Identification of barley mutants in the cultivar 'Lux' at the Dhn loci through TILLING
Identification of barley mutants in the cultivar 'Lux' at the Dhn loci through TILLING
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DOI：
10.1111/j.1439-0523.2009.01640.x
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发表时间：
2009-08-01
期刊：
影响因子：
2
通讯作者：
Jahoor, A.
中科院分区：
文献类型：
--
作者：
Lababidi, S.;Mejlhede, N.;Jahoor, A.
TILLING是一种反向遗传策略，其允许在植物突变群体中筛选具有已知序列的基因中的突变。已经为丹麦大麦品种‘Lux’（Hordeum vulgare L.）开发了TILLING群体，通过使用叠氮化钠来诱导突变。在M-3植株中，大麦幼苗的四个可见表型性状相对于亲本品种'Lux'的评分显示超过3.5%的致死率。测试了来自突变株系的混合DNA的一系列池比率，并且10倍池似乎是用于检测500-700 bp范围内的片段的实际混合比率。两个已知的13个大麦蛋白基因，Dhn 12和Dhn 13，分别进行了检查和5个独立的错义突变，从人口的9575大麦突变体植物。这对应于这两个基因的突变密度约为每250万个碱基对一个突变。由于合并率高，在短时间内对约10000个株系的突变群体进行了两个基因的突变筛查。
TILLING is a reverse genetic strategy that allows screening for mutations in genes with known sequences in a plant mutant population. A TILLING population has been developed for the Danish barley variety 'Lux' (Hordeum vulgare L.), by using sodium azide to induce mutations. Scoring of four visible phenotypic characters of barley seedling in reference to the parental cultivar 'Lux' in the M-3 plants showed over 3.5% lethality. A series of pool ratios of mixed DNA from mutant lines were tested and 10-fold pools appeared to be the practical mixing ratio for the detection of fragments in the 500-700 bp range. Two of the 13 known dehydrin genes, Dhn12 and Dhn13, respectively, were examined and five independent missense mutations were obtained from a population of 9575 barley mutant plants. This corresponds to a mutation density of approximately one mutation every two and half million base pairs for these two genes. The mutant population of approximately 10 000 lines was screened for mutations in two genes in a short time due to high pooling ratio.