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Molecular tools for marker-assisted breeding of buffelgrass.

机译：水牛草的标记辅助育种的分子工具。

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The increasing availability of molecular tools is facilitating marker-assisted selection (MAS) in plant improvement programs. The objectives of this research were to: (1) populate the framework buffelgrass genome map with additional molecular markers, (2) develop polymerase chain reaction (PCR)-based markers from selected, informative restriction fragment length polymorphism (RFLP) markers on the buffelgrass genome map, and (3) increase marker resolution near the locus conferring apomixis (PApol). Buffelgrass [Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.] (2n = 4x = 36), a highly polymorphic, apomictic, perennial forage grass, is well-suited for genetic linkage analyses. One hundred and seventy-one probes from an apomictic, spikelet-specific, complementary deoxyribonucleic acid (cDNA) library and 70 expressed sequence tag simple sequence repeats (EST-SSRs) from apomictic pistil cDNAs were evaluated and added to the framework buffelgrass genome map. The improved linkage map contains 851 markers from 11 grass species and covers approximately 80-85% of the buffelgrass genome. Two RFLPs from the buffelgrass genome map were converted to PCR-based markers for both the identification of hybrids and quantification of sexual versus apomictic reproduction. A gel-free, high-throughput technique was developed to analyze these markers directly in 96-well plates. Five additional markers were placed onto the buffelgrass linkage group with the PApo1 apomixis locus through comparative mapping of candidate orthologs from the sorghum genome map and bulked-segregant analysis of amplified-fragment-length-polymorphisms (BSA-AFLP). Increasing the mapping population size did not increase map resolution in the PApo1 region. Association mapping revealed that the recombination suppression near PApol is moderate and would complicate comparative map-based cloning efforts of the orthologous region in sorghum.
机译：分子工具可用性的提高正在促进植物改良计划中的标记辅助选择（MAS）。这项研究的目的是：（1）用其他分子标记物填充框架水牛草基因组图谱；（2）从水牛草中选定的信息丰富的限制性片段长度多态性（RFLP）标记物中开发基于聚合酶链反应（PCR）的标记物基因组图谱；以及（3）提高授予无融合生殖（PApol）的基因座附近的标志物分辨率。 Buffelgrass [Pennisetum ciliare（L.）Link syn。 Cenchrus ciliaris L.]（2n = 4x = 36）是高度多态的，无融合生殖的多年生牧草，非常适合进行遗传连锁分析。评价了来自无融合生殖的小穗特异性互补脱氧核糖核酸（cDNA）文库的171个探针和来自无融合生殖的雌蕊cDNA的70个表达的序列标签简单序列重复（EST-SSR），并将其添加到牛黄草基因组图谱中。改进的连锁图谱包含来自11种草的851个标记，覆盖了约25％的水牛草基因组。来自水牛草基因组图谱的两个RFLP被转换为基于PCR的标记，用于鉴定杂种以及定量性繁殖与无融合生殖的定量。开发了一种无凝胶的高通量技术，可直接在96孔板中分析这些标记。通过比较高粱基因组图谱中候选直向同源物的图谱以及扩增片段长度多态性（BSA-AFLP）的大片段分离分析，将五个其他标记物与PApo1无融合生殖基因座一起置于牛毛草连锁群上。增加地图人口的大小并没有增加PApo1区域中的地图分辨率。关联图谱显示，PApol附近的重组抑制是中等程度的，会使高粱直系同源区域的基于图谱的比较克隆工作变得复杂。

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