단일항체를 발현하는 형질전환 식물체의 In Vitro 화분발아에 대한 Boric Acid의 영향

Abstract

Pollen germination viability is an essential factor to produce seeds from pollination and fertilization, which are required to maintain plant generation. In this study, we tried to identify the effect of boric acid on pollen germination and tube growth in non-transgenic and transgenic plants expressing monoclonal antibodies (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55, and anti-rabies virus mAb57). The pollen of non-transgenic plant was treated with different concentration of boric acid (0, 5, 10, 15, 20, 40 μg/mL) in germination buffer to investigate its effect on in vitro pollen germination. At 20 μg/mL of boric acid, the pollen germination rate was the highest (49.5%) compared to other concentrations. In general, the germination rate significantly increased 3-10 folds in boric acid (20 μg/mL) treated group in non-transgenic and transgenic plants. Also, the pollen tube length increased in boric acid (20 μg/mL) treated groups. In the treated group, the pollen tube length increased until 3 h boric acid treatment and decreased after the 3 h, indicating that the 3 h is the most appropriate incubation time period. Western blot analysis showed that the mAb transgene expression was more stable in leaf than pollen in transgenic plants. This study suggested that 20 μg/mL of boric acid is ideal concentration to induce in vitro pollen germination of transgenic plants expressing therapeutic monoclonal antibodies, indicating stable pollination and fertilization in transgenic plants.