Department of Horticulture, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Plant Physiol. 2013 Sep;163(1):21-9. doi: 10.1104/pp.113.221903. Epub 2013 Jul 29.
Insertional mutagenesis using transfer DNA or transposable elements, which is an important tool in functional genomics and is well established in several crops, has not been developed in potato (Solanum tuberosum). Here, we report the application of the tobacco (Nicotiana tabacum) Tnt1 retrotransposon as an insertional mutagen in potato. The Tnt1 retrotransposon was introduced into a highly homozygous and self-compatible clone, 523-3, of the diploid wild potato species Solanum chacoense. Transposition of the Tnt1 elements introduced into 523-3 can be efficiently induced by tissue culture. Tnt1 preferentially inserted into genic regions in the potato genome and the insertions were stable during sexual reproduction, making Tnt1 an ideal mutagen in potato. Several distinct phenotypes associated with plant stature and leaf morphology were discovered in mutation screening from a total of 38 families derived from Tnt1-containing lines. We demonstrate that the insertional mutagenesis system based on Tnt1 and the 523-3 clone can be expanded to the genome-wide level to potentially tag every gene in the potato genome.
利用转移 DNA 或转座元件进行插入突变,这是功能基因组学的重要工具,在几种作物中已经得到很好的确立,但在马铃薯(Solanum tuberosum)中尚未开发。在这里,我们报告了烟草(Nicotiana tabacum)Tnt1 反转录转座子在马铃薯中的插入突变应用。Tnt1 反转录转座子被引入到高度纯合和自交亲和的二倍体野生马铃薯种 Solanum chacoense 的 523-3 克隆中。Tnt1 元件的转座可以通过组织培养有效地诱导。Tnt1 优先插入马铃薯基因组中的基因区域,并且在有性繁殖过程中插入是稳定的,使 Tnt1 成为马铃薯中的理想诱变剂。在总共 38 个来自含有 Tnt1 系的家族的突变筛选中,发现了与植物株高和叶片形态相关的几种不同表型。我们证明,基于 Tnt1 和 523-3 克隆的插入诱变系统可以扩展到全基因组水平,以潜在地标记马铃薯基因组中的每个基因。