Abstract
Two single nucleotide polymorphic mutations and deletion affecting Y1059V1060V1061 amino acid residues in a host translation initiation factor four gamma (eIF4G) gene in rice are reported to confer resistance to rice tungrospherical virus in resistant genotypes. A CRISPR-based adenine base editing vector was used to target these residues in a susceptible indica cultivar, ASD16.Agrobacterium-mediated transformation of ASD16 generated 16 missense mutants and two deletion mutants. Substitution mutations occurred at A5> G5 and A4> G4, where 5.5 % and 3.37 % of adenosines got converted to guanosines, respectively. The mutantsgenerated had missense mutations affecting the YVV residues and the residues immediately adjacent to YVV.Thus,these novel mutationsare promising candidates in imparting resistance against rice tungro disease.
Highlights
Rice tungro disease (RTD) causes severe yield losses in rice-growing endemic regions of South and Southeast Asia (Herdt, 1988; Azzam and Chancellor, 2002; Muralidharan et al, 2003)
Management of tungro disease via the development of broad-spectrum resistance through suppression of Rice tungro spherical virus (RTSV) has been the preferred choice since rice plants infected with RTBV exclusively are incapable of spreading the disease
BsaI adaptors, 5’ GGCG 3’ and 5’ AAAC 3’ were added to the oligos during synthesis to complement their ligation into BsaI restricted plant adenine base editing vector, pH-PABE7-esgRNA
Summary
Rice tungro disease (RTD) causes severe yield losses in rice-growing endemic regions of South and Southeast Asia (Herdt, 1988; Azzam and Chancellor, 2002; Muralidharan et al, 2003). Rice tungro spherical virus (RTSV), along with its counterpart RTBV (Rice tungro bacilliform virus, a dsDNA virus), is responsible for RTD (Bunawan et al, 2014). Management of tungro disease via the development of broad-spectrum resistance through suppression of RTSV has been the preferred choice since rice plants infected with RTBV exclusively are incapable of spreading the disease. RTSV resistant cultivars rather than RTBV could successfully reduce the incidence of tungro disease in the field (Hibino, 1996; Anjaneyulu et al, 1995; Lee et al, 2010). RTSV is one such RNA virus that leverages a host translation initiation factor four gamma (eIF4G) gene of rice to replicate and establish within rice plant (Lee et al, 2010)
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