Antonius-Klemola Kristiina, Kalendar Ruslan, Schulman Alan H
MTT Agrifood Research Finland/Crops and Biotechnology, 31600 Jokioinen, Finland.
Theor Appl Genet. 2006 Apr;112(6):999-1008. doi: 10.1007/s00122-005-0203-0. Epub 2006 Jan 11.
Retrotransposon markers have been demonstrated to be powerful tools for investigating linkage, evolution and genetics diversity in plants. In the present study, we identified and cloned three full-size TRIM (terminal-repeat retrotransposon in miniature) group retrotransposon elements from apple (Malus domestica) cv. 'Antonovka', the first from the Rosaceae. To investigate their utility as markers, we designed primers to match the long terminal repeats (LTRs) of the apple TRIM sequences. We found that PCR reactions with even a single primer produced multiple bands, suggesting that the copy number of these TRIM elements is relatively high, and that they may be locally clustered or nested in the genome. Furthermore, the apple TRIM primers employed in IRAP (inter-retrotransposon amplified polymorphism) or REMAP (retrotransposon-microsatellite amplified polymorphism) analyses produced unique, reproducible profiles for 12 standard apple cultivars. On the other hand, all seven of the sport mutations in this study were identical to their mother cultivar. Genetic similarity values calculated from the IRAP/REMAP analyses or the STMS (sequence tagged microsatellite sites) analysis were generally comparable. PAUP cluster analysis based on IRAP and REMAP markers in apple and Japanese quince generated an NJ tree that is in good accordance with both a tree based on SMTS markers and the origin of the studied samples. Our results demonstrate that, although they do not encode the proteins necessary to carry out a life cycle and are thereby non-autonomous, TRIMs are at least as polymorphic in their insertion patterns as conventional complete retrotransposons.
反转录转座子标记已被证明是研究植物连锁、进化和遗传多样性的有力工具。在本研究中,我们从苹果(Malus domestica)品种‘安托诺夫卡’中鉴定并克隆了三个全长的TRIM(微型末端重复反转录转座子)类反转录转座子元件,这是首次从蔷薇科中分离得到。为了研究它们作为标记的实用性,我们设计了引物以匹配苹果TRIM序列的长末端重复序列(LTR)。我们发现,即使只用单个引物进行PCR反应也能产生多条带,这表明这些TRIM元件的拷贝数相对较高,并且它们可能在基因组中局部聚集或嵌套。此外,用于IRAP(反转录转座子间扩增多态性)或REMAP(反转录转座子 - 微卫星扩增多态性)分析的苹果TRIM引物为12个标准苹果品种产生了独特的、可重复的图谱。另一方面,本研究中的所有七个芽变与其母本品种相同。通过IRAP/REMAP分析或STMS(序列标签微卫星位点)分析计算得到的遗传相似性值通常具有可比性。基于苹果和日本木瓜的IRAP和REMAP标记进行的PAUP聚类分析生成了一个NJ树,该树与基于SMTS标记的树以及所研究样本的起源都非常吻合。我们的结果表明,尽管TRIMs不编码完成生命周期所需的蛋白质,因此是非自主的,但它们在插入模式上至少与传统的完整反转录转座子一样具有多态性。