microRNA miR-92a-1-p5导入蚜虫条件筛选
Optimized delivery method of microRNA miR-92a-1-p5 into Acyrthosiphon pisum(Hemiptera:Aphididae)
赵许惠 1李忠蔚 1常美玲 1蔡智岩 1王云开 1阚云超 2张东海 3李丹丹1
作者信息
- 1. 南阳师范学院河南省伏牛山昆虫生物学重点实验室,河南省昆虫生物学国际联合实验室,河南南阳 473061
- 2. 南阳师范学院河南省伏牛山昆虫生物学重点实验室,河南省昆虫生物学国际联合实验室,河南南阳 473061;河南科技学院资源与环境学院,河南新乡 453003
- 3. 河南喜夫农生物科技有限公司,河南 开封 475500
- 折叠
摘要
微小RNA(microRNA,miRNA)在蚜虫发育过程中具有重要调控作用,近年来利用miRNA作为靶标进行害虫防治逐渐受到人们的重视,但miRNA导入蚜虫的方式有很大局限.为探索miRNA导入蚜虫的最适条件,以豌豆蚜(Acyrthosi-phon pisum)为研究对象,探究显微注射、饲喂、纳米系统递送三种方式对microRNA miR-92a-1-p5的导入效果;同时将不同浓度的miRNA模拟物或抑制剂通过纳米递送系统导入蚜虫体内,定量PCR(Quantitative PCR,qRT-PCR)筛选miRNA导入蚜虫的最佳浓度.结果发现,蚜虫体壁薄且体液多,显微注射方式会导致内容物流出过多而死亡,因此显微注射方式不适于蚜虫miRNA的导入.使用豌豆苗幼嫩组织吸收miRNA抑制剂后再饲喂蚜虫,发现处理48 h后miR-92a-1-p5表达量下降45.3％,其靶标基因flightin表达量上升65.6％,呈显著负调控(P＜0.01),表明该方式适合miRNA在蚜虫中的有效导入.使用纳米材料包裹miRNA模拟物或抑制剂后在蚜虫背板点滴,可实现miRNA在蚜虫中的有效过表达或干涉,且最适浓度为160 ng/μL.综上所述,植物吸收miRNA后饲喂蚜虫的方式和纳米递送系统介导miRNA导入的方式,均能实现miRNA在豌豆蚜中的有效导入,纳米递送系统操作更简单,效率更高.研究为miRNA导入蚜虫方式提供了参考,也为miRNA用于害虫防治提供了新的思路.
Abstract
MicroRNA(miRNA)plays a crucial role in the development of aphids.The potential use of miRNA as a target for aphid pest control has garnered increasing attention in recent years.However,significant limita-tions exist regarding the introduction of miRNA into aphids.In order to explore the optimal conditions for miRNA introduction into aphids,the introduction effect of microRNA miR-92a-1-p5 by microinjection,feeding and nano-system delivery was examined by taking Acyrthosiphon pisum as the study object.Simultaneously,various concen-trations of miRNA mimics or inhibitors were introduced into aphids via nanodelivery systems,and Quantitative PCR(qRT-PCR)was employed to identify the optimal concentration for effective miRNA incorporation into aphids.The results show that microinjection is not suitable for introducing miRNA due to the thin body wall and high fluid content leaking of aphids,leading to high mortality.The 2 μmmol/L miR-92a-1-p5 antagomir was uti-lized to absorb miRNA inhibitors into the seedling of Pisum sativum,which were subsequently fed to aphids.After 48 hours of treatment,it was observed that the expression level of miR-92a-1-p5 decreased by 45.3％,while the expression level of its target gene flightin increased by 65.6％(P＜0.01),suggesting that this method is suitable for effectively introducing miRNA into aphids.The effective overexpression or interference of miRNA in aphids can be achieved by using nanomaterials coated with miRNA mimics or inhibitors and dropping them on the back plane of aphids,and the optimal concentration is 160 ng/µL.In conclusion,both feeding aphids after plant-mediated absorption of miRNA and employing a nanodelivery system for introducing miRNA can effectively facilitate its introduction into pea aphids.The nanodelivery system proves to be simpler and more efficient.This study provides valuable insights for future efforts aiming at introducing miRNA into aphids and presents new strat-egies for utilizing miRNA in pest control.
关键词
豌豆蚜/miR-92a-1-p5/RNA干涉/纳米递送系统Key words
Acyrthosiphon pisum/miR-92a-1-p5/RNA interference/nanocarrier-delivery system引用本文复制引用
出版年
2025
国家科技期刊平台
NSTL
万方数据