Special origin of stem sequence influence the resistance of hairpin expressing plants against PVY
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In this study, 16 hairpin RNA (hpRNA) vectors were constructed, each harboring 50 bp viral RNA sequence as the stem. They all targeted the coat protein (CP) gene of Potato virus Y (PVY). Virus resistance assay revealed that hairpin constructs targeting the anterior 200 bp regions of the CP gene were unable to induce virus resistance, while the 12 hpRNA constructs targeting posterior 600 bp regions induced high virus resistance up to 77.78 %. Northern blot analysis revealed that 50 bp-length hpRNA constructs could be transcribed efficiently and processed into siRNAs; however, no correlation between siRNA accumulation and degree of antiviral defense was observed. Results presented here indicated that the middle and 3′ end of the CP cDNA was important for hpRNA-mediated PVY resistance, improving the design of pathogen-derived hpRNA expression cassettes for transgenic plant against viruses.
Additional key wordsAgrobacterium tumefaciens hpRNA-mediated virus resistance Nicotiana tabacum RNA silencing
cauliflower mosaic virus
double strand RNA
post-transcriptional gene silencing
Potato virus Y
sodium chloride/sodium citrate
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