图4. 铁棍山药微型块茎切片的遗传转化、植株再生及转基因植株的分子鉴定
(A) 农杆菌侵染后的切片; (B) 共培养3天后的切片; (C) 再生苗培养基上培养40天的愈伤组织; (D) 再生苗培养基上培养70天的愈伤组织; (E) 抗性再生苗转入生根培养基; (F) 生根培养基上生长30天的再生苗; (G) 转基因植株基因组DNA的PCR鉴定(M: DNA分子量标记; P: 分别以质粒pCAMBIA1301-DoSERK2 (左)和pART27-DoSERK2 (右)为模板; WT: 以野生型植株的基因组DNA为模板; OE: 以过表达转基因株系基因组DNA为模板; RNAi-1-6: 以沉默载体转基因株系1-6的基因组DNA为模板); (H) 过表达转化叶片及叶柄中的GUS表达情况, 野生型植株(WT)和转基因植株(OE)分别经GUS染色液染色; (I) 内源DoSERK2表达水平的RT-qPCR检测(WT: 以野生型植株的cDNA为模板; RNAi-1-6: 以沉默载体转基因株系1-6 cDNA为模板)。(A)-(F), (H) Bars=1 cm

Figure 4. Genetic transformation and regeneration of Dioscorea opposita cv. ‘Tiegun’ microtuber slices and molecular verification of trans- genic plants
(A) Microtuber slices infected by Agrobacterium; (B) Microtuber slices after co-cultivation with Agrobacterium for 3 days; (C) Tissues cultured on regeneration medium for 40 days; (D) Tissues cultured on regeneration medium for 70 days; (E) Antibiotic-resistant regenerated seedlings that had been transferred to rooting medium; (F) Regenerated seedlings that had been cultured on rooting medium for 30 days; (G) PCR assay of genome DNA from transgenic plants (M: DNA marker; P: Plasmid pCAMBIA1301-DoSERK2 (left panel) and pART27-DoSERK2 (right panel) as template; WT: Genome DNA of wild-type plants as template; OE: Genome DNA from overexpression transgenic plants as template; RNAi-1-6: Genome DNA from line 1-6 of transgenic plants of the silencing vector as template); (H) GUS activity assay in the leaf of an overexpression transformant, nodal or leaf segments of a wild-type plant (WT) or transgenic plant (OE) were stained with GUS staining solution; (I) RT-qPCR assay of the endogenous expression levels of DoSERK2 (WT: cDNA of wild-type plants as template; RNAi-1-6: cDNA from line 1-6 of transgenic plants of the silencing vector as template). (A)-(F), (H) Bars=1 cm