Overexpression of a RNA silencing suppressor, B2 protein encoded by Flock House virus , in tobacco plants results in tol
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ORIGINAL ARTICLE
Overexpression of a RNA silencing suppressor, B2 protein encoded by Flock House virus, in tobacco plants results in tolerance to salt stress Kumari Veena Sinha & Sudhanshu Sekhar Das & Neeti Sanan-Mishra
Received: 16 January 2020 / Accepted: 7 September 2020 # Springer Nature B.V. 2020
Abstract RNA silencing is an evolutionarily conserved defense response against virus invasion and suppression of silencing is a counter-defense mechanism acquired by viruses. The B2 protein encoded by insect Flock House virus (FHVB2) is a well-known RNA silencing suppressor (RSS). It is capable of reversing the suppression of GFP reporter gene in planta. In this study the effect of point mutation and deletions of FHVB2 on the in planta RSS activity was quantified and validated. The results showed drastic reduction in RSS activity of point mutant and deletion constructs. It is known that viruses like Cucumber mosaic virus can enhance the ability of plants to tolerate abiotic stress although the underlying mechanism is not known. Since the non-plant virus encoded FHVB2 is functionally similar to Cucumber mosaic virus encoded 2b and can act as a RSS in plants, it was
Kumari Veena Sinha and Sudhanshu Sekhar Das contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12600-020-00847-y) contains supplementary material, which is available to authorized users. K. V. Sinha : S. S. Das : N. Sanan-Mishra (*) Plant RNAi Biology Group, International Centre for Genetic Engineering and Biotechnology, Delhi, New Delhi, India e-mail: [email protected]
K. V. Sinha e-mail: [email protected] S. S. Das e-mail: [email protected]
used to understand if there is any role of host RNA silencing activity in imparting stress tolerance. FHVB2 was overexpressed in tobacco and transgenics analyzed for response to different abiotic stress conditions. The transgenic plants could tolerate high concentrations of salt and showed enhanced accumulation of proline. Transient assays using point mutant and deletion constructs indicate that functional RSS activity is not required for salt tolerance. Keywords Cucumber mosaic virus 2b . Alphanodavirus . Tobacco transgenic . Salinity stress response . High temperature stress
Introduction The mechanism of RNA silencing is known to play a central role in regulating plant development, genome integrity and response to stress (Fagard et al. 2000; Baulcombe 2004; Bucher and Prins 2006; Llave 2010). There are lot of reports on the activation of RNA silencing as a primary defense response during the initial and systemic phases of virus invasion (Aigner 2006; Llave 2010; Gupta and Mukherjee 2019). The phenomenon is mediated by primary and transitive small interfering RNAs (siRNAs) that act in a sequence-specific manner to target and degrade the viral transcripts (Kim 2005; Sharma and Singh 2016). On the other hand, virus encoded proteins such as coat proteins, replicases, movement proteins, helper components for viral transmission e
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