A high-density collection of EMS-induced mutations for TILLING in Landsberg erecta genetic background of Arabidopsis.
A high-density collection of EMS-induced mutations for TILLING in Landsberg erecta genetic background of Arabidopsis.
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DOI：
10.1186/1471-2229-9-147
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发表时间：
2009-12-14
影响因子：
5.3
通讯作者：
Alonso-Blanco C
中科院分区：
文献类型：
--
作者：
Martín B;Ramiro M;Martínez-Zapater JM;Alonso-Blanco C
拟南芥是植物分子遗传学研究的主要模式物种，全世界都在致力于确定其所有基因的功能。为此，通过在化学诱导突变的永久集合中进行TILING(靶向诱导的局部损伤)的反向遗传学在哥伦比亚大学的遗传背景中提供了一种独特的资源。在这项工作中，我们的目标是通过在第二种最常见的参考菌株中开发一个新的甲烷磺酸乙酯(EMS)诱导突变株来扩大拟南芥可用的耕作资源。此外，我们试图使EMS诱导的突变数量达到饱和，这些突变是有活力的和可育的植物可以容忍的。通过不同浓度的EMS诱变，我们在拟南芥参考菌株Landsberg erecta(LER)中获得了3712个M2/M3独立突变株系。这一种群被命名为拟南芥分蘖种群。通过使用种子育性低但足够的M1植株，每个品系的突变频率被最大化。应用TILLING在14个基因中搜索突变，每个基因识别出21到46个突变，对应于总共450个突变。所有基因都发现了错义突变，而除了一个基因外，所有基因都选择了截断。我们估计，平均而言，这些品系每89kb携带一个突变，总共提供了500多万个诱导突变。据估计，Tiller收集显示的每个个体的EMS突变密度是之前报道的A.thaliana种群的两到三倍。对分蘖样品的分析表明，在另一种参考遗传背景下，它在大规模分蘖反向遗传中是有用的。与其他生物中的耕作种群的比较表明，这个新的刺参种群携带着已知的二倍体物种中每个个体最高的化学诱导突变密度。
Arabidopsis thaliana is the main model species for plant molecular genetics studies and world-wide efforts are devoted to identify the function of all its genes. To this end, reverse genetics by TILLING (Targeting Induced Local Lesions IN Genomes) in a permanent collection of chemically induced mutants is providing a unique resource in Columbia genetic background. In this work, we aim to extend TILLING resources available in A. thaliana by developing a new population of ethyl methanesulphonate (EMS) induced mutants in the second commonest reference strain. In addition, we pursue to saturate the number of EMS induced mutations that can be tolerated by viable and fertile plants. By mutagenizing with different EMS concentrations we have developed a permanent collection of 3712 M2/M3 independent mutant lines in the reference strain Landsberg erecta (Ler) of A. thaliana. This population has been named as the Arabidopsis TILLer collection. The frequency of mutations per line was maximized by using M1 plants with low but sufficient seed fertility. Application of TILLING to search for mutants in 14 genes identified 21 to 46 mutations per gene, which correspond to a total of 450 mutations. Missense mutations were found for all genes while truncations were selected for all except one. We estimated that, on average, these lines carry one mutation every 89 kb, Ler population providing a total of more than five million induced mutations. It is estimated that TILLer collection shows a two to three fold higher EMS mutation density per individual than previously reported A. thaliana population. Analysis of TILLer collection demonstrates its usefulness for large scale TILLING reverse genetics in another reference genetic background of A. thaliana. Comparisons with TILLING populations in other organisms indicate that this new A. thaliana collection carries the highest chemically induced mutation density per individual known in diploid species.
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影响因子：
5.3
作者：
Xin, Zhanguo;Wang, Ming Li;Burke, John
通讯作者：
Burke, John
影响因子：
14.9
作者：
Oleykowski, CA;Mullins, CRB;Yeung, AT
通讯作者：
Yeung, AT
影响因子：
56.9
作者：
Alonso, JM;Stepanova, AN;Ecker, JR
通讯作者：
Ecker, JR
影响因子：
7
作者：
Winkler, S;Schwabedissen, A;González-Gaitán, M
通讯作者：
González-Gaitán, M
影响因子：
7.2
作者：
Ostergaard, L;Yanofsky, MF
通讯作者：
Yanofsky, MF