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Australasian Plant Pathology

, Volume 47, Issue 2, pp 227–230 | Cite as

RNA interference-mediated resistance to Tobacco streak virus in transgenic peanut

  • C. Senthilraja
  • M. Gurivi Reddy
  • J. Rajeswaran
  • E. Kokiladevi
  • R. Velazhahan
Research Note
  • 135 Downloads

Abstract

Peanut stem necrosis disease (PSND) caused by Tobacco streak virus (TSV) is one of the important diseases of peanut (Arachis hypogaea L.) worldwide. In this study, we investigated the possibility of developing PSND- resistant transgenic peanut plants using RNAi technique. Peanut cultivar TMV-7 was transformed with a hairpin RNA (hpRNA) vector, pART27-TSV-IR-CP, containing inverted repeats of TSV-coat protein (CP) gene sequence flanking a spacer sequence (Pdk Intron) under the control of the CaMV 35S promoter using Agrobacterium tumefaciens -mediated transformation method. The presence of the TSV CP gene sequence in T0, T1 and T2 transgenic plants was confirmed by PCR analysis using CP gene specific primers. Evaluation of the transgenic plants (T2 generation) for resistance to TSV by using sap inoculation method revealed that none of the inoculated transgenic plants showed symptoms of infection, whereas non-transgenic control plants showed necrosis of leaves 7–9 days after inoculation and stem necrosis 16–19 days after inoculation. This study suggests that resistance against TSV can be induced in peanut plants through genetic engineering by expressing dsRNA of coat protein gene of TSV as target for RNAi.

Keywords

Arachis hypogaea coat protein gene RNA interference Tobacco streak virus 

Notes

Acknowledgements

This work was supported by the University Grants Commission, New Delhi, India (Grant No. 42-200/ 2013-SR). We thank the CSIRO Plant Industry, Australia for providing the vector pHANNIBAL and pART27.

Supplementary material

13313_2018_549_MOESM1_ESM.jpg (23 kb)
Supplementary Fig. 1 Hairpin RNAi construct used to transform peanut plants. Adapted by permission from Springer Customer Service Centre GmbH: Springer Nature. Pradeep et al. 2012. Engineering resistance against Tobacco streak virus (TSV) in sunflower and tobacco using RNA interference. Biologia Plantarum 56:735-741. Copyright (2012) (JPEG 23 kb)
13313_2018_549_MOESM2_ESM.jpg (29 kb)
Supplementary Fig. 2a (JPEG 28 kb)
13313_2018_549_MOESM3_ESM.jpg (23 kb)
Supplementary Fig. 2b (JPEG 22 kb)
13313_2018_549_MOESM4_ESM.jpg (29 kb)
Supplementary Fig. 2c Molecular analysis of T 0 (a), T 1 (b) and T 2 (c) generation transgenic peanut plants. Lane PC: pHANNIBAL(TSV-CP) plasmid as a positive control; lane NC: genomic DNA isolated from a non-transformed plant as a negative control. (JPEG 28 kb)
13313_2018_549_MOESM5_ESM.jpg (53 kb)
Supplementary Fig. 3 Reaction of transgenic (a) and non-transgenic control (b & c) peanut plants to TSV. The arrows indicate lesions on the inoculated non-transgenic control plants. (JPEG 52 kb)
13313_2018_549_MOESM6_ESM.docx (47 kb)
Supplementary Table 1 Segregation of TSV CP gene in T2 progeny plants of transgenic peanut lines. (DOCX 46 kb)

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Copyright information

© Australasian Plant Pathology Society Inc. 2018

Authors and Affiliations

  • C. Senthilraja
    • 1
  • M. Gurivi Reddy
    • 1
  • J. Rajeswaran
    • 1
  • E. Kokiladevi
    • 2
  • R. Velazhahan
    • 1
    • 3
  1. 1.Department of Plant PathologyTamil Nadu Agricultural UniversityCoimbatoreIndia
  2. 2.Department of Biotechnology, Agricultural College and Research InstituteTamil Nadu Agricultural UniversityMaduraiIndia
  3. 3.Department of Crop Sciences, College of Agricultural and Marine SciencesSultan Qaboos UniversityMuscatSultanate of Oman

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