Clay nanosheet-mediated delivery of recombinant plasmids expressing artificial miRNAs via leaf spray to prevent infection by plant DNA viruses

查看全文 作　　者：Qili [1,2,3]Liu;Yanpeng [4]Li;Kedong [5]Xu;Dongxiao [3]Li;Haiyan [3]Hu;Feng [3]Zhou;Puwen [3]Song;Yongang [3]Yu;Qichao [3]Wei;Qian [1]Liu;Weipeng [1]Wang;Ruifang [3]Bu;Haili [3]Sun;Xiaohui [4]Wang;Jianjun [6]Hao;Honglian [2]Li;Chengwei [1,3,5]Li

机构地区：[1]Postdoctoral Research Base,Henan Institute of Science and Technology,Xinxiang,China;[2]College of Plant Protection,Henan Agricultural University,Zhengzhou,Henan,China;[3]Henan Engineering Research Center of Crop Genome Editing,Henan Institute of Science and Technology,Xinxiang,China;[4]State Key Laboratory of Pulp&Paper Engineering,South China University of Technology,Guangzhou,China;[5]Key Laboratory of Plant Genetics and Molecular Breeding,Zhoukou Normal University,Zhoukou,China;[6]School of Food and Agriculture,The University of Maine,Orono,ME 04469,USA

出　　处：《Horticulture Research》索引2020年第7卷第1期,共12页

基　　金：funded by the National Natural Science Foundation of China(31801707);the Key Projects of Science and Technology Research in Henan Province(182102110470);the Plant Protection of Key Discipline Project of Henan province(107020219001/005);the National Key Research and Development Program of China(2016YFD0300203-3);First-Class Postdoctoral Research Grant in Henan Province(001701038);the Science-Technology Foundation for High Level Talent of Henan Institute of Science and Technology(2015028). 摘　　要：Whitefly-transmitted begomoviruses are economically important plant pathogens that cause severe problems in many crop plants,such as tomato,papaya,cotton,and tobacco.Tomato yellow leaf curl virus(TYLCV)is a typical monopartite begomovirus that has been extensively studied,but methods that can efficiently control begomoviruses are still scarce.In this study,we combined artificial microRNA(amiRNA)-mediated silencing technology and clay nanosheetmediated delivery by spraying and developed a method for efficiently preventing TYLCV infection in tomato plants.We designed three amiRNAs that target different regions of TYLCV to silence virus-produced transcripts.Three plant expression vectors expressing pre-amiRNAs were constructed,and recombinant plasmid DNAs(pDNAs)were loaded onto nontoxic and degradable layered double hydroxide(LDH)clay nanosheets.LDH nanosheets containing multiple pDNAs were sprayed onto plant leaves.We found that the designed amiRNAs were significantly accumulated in leaves 7 days after spraying,while the pDNAs were sustainably detected for 35 days after the spray,suggesting that the LDH nanosheets released pDNAs in a sustained manner,protected pDNAs from degradation and efficiently delivered pDNAs into plant cells.Importantly,when the LDH nanosheets coated with pDNAs were sprayed onto plants infected by TYLCV,both the disease severity and TYLCV viral concentration in sprayed plants were significantly decreased during the 35 days,while the levels of H_(2)O_(2) were significantly increased in those plants.Taken together,these results indicate that LDH nanosheets loaded with pDNAs expressing amiRNAs can be a sustainable and promising tool for begomovirus control. 关键词：EXPRESSING artificial SUSTAINED

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Clay nanosheet-mediated delivery of recombinant plasmids expressing artificial miRNAs via leaf spray to prevent infection by plant DNA viruses

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