作物杂志,2023, 第2期: 4650 doi: 10.16035/j.issn.1001-7283.2023.02.007
郭燕1(), 刘志达1(), 康立茹2, 包婷婷1, 杨旭1, 赵君1, 张之为1()
Guo Yan1(), Liu Zhida1(), Kang Liru2, Bao Tingting1, Yang Xu1, Zhao Jun1, Zhang Zhiwei1()
摘要:
病毒诱导的基因沉默(VIGS)具有独特优势,被广泛应用于基因功能验证。高效基因沉默体系的建立和优化是对目标基因功能进行研究的前提和基础。以番茄品种money maker和携带八氢番茄红素脱氢酶(PDS)基因序列的TRV病毒载体为材料,研究了农杆菌不同OD值、不同侵染方法以及侵染后不同培养温度对PDS基因沉默株率的影响。结果表明,农杆菌OD值分别为0.50、1.00和1.50时,PDS基因的沉默株率分别为5%、13%和6%;当农杆菌OD值为1.0时,采用注射法、真空浸润法和注射处理24h后再进行真空浸润后的PDS基因沉默株率分别为13%、30%和15%;侵染后的培养温度对于基因沉默株率也有显著影响,在25℃时的沉默株率为13%,22℃时沉默株率达到46%。以上研究表明,农杆菌OD值为1.00时,注射24h后再进行真空浸润,接种后植株在22℃下生长能够提高番茄目标基因的沉默株率。
[1] | 李君明, 项朝阳, 王孝宣, 等. “十三五”我国番茄产业现状及展望. 中国蔬菜, 2021(2):13-20. |
[2] |
Sharma M K, Solanke A U, Singh Y, et al. A simple and efficient Agrobacterium-mediated procedure for transformation of tomato. Journal of Biosciences, 2009, 34(3):423-433.
doi: 10.1007/s12038-009-0049-8 pmid: 19805904 |
[3] |
Becker A, Lange M. VIGS-genomics goes functional. Trends in Plant Science, 2010, 15(1):1-4.
doi: 10.1016/j.tplants.2009.09.002 |
[4] |
Burch-Smith T M, Anderson J C, Martin G B, et al. Applications and advantages of virus-induced gene silencing for gene function studies in plants. The Plant Journal, 2004, 39(5):734-746.
doi: 10.1111/tpj.2004.39.issue-5 |
[5] | 宋震, 李中安, 周常勇. 病毒诱导的基因沉默(VIGS)研究进展. 园艺学报, 2014, 41(9):1885-1894. |
[6] | 蔡文, 陈长明, 陈国菊, 等. 病毒诱导基因沉默技术在茄科植物功能基因组学研究中的应用进展. 辣椒杂志, 2017, 15(1):1-12. |
[7] |
Ruiz M T, Voinnet O, Baulcombe D C. Initiation and maintenance of virus-induced gene silencing. The Plant Cell, 1998, 10(6):937- 946.
doi: 10.1105/tpc.10.6.937 pmid: 9634582 |
[8] | 刘冠, 赵婷婷, 杨欢欢, 等. VIGS在几种番茄真菌性病害的应用. 分子植物育种, 2016, 14(9):2377-2382. |
[9] |
Muthappa S K, Mysore K S. Virus-induced gene silencing can persist for more than 2 years and also be transmitted to progeny seedlings in Nicotiana benthamiana and tomato. Plant Biotechnology Journal, 2011, 9(7):797-806.
doi: 10.1111/pbi.2011.9.issue-7 |
[10] |
Thomas C L, Jones L, Baulcombe D C, et al. Size constraints for targeting post-transcriptional gene silencing and for RNA- directed methylation in Nicotiana benthamiana using a potato virus X vector. The Plant Journal, 2010, 25(4):417-425.
doi: 10.1046/j.1365-313x.2001.00976.x |
[11] |
Liu E, Page J E. Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus. Plant Methods, 2008, 4(1):1-13.
doi: 10.1186/1746-4811-4-1 |
[12] |
Tuttle J R, Idris A M, Brown J K, et al. Geminivirus-mediated gene silencing from cotton leaf crumple virus is enhanced by low temperature in cotton. Plant Physiology, 2008, 148(1):41-50.
doi: 10.1104/pp.108.123869 pmid: 18621976 |
[13] | Fu D Q, Zhu B Z, Zhu H L, et al. Enhancement of virus-induced gene silencing in tomato by low temperature and low humidity. Molecules and Cells, 2006, 21(1):153-160. |
[14] | 王鹏摇, 刘鹏, 耿丹丹, 等. 牡丹授粉受精基因PoMYBs的VIGS体系的建立. 青岛农业大学学报(自然科学版), 2021, 38(3):199-204,217. |
[15] |
Ryu C M, Anand A, Li K, et al. Agrodrench:a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse solanaceous species. Plant Journal, 2010, 40(2):322-331.
doi: 10.1111/tpj.2004.40.issue-2 |
[16] |
Ding X S, Schneider W L, Chaluvadi S R, et al. Characterization of a brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Molecular Plant-Microbe Interactions, 2006, 19(11):1229-1239.
doi: 10.1094/MPMI-19-1229 pmid: 17073305 |
[17] | 于国红, 马雪峰, 徐娜, 等. VIGS介导的番茄转录因子SIDREB2基因的耐旱特征. 生物技术通报, 2012(12):93-100. |
[18] |
Chen X, Duan X, Wang S, et al. Virus-induced gene silencing (VIGS) for functional analysis of MYB80 gene involved in solanum lycopersicum cold tolerance. Protoplasma, 2019, 256:409-418.
doi: 10.1007/s00709-018-1302-5 |
[19] | 刘冠, 赵婷婷, 杨欢欢, 等. VIGS在几种番茄真菌性病害的应用. 分子植物育种, 2016, 14(9):2377-2382. |
[20] | 杨同文, 武安泉, 盛东峰, 等. 病毒诱导的基因沉默在番茄功能基因组学研究中的应用进展. 园艺学报, 2014, 41(3):564- 576. |
[21] | 徐幼平, 徐秋芳, 宋晓毅, 等. 病毒诱导的基因沉默. 浙江大学学报(农业与生命科学版), 2008, 32(2):119-131. |
[22] |
Fei Y, Pyott D E, Molnar A. Temperature modulates virus‐ induced transcriptional gene silencing via secondary small RNAs. New Phytologist, 2021, 232:356-371.
doi: 10.1111/nph.v232.1 |
[23] |
Zhang X, Zhang X, Singh J, et al. Temperature-dependent survival of turnip crinkle virus-infected arabidopsis plants relies on an rna silencing-based defense that requires DCL2,AGO2,and HEN1. Journal of Virology, 2012, 86(12):6847-6854.
doi: 10.1128/JVI.00497-12 |
[24] |
Qu F, Ye X H, Hou G C, et al. RDR6 has a broad-spectrum but temperature-dependent antiviral defense role in Nicotiana benthamiana. Journal of Virology, 2005, 79(24):15209-15217.
doi: 10.1128/JVI.79.24.15209-15217.2005 |
[25] |
Ma L, Huang X, Yu R, et al. Elevated ambient temperature differentially affects virus resistance in two tobacco species. Phytopathology, 2016, 106(1):94-100.
doi: 10.1094/PHYTO-11-14-0300-R pmid: 26474332 |
[26] |
Yang J, Zhang T Y, Liao Q S, et al. Chinese wheat mosaic virus- induced gene silencing in monocots and dicots at low temperature. Frontiers in Plant Science, 2018, 9:1627.
doi: 10.3389/fpls.2018.01627 |
[27] | 周婷. TRV介导的马铃薯基因瞬时沉默体系建立及部分抗病相关基因功能初步鉴定. 武汉:华中农业大学, 2010. |
[28] |
Tuttle J R, Idris A M, Brown J K, et al. Geminivirus-mediated gene silencing from cotton leaf crumple virus is enhanced by low temperature in cotton. Plant Physiology, 2008, 148(1):41-50.
doi: 10.1104/pp.108.123869 pmid: 18621976 |
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