表征,以期通过RNAi破坏其正常的肌肉发育。系统发育分析表明,与其他昆虫物种的对应基因相比,DcMP20与果蝇的MP20关系更为密切。发育表达分析显示,DcMP20的转录依赖于发育过程,在若虫期的最后一龄(第四至五龄)达到最高水平。亚洲柑橘木虱中RNAi的程度呈剂量依赖性,75 ng μL的dsRNA-DcMP20足以敲低内源性DcMP20的表达,这导致了显著的死亡率和体重减轻,且与DcMP20的沉默呈正相关。使用dsRNA-GFP或水时未发现效果,表明dsRNA-DcMP20具有特异性作用。
我们的结果表明,dsRNA可通过浸泡法导入亚洲柑橘木虱体内,DcMP20是用于管理亚洲柑橘木虱的有效RNAi靶标。© 2017化学工业协会。