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Molecular Plant 2009年第5期2卷 1084-1094页

作者： Tanya Renner Jennifer Bragg Heather E. Driscoll Juliana Cho Andrew O. Jackson Chelsea D. Specht Department of Plant and Microbial Biology University of California Berkeley CA 94720 USA Present address: Genornics and gene Discovery USDA ARS PWA WRRC-GGD 800 Buchanan Street Albany CA 94710 USA

virus-induced gene silencing （VIGS） has been shown to be effective for transient knockdown of gene expression in plants to analyze the effects of specific genes in development and stress-related responses. VlGS is well established for studies of model systems and crops within the Solanaceae, Brassicaceae, Leguminaceae, and Poaceae, but only recently has been applied to plants residing outside these families. Here, we have demonstrated that barley stripe mosaic virus （BSMV） can infect two species within the Zingiberaceae, and that BSMV-VIGS can be applied to specifically down-regulate phytoene desaturase in the culinary ginger Zingiber officinale. These results suggest that extension of BSMV-VlGS to monocots other than cereals has the potential for directed genetic analyses of many important temperate and tropical crop species.

关键词： Barley stripe mosaic virus virus-induced gene silencing VIGS Zingiber officinale Monocot.

virus-induced gene silencing in Eggplant(Solanum melongena)

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Journal of Integrative Plant Biology 2012年第6期54卷 422-429页

作者： Haiping Liu Daqi Fu Benzhong Zhu Huaxue Yan Xiaoying Shen Jinhua Zuo Yi Zhu Yunbo Luo Laboratory of Fruit Biology College of Food Science&Nutritional EngineeringChina Agricultural UniversityBeijing 100083China

Eggplant （Solanum melongena） is an economically important vegetable requiring investigation into its various genomic functions. The current limitation in the investigation of genomic function in eggplant is the lack of effective tools available for conducting functional assays. virus-induced gene silencing （VIGS） has played a critical role in the functional genetic analyses. In this paper, TRV-mediated VIGS was successfully elicited in eggplant. We first cloned the CDS sequence of PDS （PHYTOENE DESATURASE） in eggplant and then silenced the PDS gene. Photo-bleaching was shown on the newly-developed leaves four weeks after agroinoculation, indicating that VIGS can be used to silence genes in eggplant. To further illustrate the reliability of VIGS in eggplant, we selected Chl H, Su and CLA1 as reporters to elicit VIGS using the high-pressure spray method. Suppression of Chl H and Su led to yellow leaves, while the depletion of CLA1 resulted in albino. In conclusion, four genes, PDS, Chl H, Su （Sulfur）, CLA1, were down-regulated significantly by VIGS, indicating that the VIGS system can be successfully applied in eggplant and is a reliable tool for the study of gene function.

关键词： virus-induced gene silencing tobacco rattle virus PHYTOENE DES, ATURASE Solanum melongena

virus-induced gene silencing and its application in plant functional genomics

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Science China(Life Sciences) 2012年第2期55卷 99-108页

作者： HUANG ChangJun QIAN YaJuan LI ZhengHe ZHOU XuePing State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou 310058China

virus-induced gene silencing is regarded as a powerful and efficient tool for the analysis of gene function in plants because it is simple, rapid and transformation-free. It has been used to perform both forward and reverse genetics to identify plant functional genes. Many viruses have been developed into virus-induced gene silencing vectors and gene functions involved in development, biotic and abiotic stresses, metabolism, and cellular signaling have been reported. In this review, we discuss the development and application of virus-induced gene silencing in plant functional genomics.

关键词： virus-induced gene silencing gene function gene expression

Establishment and Verification of An Efficient virus-induced gene silencing System in Forsythia

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Horticultural Plant Journal 2021年第1期7卷 81-88页

作者： Jianshuang Shen Weijia Si Yutong Wu Yang Xu Jia Wang Tangren Cheng Qixiang Zhang Huitang Pan Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular Breeding National Engineering Research Center for FloricultureBeijing Laboratory of Urban and Rural Ecological Environment of Beijing Municipal Education CommissionCollege of Landscape ArchitectureBeijing Forestry UniversityBeijing 100083China Departmentof Landscape Architectrue Jiyang College of Zhejiang A&F UniversityZhujiZhejiang 311800China

To understand the functional identification of large-scale genomic sequences in Forsythia,tobacco rattle virus(TRV)-mediated virus-induced gene silencing(VIGS),suitable for the plant,was explored in this *** results showed that the TRV-mediated VIGS system could be successfully used in Forsythia for silencing the reporter gene FsPDS(Forsythia phytoene desaturase)using stem infiltration and leaf *** the treated plants were pruned below the injection site after 7–15 d infection;the FsPDS was silenced and typical photobleaching symptoms were observed in newly sprouted leaves at the whole-plant ***,this system has been successfully tested and verified through virus detection and qRT-PCR *** the optimization,Forsythia magnesium chelatase subunit H(FsChlH)was silenced successfully in Forsythia using this system,resulting in yellow leaveswith decreased chlorophyll *** system was stable,highly efficient and had greater rapidity and convenience,which made it suitable to study the function of genes related to physiological pathways such as growth and development,and metabolic regulation in Forsythia.

关键词： Forsythia virus-induced gene silencing phytoene desaturase TRV whole-plant level

Applications of virus-induced gene silencing for identification of gene function in fruit

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Food Quality and Safety 2021年第3期5卷 222-230页

作者： Gangshuai Liu Hongli Li Daqi Fu Laboratory of Fruit Biology College of Food Science&Nutritional EngineeringChina Agricultural UniversityBeijingChina

With the development of bioinformatics,it is easy to obtain information and data about thousands of genes,but the determi nation of the functions of these genes depends on methods for rapid and effective functi on al ***-induced gene sile ncing(VIGS)is a mature method of gene functional identification developed over the last 20 years,which has been widely used in many research fields involving many *** quality formation is a complex biological process,which is closely related to ***,we review the progress and contribution of VIGS to our understanding of fruit biology and its advantages and disadvantages in determining gene function.

关键词： gene function virus-induced gene silencing fruit viral vector.

Heterologous virus-induced gene silencing is an efficient approach for functional genomics in woody plants

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Heterologous virus-induced gene silencing is an efficient ap...

Application of virus-induced gene silencing approach in Camptotheca acuminata

第十四届全国植物基因组学大会

作者： Yuanzhong Jiang Shenglong Ye Lijun Wang Yanjiao Duan Keming Luo Key Laboratory of Eco-environments of Three Gorges Reservoir Region Ministry of EducationChongqing Key Laboratory of Transgenic Plant and Safety ControlInstitute of Resources BotanySchool of Life SciencesSouthwest University

virus-induced gene silencing(VIGS) is an effective tool for studying the functions of plant genes,but only a few VIGS vectors available for woody plants were reported so *** we present an effective heterologous VIGS s... 详细信息

关键词： virus-induced gene silencing functional genomics Vernicia fordii Populus Nicotiana benthamiana Camellia oleifera

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Application of virus-induced gene silencing approach in Camp...

A small knottin-like peptide negatively regulates in wheat to stripe rust resistance during early infection of wheat

2016年全国植物生物学大会

作者： Yanyan Wang Fang Yu School of Biological Engineering Dalian Polytechnic University

Camptotheca acuminata is the major plant for the production of camptothecin(CPT),an important anti-cancer drug used for the treatment of various cancers throughout the world. The low accumulation of CPT in plants limited its supply in the market and led to urgent need of promoting its accumulation by means of plant metabolic engineering,which relied on deep understanding of CPT biosynthesis pathway. However,missing of most of pathway genes restricted the attempts for regulating CPT biosynthesis. To unveil the CPT biosynthesis pathway,many efforts have been done for pathway gene identification and the application of large-scale RNA-sequencing technique accelerated screening of candidate genes which could be involved in CPT biosynthesis. To identify the function of these candidate genes in planta,it needs to develop an effective approach,such as virusinduced gene silencing(VIGS) in C. acuminata. In this work,a Tobacco Rattle virus-based VIGS method was developed and the application of this method successfully silenced the expression of four known genes involved in the early steps of CPT biosynthesis and resulted in clear decrease of CPT and 10-hydroxycamptothecin(10-HCPT) accumulation. This VIGS approach could be further applied to functional identification of candidate genes to elucidate CPT biosynthesis in C. acuminata.

关键词： Camptotheca acuminata virus-induced gene silencing camptothecin biosynthesis pathway alkaloids

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The Crop Journal 2023年第2期11卷 457-467页

作者： Shuangyuan Guo Min Li Huankun Li Feng Zhang Qiong Zhang Xueling Huang Xing Li Xiaojie Wang Zhensheng Kang Xinmei Zhang College of Life Sciences Northwest A&F UniversityYangling 712100ShaanxiChina College of Plant Protection Northwest A&F UniversityYangling 712100ShaanxiChina State Key Laboratory of Crop Stress Biology for Arid Areas Yangling 712100ShaanxiChina

Although Blufensins(Bln)have important functions in the response of plants to biotic stress the precise functioning of Bln in wheat remains largely *** we isolated a Bln gene(TaBln4)from Suwon 11 infected by Puccinia striiformis ***(Pst).Expression of TaBln4 increased in host plants at the early stage of infection with a virulent Pst race(CYR31)but was unchanged in response to infection by an avirulent race(CYR23).Transcription levels of TaBln4 were also regulated by hormone and abiotic *** of TaBln4 in tobacco leaves suppressed Bax-induced programmed cell *** of TaBln4 by virus-induced gene silencing inhibited colonization of race CYR31 by increasing the accumulation of H2O2 and formation of hypersensitive responses(HR).Transient overexpression of TaBln4 by a transient overexpression system(BSMV-VOX)increased the susceptibility of wheat to *** from bimolecular fluorescence complementation and pull-down assays demonstrated that TaBLN4 interacted with *** together,our results suggest that TaBln4 negatively regulates resistance in wheat to Pst in a reactive oxygen species(ROS)-and HR-dependent manner.

关键词： HR Hypersensitivity Puccinia striiformis f.sp.tritici Triticum aestivum virus-induced gene silencing

Cloning and Function Identification of a Phytoene Desaturase gene from Eucommia ulmoides

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Phyton-International Journal of Experimental Botany 2023年第5期92卷 1377-1389页

作者： Jiali Wang Xiangmei Chen Xiaozhen Huang Yichen Zhao Degang Zhao The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education) College of Tea SciencesCollege of Life SciencesGuizhou UniversityGuiyang550000China Plant Conservation Technology Center Guizhou Key Laboratory of Agricultural BiotechnologyGuizhou Academy of Agricultural SciencesGuiyang550000China

The phytoene desaturase(PDS)encodes a crucial enzyme in the carotenoid biosynthesis *** or inhibiting PDS expression leads to the appearance of mottled,chlorosis,or albino *** this study,the CDS sequence of EuPDS(Eucommia ulmoides Phytoene Desaturase)was first cloned and then PDS was silenced in Nicotiana *** showed the expression level of EuPDS in leaves was higher than that in the roots and *** *** leaves,which were treated by Agrobacterium for 24 h,photo-bleaching was shown on the fresh leaves one week after injection and the transcript level of PDS was down-regulated during the period of *** suggested that EuPDS could silence PDS of ***,so as to cause the phenotype of leaf *** is the main reporter gene involved in virus-induced gene silencing(VIGS).This study offered molecular evidence for identifying PDS gene involved in Carotenoid’s biosynthesis pathway and the regulation networks in *** also laid a useful foundation for study on leaf discoloration mechanism of other woody plants.

关键词： Eucommia ulmoides EuPDS virus-induced gene silencing TRV relative expression

Tomato Fruit Development and Ripening Are Altered by the silencing of LeEIN2 gene

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Journal of Integrative Plant Biology 2006年第12期48卷 1478-1485页

作者： Hong-Liang Zhu Ben-Zhong Zhu Yi Shao Xiao-Guang Wang Xi-Jin Lin Yuan-Hong Xie Ying-Cong Li Hong-Yan Gao Yun-Bo Luo Laboratory of Fruit Biology College of Food Science and Nutritional Engineering China Agricultural University Beijing 100083 China

Loss-of-function ethylene insensitive 2 （EIN2） mutations showed ethylene insensitivity in Arabidopsis, which indicated an essential role of EIN2 in ethylene signaling. However, the function of EIN2 in fruit ripening has not been investigated. To gain a better understanding of EIN2, the temporal regulation of LeEIN2 expres- sion during tomato fruit development was analyzed. The expression of LeEIN2 was constant at different stages of fruit development, and was not regulated by ethylene. Moreover, LeEIN2-silenced tomato fruits were developed using a virus-induced gene silencing fruit system to study the role of LeEIN2 in tomato fruit ripening. Silenced fruits had a delay in fruit development and ripening, related to greatly descended expression of ethylene-related and ripening-related genes in comparison with those of control fruits. These results suggested LeEIN2 positively mediated ethylene signals during tomato development. In addition, there were fewer seeds and Iocules in the silenced fruit than those in the control fruit, like the phenotype of parthenocarpic tomato fruit. The content of auxin and the expression of auxin-regulated gene were declined in silenced fruit, which indicated that EIN2 might be important for crosstalk between ethylene and auxin hormones.

关键词： ethylene fruit development LeEIL1 LeEIN2 tomato virus-induced gene silencing