AI Article Synopsis
- - A citrus tristeza virus-based vector, pT36CA-V1.3, was modified for gene silencing in California citrus, showing effective silencing with both sense and antisense constructs of a specific gene.
- - Modifications were made to the vector, resulting in variants T36CA-V1.4 and T36CA-V1.5, which showed higher viral accumulation than the original vector in host plants after 8-12 months, although levels were similar after 36 months.
- - Grafting experiments indicated that plants infected with these viruses were able to deliver the T36CA virus to some commercial citrus varieties, marking a preliminary success in using this vector for citrus research.
Article Abstract
A protein-expressing citrus tristeza virus-based vector construct, pT36CA-V1.3, obtained from a California isolate of the T36 strain (T36CA), was retooled into a virus-induced gene silencing system intended for use with studies of California citrus. Virus-induced gene silencing constructs engineered with a truncated () gene sequence in the sense or antisense orientation worked equally well to silence the endogenous gene. In a parallel effort to optimize vector performance, two nonsynonymous nucleotides in open reading frame 1a of pT36CA-V1.3 were replaced with those conserved in the reference sequences from the T36CA cDNA library. The resulting viruses, T36CA-V1.4 (with one amino acid modification: D760N) and T36CA-V1.5 (with two amino acid modifications: D760N and P1174L), along with T36CA-V1.3, were individually propagated in and plants. Enzyme-linked immunosorbent assay (ELISA) measurements of extracts of the newly emerged leaves suggested that all three viruses accumulated to similar levels in plants at 5 weeks postinoculation. ELISA values of T36CA-V1.4- and -V1.5-infected samples were significantly higher than that of T36CA-V1.3-infected samples within an 8- to 12-month postinoculation window, suggesting a higher accumulation of T36CA-V1.4 and -V1.5 than T36CA-V1.3. However, at 36 months postinoculation, the ELISA values suggested that all three viruses accumulated to similar levels. When plants infected with each of the three viruses were grafted to commercial citrus varieties, a limited number of receptor plants became infected, demonstrating a weak but nonetheless (the first) successful delivery of T36CA to California-grown commercial citrus.
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| http://dx.doi.org/10.1094/PHYTO-05-24-0167-R | DOI Listing |
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