Madsen Lene Heegaard, Fukai Eigo, Radutoiu Simona, Yost Christopher Karl, Sandal Niels, Schauser Leif, Stougaard Jens
Laboratory of Gene Expression, Department of Molecular Biology, University of Aarhus, Gustav Wieds Vej 10, 8000 Aarhus C, Denmark.
Plant J. 2005 Nov;44(3):372-81. doi: 10.1111/j.1365-313X.2005.02534.x.
We have identified a low-copy-number retrotransposon family present in nine to 10 copies in the Lotus japonicus model legume genome, and characterized its activity. LORE1 (Lotus retrotransposon 1) belongs to the Ty3-gypsy group of elements, and is a long terminal repeat (LTR) retrotransposon. Genetic mapping located LORE1 elements in gene-rich regions of Lotus chromosomes, and analysis of native as well as new insertion sites revealed integration outside the highly repetitive sequences of centromeres and telomeres. Sequencing of individual LORE1 family members identified several intact elements, and analysis of new insertions showed that at least one member is active and reinserts into functional genes, creating gene-disruption mutations. Southern blot analysis and SSAP on a selection of symbiotic mutants revealed up to 12 new insertion sites in individual mutant lines and a Mendelian segregation of new inserts. Expression analysis showed that LORE1 elements are transcribed in all organs analysed and, in contrast to other active retrotransposons, LORE1 appears not to be transcriptionally upregulated during in vitro tissue culture. Activity of LORE1 in callus and whole plants suggests that a simple insertion mutagenesis based on endogenous LORE1 elements can be established for Lotus.
我们在百脉根属模式豆科植物基因组中鉴定出一个低拷贝数反转录转座子家族,其拷贝数为9至10个,并对其活性进行了表征。LORE1(百脉根反转录转座子1)属于Ty3-gypsy元件组,是一种长末端重复(LTR)反转录转座子。遗传定位将LORE1元件定位在百脉根染色体的基因丰富区域,对天然以及新插入位点的分析表明,其整合发生在着丝粒和端粒的高度重复序列之外。对单个LORE1家族成员进行测序鉴定出了几个完整元件,对新插入序列的分析表明,至少有一个成员具有活性,并重新插入功能基因中,从而产生基因破坏突变。对一系列共生突变体进行Southern印迹分析和SSAP分析,在单个突变体品系中发现了多达12个新插入位点,并且新插入序列呈孟德尔分离。表达分析表明,LORE1元件在所有分析的器官中均有转录,并且与其他活性反转录转座子不同,LORE1在体外组织培养过程中似乎没有转录上调。LORE1在愈伤组织和整株植物中的活性表明,可以基于内源性LORE1元件为百脉根建立一种简单的插入诱变方法。