Department of Biology, Campus Box 1137, Washington University in St Louis, St Louis, MO 63130, USA.
Centre for Plant Sciences, Leeds University, Leeds, LS2 9JT, UK.
New Phytol. 2011 Jul;191(1):301-306. doi: 10.1111/j.1469-8137.2011.03668.x. Epub 2011 Mar 2.
• The sequencing of the Physcomitrella patens genome, combined with the high frequency of gene targeting in this species, makes it ideal for reverse genetic studies. For forward genetic studies, experimental crosses and genetic analysis of progeny are essential. • Since P. patens is monoicous, producing both male and female gametes on the same gametophore, and undergoing self-fertilization at a high frequency, the identification of crossed sporophytes is difficult. Usually spores from many sporophytes from a mixed culture must be tested for the production of recombinant progeny. • Here, we describe the use of transgenic lines that express a fluorescent transgene constitutively, to provide a direct visual screen for hybrid sporophytes. • We show that segregations in crosses obtained with this technique are as expected, and demonstrate its utility for the study of the rate of outcrossing between three isolates of P. patens.
• 由于泡叶藻的基因组测序工作已经完成，再加上该物种的基因靶向频率很高，因此它非常适合进行反向遗传学研究。对于正向遗传学研究，实验杂交和后代的遗传分析是必不可少的。• 由于泡叶藻是雌雄同体的，在同一个配子体上同时产生雄性和雌性配子，并以很高的频率进行自交，因此杂交孢子体的鉴定很困难。通常必须测试来自混合培养物的许多孢子体的孢子，以确定是否产生重组后代。• 在这里，我们描述了使用稳定表达荧光转基因的转基因系，为杂交孢子体提供直接的可视化筛选。• 我们表明，通过该技术获得的杂交种的分离情况符合预期，并证明了其在研究三种泡叶藻之间的异交率方面的实用性。
