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六倍体小黑麦中 secaloindoline 等位基因形式的 CE 测定。

CE determination of secaloindoline allelic forms in hexaploid triticale (x Triticosecale Wittmack).

机构信息

Institute of Plant Genetics, Polish Academy of Sciences, Poznań, Poland.

出版信息

J Sep Sci. 2010 Mar;33(4-5):643-50. doi: 10.1002/jssc.200900601.

Abstract

Differences in kernel texture are mainly caused by specific secaloindoline (SIN) proteins occurring in friabilin fraction of hexaploid triticale (x Triticosecale Wittmack) grain. SINs were isolated using Triton X-114 partitioning from either kernels/flour or starch of five triticale cultivars with wide range of different hardness. Crude SIN fraction was obtained by size-exclusion HPLC. SINs were separated on an uncoated fused-silica capillary using the iminodiacetic (IDA) buffer in conjunction with lower-concentrated poly(ethylene oxide) and ACN. A low-concentrate mixture of hydrophilic polymers, PVP and hydroxypropylmethylcellulose in IDA buffer was employed for dynamic coating of capillary inner wall. In total, on the basis of CZE profiles, two SIN-a proteins and two SIN-b proteins were identified. Allelic forms SIN-a1 and SIN-b1 have both two soft and one medium hard genotypes, however other allelic forms, designed as SIN-a2 and SIN-b2, were identified in hard and other medium hard cultivars. The CZE profiles showed that the ratio of the peak areas of SIN-b proteins isolated from triticale starch can be preliminarily used to distinguish cultivars with soft and hard grain.

摘要

内核质地的差异主要是由六倍体黑小麦(x Triticosecale Wittmack)籽粒中脆性素(friabilin)级分中特定的 secaloindoline(SIN)蛋白引起的。使用 Triton X-114 从五种硬度差异很大的黑小麦品种的籽粒/面粉或淀粉中进行分配,从而分离出 SIN。使用尺寸排阻 HPLC 获得粗 SIN 级分。使用未涂覆的熔融硅毛细管,在 iminodiacetic(IDA)缓冲液中结合较低浓度的聚(乙二醇)和 ACN 分离 SIN。在 IDA 缓冲液中使用低浓度的亲水性聚合物聚乙烯吡咯烷酮和羟丙基甲基纤维素的混合物对毛细管内壁进行动态涂层。总共,根据 CZE 图谱,鉴定出两种 SIN-a 蛋白和两种 SIN-b 蛋白。等位基因形式 SIN-a1 和 SIN-b1 均具有两个软基因型和一个中硬基因型,而其他等位基因形式,设计为 SIN-a2 和 SIN-b2,则在硬粒和其他中硬粒品种中被鉴定出来。CZE 图谱表明,从小麦淀粉中分离出的 SIN-b 蛋白的峰面积比值可初步用于区分软粒和硬粒品种。

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