항바이러스성 고추 품종 육성을 위한 TMV-OM 외피단백질유전자 형질전환

Transformation of TMV-OM strain coat protein gene in Capsicum annuum L. for viral disease resistance

The goal of the present experiments was to develop a tobacco mosaic virus (TMV) resistant hot papper (Capsicum annuum L.) by transformation of TMV-OM strain coat protein (Cp) gene. The optimum condition of tissue culture for shoot formation was NAA (0.01~0.25㎎/ℓ) +Zeatin (2~8㎎/ℓ) in ‘Gersung’ and NAA (0.01~0.1㎎/ℓ) +Zeatin (2~4㎎/ℓ) in ‘Seoul Line 7’, respectively. The number of shoot formations per cotyledonous explant in ‘Gersung’ (3.78) was much higher than that of ‘Seoul Line 7’(1.09), but plant viability of the former (0.59) was nearly same with the latter (0.61). Cotyledons of ‘Seoul Line 7’ and ‘Gersung’ were used as a plant materials. Expression vector, pGA1141, containing TMV-OM strain Cp gene was mobilized into Agrobacterium tumefaciens strain LBA4404. The resulting bacteria and both cotyledons, ‘Seoul Line 7’ and ‘Gersung’, were cocultured, respectively. Selecting of npt II resistants was performed on MS medium containing kanamycin (100㎎/ℓ) and Carbenicillin (250㎎/ℓ) from two days after cocultivation by eight time subsequent culture with one week intervel. The results of TAIV-OM Cp screening indicated that one plants by Northern blotting and three plants by PCR out of 46 cotyledons had kanamycin resistance, in Seoul line 7, and three plants by Northern blotting and one plants by PCR out of 20 cotyledons had kanamycin resistance in Gersung. These results were demonstrated by Northern anaysis that exogenic TMV-OM strain Cp gene was integrated and stably expressed two transformed plants of C, annuum. In future we will check the transformed plants maintaining TMV disease resistance effectively in the field.