A mutant Brassica napus (canola) population for the identification of new genetic diversity via TILLING and next generation sequencing.
A mutant Brassica napus (canola) population for the identification of new genetic diversity via TILLING and next generation sequencing.
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DOI：
10.1371/journal.pone.0084303
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Haughn GW
中科院分区：
文献类型：
--
作者：
Gilchrist EJ;Sidebottom CH;Koh CS;Macinnes T;Sharpe AG;Haughn GW
我们已经产生了3，158个EMS诱变品系的甘蓝型油菜（油菜）群体，并使用TILLING来证明该群体具有足够高的突变密度，其将用于鉴定该重要作物物种中感兴趣的基因中的突变。TILLING是一种反向遗传学技术，已成功用于许多植物和动物物种。经典的TILLING涉及诱变群体的产生，然后使用仅切割携带突变的那些PCR产物的错配特异性内切核酸酶筛选DNA样品。然后使用聚丙烯酰胺凝胶检测来可视化任何感兴趣的基因中的突变。我们使用TILLING技术鉴定了B中26个不同基因的432个独特突变。欧洲油菜（canola cv. DH12075）。这反映了突变密度范围为1/56 kb至1/308 kb（取决于基因座），平均为1/109 kb。我们还成功地验证了下一代测序技术作为鉴定植物群体中罕见突变的强有力方法的实用性，即使是在多倍体物种如B中。油菜。我们发现的大多数突变体都是可利用的。
We have generated a Brassica napus (canola) population of 3,158 EMS-mutagenised lines and used TILLING to demonstrate that the population has a high enough mutation density that it will be useful for identification of mutations in genes of interest in this important crop species. TILLING is a reverse genetics technique that has been successfully used in many plant and animal species. Classical TILLING involves the generation of a mutagenised population, followed by screening of DNA samples using a mismatch-specific endonuclease that cleaves only those PCR products that carry a mutation. Polyacrylamide gel detection is then used to visualise the mutations in any gene of interest. We have used this TILLING technique to identify 432 unique mutations in 26 different genes in B. napus (canola cv. DH12075). This reflects a mutation density ranging from 1/56 kb to 1/308 kb (depending on the locus) with an average of 1/109 kb. We have also successfully verified the utility of next generation sequencing technology as a powerful approach for the identification of rare mutations in a population of plants, even in polyploid species such as B. napus. Most of the mutants we have identified are publically available.
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影响因子：
5.3
作者：
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