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Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus

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Abstract

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Virus-induced gene silencing (VIGS) system could be performed successfully in Gladiolus hybridus with vacuum infiltration of cormels and young plants.

Abstract

Functional analysis of genes in gladiolus has previously been impractical due to the lack of an efficient stable genetic transformation method. However, virus-induced gene silencing (VIGS) is effective in some plants which are difficult to transform through other methods. Although the Tobacco rattle virus (TRV)-based VIGS system has been developed and used for verifying gene functions in diverse plants, an appropriate TRV-VIGS approach for gladiolus has not been established yet. In this report we describe the first use of the TRV-VIGS system for gene silencing in gladiolus. Vacuum infiltration of cormels and young plants with the GhPDS-VIGS vector effectively down-regulated the PHYTOENE DESATURASE ortholog GhPDS gene and also resulted in various degrees of photobleaching in Gladiolus hybridus. The reduction in GhPDS expression was tested after TRV-based vector infection using real-time RT-PCR. In addition, the progress of TRV infection was detected by fluorescence visualization using a pTRV2: CP-GFP vector. In conclusion, the TRV-mediated VIGS described here will be an effective gene function analysis mechanism in gladiolus.

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Acknowledgments

We are grateful to Dr. Michael Reid and Dr. Cai-Zhong Jiang (University of California, Davis) for kindly offering pTRV1 and pTRV2 vectors. Our thanks also go to Charles Copeland (University of British Columbia) for helpful editing of this manuscript and to Lin Xi PhD (China Agricultural University) for helpful comments. This study has been supported by the Science and Technology Specific Project Foundation of Ministry of Agriculture, PR China (No. 200903020), National Natural Science Foundation of China (No. 31171991).

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Authors and Affiliations

  1. Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University, Yuan Mingyuan Western Road 2#, Beijing, 100193, China

    Xionghui Zhong, Ze Wu, Muhammad Ali Khan, Jin Chen, Xiaoxin Li, Benhe Gong, Yang Zhao, Jian Wu, Chenyu Wu & Mingfang Yi

  2. National Library of China, Zhongguancun South Street 33#, Hai Dian District, Beijing, 100081, China

    Xue Yuan

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  1. Xionghui Zhong
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  2. Xue Yuan
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Correspondence to Mingfang Yi.

Additional information

Communicated by K. Kamo.

Electronic supplementary material

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Supplementary Figure S1 PDS sequence similarity analysis. (a) Sequence comparison of the coding region of PDS amplified from Gladiolus hybridus. Gladiolus hybridus (GhPDS, KC344859), Nicotiana benthamiana (NbPDS, DQ469932), Crocus sativus (CsPDS, AY183118), Oryza sativa (OsPDS, AF049356), Narcissus tazetta var. chinensis cultivar yellow (NtPDS, JQ797377) and Lilium hybrid division I (LiPDS, AB445118) (b) Phylogenetic tree.

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Zhong, X., Yuan, X., Wu, Z. et al. Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus . Plant Cell Rep 33, 301–312 (2014). https://doi.org/10.1007/s00299-013-1530-2

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  • DOI: https://doi.org/10.1007/s00299-013-1530-2

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