青枯病 (Pseudomonas solanacearum) 抵抗性付与のための抗細菌性遺伝子のAgrobacterium法による4倍体バレイショ品種への形質転換

継続後誌:近畿大学先端技術総合研究所紀要 = Memoirs of Institute of Advanced Technology, Kinki UniversitySeven tetraploid potato genotypes were employed to put artificial antibacterial genes against bacterial wilt (Pseudomonas solanacearum) . The antibacterial genes derived from giant silk moth (Hyalophora cecropia). These genes are associated with the antibacterial peptides, cecropin, attacin, lysozyme or shiva. These genes were constructed with either cauliflower mosaic virus (CaMV35S) or wound-inducible (WI) promoter, and with franking gene markers in a binary vector pBI121. Agrobacterium rhizogenes (R1000) which contains the binary vector was used to transform the potato genotypes. Hairy roots obtained were cultured in MS medium with Claforan then, in a regeneration medium containing 100ppm kanamycin. Regenerated plantlets were evaluated for the expression of flanking marker genes: kanamycin resistance and GUS reaction. After selection, those plantlets were used for Southern hybridization to confirm the presence of the target gene(s) for antibacteria. Phenotypic resistance evaluation on them was made using Pseudomonas solanacearum, Race 3 , Biovar II, CIP isolate 204 at greenhouse. Some improvement on the level of resistance was seen on regenerates from three original genotypes, while there was variation on the level of resistance among regenerates from the same original genotype. RT-PCR revealed substantial level of gene expression, however, the amount of resulting lytic peptides was not detected well by ELISA. Thus, it was speculated that a post-transcriptional degradation and/or a post-translational degeneration by endogenious xenobiotic metabolizing mechanisms or endogenous proteinases. （和文） バレイショ品種における青枯病 (Pseudomonas solanacearum)に対する抵抗性を飛躍的に向上させるため、カイコ由来の溶細菌性タンパクを生産する遺伝子群を用いた。溶細菌性遺伝子を含むbinary vectorをそれぞれ導入した、Agrobacterium rhizogenes, R1000を用いて、バレイショ品種に遺伝子の導入・発現を試みた。組み換えバレイショ系統は多数得られたものの、閉鎖系温室での抵抗性の程度は組み換え個体問で大きな違いがあり、遺伝子構築や発現様式の調整が今後必要であることが解った