In vitro Propagation and Agrobacterium-mediated Transformation of Safflower ('Carthamus tinctorius' L.) Using a Bacterial Mutated Aroa Gene
In vitro Propagation and Agrobacterium-mediated Transformation of Safflower ('Carthamus tinctorius' L.) Using a Bacterial Mutated Aroa Gene
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发表时间：
2011-04
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通讯作者：
J. Motamedi;A. Zebarjadi;D. Kahrizi;A. Salmanian
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作者：
J. Motamedi;A. Zebarjadi;D. Kahrizi;A. Salmanian
以红花（Carthamus tinctorius L.）两个品种为材料，通过农杆菌介导的aroA基因转化，获得了红花愈伤组织诱导和植株再生体系。平均比较表明，在MS +1 mg l ~（-1）NAA + 1 mg l ~（-1）BAP的培养基上愈伤组织诱导率最高。Dincer（94.33%）和0.5mgl ~（-1）NAA +0.5mgl ~（-1）BAP（97%）。此外，Dincer品种的子叶外植体在附加0.1 mg l ~（-1）NAA + 2 mg l ~（-1）BAP的培养基上再生率最高（35.1%）。在转化效率方面，在含有50 mg l-1卡那霉素的选择性培养基上，cv. Dincer和LBA 4404菌株（20.61%）。用特异引物EPS 1F/EPS 2 R和EPS 2 R/35 SF对16株Dincer试管苗进行PCR扩增，结果表明，扩增出的aroA基因片段大小分别为1300 bp和1800 bp，而Sina试管苗中未检测到该基因。本研究表明，黄瓜子叶外植体。红花种子具有良好的直接芽再生潜力，A.根癌农杆菌LBA 4404为aroA基因突变体转化该油料作物奠定了基础。
Callus induction and in vitro plantlet regeneration systems following Agrobacterium-mediated transformation using a mutated bacterial aroA gene were obtained in two cultivars of safflower (Carthamus tinctorius L.). Mean comparison showed that the highest percentage of induced callus was occurred on MS medium containing (1 mg l-1 NAA + 1 mg l-1 BAP) in cv. Dincer (94.33%) and (0.5 mg l-1 NAA + 0.5 mg l-1 BAP) in hypocotyl explant (97%), respectively. In addition, the highest percentage of shoot regeneration also was achieved on a range of media supplemented with 0.1 mg l-1 NAA + 2 mg l-1 BAP from cotyledon explant of Dincer cultivar (35.1%). In respect of transformation efficiency, the highest percentage of putative regenerated shoots on selective medium containing 50 mg l-1 kanamycin was achieved in cv. Dincer and LBA4404 strain (20.61%). PCR analysis of sixteen Dincer plantlets for both strains confirmed that mutated aroA gene was amplified using specific primers (EPS1F/EPS2R) and (EPS2R/35SF) yielded fragments of 1300 bp and 1800 bp, respectively whereas no plantlets contained this gene in Sina cultivar. This study revealed that cotyledon explant of cv. Dincer of safflower have a good potential for direct shoot regeneration, and A. tumefaciens LBA4404 can be established a beneficial method for the transformation of this oilseed crop by mutated aroA gene.