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AFLP Genetic Characterization in Asparagus officinalis Doubled Haploid (DH) Clones Collection

机译:Aflp在芦笋遗传表征中officinalis双倍单倍体(DH)克隆收集

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This work focused on the genetic characterization of several double haploid lines, part of a large collection present at Montanaso Lombardo C.R.A. Institute (Italy). This DH clones were used as parentals for obtaining hybrids, of commercial (Asparagus officinalis), wild species and some genotypes from interspecific crosses named OMAO {[(officinalis × maritimus) × acutifolius] × officinalis}. Sampling was completed in Autumn 2007-Spring 2008 by taking materials from six different Asparagus species (A. acutifolius, scaber, verticillus, prostratus, stipularis and officinalis) and A. officinalis is the larger group among them with 182 samples divided in local population, commercial cultivars and DH clones. Genetic diversity was analyzed using AFLP molecular markers strategy; forward primers of the nine primer-pairs used were labelled with ABI PRISM fluorescent dye 6-FAM, and the PCR reactions were run on ABI PRISM. The automated sequence system ABI PRISM combined with fluorescent labelling of expected fragments has been applied as an alternative to radioactivity detection using [~(32)P] or [~(33)P]-labelling. This technology provides an automatic and rapid sizing of the fragments through the use of specific internal size standard (GS500-Liz) and allows analysis of fragments. The data collected from each sample were automatically analysed by GeneMapper Analysis Software. The genetic diversity was estimated, and multivariate analysis methods were applied for the analysis of genetic relationships among double haploid accessions. Our results demonstrates that cultivated cultivars and DH genotypes form only one cluster instead of wild species and interspecific genotypes (forming another separate specific cluster). The only two tetraploid cultivated cultivar (the Italian 'Violetto d'Albenga' and the Spanish 'Morado'), are in the cultivated cultivars cluster but they show an high genetic similarity with the only tetraploid wild specie in our panel: A. acutifolius.
机译:这项工作的重点是几种双倍倍增管的遗传表征,在Montanaso Lombardo C.R.A的一部分大型系列的一部分。研究所(意大利)。这种DH克隆用作获得杂种的母体,用于从名为OMAO的三角形交叉的商业(芦笋型),野生物种和一些基因型,其中包括omao {[(officinalis×maritimus)×acutifolius]×officinalis}。通过从六种不同的芦笋物种(A. Acutifolius,Verticillus,Prostratus,Pripularis和Officinalis)和A. Officinalis中,通过从六种不同的芦笋物种(A. Acutifolius,Verticillus,Prostratus,Persformis)以及其中182个样本分为当地人口中的较大组,商业品种和DH克隆。使用AFLP分子标记策略分析遗传多样性;使用的九个引物对的前后引物用ABI棱镜荧光染料6-FAM标记,并且PCR反应在ABI棱镜上进行。使用[〜(32)p]或[〜(33)p] - 标记,将自动序列系统与预期片段的荧光标记联合的预期片段的荧光标记相结合。该技术通过使用特定的内部尺寸标准(GS500-LIZ)提供碎片的自动快速尺寸,并允许分析片段。通过GeneMapper分析软件自动分析从每个样本收集的数据。估计遗传多样性,应用多变量分析方法用于分析双倍单倍体涂层之间的遗传关系。我们的研究结果表明,培养的品种和DH基因型仅形成一个簇而不是野生物种和三种基因型(形成另一个单独的特异性簇)。唯一两种四倍体栽培品种(意大利'Violetto d'Albenga'和西班牙语“Morado”)在培养品种群中,但它们与我们小组中唯一的四倍体野生物种展现出高遗传相似性:A. Acutifolius。

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