Introduction of CyNAC3 to Protocorm-like Bodies in Cymbidium Mediated by Agrobacterium tumefaciens

Inflorescence growth of Cymbidium with shorter than 7.5 mm was blasted under high temperature condition (over 25 degrees C) in the night. Growers want to form flower buds from May to June, but young flower buds are encountered in summer in Japan. Therefore, we tried to introduce CyNAC3 to PLBs by A. tumefaciens to produce high temperature resistant Cymbidium in this study. PLBs pre-cultured with 100 mu M acetosyringone were cultured in mixed liquid (A. tumefaciens suspension solution and pre-culture solution) for infection. After co-cultivation for 3 days, PLBs washed with 40 mg/L meropenem were cultured on Kyoto medium with 20 mg/L hygromycin for selection. Regenerated plants were checked the transformation and the expression of CyNAC3 by PCR and RT-PCR respectively. New PLBs were produced within 4 weeks after selection. As a result of PCR and RTPCR, we confirmed the introduction and the expression of CyNAC3 in all regenerated plants. However, more research is needed to confirm whether these transformants blast or grow normally under high temperature condition in the night.