A Codon-Optimized Nucleic Acid Hydrolyzing Single-Chain Antibody Confers Resistance to Chrysanthemums Against Chrysanthemum Stunt Viroid Infection

Abstract

Chrysanthemum stunt viroid (CSVd), the smallest plant pathogen known to infect chrysanthemums, is a single-stranded circular RNA viroid that induces stunting that results in an overall height reduction of 30-50 % in mature plants. A catalytic single-chain variable antibody, 3D8 scFv, which exhibits intrinsic DNase and RNase activities, was expressed in chrysanthemums to generate transgenic plant resistance to CSVd infection. Moreover, a codon-optimized version of the 3D8 scFv gene for chrysanthemums was also transformed into plants; these codon-optimized transgenic chrysanthemums expressed twice as much 3D8 scFv and displayed 60 % more resistance to CSVd infection, compared with transgenic chrysanthemums harboring the original 3D8 scFv gene. CSVd challenge experiments with codon-optimized and original 3D8 scFv-transgenic chrysanthemums showed that CSVd in newly produced leaves of both codon-optimized and original 3D8 scFv-transgenic plants was not detected by RT-PCR. This is the first report describing the development of a CSVd-resistant chrysanthemum harboring a catalytic single-chain antibody, 3D8 scFv, which has intrinsic RNase activity.