Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (06): 844-849.doi: 10.3724/SP.J.1006.2016.00844


Establishment and Optimisation of Virus-Induced Gene Silencing in System Hydroponic Cotton

MU Chun,ZHOU Lin,LI Mao-Ying,DU Ming-Wei,ZHANG Ming-Cai,TIAN Xiao-Li,LI Zhao-Hu*   

  1. Engineering Research Center of Plant Growth Regulator, Ministry of Education / College of Agronomy, China Agricultural University, Beijing 100193, China
  • Received:2016-01-02 Revised:2016-03-14 Online:2016-06-12 Published:2016-03-21
  • Contact: 李召虎, E-mail: lizhaohu@cau.edu.cn, Tel: 010-62733049 E-mail:muc320@163.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31271628).


This experiment using GhCLA1 as a marker gene and cotton variety Guoxinmian 3 plants as material was conducted to explore effects of temperature, syringe-infiltrated concentrations and time, cultivation patterns, and cotton varieties on efficiency of tobacco rattle virus (TRV)-induced gene silencing (VIGS) under hydroponic condition. The results showed that higher silencing efficiency was induced by syringe-infiltrated time at 3 to 5 days after emergence and optimum growth temperature at 24 ºC under hydroponic condition, but syringe-infiltrated concentrations could not affect VIGS silence efficiency. Moreover, pTRV-GFP as null fragment could alleviate the adverse effect of inserted fragment for plant growth. Silencing phenotype could be visible earlier in hydroponics culture than in soil culture, and the experimental period was significantly shortened under hydroponic condition. In addition, GhCLA1 could be silenced in all tested varieties (lines) under hydroponic condition. Cotton plants with silenced GhCTR1 were severely dwarfed, which indicated TRV-VIGS system can be applied widely in hydroponic cotton.

Key words: Cotton, VIGS, GhCLA1, Hydroponic

[1] Cai X Z, Xu Q F, Wang C C, Zheng Z. Development of a virus-induced gene-silencing system for functional analysis of the RPS2-dependent resistance signalling pathways in Arabidopsis. Plant Mol Biol, 2006, 62: 223–232
[2] Hou P Q, Lee Y I, Hsu K T, Lin Y T, Wu W Z, Lin J Y, Nam T N, Fu S F. Functional characterization of Nicotiana benthamiana chromomethylase 3 in developmental programs by virus-induced gene silencing. Physiol Plant, 2014, 150: 119–132
[3] 黄赛花, 郑桂杰, 杨永庆,智海剑. 利用VIGS技术对抗SMV候选基因GmZ15的功能分析. 大豆科学, 2015, 34: 582–587
Huang S H, Zheng G J, Yang Y Q, Zhi H J. Analysis on the candidate resistance gene GmZ15 to soybean mosaic virus by VIGS. Soybean Sci, 2015, 34: 582–587 (in Chinese with English abstract)
[4] Becker A, Lange M. VIGS–genomics goes functional. Trends Plant Sci , 2010, 15: 1–4
[5] Ratcliff F G, MacFarlane S A, Baulcombe D C. Gene silencing without DNA: RNA-mediated cross protection between viruses. Plant Cell, 1999, 11: 1207–1215
[6] Gao X Q, Wheeler T, Li Z H, Kenerley C M, He P, Shan L B. Silencing GhNDR1 and GhMKK2 compromises cotton resistance to Verticillium wilt. Plant J, 2011, 66: 293–305
[7] 王丽, 穆春, 张明才, 杜明伟, 田晓莉, 李召虎.GhCPS基因沉默对棉花幼苗生长和内源激素含量的影响. 棉花学报, 2014, 26: 189–196
Wang L, Mu C, Zhang M C, Du M W, Tian X L, Li Z H. Effect of silencing GhCPS on the growth and endogenous hormone content of cotton seedlings(Gossypium himutum L.) Cotton Sci, 2014, 26: 189–196 (in Chinese with English abstract)
[8] 王心宇, 吕坤, 蔡彩平, 徐君, 郭旺珍. TRV病毒介导的基因沉默体系在棉花中的建立及应用. 作物学报, 2014, 40: 1356–1363
Wang X Y, Lü K, Cai C P, Xu J, Guo W Z. Establishment and application of TRV-mediated virus-induced gene silencing in cotton. Acta Agron Sin, 2014, 40: 1356–1363 (in Chinese with English abstract)
[9] Faivre-Rampant O, Gilroy E M, Hrubikova K, Hein I, Millam S, Loake G J, Birch P, Taylor M, Lacomme C. Potato virus X-induced gene silencing in leaves and tubers of potato. Plant Physiol, 2004, 134:1308–1316
[10] Liu E, Page J E. Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus. Plant Methods, 2008, 4: 1–13
[11] Lichtenthaler H K, Wellbum A R. Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans, 1983, 11: 591–592
[12] Fu D Q, Zhu B Z, Zhu H L, Zhang H X, Xie Y H, Jiang W B, Zhao X D, Luo Y B. Enhancement of virus-induced gene silencing in tomato by low temperature and low humidity. Mol. Cells, 2006, 21: 153–160
[13] Tuttle J R, Idris A M, Brown J K, Haigler C H, Robertson D. Geminivirus-mediated gene silencing from cotton leaf crumple virus is enhanced by low temperature in cotton. Plant Physiol. 2008, 148: 41–50
[14] Szittya G, Silhavy D, Molnar A, Havelda Z, Lovas A, Lakatos L, Banfalvi Z, Burgyan J. Low temperature inhibits RNA silencing-mediated defence by the control of siRNA generation. Embo J, 2003, 22: 633–640
[15] Burch‐Smith T M, Anderson J C, Martin G B, Dinesh‐Kumar S P. Applications and advantages of virus‐induced gene silencing for gene function studies in plants. Plant J , 2004, 39: 734–746
[16] Nethra P, Nataraja K N, Rama N, Udayakumar M. Standardization of environmental conditions for induction and retention of post-transcriptional gene silencing using tobacco rattle virus vector. Curr Sci, 2006, 90: 431–435
[17] Ekengren S K, Liu Y L, Schiff M, Dinesh-Kumar S P, Martin G B, Two MAPK cascades, NPR1, and TGA transcription factors play a role in Pto-mediated disease resistance in tomato. Plant J, 2003, 36: 905–917
[18] Hartl M, Merker H, Schmidt D D, Baldwin I T. Optimized virus-induced gene silencing in Solanum nigrum reveals the defensive function of leucine aminopeptidase against herbivores and the shortcomings of empty vector controls. New Phytol, 2008, 179: 356–365
[19] Wang C C, Cai X Z, Wang X M, Zheng Z. Optimisation of tobacco rattle virus-induced gene silencing in Arabidopsis. Funct Plant Biol, 2006, 33: 347–355
[20] Kieber J J, Rothenberg M, Roman G, Feldmann K A, Ecker J R. CTR1, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the Raffamily of protein kinases. Cell, 1993, 72: 427–441

[1] ZHOU Jing-Yuan, KONG Xiang-Qiang, ZHANG Yan-Jun, LI Xue-Yuan, ZHANG Dong-Mei, DONG He-Zhong. Mechanism and technology of stand establishment improvements through regulating the apical hook formation and hypocotyl growth during seed germination and emergence in cotton [J]. Acta Agronomica Sinica, 2022, 48(5): 1051-1058.
[2] SUN Si-Min, HAN Bei, CHEN Lin, SUN Wei-Nan, ZHANG Xian-Long, YANG Xi-Yan. Root system architecture analysis and genome-wide association study of root system architecture related traits in cotton [J]. Acta Agronomica Sinica, 2022, 48(5): 1081-1090.
[3] YAN Xiao-Yu, GUO Wen-Jun, QIN Du-Lin, WANG Shuang-Lei, NIE Jun-Jun, ZHAO Na, QI Jie, SONG Xian-Liang, MAO Li-Li, SUN Xue-Zhen. Effects of cotton stubble return and subsoiling on dry matter accumulation, nutrient uptake, and yield of cotton in coastal saline-alkali soil [J]. Acta Agronomica Sinica, 2022, 48(5): 1235-1247.
[4] ZHENG Shu-Feng, LIU Xiao-Ling, WANG Wei, XU Dao-Qing, KAN Hua-Chun, CHEN Min, LI Shu-Ying. On the green and light-simplified and mechanized cultivation of cotton in a cotton-based double cropping system [J]. Acta Agronomica Sinica, 2022, 48(3): 541-552.
[5] ZHANG Yan-Bo, WANG Yuan, FENG Gan-Yu, DUAN Hui-Rong, LIU Hai-Ying. QTLs analysis of oil and three main fatty acid contents in cottonseeds [J]. Acta Agronomica Sinica, 2022, 48(2): 380-395.
[6] ZHANG Te, WANG Mi-Feng, ZHAO Qiang. Effects of DPC and nitrogen fertilizer through drip irrigation on growth and yield in cotton [J]. Acta Agronomica Sinica, 2022, 48(2): 396-409.
[7] ER Chen, LIN Tao, XIA Wen, ZHANG Hao, XU Gao-Yu, TANG Qiu-Xiang. Coupling effects of irrigation and nitrogen levels on yield, water distribution and nitrate nitrogen residue of machine-harvested cotton [J]. Acta Agronomica Sinica, 2022, 48(2): 497-510.
[8] ZHAO Wen-Qing, XU Wen-Zheng, YANG Liu-Yan, LIU Yu, ZHOU Zhi-Guo, WANG You-Hua. Different response of cotton leaves to heat stress is closely related to the night starch degradation [J]. Acta Agronomica Sinica, 2021, 47(9): 1680-1689.
[9] YUE Dan-Dan, HAN Bei, Abid Ullah, ZHANG Xian-Long, YANG Xi-Yan. Fungi diversity analysis of rhizosphere under drought conditions in cotton [J]. Acta Agronomica Sinica, 2021, 47(9): 1806-1815.
[10] ZENG Zi-Jun, ZENG Yu, YAN Lei, CHENG Jin, JIANG Cun-Cang. Effects of boron deficiency/toxicity on the growth and proline metabolism of cotton seedlings [J]. Acta Agronomica Sinica, 2021, 47(8): 1616-1623.
[11] GAO Lu, XU Wen-Liang. GhP4H2 encoding a prolyl-4-hydroxylase is involved in regulating cotton fiber development [J]. Acta Agronomica Sinica, 2021, 47(7): 1239-1247.
[12] MA Huan-Huan, FANG Qi-Di, DING Yuan-Hao, CHI Hua-Bin, ZHANG Xian-Long, MIN Ling. GhMADS7 positively regulates petal development in cotton [J]. Acta Agronomica Sinica, 2021, 47(5): 814-826.
[13] XU Nai-Yin, ZHAO Su-Qin, ZHANG Fang, FU Xiao-Qiong, YANG Xiao-Ni, QIAO Yin-Tao, SUN Shi-Xian. Retrospective evaluation of cotton varieties nationally registered for the Northwest Inland cotton growing regions based on GYT biplot analysis [J]. Acta Agronomica Sinica, 2021, 47(4): 660-671.
[14] ZHOU Guan-Tong, LEI Jian-Feng, DAI Pei-Hong, LIU Chao, LI Yue, LIU Xiao-Dong. Efficient screening system of effective sgRNA for cotton CRISPR/Cas9 gene editing [J]. Acta Agronomica Sinica, 2021, 47(3): 427-437.
[15] HAN Bei, WANG Xu-Wen, LI Bao-Qi, YU Yu, TIAN Qin, YANG Xi-Yan. Association analysis of drought tolerance traits of upland cotton accessions (Gossypium hirsutum L.) [J]. Acta Agronomica Sinica, 2021, 47(3): 438-450.
Full text



No Suggested Reading articles found!