Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AB, UK.
Ann Bot. 2012 Nov;110(6):1185-93. doi: 10.1093/aob/mcs185. Epub 2012 Aug 23.
Large-scale ploidy surveys using flow cytometry have become an essential tool to study plant genome dynamics and to gain insight into the mechanisms and genetic barriers framing ploidy diversity. As an ideal complement to traditional techniques such as chromosome counting, the analysis of cytotype diversity in plant systems such as Sorbus provides primary investigation into the potential patterns and evolutionary implications of hybrid speciation.
Ploidy was assessed by means of relative nuclear DNA content using propidium iodide flow cytometry in 474 Sorbus samples collected from 65 populations in southern Wales and South-West England. Statistical tests were applied to evaluate the utility of this technique to confidently discriminate ploidy in the genus.
Flow cytometric profiles revealed the presence of four cytotypes (2x, 3x, 4x and 5x), confirming in many cases chromosome counts previously reported and demonstrating cytotype heterogeneity within specific Sorbus aggregates. Diploid cytotypes were restricted to the potential parental species and homoploid hybrids. Most of the samples processed were polyploid. The occurrence of the pentaploid cytotype had previously only been reported from a single specimen; it is now confirmed for two taxa occurring at different sites.
Flow cytometry results obtained have proved useful in shedding light on the taxonomy of several controversial taxa and in confirming the presence of cytoypes which occur at very low frequencies. Notably, the coexistence of several cytotypes in Sorbus populations has probably been facilitated by the overlapping distribution of many of the species studied, which might also explain the high incidence of potential hybrid apomictic polyploids. These results will provide a solid baseline for molecular research aiming to better understand the genetic pathways controlling the formation and establishment of polyploid Sorbus.
利用流式细胞术进行大规模倍性调查已成为研究植物基因组动态、深入了解构成倍性多样性的机制和遗传障碍的重要工具。作为传统技术(如染色体计数)的理想补充,在像苹果属这样的植物系统中分析细胞型多样性,为杂种形成的潜在模式和进化意义提供了初步研究。
使用碘化丙啶流式细胞术,对从威尔士南部和英格兰西南部 65 个种群中采集的 474 个苹果属样本的相对核 DNA 含量进行了倍性评估。应用统计检验来评估该技术在准确鉴别该属倍性方面的有效性。
流式细胞术图谱显示存在四种细胞型(2x、3x、4x 和 5x),这在许多情况下证实了先前报道的染色体计数,并证明了特定苹果属聚集体内的细胞型异质性。二倍体细胞型仅限于潜在的亲本种和同倍体杂种。处理的大多数样本都是多倍体。五倍体细胞型的发生以前仅在一个标本中报道过;现在已在两个不同地点出现的两个分类群中得到确认。
流式细胞术结果已被证明有助于阐明几个有争议的分类群的分类学,并确认了在非常低频率下发生的细胞型。值得注意的是,在苹果属种群中几种细胞型的共存可能是由于许多研究物种的分布重叠所致,这也可能解释了潜在的杂种无融合生殖多倍体的高发生率。这些结果将为旨在更好地理解控制多倍体苹果属形成和建立的遗传途径的分子研究提供坚实的基础。