Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou, China.
Plant Cell Rep. 2012 Jan;31(1):121-31. doi: 10.1007/s00299-011-1145-4. Epub 2011 Sep 20.
A resveratrol synthase gene was cloned from the peanut plant (Arachis hypogaea) by RT-PCR and was transformed into purple sweet potato (Ipomoea batatas) by Agrobacterium-mediated transformation. Stem sections were infected with bacterial solution of OD(600) = 0.4 for 20 min and then cocultured for 2 days. Infected explants were cultured on MS media containing 50 mg/l kanamycin, 0.02 mg/l NAA and 1 mg/l 6-BA for bud induction or containing 75 mg/l kanamycin, 1.0 mg/l NAA and 0.1 mg/l 6-BA for root formation. The bud and root induction rates were 37.5 and 25.0%, respectively. 105 regenerated plants were obtained, with 11 positive plants by PCR and Southern blotting analyses. A high level of resveratrol glucoside (340 μg/g dry weight), but no resveratrol, was detected in the transformed plants by HPLC. This study also provides a stable genetic transformation and plant regeneration method for metabolic modification of purple sweet potato.
通过 RT-PCR 从落花生（Arachis hypogaea）中克隆了白藜芦醇合酶基因，并通过农杆菌介导的转化将其转化到紫色番薯（Ipomoea batatas）中。茎段用 OD（600）= 0.4 的细菌溶液感染 20 分钟，然后共培养 2 天。将感染的外植体在含有 50mg/L 卡那霉素、0.02mg/L NAA 和 1mg/L 6-BA 的 MS 培养基上进行芽诱导，或在含有 75mg/L 卡那霉素、1.0mg/L NAA 和 0.1mg/L 6-BA 的 MS 培养基上进行生根。芽诱导率和生根诱导率分别为 37.5%和 25.0%。通过 PCR 和 Southern 印迹分析，获得了 105 株再生植株，其中 11 株为阳性植株。通过 HPLC 检测，转化植株中检测到高水平的白藜芦醇葡萄糖苷（340μg/g 干重），但未检测到白藜芦醇。该研究还为紫色番薯的代谢修饰提供了一种稳定的遗传转化和植株再生方法。
