Thieme Mercedes, Hochmuth Anton, Ilse Theresa Elisabeth, Cuesta-Seijo Jose A, Stoma Szymon, Meier Roger, Nørrelykke Simon Flyvbjerg, Pedas Pai Rosager, Braumann Ilka, Zeeman Samuel C
Institute of Molecular Plant Biology, ETH Zurich, 8092 Zurich, Switzerland; Carlsberg Research Laboratory, J.C. Jacobsens Gade 4, 1799 Copenhagen V, Denmark.
Institute of Molecular Plant Biology, ETH Zurich, 8092 Zurich, Switzerland.
Carbohydr Polym. 2023 Jan 1;299:120169. doi: 10.1016/j.carbpol.2022.120169. Epub 2022 Sep 29.
Starch forms semi-crystalline, water-insoluble granules, the size and morphology of which vary according to biological origin. These traits, together with polymer composition and structure, determine the physicochemical properties of starch. However, screening methods to identify differences in starch granule size and shape are lacking. Here, we present two approaches for high-throughput starch granule extraction and size determination using flow cytometry and automated, high-throughput light microscopy. We evaluated the practicality of both methods using starch from different species and tissues and demonstrated their effectiveness by screening for induced variation in starch extracted from over 10,000 barley lines, yielding four with heritable changes in the ratio of large A-granules to small B-granules. Analysis of Arabidopsis lines altered in starch biosynthesis further demonstrates the applicability of these approaches. Identifying variation in starch granule size and shape will enable identification of trait-controlling genes for developing crops with desired properties, and could help optimise starch processing.
淀粉形成半结晶的、水不溶性颗粒，其大小和形态因生物来源而异。这些特性，连同聚合物组成和结构，决定了淀粉的物理化学性质。然而，目前缺乏用于鉴定淀粉颗粒大小和形状差异的筛选方法。在此，我们提出了两种使用流式细胞术以及自动化高通量光学显微镜进行高通量淀粉颗粒提取和大小测定的方法。我们使用来自不同物种和组织的淀粉评估了这两种方法的实用性，并通过筛选从10000多个大麦品系中提取的淀粉的诱导变异证明了它们的有效性，获得了4个大A颗粒与小B颗粒比例具有可遗传变化的品系。对淀粉生物合成发生改变的拟南芥品系的分析进一步证明了这些方法的适用性。鉴定淀粉颗粒大小和形状的变异将有助于鉴定控制性状的基因，从而培育出具有所需特性的作物，并有助于优化淀粉加工。
