Ghorbanzade Zahra, Ahmadabadi Mohammad
Department of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran.
Iran J Biotechnol. 2015 Mar;13(1):11-16. doi: 10.15171/ijb.1037.
A highly efficient genetic transformation system is essential for a successful genetic manipulation of the African violet ( Wendl.).
Developing a particle bombardment-based genetic transformation system for the African violet.
A local cultivar of the African violet from Guilan province was used for transformation experiments. The pFF19G and pBin61-Ech42 vectors were used for transient and stable transformation experiments, respectively. The PCR and RT-PCR techniques were used to verify transgene presence and transcript levels in candidate transgenic lines, respectively.
Using leaf explants as target tissues, we transferred an endochitinase gene cDNA into African violet. Transgenic plants were regenerated on selection medium at a reasonable frequency (in average, one stable transgenic line per shot). Molecular analysis of transgenic plants by PCR and RT-PCR techniques confirmed successful integration and expression of transgene in several independent transgenic lines.
Our results provide an efficient stable transformation system for genetic transformation of African violet.
高效的遗传转化系统对于成功对非洲紫罗兰(Wendl.)进行遗传操作至关重要。
为非洲紫罗兰开发一种基于粒子轰击的遗传转化系统。
使用来自吉兰省的一个非洲紫罗兰地方品种进行转化实验。分别使用pFF19G和pBin61-Ech42载体进行瞬时和稳定转化实验。分别使用PCR和RT-PCR技术验证候选转基因株系中转基因的存在和转录水平。
以叶片外植体作为靶组织,我们将一个内切几丁质酶基因cDNA转入非洲紫罗兰。转基因植株在选择培养基上以合理的频率再生(平均每次轰击获得一个稳定的转基因株系)。通过PCR和RT-PCR技术对转基因植株进行分子分析,证实了转基因在几个独立的转基因株系中成功整合和表达。
我们的结果为非洲紫罗兰的遗传转化提供了一种高效的稳定转化系统。