Study on Chinese Tomato Yellow Leaf Curl Virus-induced Gene Silencing(VIGS)via Root Absorption

Abstract

Abstract: The tomato yellow leaf curl virus satellite DNA was used as vector which containing the Su gene, and transformed the plasmid into Agrobacterium by freeze thawing method. Agrobacterium infected plants via root absorption, and virus-induced gene silencing system was optimized, twexplore the influence to virus-induced gene silencing efficiency. The results showed infected 22-25 days tomato seedling via OD600 1.5 Agrobacterium-mediated root-absorption which containing plasmid pBinPLUS-2mβ-Su and pBinPLUS-1.7A, the silencing of Su gene resulted in photo-blcached young leaves of tomato plants. Semi-quantitative RT-PCR analysis was employed to test the effect of Su gene silencing, which showed that Su gene was degraed significantly. This gene-silencing system would benefit for the high-throughput analysis of plant gene.

Key words: Virus-induced gene silencing, Su gene, Root absorption method

Zhang Zhaojun, Wang Xiaobin, Wang Hui, Liu Lin, Zhang Xinge, He Xiuxia. Study on Chinese Tomato Yellow Leaf Curl Virus-induced Gene Silencing(VIGS)via Root Absorption[J]. Biotechnology Bulletin, 2014, 0(1): 143-146.

References

[1] Baulcombe DC. Fast forward genetics based on virus-induced gene silencing[J].Curr Opin Plant Biol, 1999, 2(2): 109-113.
[2] Dinesh-kumar SP, Anandalakshmi R, Marathe R, et al. Virus-induced gene silencing[J].Methods Mol Biol, 2003, 236: 287-293.
[3] Fu DQ, Zhu BZ, Zhu HL, et al. Virus-induced gene silencing in tomato fruit[J].The Plant Journal, 2005, 43(2): 299-308.
[4] 陶小荣, 周雪平, 崔晓峰, 等. 病毒诱导的基因沉默及其在植物基因功能研究中的应用[J]. 生物化学与生物物理进展, 2004,31(9): 777-783.
[5] 周晓馥, 王兴智. 病毒载体诱导转录后基因沉默系统的建立及烟草基因功能的研究[J]. 分子植物育种, 2003, 1(3): 423- 424.
[6] 傅达奇, 朱本忠, 赵晓丹, 等. 病毒诱导烟草的基因沉默[J].西北植物学报, 2005, 25(8): 1517-1520.
[7] Brigneti G, Martin-Hernandez AM, et al.Virus-induced gene silencing in Solanum species[J].Plant J, 2004, 39(2): 264-272.
[8] 程维舜, 徐秋芳, 等. 适于烟草脆裂病毒诱导的本氏烟基因沉默分析的对照载体构建[J]. 浙江大学学报, 2012, 38(1): 10-20.
[9] Brigneti G, Martín-Hernández AM, Jin H, et al. Virus-induced gene silencing in Solanum species[J].Plant J, 2004, 39(2): 264-272.
[10] Chung E, Seong E, Kim YC, et al. A methodof high frequency virus-induced gene silencing in chili pepper Capsicum annuum Lcvbukang[J]. Mol Cell, 2004, 17(23): 377-380.
[11] Constantin GD, Krath BN, MacFarlane SA, et al. Virus-induced gene silencing as a tool for functional genomics in a legume species[J]. Plant J, 2004, 40(4): 622-631.
[12] 燕飞, 宋雪梅, 成卓敏. 番茄丛矮病毒p19 蛋白抑制转录后基因沉默作用机制[J]. 病毒学报, 2005, 21(5): 403-405.
[13] 杨美英, 贺红霞, 王艳, 等. 番茄茄红素β-环化酶基因高效沉默载体构建[J]. 分子植物育种, 2007, 5(1): 43-46.
[14] 陶小荣, 钱亚娟, 等. 卫星沉默载体的改良及其改变矮牵牛叶色和花色的研究[J]. 科学通报, 2006, 51(17): 2041-2044.
[15] 黄昌军, 钱亚娟, 李正和, 等. 病毒诱导的基因沉默及其在植物功能基因组研究中的应用[J]. 中国科学, 2012, 42(1): 3-15.
[16] Fu DQ, Zhu BZ, Zhu HL, et al. Enhancement of virus-induced gene silencing through low temperature and low humidity in tomato[J]. Mol Cells, 2006, 21(2): 153-160.
[17] Ryu CM, Anand A, Kang L, et al. Agrodrench: a novel and effective agroinoculation method for virus-induced gene silencing in roots and diverse Solanaceous species[J].Plant J, 2004, 40(2): 322-331.