Alam Pravej, Mohammad Anis, Ahmad M M, Khan Mather Ali, Nadeem Mohd, Khan Riyazuddeen, Akmal Mohd, Ahlawat Seema, Abdin M Z
Centre for Transgenic Plant Development, Department of Biotechnology, Faculty of Science, Jamia Hamdard, New Delhi, India 62.
Recent Pat Biotechnol. 2014;8(1):102-7. doi: 10.2174/18722083113079990001.
Artemisinin, a potent antimalarial natural products isolated from aerial parts of Artemisia annua L. Many patents have been reported that the demand for artemisinin is exponentially increasing year after year due to increased incidences of drug resistant malaria throughout the world. Leaf explants were used frequently as target tissue to generate transgenic of Artemisia. annua L. However, obtaining a large number of transgenic lines through out the year is a laborious and delicate process. To circumvent this, we have developed a highly efficient leaf explant based Agrobacterium mediated transformation of A. annua L. plant. The gus gene was used as screenable marker to assess and optimize the performance of T-DNA delivery. The age of explant, kind of bacterial inoculation, suspension duration, infection times and co-culture conditions were optimized. The co-culture was carried out with Agrobacterium tumefaciens strain EHA105 under desiccation condition in the dark at 25-28 0C for 2-4 days. Complete analysis of transgene insertion demonstrated that the optimized method of transformation from leaf explants of A. annua L. was efficient and highly reproducible.
青蒿素是一种从黄花蒿地上部分分离得到的强效抗疟天然产物。许多专利报告指出，由于全球耐药性疟疾发病率上升，对青蒿素的需求逐年呈指数增长。叶片外植体常被用作产生黄花蒿转基因植株的靶组织。然而，全年获得大量转基因株系是一个费力且精细的过程。为了规避这一问题，我们开发了一种基于叶片外植体的高效农杆菌介导的黄花蒿植株转化方法。gus基因用作筛选标记，以评估和优化T-DNA传递的性能。对外植体的年龄、细菌接种种类、悬浮时间、感染次数和共培养条件进行了优化。共培养在黑暗中于25-28℃干燥条件下与根癌农杆菌菌株EHA105进行2-4天。对转基因插入的全面分析表明，从黄花蒿叶片外植体转化的优化方法是高效且高度可重复的。
