Silencing AC1 of Tomato leaf curl virus using artificial microRNA confers resistance to leaf curl disease in transgenic
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ORIGINAL ARTICLE
Silencing AC1 of Tomato leaf curl virus using artificial microRNA confers resistance to leaf curl disease in transgenic tomato Namisha Sharma1 · Manoj Prasad1 Received: 24 May 2020 / Accepted: 20 August 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Key message Expression of artificial microRNA targeting ATP binding domain of AC1 in transgenic tomato confers resistance to Tomato leaf curl disease without impacting the yield of tomato. Abstract Tomato curl leaf disease caused by Tomato leaf curl virus (ToLCV) is a key constraint to tomato cultivation worldwide. Engineering transgenic plants expressing artificial microRNAs (amiRNAs) against the AC1 gene of Tomato leaf curl New Delhi virus (ToLCNDV), which is important for virus replication and pathogenicity, would consequently confer virus resistance and reduce crop loss in the economically important crops. This study relates to an amiRNA developed on the sequence of Arabidopsis miRNA319a, targeting the ATP/GTP binding domain of AC1 gene of ToLCNDV. The AC1amiR was found to regulate the abundance of AC1, providing an excellent strategy in providing defense against ToLCNDV. Transgenic lines over-expressing AC1-amiR, when challenged with ToLCNDV, showed reduced disease symptoms and high percentage resistance ranging between ∼ 40 and 80%. The yield of transgenic plants was significantly higher upon ToLCNDV infection as compared to the non-transgenic plants. Although the natural resistance resources against ToLCNDV are not available, this work streamlines a novel amiRNA-based mechanism that may have the potential to develop viral resistance strategies in tomato, apart from its normal symptom development properties as it is targeting the conserved region against which higher accumulation of small interfering RNAs (siRNA) occurred in a naturally tolerant tomato cultivar. Keywords Artificial microRNA · Tomato leaf curl disease · Tomato leaf curl New Delhi virus · Virus resistance · Replication initiator protein
Introduction Tomato (Solanum lycopersicum) has become an inevitable component of human diet since its domestication, and it has been a major factor driving the growth and economy of several nations that contribute to the livelihood of millions of farmers. As any other crops, tomato is also challenged by several diseases among which viral infections are devastative. Particularly, Tomato leaf curl disease (ToLCD) caused Communicated by Neal Stewart. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00299-020-02584-2) contains supplementary material, which is available to authorized users. * Manoj Prasad [email protected] 1
National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi 110067, India
by different strains of Tomato leaf curl virus (ToLCV) is a major constraint to tomato production worldwide as they are difficult to control or eradicate due to the rapid rates at which these viruses evolve (Chakraborty 2008; Sharma and Prasad 2017; P
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