scholarly journals An efficient virus-induced gene silencing vector for maize functional genomics research

2016 ◽  
Vol 86 (1) ◽  
pp. 102-115 ◽  
Author(s):  
Rong Wang ◽  
Xinxin Yang ◽  
Nian Wang ◽  
Xuedong Liu ◽  
Richard S. Nelson ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Virus Induced Gene Silencing ◽  
Genomics Research

scholarly journals An efficient virus-induced gene silencing (VIGS) system for functional genomics in Brassicas using a cabbage leaf curl virus (CaLCuV)-based vector

2020 ◽  
Author(s):  
Zhiliang Xiao ◽  
Miaomiao Xing ◽  
Xing Liu ◽  
Zhiyuan Fang ◽  
Limei Yang ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Incubation Temperature ◽  
Virus Induced Gene Silencing ◽  
Posttranscriptional Gene Silencing ◽  
Cabbage Leaf Curl Virus ◽  
Brassica Crops ◽  
Genomics Research ◽  
Lobed Leaf ◽  
Leaf Curl Virus

Abstract Background: Virus-induced gene silencing (VIGS), a posttranscriptional gene silencing method, represents an effective technology for the analysis gene functions in plants. However, no VIGS vectors are available for Brassica oleracea till now. The gene silencing vectors TRV, pTYs and CaLCuV were chose to improve the VIGS system in cabbage, using phytoene desaturase (PDS) gene as an efficient visual indicator for VIGS.Results: We successfully silenced the expression of PDS and observed the photobleaching phenomenon in cabbage by pTYs and CaLCuV, with the latter displaying higher efficiency and lower cost. Then, the factors potentially affecting the silencing efficiency of VIGS by CaLCuV in cabbage, including targeting fragment strategy, inoculation method and incubation temperature, were compared. The optimized CaLCuV-based VIGS system is as follows: approximately 500 bp insert sequence, Agrobacterium OD600=1.0, vacuum osmosis method at the bud stage, and the incubation temperature of 22 °C, through which we achieved a stable silencing efficiency of 65%. To further test the effectiveness of the system, we selected two other genes in cabbage including Mg-chelataseH subunit (ChlH) and lobed-leaf 1 (LL1) to knock down their expression, and we observed the expected yellow or lobed leaves. In addition, we successfully applied the CaLCuV-based VIGS system in two other representative Brassica crops including B. rapa and B. nigra and thus expanded its application scope.Conclusion: These results demonstrate that CaLCuV-based VIGS effectively work in cabbage and can be used for analyzing gene function. Our VIGS system described here will contribute to efficient functional genomics research in Brassica crops.

scholarly journals Agrobacterium-Mediated Gene Transient Overexpression and Tobacco Rattle Virus (TRV)-Based Gene Silencing in Cassava

2019 ◽  
Vol 20 (16) ◽  
pp. 3976 ◽  
Author(s):  
Hongqiu Zeng ◽  
Yanwei Xie ◽  
Guoyin Liu ◽  
Yunxie Wei ◽  
Wei Hu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Transient Expression ◽  
Fluorescent Protein ◽  
Tobacco Rattle Virus ◽  
Mosaic Disease ◽  
African Cassava Mosaic Virus ◽  
Cassava Mosaic Disease ◽  
Virus Induced Gene Silencing ◽  
Transient Expression Assay

Agrobacterium-mediated transient expression and virus-induced gene silencing (VIGS) are very useful in functional genomics in plants. However, whether these methods are effective in cassava (Manihot esculenta), one of the most important tropical crops, remains elusive. In this study, we used green fluorescent protein (GFP) and β-glucuronidase (GUS) as reporter genes in a transient expression assay. GFP or GUS could be detected in the infiltrated leaves at 2 days postinfiltration (dpi) and were evidenced by visual GFP and GUS assays, reverse-transcription PCR, and Western blot. In addition, phytoene desaturase (PDS) was used to show the silencing effect in a VIGS system. Both Agrobacterium GV3101 and AGL-1 with tobacco rattle virus (TRV)-MePDS-infiltrated distal leaves showed an albino phenotype at 20 dpi; in particular, the AGL-1-infiltrated plants showed an obvious albino area in the most distal leaves. Moreover, the silencing effect was validated by molecular identification. Notably, compared with the obvious cassava mosaic disease symptom infiltrated by African-cassava-mosaic-virus-based VIGS systems in previous studies, TRV-based VIGS-system-infiltrated cassava plants did not show obvious virus-induced disease symptoms, suggesting a significant advantage. Taken together, these methods could promote functional genomics in cassava.

scholarly journals An efficient virus-induced gene silencing (VIGS) system for functional genomics in Brassicas using a cabbage leaf curl virus (CaLCuV)-based vector

Planta ◽  
2020 ◽  
Vol 252 (3) ◽  
Author(s):  
Zhiliang Xiao ◽  
Miaomiao Xing ◽  
Xing Liu ◽  
Zhiyuan Fang ◽  
Limei Yang ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Virus Induced Gene Silencing ◽  
Cabbage Leaf Curl Virus ◽  
Leaf Curl Virus ◽  
Leaf Curl

Virus-induced gene silencing using a modified betasatellite: a potential candidate for functional genomics of crops

2014 ◽  
Vol 159 (8) ◽  
pp. 2109-2113 ◽  
Author(s):  
Jitendra Kumar ◽  
Samatha Gunapati ◽  
Jitesh Kumar ◽  
Anita Kumari ◽  
Abhinav Kumar ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Potential Candidate ◽  
Virus Induced Gene Silencing

scholarly journals Virus-induced gene silencing for Asteraceae-a reverse genetics approach for functional genomics in Gerbera hybrida

2012 ◽  
Vol 10 (8) ◽  
pp. 970-978 ◽  
Author(s):  
Xianbao Deng ◽  
Paula Elomaa ◽  
Cuong X. Nguyen ◽  
Timo Hytönen ◽  
Jari P. T. Valkonen ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Reverse Genetics ◽  
Virus Induced Gene Silencing ◽  
Gerbera Hybrida

Graft-accelerated virus-induced gene silencing facilitates functional genomics in rose flowers

2017 ◽  
Vol 60 (1) ◽  
pp. 34-44 ◽  
Author(s):  
Huijun Yan ◽  
Shaochuan Shi ◽  
Nan Ma ◽  
Xiaoqian Cao ◽  
Hao Zhang ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Virus Induced Gene Silencing

Virus-induced gene silencing as a tool for functional genomics in a legume species

2004 ◽  
Vol 40 (4) ◽  
pp. 622-631 ◽  
Author(s):  
Gabriela D. Constantin ◽  
Britta N. Krath ◽  
Stuart A. MacFarlane ◽  
Mogens Nicolaisen ◽  
Ida Elisabeth Johansen ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Legume Species ◽  
Virus Induced Gene Silencing
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