[1]莫亦琳,陈伟钊,黄丽娟,等.ShRNA-LDH纳米颗粒的制备及薇甘菊生物防治[J].深圳大学学报理工版,2023,40(2):163-170.[doi:10.3724/SP.J.1249.2023.02163]
　MO Yilin,CHEN Weizhao,et al.Preparation of ShRNA-LDH nanoparticle and its application in biological control of Mikania micrantha[J].Journal of Shenzhen University Science and Engineering,2023,40(2):163-170.[doi:10.3724/SP.J.1249.2023.02163]
点击复制

ShRNA-LDH纳米颗粒的制备及薇甘菊生物防治()
分享到：

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第40卷
期数:
2023年第2期
页码:
163-170
栏目:
生命与健康
出版日期:
2023-03-15

文章信息/Info

Title:
Preparation of ShRNA-LDH nanoparticle and its application in biological control of Mikania micrantha
文章编号:
202302005
作者:
莫亦琳1; 2;  陈伟钊1;  黄丽娟1;  吴飞燕1; 2;  肖念1; 2;  于宇1;  刘学东1
1）深圳大学生命与海洋科学学院，广东省植物表观遗传学重点实验室，广东深圳 518071
2）加州大学伯克利分校分子与细胞生物学系，美国伯克利 94720
Author(s):
MO Yilin1;  2;  CHEN Weizhao1;  HUANG Lijuan1;  WU Feiyan1;  2;  XIAO Nian1;  2;  YU Yu1;  and LIU Xuedong1
1) College of Life Sciences and Oceanography, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen University, Shenzhen 518071, Guangdong Province, P.R.China
2) Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley 94720, USA
关键词:
核糖核酸干扰; 植物保护; 薇甘菊; 叶绿素a/b结合蛋白; 黏土纳米片; 短发夹核糖核酸
Keywords:
RNA interference;  plant protection;  Mikania micrantha;  light harvesting chlorophyl a/b binding protein;  clay nanosheets;  short hairpin RNA
分类号:
S476+.9
DOI:
10.3724/SP.J.1249.2023.02163
文献标志码:
A
摘要:
核糖核酸干扰（ribonucleic acid interference， RNAi）技术在植物保护中具有高效、安全和环保的特点．外源喷施短发夹RNA（short hairpin RNA， shRNA）可以有效沉默薇甘菊（Mikania micrantha）叶绿素a /叶绿素b结合蛋白相关基因．为提高喷施shRNA对薇甘菊的防治效果，制备直径为10～100 nm的层状双氢氧化物（layered double hydroxide， LDH）黏土纳米片为shRNA的载体，该载体具有良好的生物相容性，具环境友好．研究发现，LDH结合shRNA的最佳质量比为1∶10．shRNA喷施实验表明，有轻微创伤的薇甘菊叶片对shRNA更敏感，并且LDH直接喷施对薇甘菊的生长几乎没有任何影响．为比较LDH-shRNA和单独的shRNA对薇甘菊的控制效果，利用分别搭载了6个shRNA的LDH纳米颗粒或单独的shRNA喷施薇甘菊叶片，在喷施6 d后观察到叶片喷施LDH-shRNA有更加明显的坏死表型．研发的基于LDH的shRNA携带系统可显著提高RNAi在防控薇甘菊中的有效性．
Abstract:
Ribonucleic acid interference (RNAi) is a highly efficient, safe, and environmentally friendly method in plant protection. Exogenous spraying of short hairpin RNAs (shRNAs) targeting light-harvesting chlorophyll a or chlorophyll b binding protein genes onto the leaves of Mikania micrantha (M.micrantha) H. B.K.can efficiently silence the expression of these genes. To improve the effectiveness of shRNA-mediated control of M.micrantha, layered double hydroxide (LDH) clay nanosheets with a diameter between 10 to 100 nm and good biocompatibility and environmental friendliness were prepared as shRNA carriers in this study. We found that the optimal binding mass ratio of LDH to shRNA was 1∶10. ShRNA spraying assays showed that M.micrantha leaves with micro-wound were more sensitive to shRNA, and spraying LDH directly to M. micrantha had no effect on plant growth. To compare the effects of LDH-shRNA and shRNA alone on the control of M.micrantha, we sprayed LDH loaded with six different shRNA respectively or shRNA alone onto M.micrantha leaves, and a more severe necrotic phenotype was observed on the 6th day after LDH-shRNA-sprayed. Taken together, our study successfully developed an LDH-based shRNA carrying system to significantly improve the effectiveness of RNAi in the control of M.micrantha.

参考文献/References:

［1］ LIU Bo, YAN Jian, LI Weihua, et al. Mikania micrantha genome provides insights into the molecular mechanism of rapid growth [J]. Nature Communications, 2020, 11(1): 340.
［2］ WANG Bosun, LIAO Wenbo, ZAN Qijie, et al. The spreads of Mikania micrantha in China [J]. Acta Scientiarum Naturalium Universitatis Sunyatseni, 2003, 42(4):47-50, 54.(in Chinese)
王伯荪，廖文波，昝启杰，等．薇甘菊Mikania micrantha在中国的传播［J］．中山大学学报自然科学版，2003，42（4）：47-50，54.
［3］ GUO Wuxia, LIU Ying, NG Weilun, et al. Comparative transcriptome analysis of the invasive weed Mikania micrantha with its native congeners provides insights into genetic basis underlying successful invasion [J]. BMC Genomics, 2018, 19(1): 392.
［4］ ZHONG Xiaoqing, HUANG Zhuo, SI Huan, et al. Analysis of ecological-economic loss caused by weed Mikania micrantha in Neilingding Island, Shenzhen, China [J]. Journal of Tropical and Subtropical Botany, 2004, 12(2): 167-170.(in Chinese)
钟晓青，黄卓，司寰，等．深圳内伶仃岛薇甘菊危害的生态经济损失分析［J］．热带亚热带植物报，2004，12（2）：167-170.
［5］ MAO Zilong, LAI Meidong, ZHAO Zhenye, et al, Effects of Mikania micrantha invasion on carbon storage in mangrove ecosystems in Shenzhen Bay [J]. Ecology and Environmental Sciences, 2011, 20(12): 1813-1818.(in Chinese)
毛子龙，赖梅东，赵振业，等．薇甘菊入侵对深圳湾红树林生态系统碳储量的影响［J］．生态环境学报，2011，20（12）：1813-1818.
［6］ LI Zhijie, HUANG Jianghua. Research progress on the control of M. micrantha [J]. Journal of Zhongkai University of Agriculture and Engineering, 2018, 31(1): 66-71.(in Chinese)
李志杰，黄江华．薇甘菊防治研究进展［J］．仲恺农业工程学院学报，2018，31（1）：66-71.
［7］ HERNANDEZ-SOTO A, CHACON-CERDAS R. RNAi crop protection advances [J]. International Journal of Molecular Sciences, 2021, 22(22): 12148.
［8］ ZHANG Lingxiao, HU Jing, JIA Yingbo, et al. Two-dimensional layered double hydroxide nanoadjuvant: recent progress and future direction [J]. Nanoscale, 2021, 13(16): 7533-7549.
［9］ GU Zi, ATHERTON J J, XU Zhiping. Hierarchical layered double hydroxide nanocomposites: structure, synthesis and applications [J]. Chemical Communications, 2015, 51(15): 3024-3036.
［10］ MITTER N, WORRALL E A, ROBINSON K E, et al. Clay nanosheets for topical delivery of RNAi for sustained protection against plant viruses [J]. Nature Plants, 2017, 3: 16207.
［11］ LIU Qili, LI Yanpeng, XU Kelong, et al. Clay nanosheet-mediated delivery of recombinant plasmids expressing artificial miRNAs via leaf spray to prevent infection by plant DNA viruses [J]. Horticulture Research, 2020, 7(1): 179.
［12］ YONG Jiaxi, ZHANG Run, BI Shengnan, et al. Sheet-like clay nanoparticles deliver RNA into developing pollen to efficiently silence a target gene [J]. Plant Physiology, 2021, 187(2): 886-899.
［13］ MAI Jiantao, LIAO Lingling, LING Rongsong, et al. Study on RNAi-based herbicide for Mikania micrantha. Synthetic and Systems Biotechnology, 2021, 6(4): 437-445.
［14］ RUAN Xiaoxian, WANG Zhen, SU Yingjuan, et al. Population genomics reveals gene flow and adaptive signature in invasive weed Mikania micrantha [J]. Genes, 2021, 12(8): 1279.
［15］ JIN Mengjiao, LIU Bo, WANG Kangkang, et al. Responses of light energy utilization and chlorophyll synthesis in Mikania micrantha under different light intensities [J]. Scientia Agricultura Sinica, 2022, 55(12): 2347-2359.(in Chinese)
金梦娇，刘博，王抗抗，等．薇甘菊光能利用及叶绿素合成在不同光照强度下的响应［J］．中国农业科学，2022，55（12）：2347-2359.
［16］ CUI Can, WANG Zhen, SU Yingjuan, et al. New insight into the rapid growth of the Mikania micrantha stem based on DIA proteomic and RNA-Seq analysis [J]. Journal of Proteomics, 2021, 236: 104126.
［17］ RANK A P, KOCH A. Lab-to-field transition of RNA spray applications - How far are we? [J]. Frontiers in Plant Science, 2021, 12: 755203.
［18］ DALAKOURAS A, WASSENEGGER M, DADAMI E, et al. Genetically modified organism-free RNA interference: exogenous application of RNA molecules in plants [J]. Plant Physiology, 2020, 182(1): 38-50.
［19］ DAS P R, SHERIF S M. Application of exogenous dsRNAs-induced RNAi in agriculture: challenges and triumphs [J]. Frontiers in Plant Science, 2020, 11: 946.
［20］ WU Honghong, LI Zhaohu. Nano-enabled agriculture: How do nanoparticles cross barriers in plants? [J]. Plant Communication, 2022, 100346.
［21］ HOANG B T L, FLETCHER S J, BROSNAN C A, et al. RNAi as a foliar spray: efficiency and challenges to field applications [J]. International Journal of Molecular Sciences, 2022, 23(12): 6639.
［22］ SCHWARTZ S H, HENDRIX B, HOFFER P, et al. Carbon dots for efficient small interfering RNA delivery and gene silencing in plants [J]. Plant Physiology, 2020, 184(2): 647-657.

备注/Memo

备注/Memo:
Received: 2022- 09-19; Accepted: 2022-09-30; Online (CNKI): 2022-11-17
Foundation: National Key R & D Program of China (2019YFA0903902); National Natural Science Foundation of China (32270595); Shenzhen Science and Technology Program (JCYJ20190808115005598)
Corresponding author: Postdoctoral researcher LIU Xuedong. E-mail: xuedong007@126.com
Citation: MO Yilin, CHEN Weizhao, HUANG Lijuan, et al. Preparation of ShRNA-LDH nanoparticle and its application in biological control of Mikania micrantha [J]. Journal of Shenzhen University Science and Engineering, 2023, 40(2): 163-170.
基金项目：国家重点研发计划资助项目（2019YFA0903902）；国家自然科学基金资助项目（32270595）；深圳市基础研究计划资助项目（JCYJ20190808115005598）
作者简介：莫亦琳（2002—），深圳大学和加州大学伯克利分校联合培养本科生. 研究方向：shRNA在生物防治中的应用. E-mail: Lindsay_m@berkeley.edu
引文：莫亦琳，陈伟钊，黄丽娟，等.ShRNA-LDH纳米颗粒的制备及薇甘菊生物防治［J］.深圳大学学报理工版，2023，40（2）：163-170.（英文版）
更新日期/Last Update: 2023-03-30