Development of a simple PCR-based marker for the determination of growth habit in Vicia faba L. using a candidate gene approach
Development of a simple PCR-based marker for the determination of growth habit in Vicia faba L. using a candidate gene approach
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DOI：
10.1007/s11032-005-4075-4
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发表时间：
2006-04-01
影响因子：
3.1
通讯作者：
Torres, AM
中科院分区：
文献类型：
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作者：
Avila, CM;Nadal, S;Torres, AM
我们利用候选基因方法开发了第一个适合蚕豆生长习性选择的分子标记。我们从公共数据库中获得TFL1/CEN样基因序列，并在保守结构域上设计引物。选用3个生长习性确定的品种和4个生长习性不确定的材料（2个品种和2个品系）。所有这些基因型都用于我们的蚕豆育种计划。获得了一个单态PCR片段。测定了一组限制性内切酶。酶Hind1II在确定基因型和不确定基因型之间产生明显的多态性。利用杂交品种‘Verde Bonita’× 2N52的生长习性F-2群体对比，对这一新的cleaved amplification polymorphism （CAP）标记进行了检验。该标记对两种基因型的鉴别效率均为100%。此外，该标记的共显性允许检测杂合个体，从而促进不同基因的锥体育种过程。该标记与该性状的完美共分离表明，TFL1/CEN同源基因控制着蚕豆的生长习性。作为确定生长习性的供体，该标记已在我们蚕豆育种计划中使用的所有基因型中进行了测试。因此，它有望在与这些亲本系进行的所有杂交中表现良好，就像在F-2试验中发生的那样。在这项工作中开发的CAPS标记将有助于标记辅助选择程序。此外，该标记可用于质量控制，以确定商业种子批次中外来种子的百分比。此外，它是育种者提交蚕豆新品种注册时的一个有价值的工具，因为该方法可以保证外来植物保持在要求的注册限制之内。
We have developed the first molecular marker suitable for the selection of determinate growth habit in faba bean using the candidate gene approach. We obtained the sequences of TFL1/CEN like genes from public databases and designed primers on conserved domains. We used three cultivars with determinate growth habit and four accessions (two cultivars and two lines) with indeterminate growth habit. All these genotypes are used in our faba bean breeding program. A single monomorphic PCR fragment was obtained. A set of restriction enzymes was assayed. The enzyme Hind1II produced a clear polymorphism between determinate and indeterminate genotypes. This new cleaved amplified polymorphism (CAP) marker was tested using an F-2 population contrasting for growth habit derived from the cross 'Verde Bonita' x 2N52. This marker showed 100% efficiency in discriminating both types of genotypes. Moreover, the codominancy of this marker allows the detection of heterozygous individuals facilitating the breeding process when pyramiding different genes. The perfect cosegregation of the marker with the trait indicates that an orthologue of TFL1/CEN controls the growth habit in faba bean. This marker has been tested in all the genotypes used in our faba bean breeding program as donors of the determinate growth habit. Therefore, it is expected to work well in all the crosses performed with these parental lines as happens in the F-2 tested. The CAPS marker developed in this work will be useful for Marker Assisted Selection programs. In addition, this marker is useful for quality control to determinate the percentage of outsider seeds in commercial seed lots. Moreover, it is a valuable tool to breeders when submitting new faba bean varieties for registration since the method allows guaranteeing that outsider plants remains under the requested limit for registration.