Construction of Virus-Mediated Genetic Transformation System of Camellia sinensis

doi:10.16420/j.issn.0513-353x.2024-0075

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (11): 2710-2724.doi: 10.16420/j.issn.0513-353x.2024-0075

• New Technology and New Method • Previous Articles Next Articles

Construction of Virus-Mediated Genetic Transformation System of Camellia sinensis

HU Jinyu¹, LIU Guizhi¹, CHEN Lan¹, HUANG Mengdi¹, SU Qin¹, TAN Yueping³, LIU Shuoqian¹^,²^,^*(), TIAN Na¹^,²^,^*()

¹ Key Laboratory of Tea Science of Ministry of Education，College of Horticulture, Hunan Agricultural University，Changsha 410128，China
² Department of Tea Science，College of Horticulture，Hunan Agricultural University，Changsha 410128，China
³ Hunan Tea Group Co.，Ltd.，Changsha 410126，China

Received:2024-05-30 Revised:2024-07-16 Online:2024-12-12 Published:2024-11-26
Contact: LIU Shuoqian, TIAN Na

Abstract

Abstract:

In this study，Camellia sinensis phytoene desaturase gene（CsPDS）and tea caffeine synthase gene（TCS1）as indicator genes and Nicotiana benthamiana as a vector were employed to enrich tobacco rattle virus. The effects of the concentration of infiltration solution，incubation temperature，different vectors and incubation time on the enriched viruses were investigated，with N. benthamiana used as the vector to enrich the viruses. The effects of different inoculation methods and cultivars on the construction of a virus-induced gene silencing（VIGS）system were also investigated in tea plant. The results demonstrated that inoculation of tobacco with acetosyringone（AS）at a concentration of 200 μmol · L^-1 bacteriophage with an optical density（OD₆₀₀）of 1.0 was conducive to virus enrichment in tobacco when incubated at 23 ℃. The leaves of‘Shuchazao’were used to construct the VIGS system of tea plant by injection inoculation, and the system was successfully used to explore the possibility of foreign gene expression with GFP as the indicator gene.

Key words: Camellia sinensis, virus-induced gene silencing, tobacco rattle virus, phytoene desaturase gene, caffeine synthase 1 gene, green fluorescent protein gene

HU Jinyu, LIU Guizhi, CHEN Lan, HUANG Mengdi, SU Qin, TAN Yueping, LIU Shuoqian, TIAN Na. Construction of Virus-Mediated Genetic Transformation System of Camellia sinensis[J]. Acta Horticulturae Sinica, 2024, 51(11): 2710-2724.

Figures/Tables 12

Table 1 Information on primers used in this study

作用 Function	引物名称 Primer name	引物序列 Primer sequence
载体构造 Tectonic carrier	TRV2-CsPDS2	F：5′-gtgagtaaggttaccgaattcTGCTGACATGTCAGTGGCATG-3′ R：5′-cgtgagctcggtaccggatccGTCACCAGACAAATAGAACCCTTCT-3′
	TRV2-TCS1-2	F：5′-gtgagtaaggttaccgaattcGAAATGATTTCAATACCCTCTTCAAA-3′ R：5'-cgtgagctcggtaccggatccCATTGTGAAATCTTCATGAAATTGAG-3′
	C2b-GFP2	F：5′-gttgacagtagtggtgtcgacATGGTGAGCAAGGGCGAGG-3′ R：5′-ttcgggacatgcccggtcgacTTACTTGTACAGCTCGTCCATGCC -3′
	pTRV2-GFP2	F：5′-atttcctttaccattgacgtcATGGTGAGCAAGGGCGAGG -3′ F：5′-gctaccaacgacactgacgtcTTACTTGTACAGCTCGTCCATGCC-3′
病毒检测 Virus detection	pTRV1	F：5′-GGTATTTGGTGGCAACGACT-3′ R：5'-ACCGCTGTGCTTTGATTTCT-3′
	TRV2-CsPDS1	F：5′-TTGTTACTCAAGGAAGCACGAT-3′ R：5′-cgtgagctcggtaccggatccGTCACCAGACAAATAGAACCCTTCT-3′
	TRV2-TCS1-1	F：5′- TTGTTACTCAAGGAAGCACGAT-3′ R：5′-cgtgagctcggtaccggatccCATTGTGAAATCTTCATGAAATTGAG-3′
	C2b-GFP1	F：5′- TTGTTACTCAAGGAAGCACGAT-3′ F：5′-gttgacagtagtggtgtcgacATGGTGAGCAAGGGCGAGG-3′
	pTRV2-GFP1	F：5′- TTGTTACTCAAGGAAGCACGAT-3′ F：5′-gctaccaacgacactgacgtcTTACTTGTACAGCTCGTCCATGCC-3′
qRT-PCR	qCsPDS	F：5′-GCTACGATGAAGGAACTGGCAA-3′ R：5′-TGCAATGGACGACAAGGTTCAC-3′
	qTCS1	F：5′- CCTCTTCAAAGGCCTGTCGTCT-3′ R：5′- CAAGCCTTCTCTGCTTGTGA-3′
	CsGAPDH	F：5′-TTGGCATCGTTGAGGGTCT-3′ R：5′-CAGTGGGAACACGGAAAGC-3′

Fig. 1 Effects of different concentrations of As（μmol · L-1）and Agrobacterium tumefaciens（OD600）in infected solution containing pTRV2-CsPDS vector on tobacco infection

Fig. 2 Effect of temperature on the proliferation of tobacco virus inoculated with infection solution containing pTRV2-CsPDS vector

Fig. 3 Phenotypic changes of tobacco in different days after inoculation

Fig. 4 Effects of different inoculation methods of tobacco grinding liquid carrying pTRV2-CsPDS and pTRV2-c2b-CsPDS on tea plants growth

Fig. 5 Electrophoresis of different tea varieties inoculated with tobacco grinding liquid carrying pTRV2-CsPDS and pTRV2-c2b-CsPDS M：DL2000 marker. 1：Longjing 43；2：Shuchazao；3：Longjing Changye；4：Zhuyeqi；5：Tieguanyin；6：Fuding Dabai；7：Bixiangzao.

Fig. 6 RT-PCR electropherogram for the detection of viruses in the VIGS system of tea plants M：DL2000 marker；WT：Virus not injected. 1-7：Forward primer and reverse primer of pTRV1；8-13：Forward primer of RNA2 vector and reverse primer of target gene.

Table 2 Infection rates of tea plants

接种的载体 Inoculated vector	接种茶树数 Number of vaccinations	感染数 Number of infections	感染率/% Infection
pTRV1 + pTRV2-CsPDS	50	16	32
pTRV1 + pTRV2-c2b-CsPDS	50	21	42
pTRV1 + pTRV2-TCS1	50	15	30
pTRV1 + pTRV2-c2b-TCS1	50	19	38

Fig. 7 Changes of caffeine content in new buds and leaves of different parts at different time and 40 d after tea seedlings were inoculated with TCS1 gene virus silencing vector WT：Unvaccinated；**indicates extremely significant difference（P < 0.01）.

Fig. 8 Changes in the relative expression of TCS1 and CsPDS in the VIGS system of tea plants WT：Unvaccinated；** indicates extremely significant difference（P < 0.01）.

Fig. 9 Phenotypic changes of tea plants inoculated with VIGS system for 40 days

Fig. 10 Fluorescence diagram（a）and RT-PCR electrophoresis diagram（b）of tobacco inoculated with infection solution containing pTRV2-GFP

References 66

[1]	Ashihara H, Crozier A. 2001. Caffeine:a well known but little mentioned compound in plant science. Trends in Plant Science, 9:407-413.
[2]	Broderick S R, Jones M L. 2014. An optimized protocol to increase virus-induced gene silencing efficiency and minimize viral symptoms in Petunia. Plant Molecular Biology Reporter, 32:219-233. pmid: 24465085
[3]	Cai Wen,Chen Changming,Chen Guoju,Cao Bihao,Lei Jianjun. 2017. Application progress of virus-induced gene silencing technology in functional genomics research of Solanaceae plants. Journal of China Capsicum,(1):1-12. (in Chinese)
	蔡文, 陈长明, 陈国菊, 曹必好, 雷建军. 2017. 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,(1):1-12.
[4]	Chen Y X, Guo X Y, Gao T, Zhang N, Song C K. 2020. UGT74AF3 enzymes specifically catalyze the glucosylation of 4-hydroxy-2,5-dimethylfuran-3(2H)-one,an important volatile compound in Camellia sinensis. Horticultural research,7 (1):11.
[5]	Chen Zheng, Xin Liqin, Wu Chaoqun, Wang Qingqing, He Shaoyun. 2016. Research progress on the application of VIGS technology in ornamental plants. Northern Horticulture,(13):196-201. (in Chinese)
	陈峥, 辛丽琴, 吴超群, 王青青, 何少云. 2016. VIGS技术在观赏植物中应用的研究进展. 北方园艺,(13):196-201.
[6]	Dommes A B, Gross T, Herbert D B, Kivivirta K I, Becker A. 2019. Virus-induced gene silencing:empowering genetics in non-model organisms. Journal of Experimental Botany, 70 (3):757-770. doi: 10.1093/jxb/ery411 pmid: 30452695
[7]	Fu D Q, Zhu B Z, Zhu H L, Jiang W B, Luo Y B. 2005. Virus-induced gene silencing in tomato fruit. The Plant Journal, 43 (2):299-308.
[8]	Fu D Q, Zhu B Z, Zhu H L, Zhang H X, Xie Y H, Jiang W B, Zhao X D, Luo K B. 2006. Enhancement of virus-induced gene silencing in tomato by low temperature and low humidity. Molecules & Cells, 21(1):153-160.
[9]	Gedling C R, Ali E M, Gunadi A, Finer J J, Xie K, Liu Y, Yoshikawa N, Qu F, Dorrance A E. 2018. Improved apple latent spherical virus-induced gene silencing in multiple soybean genotypes through direct inoculation of agro-infiltrated Nicotiana benthamiana extract. Plant Methods, 14:1-10.
[10]	Guo H S, Ding S W. 2002. A viral protein inhibits the long range signaling activity of the gene silencing signal. The EMBO Journal, 21 (3):398-407.
[11]	Hao Mengyuan, Hang Qi, Shi Gongyao. 2022. Application and prospect of VIGS gene silencing technology in crop gene function research. China Agricultural Science and Technology Herald, 24 (1):1-13. (in Chinese)
	郝梦媛, 杭琦, 师恭曜. 2022. VIGS基因沉默技术在作物基因功能研究中的应用与展望. 中国农业科技导报, 24 (1):1-13. doi: 10.13304/j.nykjdb.2020.0781
[12]	Holzberg S, Brosio P, Gross C, Pogue G P. 2002. Barley stripe mosaic virus—induced gene silencing in a monocot plant. The Plant Journal, 30 (3):315-327.
[13]	Huang C J, Qian Y J, Li Z H, Zhou X P. 2012. Virus-induced gene silencing and its application in plant functional genomics. Science China:Life Sciences, 55 (2):99-108.
[14]	Huang Shoubo. 1981. Agrometeorological indicators of tea tree growth. Agrometeorology, 2 (3):54-58. (in Chinese)
	黄寿波. 1981. 茶树生长的农业气象指标. 农业气象, 2 (3):54-58.
[15]	Huang Shoubo. 1986. Relationship between meteorological factors and growth,yield and quality of tea trees. Scientia Agricultura Sinica, 19 (4):96. (in Chinese)
	黄寿波. 1986. 气象因子与茶树生育、产量、品质的关系. 中国农业科学, 19 (4):96.
[16]	John K M, Joshi S D, Mandal A K, Kumar S R, Kumar R R. 2009. Agrobacterium rhizogenes-mediated hairy root production in tea leaves[Camellia sinensis(L.) O. Kuntze]. Indian Journal of Biotechnology, 8 (4):430-434.
[17]	Khan A H, Akram A, Saeed M, Rahman M, Rehman A, Mansoor S, Amin I. 2022. Establishment of transcriptional gene silencing targeting the promoter regions of GFP,PDS,and PSY genes in cotton using virus-induced gene silencing. Molecular Biotechnology, 65:1052-1061.
[18]	Krenz B, Christina W, Holger J. 2010. Cell-free construction of disarmed Abutilon mosaic virus-based gene silencing vectors. Journal of Virological Methods, 169 (1):129-137. doi: 10.1016/j.jviromet.2010.07.010 pmid: 20638413
[19]	Kumagai M H, Donson J, Della C G, Harvey D, Hanley K, Grill L K. 1995. Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proceedings of the National Academy of Sciences of the United States of America, 92 (5):1679-1683.
[20]	Lee W S, Jason J R, Kostya K. 2015. Virus induced gene silencing(VIGS)for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance. Fungal Genetics & Biology, 79:84-88.
[21]	Li G D, Li Y, Yao X Z, Lu L T. 2022. Establishment of a virus-induced gene-silencing(VIGS)system in tea plant and its use in the functional analysis of CsTCS1. International Journal of Molecular Sciences, 24 (1):392.
[22]	Li Lingling, Jiang Changjun, Fang Wanping, Deng Weiwei. 2007. Preliminary study on dsTCS gene transformation in tea plants by pollen tube pathway method. Journal of Anhui Agricultural University, 34 (1):20-22. (in Chinese)
	李玲玲, 江昌俊, 房婉萍, 邓威威. 2007. 花粉管通道法对茶树进行dsTCS基因转化的初步研究. 安徽农业大学学报, 34 (1):20-22.
[23]	Li Ranran, Wang Shubin, Pan Baogui, Diao Weiping, Guo Guangjun, Liu Jinbing, Ge Wei. 2017. Optimization of gene silencing inoculation method induced by pepper virus. Jiangsu Agricultural Sciences, 45 (14):82-84. (in Chinese)
	李然然, 王述彬, 潘宝贵, 刁卫平, 郭广君, 刘金兵, 戈伟. 2017. 辣椒病毒诱导基因沉默接种方法的优化. 江苏农业科学, 45 (14):82-84.
[24]	Liao Shujuan, Li Huajun, Ji Dangling. 2004. Application of genetic engineering in tea breeding. Journal of Tea Science,(4):6-8. (in Chinese)
	廖书娟, 李华钧, 吉当玲. 2004. 基因工程在茶树育种中的应用. 茶叶科学技术,(4):6-8.
[25]	Lin S J, Chen Z P, Chen T T, Deng W W, Wan X C, Zhang Z L. 2023. Theanine metabolism and transport in tea plants(Camellia sinensis L.):advances and perspectives. Critical Reviews in Biotechnology, 43 (3):327-341.
[26]	Liu Fan,Wang Guoying,Cao Mingqing. 2003. Advances in Agrobacterium-mediated in-situ transgenic plants. Molecular Plant Breeding, 1 (1):108-116. (in Chinese)
	刘凡, 王国英, 曹鸣庆. 2003. 农杆菌介导的植物原位转基因方法研究进展. 分子植物育种, 1 (1):108-116.
[27]	Liu Y L, Schiff M, Marathe R, Dinesh-Kumar S P. 2002. Tobacco Rar1,EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. The Plant Journal, 30 (4):415-429.
[28]	Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2^{- ΔΔCT} method. Methods, 25 (4):402-408. doi: 10.1006/meth.2001.1262 pmid: 11846609
[29]	Mainaak M, Tapan K, Mondal P, Chand K. 2016. Biotechnological advances in tea(Camellia sinensis[L.]O. Kuntze):a review. Plant Cell Reports, 35 (2):255-287.
[30]	Mei Y, Whitham S A. 2018. Virus-induced gene silencing in maize with a foxtail mosaic virus vector. Methods in Molecular Biology, 1676:129-139. doi: 10.1007/978-1-4939-7315-6_7 pmid: 28986907
[31]	Mondal T K, Bhattacharya A, Laxmikumaran M. 2004. Recent advances of tea(Camellia sinensis)biotechnology. Plant Cell Tissue & Organ Culture, 76 (3):195-254.
[32]	Mao Qingli, Shi Zhaopeng, Li Ling, Liu Zhonghua, Zhu Qi. 2007. Inhibitory effect of tea catechin on Agrobacterium rhizogenes and screening of phenol-resistant strains. Journal of Tea Science, 27 (3):243-247. (in Chinese)
	毛清黎, 施兆鹏, 李玲, 刘仲华, 朱旗. 2007. 茶叶儿茶素对发根农杆菌的抑制作用及抗酚菌株筛选研究. 茶叶科学, 27 (3):243-247.
[33]	Mikic D. 2012. The question of autonomy of Bosnia and Herzegovina as Serbian self-defence. Politea, 2 (4):89-99.
[34]	Navarro J A, Jesus A, Sanchez N, Vicente P. 2019. Key checkpoints in the movement of plant viruses through the host. Advances in Virus Research, 104:1-64. doi: S0065-3527(19)30011-9 pmid: 31439146
[35]	Pan Jiajia, Zhang Dongmei, Meng Jian, Gao Sunan, Zhu Kaijie, Liu Junwei, Li Guohuai. 2023. Optimization and validation of PpPDS gene silencing induced by Prunus necrotic ring spot virus in Prunus persica. Acta Horticulturae Sinica, 50 (7):1587-1600. (in Chinese)
	潘佳佳, 张东梅, 孟建, 高苏南, 朱凯杰, 刘军伟, 李国怀. 2023. 李坏死环斑病毒诱导的桃 PpPDS 基因沉默体系的优化及验证. 园艺学报, 50 (7):1587-1600.
[36]	Qi Dongxia, Zhang Ying, Zhao Zhen, Zhang Weiwei, Chen Yuhui, Lian Yong, Tian Shibing, Liu Fuzhong. 2018. Optimization of virus-induced gene silencing system and function identification of SmIAA19 gene in eggplant. Acta Horticulturae Sinica, 45 (4):691-701. (in Chinese)
	齐东霞, 张映, 赵祯, 张伟伟, 陈钰辉, 连勇, 田时炳, 刘富中. 2018. 茄子TRV-VIGS体系的优化及SmIAA19基因功能初步分析. 园艺学报, 45 (4):691-701. doi: 10.16420/j.issn.0513-353x.2017-0643
[37]	Qu F, Ye X H, Hou C H, Shirley S, Thomas E C, MorrisT J. 2005. RDR6 has a broad-spectrum but temperature-dependent antiviral defense role in Nicotiana benthamiana. Journal of Virology, 79 (24):15209-15217.
[38]	Rahman J, Baldwin I T, Klaus G. 2021. California TRV-based VIGS vectors mediate gene silencing at elevated temperatures but with greater growth stunting. BMC Plant Biology, 21 (1):1-17.
[39]	Ratcliff F, Martin‐Hernandez A M, Baulcombe D C. 2001. Technical advance:tobacco rattle virus as a vector for analysis of gene function by silencing. The Plant Journal, 25 (2):237-245.
[40]	Ren Hengze, Li Danying, Yu Yating, Lü Wuyun, Hao Xinyuan, Wang Xinchao, Wang Yuchun. 2024. Research advances of strategies to engineer VIGS vectors and its application in plants. Acta Horticulturae Sinica, 51 (7):1455-1473. (in Chinese) doi: 10.16420/j.issn.0513-353x.2023-0753
	任恒泽, 李丹莹, 余亚婷, 吕务云, 郝心愿, 王新超, 王玉春. 2024. 植物VIGS载体构建策略研究与应用进展. 园艺学报, 51 (7):1455-1473.
[41]	Ruiz M T, Voinnet O, Baulcomb D C. 1998. Initiation and maintenance of virus-induced gene silencing. The Plant Cell, 10 (6):937-946. doi: 10.1105/tpc.10.6.937 pmid: 9634582
[42]	Senthil-Kumar M, Anand A, Uppalapati S R, Kirankumar S K. 2008. Virus-induced gene silencing and its applications. Cab Reviews Perspectives in Agriculture Veterinary Science Nutrition and Natural Resources, 3 (11):1-18.
[43]	Senthil-Kumar M, Hema R, Anand A, Li K, Udayakumar M, Mysore K S. 2007. A systematic study to determine the extent of gene silencing in Nicotiana benthamiana and other Solanaceae species when heterologous gene sequences are used for virus-induced gene silencing. New Phytologist, 176 (4):782-791. doi: 10.1111/j.1469-8137.2007.02225.x pmid: 17997764
[44]	Seonghoe J, Choi S C, Li H Y, An G, Schmelzer E, Bendahmane M. 2015. Functional characterization of Phalaenopsis aphrodite flowering genes PaFT1 and PaFD. PLoS ONE, 10 (8):e0134987.
[45]	Singh D K, Hee-Kyung L, Ismail D, Kirankumar S M. 2018. An efficient and improved method for virus-induced gene silencing in sorghum. BMC Plant Biology, 18 (1):1-12.
[46]	Singh H R, Bhattacharyya N, Agarwala N, Bhagawati P, Deka M, Das S. 2014. Exogenous gene transfer in Assam tea[Camellia assamica(Masters)] by Agrobacterium-mediated transformation using somatic embryo. Imedpub, 4 (3):166-175.
[47]	Tan Heping, Zhou Lihua, Qian Shanshan, Li Huaiping, Ye Deping. 2009. Progress in transgenic technology of tea plants. Wuhan Botanical Research, 27 (3):4. (in Chinese)
	谭和平, 周李华, 钱杉杉, 李怀平, 叶德萍. 2009. 茶树转基因技术研究进展. 武汉植物学研究, 27 (3):4.
[48]	Tan Y, Alfredas B, Attila M, Andrew H. 2020. Rapid,high efficiency virus-mediated mutant complementation and gene silencing in Antirrhinum. Plant Methods, 16 (1):1-12.
[49]	Tiedge K, Destremps J, Solano-Sanchez J. 2022. Foxtail mosaic virus-induced gene silencing(VIGS)in switchgrass(Panicum virgatum L.). Plant Methods, 18 (1):71. doi: 10.1186/s13007-022-00903-0 pmid: 35644680
[50]	Tuttle J R, Idris A M, Brown J K, Haigler C H, Robertson D. 2008. Geminivirus-mediated gene silencing from cotton leaf crumple virus is enhanced by low temperature in cotton. Plant Physiology, 148 (1):41-50. doi: 10.1104/pp.108.123869 pmid: 18621976
[51]	Wang C H, Cai X Z, Wang X M, Zheng Z. 2006. Optimisation of tobacco rattle virus-induced gene silencing in Arabidopsis. Functional Plant Biology, 33 (4):347-355.
[52]	Wang Q, Wu Y, Peng A Q, Cui J L, Zhao M Y, Pan Y T, Zhang M T, Tian K, Wilfried S, Song C K. 2022. Single-cell transcriptome atlas reveals developmental trajectories and a novel metabolic pathway of catechin esters in tea leaves. Plant Biotechnology Journal, 20 (11):2089-2106. doi: 10.1111/pbi.13891 pmid: 35810348
[53]	Wang R, Yang X X, Wang N, Liu X D, Richard S N, Li W M, Fan Z F, Zhou T. 2016. An efficient virus-induced gene silencing vector for maize functional genomics research. The Plant Journal, 86 (1):102-115. doi: 10.1111/tpj.13142 pmid: 26921244
[54]	Wang Y Q, Chai C L, Behnam K, Wolf-Rüdiger S, Michael K U, Malay C S, Yun K, Richard S N. 2021. An Efficient Brome mosaic virus-based gene silencing protocol for hexaploid wheat(Triticum aestivum L.). Frontiers in Plant Science,https://doi.org/10.3389.
[55]	Wei C L, Yang H, Wang S B, Zhao J, Liu C, Gao L P, Xia E H, Lu Y, Tai Y L, She G B. 2018. Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality. Proceedings of the National Academy of Sciences, 115 (18):E4151-E4158.
[56]	Wu Shan, Liang Yuerong, Lu Jianliang, Li Haoyan. 2005. Gene gun and optimization of transformation conditions of exogenous genes in tea plants combined with Agrobacterium. Journal of Tea Science, 25 (4):255-264. (in Chinese)
	吴姗, 梁月荣, 陆建良, 黎昊雁. 2005. 基因枪及其与农杆菌相结合的茶树外源基因转化条件优化. 茶叶科学, 25 (4):255-264.
[57]	Xi Biao, Liu Zusheng, Liang Yuerong, Yang Xiufang, Huang Weihong, Xu Jian, He Yuke, Bai Yongyan. 1997. Agrobacterium-mediated genetic transformation of tea plants. Journal of Tea Science,1 (Supplement):155-156. (in Chinese)
	奚彪, 刘祖生, 梁月荣, 杨秀芳, 黄卫红, 须健, 何玉科, 白永延. 1997. 发根农杆菌介导的茶树遗传转化. 茶叶科学,1 (增刊):155-156.
[58]	Xia E H, Zhang H B, Sheng J, Li K, Zhang Q J, Changhoon K, Zhang Y,LiuY, Zhu T, Li W. 2017. The tea tree genome provides insights into tea flavor and independent evolution of caffeine biosynthesis. Molecular Plant, 10 (6):866-877.
[59]	Xiang Wei, Shi Chengying, He Zhirong, Xu Yan. 2012. Establishment of Agrobacterium tumefaciens-mediated transgenic system for tea plants. Journal of Anhui Agricultural University, 39 (5):721-724. (in Chinese)
	项威, 史成颖, 贺志荣, 徐燕. 2012. 根癌农杆菌介导茶树转基因体系的建立. 安徽农业大学学报, 39 (5):721-724.
[60]	Xu P, ZhangY, Kang L, Roossinck M J, Mysore K S. 2006. Computational estimation and experimental verification of off-target silencing during posttranscriptional gene silencing in plants. Plant Physiology, 142 (2):429-440. doi: 10.1104/pp.106.083295 pmid: 16920874
[61]	Yang W, Chen X Y, Chen J H, Zheng P, Liu S Q, Tan X D, Sun B M. 2023. Virus-induced gene silencing in the tea plant(Camellia sinensis). Plants, 2 (17):3162.
[62]	Zhang Guanghui, Lü Youcai, Duan Hongxing. 2010. In vitro plant regeneration and genetic transformation of tea plants. Molecular Plant Breeding, 8 (2):345-349. (in Chinese)
	张广辉, 吕才有, 段红星. 2010. 茶树离体植株再生与遗传转化. 分子植物育种, 8 (2):345-349.
[63]	Zhao M Y, Zhang N, Gao T, Jin J Y, Jing T T, Wang M, Wu Y, Wan X C, Wilfried S, Song C K. 2020. Sesquiterpene glucosylation mediated by glucosyltransferase UGT91Q 2 is involved in the modulation of cold stress tolerance in tea plants. The New Phytologist, 226 (2):362-372.
[64]	Zhou Chengzhe, Chang Xiaojun, Zhu Chen, Cheng Chunzhen, Chen Yukun, Lai Zhongxiong, Lin Yuling, Guo Yuqiong. 2022. Establishment of in-situ transformation method of tea plants. Biotechnology Bulletin, 38 (2):263-268. (in Chinese) doi: 10.13560/j.cnki.biotech.bull.1985.2021-0635
	周承哲, 常笑君, 朱晨, 程春振, 陈裕坤, 赖钟雄, 林玉玲, 郭玉琼. 2022. 茶树原位转化方法的建立. 生物技术通报, 38 (2):263-268. doi: 10.13560/j.cnki.biotech.bull.1985.2021-0635
[65]	Zhou T, Liu X D, Fan Z F. 2018. Chapter 8 use of a virus gene silencing vector for maize functional genomics research. Methods and Protocols, 1676:141-150.
[66]	Zhou Y J, Deng Y J, Liu D, Wang H Z, Zhang X, Liu T T, Wang J B, Li Y, Ou L J, Liu F. 2021. Promoting virus-induced gene silencing of pepper genes by a heterologous viral silencing suppressor. Plant Biotechnology Journal, 19 (12):2398.

Construction of Virus-Mediated Genetic Transformation System of Camellia sinensis

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 12

References 66

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	REN Hengze, LI Danying, YU Yating, LÜ Wuyun, HAO Xinyuan, WANG Xinchao, and WANG Yuchun, . Research Advances of Strategies to Engineer VIGS Vectors and Its Application in Plants [J]. Acta Horticulturae Sinica, 2024, 51(7): 1455-1473.
[2]	ZHANG Lilan, YANG Jun, WANG Rangjian. Mining of Genetic Locus of Phenylethyl Alcohol Primrose Glycoside Abundance in Tea Plants [J]. Acta Horticulturae Sinica, 2024, 51(1): 77-90.
[3]	YE Yufan, WANG Yujie, FU Qianyuan, WANG Lu, HAO Xinyuan, DING Changqing, WANG Xinchao, CAO Hongli, LI Nana. Cloning and Expression Analysis of Mg-Chelatase H Subunit Gene CsChlH in Tea Plant（Camellia sinensis） [J]. Acta Horticulturae Sinica, 2024, 51(1): 91-102.
[4]	LI Yuteng, CHEN Yao, REN Hengze, LI Congcong, WANG Haoqian, CAO Hongli, YUE Chuan, HAO Xinyuan, WANG Xinchao. Identification，Expression Analysis and Interaction Validation of CsIDM in Tea Plants [J]. Acta Horticulturae Sinica, 2023, 50(8): 1679-1696.
[5]	HOU Binghao, GAO Ting, WEI Yuede, GAO Shuilian, CAI Yinbi, CHEN Zhiming, JIN Shan, ZHANG Xingtan, WANG Pengjie, YE Naixing. Studies on Genetic Variation of‘Tieguanyin’Tea Trees Asexual Reproduction Based on High-depth Genome Resequencing [J]. Acta Horticulturae Sinica, 2023, 50(7): 1505-1517.
[6]	PAN Jiajia, ZHANG Dongmei, MENG Jian, GAO Sunan, ZHU Kaijie, LIU Junwei, LI Guohuai. Optimization and Validation of PpPDS Gene Silencing Induced by Prunus Necrotic Ring Spot Virus in Prunus persica [J]. Acta Horticulturae Sinica, 2023, 50(7): 1587-1600.
[7]	WANG Zehan, YU Wentao, WANG Pengjie, LIU Caiguo, FAN Xiaojing, GU Mengya, CAI Chunping, WANG Pan, YE Naixing. WGCNA Analysis of Differentially Expressed Genes in Floral Organs of Tea Germplasms with Ovary-glabrous and Ovary-trichome [J]. Acta Horticulturae Sinica, 2023, 50(3): 620-634.
[8]	WEN Ting, LI Jing, ZHANG Jiaqi, WEI Yi, ZHU Wenrui, NI Dejiang, WANG Mingle. Expression and Functional Identification of Magnesium Transporter Gene CsMGT6 in Camellia sinensis [J]. Acta Horticulturae Sinica, 2023, 50(10): 2171-2182.
[9]	QIU Keli, WANG Yumin, HE Jinling, YU Hong, PAN Haifa, SHENG Yu, XIE Qingmei, CHEN Hongli, ZHOU Hui, ZHANG Jinyun. Identification of Peach Laccase Family Genes and Function Analysis of PpLAC21 [J]. Acta Horticulturae Sinica, 2022, 49(6): 1351-1362.
[10]	GONG Changyi, LIU Jiaojiao, DENG Qiang, ZHANG Lixin. Identification and Pathogenicity of Colletotrichum Species Causing Anthracnose on Camellia sinensis [J]. Acta Horticulturae Sinica, 2022, 49(5): 1092-1101.
[11]	XIE Siyi, ZHOU Chengzhe, ZHU Chen, ZHAN Dongmei, CHEN Lan, WU Zuchun, LAI Zhongxiong, GUO Yuqiong. Genome-wide Identification and Expression Analysis of CsTIFY Transcription Factor Family Under Abiotic Stress and Hormone Treatments in Camellia sinensis [J]. Acta Horticulturae Sinica, 2022, 49(1): 100-116.
[12]	QI Dongxia1,，ZHANG Ying1,，ZHAO Zhen1，ZHANG Weiwei1，CHEN Yuhui1，LIAN Yong1，TIAN Shibing2，and LIU Fuzhong1,**. Optimization of Virus-induced Gene Silencing System and Function Identification of SmIAA19 Gene in Eggplant [J]. ACTA HORTICULTURAE SINICA, 2018, 45(4): 691-701.
[13]	YANG Sha，ZHANG Bin，HAN Yao，YANG Xia，LI Mingyang，and GUO Yulong**. Cloning of Petunia PDS Gene and Its Application in shRNA-mediated Gene Silencing [J]. ACTA HORTICULTURAE SINICA, 2017, 44(2): 315-322.
[14]	ZHANG Lei, ZHU Li-Xin, XU Chuan, CUI Chun-Mei, SHENG Hong-Ya, LI Rui, WANG Hong-Qing. The Effect of Silencing Chalcone Synthase on Anthocyanin Metabolism in Peach [J]. ACTA HORTICULTURAE SINICA, 2015, 42(1): 31-37.
[15]	SONG Zhen, LI Zhong-An, ZHOU Chang-Yong. Research Advances of Virus-induced Gene Silencing（VIGS） [J]. ACTA HORTICULTURAE SINICA, 2014, 41(9): 1885-1894.