木薯抗叶片早衰的基因工程育种

Towards Genetic Engineering Cassava(Manihot esculenta Crantz) for Preventing Premature Leaf Shed

以农杆菌介导的转化方法将一个抗叶片早衰的基因成功地导入木薯基因组 .将成熟培养 1 5天后的胚状体的子叶切碎 ,与农杆菌LBA44 0 4共培养 3天 ,然后转入器官发生培养基 (MS基本培养基附加BAP 1mg/L、IBA 0 .5mg/L)附加 3 0mg/LG41 8进行筛选 .三个星期后出现抗性的愈伤、芽 .将这些抗性芽连同外植体转入茎轴生长培养基促进苗的生长 ,最后转入生根培养基长成植株 .所得抗性株经PCR、Southern分析证实部分植株为转基因株 .

To prevent premature leaf shed in cassava, a construct containing an ipt gene driven by a senescence specific promoter ( SAG12 ) was successfully transferred into cassava ( M Col 22 ) via Agrobacterium mediated transformation method . Mature cotyledons from somatic embryos ,which had been cultivated to be maturative for 15 days, were cut into pieces and used as explants for transformation. 3 days after inoculation with Agrobacterium , the explants were transferred onto the organogenesis induction medium ( MS basal medium supplemented with 1 mg/?L BAP and 0.5 mg/?L IBA ) with 30 mg/?L G418 for selection. Resistant organ structures and shoots appeared in about 3 weeks, the cultures were later transferred to shoot elongation medium and the resistant shoots were finally rooted on MS basal medium without hormones. PCR and Southern analysis confirmed the transgenic nature of some resistant plants.