Fojtová Miloslava, Sýkorová Eva, Najdekrová Lucie, Polanská Pavla, Zachová Dagmar, Vagnerová Radka, Angelis Karel J, Fajkus Jiří
Faculty of Science and CEITEC - Central European Institute of Technology, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic.
Plant Mol Biol. 2015 Apr;87(6):591-601. doi: 10.1007/s11103-015-0299-9. Epub 2015 Feb 21.
A comparative approach in biology is needed to assess the universality of rules governing this discipline. In plant telomere research, most of the key principles were established based on studies in only single model plant, Arabidopsis thaliana. These principles include the absence of telomere shortening during plant development and the corresponding activity of telomerase in dividing (meristem) plant cells. Here we examine these principles in Physcomitrella patens as a representative of lower plants. To follow telomerase expression, we first characterize the gene coding for the telomerase reverse transcriptase subunit PpTERT in P. patens, for which only incomplete prediction has been available so far. In protonema cultures of P. patens, growing by filament apical cell division, the proportion of apical (dividing) cells was quantified and telomere length, telomerase expression and activity were determined. Our results show telomere stability and demonstrate proportionality of telomerase activity and expression with the number of apical cells. In addition, we analyze telomere maintenance in mre11, rad50, nbs1, ku70 and lig4 mutants of P. patens and compare the impact of these mutations in double-strand-break (DSB) repair pathways with earlier observations in corresponding A. thaliana mutants. Telomere phenotypes are absent and DSB repair kinetics is not affected in P. patens mutants for DSB factors involved in non-homologous end joining (NHEJ). This is compliant with the overall dominance of homologous recombination over NHEJ pathways in the moss, contrary to the inverse situation in flowering plants.
生物学中需要一种比较方法来评估支配该学科的规则的普遍性。在植物端粒研究中,大多数关键原则是仅基于对单一模式植物拟南芥的研究而确立的。这些原则包括植物发育过程中端粒不会缩短以及端粒酶在分裂(分生组织)植物细胞中的相应活性。在这里,我们以小立碗藓作为低等植物的代表来检验这些原则。为了追踪端粒酶的表达,我们首先对小立碗藓中端粒酶逆转录酶亚基PpTERT的编码基因进行了表征,到目前为止,该基因仅有不完整的预测。在通过丝状体顶端细胞分裂生长的小立碗藓原丝体培养物中,对顶端(分裂)细胞的比例进行了量化,并测定了端粒长度、端粒酶表达和活性。我们的结果显示了端粒的稳定性,并证明了端粒酶活性和表达与顶端细胞数量成比例。此外,我们分析了小立碗藓mre11、rad50、nbs1、ku70和lig4突变体中的端粒维持情况,并将这些突变在双链断裂(DSB)修复途径中的影响与在相应拟南芥突变体中的早期观察结果进行了比较。对于参与非同源末端连接(NHEJ)的DSB因子,小立碗藓突变体中不存在端粒表型,并且DSB修复动力学不受影响。这与苔藓中同源重组相对于NHEJ途径的总体优势相符,与开花植物中的相反情况形成对比。