Tomato TILLING technology: development of a reverse genetics tool for the efficient isolation of mutants from Micro-Tom mutant libraries.
Tomato TILLING technology: development of a reverse genetics tool for the efficient isolation of mutants from Micro-Tom mutant libraries.
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DOI：
10.1093/pcp/pcr134
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发表时间：
2011-11
影响因子：
4.9
通讯作者：
Ezura H
中科院分区：
文献类型：
--
作者：
Okabe Y;Asamizu E;Saito T;Matsukura C;Ariizumi T;Brès C;Rothan C;Mizoguchi T;Ezura H
为了加速番茄功能基因组研究，我们开发了一个Micro-Tom TILLING（靶向诱导基因组局部病变）平台。从用0.5或1.0%EMS处理的3，052个甲基磺酸乙酯（EMS）突变株系构建DNA池。通过筛选10个基因计算突变频率。0.5%EMS群体具有每1，710 kb一个突变的轻度突变频率，而1.0%EMS群体具有每737 kb一个突变的频率，该频率适合于在靶基因中产生等位基因系列突变。总体突变频率为每1，237 kb一个突变，这影响了筛选的每个酶平均三个等位基因。为了评估Micro-Tom TILLING平台是否可用于有效的突变体分离，筛选番茄中的六个乙烯受体基因（SlETR 1-SlETR 6）。鉴定了导致乙烯反应降低的SlETR 1的两个等位基因突变体（Sletr 1 -1和Sletr 1 -2），表明我们的Micro-Tom TILLING平台为快速检测EMS突变体库中的突变提供了强大的工具。这项工作提供了一个实用的和公开的工具，水果生物学的研究，并获得新的遗传物质，可用于改善番茄的重要农艺性状。
To accelerate functional genomic research in tomato, we developed a Micro-Tom TILLING (Targeting Induced Local Lesions In Genomes) platform. DNA pools were constructed from 3,052 ethyl methanesulfonate (EMS) mutant lines treated with 0.5 or 1.0% EMS. The mutation frequency was calculated by screening 10 genes. The 0.5% EMS population had a mild mutation frequency of one mutation per 1,710 kb, whereas the 1.0% EMS population had a frequency of one mutation per 737 kb, a frequency suitable for producing an allelic series of mutations in the target genes. The overall mutation frequency was one mutation per 1,237 kb, which affected an average of three alleles per kilobase screened. To assess whether a Micro-Tom TILLING platform could be used for efficient mutant isolation, six ethylene receptor genes in tomato (SlETR1–SlETR6) were screened. Two allelic mutants of SlETR1 (Sletr1-1 and Sletr1-2) that resulted in reduced ethylene responses were identified, indicating that our Micro-Tom TILLING platform provides a powerful tool for the rapid detection of mutations in an EMS mutant library. This work provides a practical and publicly accessible tool for the study of fruit biology and for obtaining novel genetic material that can be used to improve important agronomic traits in tomato.
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影响因子：
5.1
作者：
Gady AL;Hermans FW;Van de Wal MH;van Loo EN;Visser RG;Bachem CW
通讯作者：
Bachem CW
影响因子：
3.7
作者：
Dahmani-Mardas F;Troadec C;Boualem A;Lévêque S;Alsadon AA;Aldoss AA;Dogimont C;Bendahmane A
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影响因子：
4.9
作者：
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通讯作者：
Ezura, Hiroshi
影响因子：
4.8
作者：
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通讯作者：
Tipper, DJ
影响因子：
6.9
作者：
Carrari, Fernando;Fernie, Alisdair R.
通讯作者：
Fernie, Alisdair R.