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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 3, pp 609–613 | Cite as

Use of TRV-mediated VIGS for functional genomics research in citrus

  • Fusheng Wang
  • Yuanyuan Xu
  • Xiaona Liu
  • Wanxia Shen
  • Shiping Zhu
  • Xiaochun ZhaoEmail author
Research Note
  • 119 Downloads

Abstract

Virus induced gene silencing (VIGS) is a powerful tool for identifying gene functions in plants. Tobacco rattle virus (TRV)-mediated VIGS has been reported for silencing of endogenous genes in many plants. In citrus, analysis of gene function is mainly relied on the table genetic transformation, even though this technique is low-efficiency and time-consuming. In this study, a TRV-mediated VIGS was successfully applied for verification of gene function in citrus. A reporter gene, Magnesium chelatase subunit I (ChlI) from C. grandis was constructed into TRV2 vector to induce gene silencing with visible photobleaching symptom. TRV VIGS induced photobleaching symptom was observed on the leaves of Nicotiana benthamiana and Citrus after infiltration. The susceptibility to VIGS vector was found to be genotype dependent. Mexican lemon is the most susceptible one. The chlorosis area per leaf was 65.5% of the whole leaf area. GXSTY pummelo exhibited less susceptible to VIGS with only tiny patches of chlorosis appeared on the leaves of treated plants. The parameters were optimized to improve the efficiency. The tiny young leaves and half expanded leaves were more susceptible to VIGS infection. Young seedlings were highly susceptible to vacuum infiltration. The expression of fluorescence from GFP reporter gene was observed in the young seedlings of GXSTY after infiltration with TRV. Close to 100% of infiltration efficiency was achieved by vacuum infiltration. The results indicated that TRV mediated VIGS with vacuum infiltration could be an ideal tool for gene function studies in citrus.

Key message

A TRV mediated VIGS has been optimized to use in the citrus for verification of gene function. The sensitivity to VIGS vector was genotype dependent. Young seedling and young leaf were the suitable materials for VIGS infection. Vacuum infiltration was a high efficiency infiltration method.

Keywords

TRV-mediated VIGS Vacuum infiltration Gene silencing Citrus 

Notes

Acknowledgements

This study was supported by the Earmarked Fund for China Agriculture Research System (CARS-26); the Earmarked Fund for Chongqing Special & Economic Agriculture Research System on Late Maturation Citrus; Cooperative Research Program for Agriculture Science & Technology Development (Grant No. PJ01211201); the “Double World-classes” Development Plan of Southwest University; China Postdoctoral Science Foundation (Grant No. 2017M622946); Chongqing Special Postdoctoral Science Foundation under Grant (No. XmT2018062); the National Natural Science Foundation of China (Grant No. 31901995). We thank Deqiang Tai (Shanxi Agricultural University) for technical assistant on vaccum infiltration method of VIGS.

Author contributions

XZ and FW designed this research. FW, YX, XL, WS, SZ carried out the experiments and data analyses. FW prepared manuscript. XZ revised manuscript.

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest.

Supplementary material

11240_2019_1698_MOESM1_ESM.doc (2.2 mb)
Supplementary material 1 (DOC 2217 kb)
11240_2019_1698_MOESM2_ESM.tif (3.2 mb)
Supplementary material 2 (TIFF 3244 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Citrus Research Institute, Southwest University/Chinese Academy of Agricultural SciencesChongqingChina
  2. 2.National Citrus Engineering Research CenterChongqingChina
  3. 3.College of Horticulture and Landscape ArchitectureSouthwest UniversityChongqingChina

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