The design and partial analysis of RNAseIII‐anti‐PVS antisense complex system to induce plant resistance

A system combining antisense RNA and RNAseIII‐binding activity was designed. Two antisense RNA vehicles, 7SL RNA and tRNA genes containing the coat protein leader domain (2245–2316) and “zinc finger”; domain (3325–3411) of PVS, respectively were prepared. Both, antisense RNA vehicles were shown to be efficiently transcribed by polymerase III using in vitro assay. It was shown by specific RT PCR, that these genes are also transcribed in potato tissues providing that no polymerase II genes are closely linked to them, suggesting some interaction between polII and polIII transcription complexes. Transgenic potato (S. tuberosum cv. Kamýk) lines expressing mutated bacterial RNAseIII, were prepared. From 25 original lines four had unchanged morphology and ability to grow vigorously in greenhouse conditions and two of them showed lower infection rate than corresponding untransformed control plants after primary infection with PVS. These differences were not observed on plants after the secondary PVS infection. Se...