Modifying fatty acid profiles through a new cytokinin-based plastid transformation system.
Modifying fatty acid profiles through a new cytokinin-based plastid transformation system.
复制标题
通过新的基于细胞分裂素的质体转化系统修改脂肪酸谱。
DOI：
10.1111/tpj.12684
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Dunne A
中科院分区：
文献类型：
--
作者：
Dunne A
The widespread use of herbicides and antibiotics for selection of transgenic plants has not been very successful with regard to commercialization and public acceptance. Hence, alternative selection systems are required. In this study, we describe the use ofipt, the bacterial gene encoding the enzyme isopentenyl transferase fromAgrobacterium tumefaciens, as a positive selectable marker for plastid transformation. A comparison between the traditional spectinomycin‐basedaadAselection system and theiptselection system demonstrated that selection of transplastomic plants on medium lacking cytokinin was as effective as selection on medium containing spectinomycin. Proof of principle was demonstrated by transformation of thekasIIIgene encoding 3‐ketoacyl acyl carrier protein synthase III into tobacco plastids. Transplastomic tobacco plants were readily obtained using theiptselection system, and were phenotypically normal despite over‐expression of isopentenyl transferase. Over‐expression of KASIII resulted in a significant increase in 16:0 fatty acid levels, and a significant decrease in the levels of 18:0 and 18:1 fatty acids. Our study demonstrates use of a novel positive plastid transformation system that may be used for selection of transplastomic plants without affecting the expression of transgenes within the integrated vector cassette or the resulting activity of the encoded protein. This system has the potential to be applied to monocots, which are typically not amenable to traditional antibiotic‐based selection systems, and may be used in combination with a negative selectable marker as part of a two‐step selection system to obtain homoplasmic plant lines.
