Salmanowicz Bolesław P
Institute of Plant Genetics, Polish Academy of Sciences, Strzeszynska Street 34, PL 60-479 Poznań, Poland.
J Agric Food Chem. 2008 Oct 22;56(20):9355-61. doi: 10.1021/jf8016546. Epub 2008 Sep 23.
An improved method for separating and characterizing high molecular weight glutenin subunits (HMW-GS) in hexaploid triticale by capillary zone electrophoresis (CZE) was developed. A low-concentrate mixture of hydrophilic polymers, poly(vinylpyrrolidone) (PVP) and hydroxypropylmethylcellulose (HPMC), in an isoelectric buffer was employed for dynamic coating of the capillary inner wall. In separation buffer PVP with lower concentrated poly(ethylene oxide) (PEO) was replaced. The CZE electropherograms of HMW-GS showed two group peaks in accordance with x- and y-type subunits with migration times of 6.8-7.8 and 8.4-11.5 min, respectively. In total, 14 HMW subunits (2 subunits encoded by Glu-A1 locus and 12 by Glu-B1) were identified. The CZE analyses revealed that each of the subunits Bx7 and By8 determined by SDS-PAGE makes up three subunits (Bx6.8, Bx7, and Bx7* and By8, By8*, and new By8**, respectively), with different migration times. It was also shown that the subunits By18 and By20 in triticale determined by SDS-PAGE have different migration times in comparison with the same subunits in bread wheat. For these new HMW-GS, the following names were assigned: By18* instead of By18 and By20* instead of By20. The presented CZE method is an efficient alternative to the SDS-PAGE procedure for early selection of useful triticale genotypes with good breadmaking quality.
开发了一种通过毛细管区带电泳(CZE)分离和鉴定六倍体小黑麦中高分子量谷蛋白亚基(HMW-GS)的改进方法。在等电缓冲液中使用低浓度的亲水性聚合物混合物,即聚乙烯吡咯烷酮(PVP)和羟丙基甲基纤维素(HPMC),对毛细管内壁进行动态涂层。在分离缓冲液中,用较低浓度的聚环氧乙烷(PEO)替代了PVP。HMW-GS的CZE电泳图显示出两组峰,分别对应x型和y型亚基,迁移时间分别为6.8 - 7.8分钟和8.4 - 11.5分钟。总共鉴定出14个HMW亚基(2个由Glu-A1位点编码,12个由Glu-B1位点编码)。CZE分析表明,通过SDS-PAGE确定的每个亚基Bx7和By8分别由三个亚基组成(分别为Bx6.8、Bx7和Bx7以及By8、By8和新的By8**),迁移时间不同。还表明,通过SDS-PAGE确定的小黑麦中的亚基By18和By20与面包小麦中的相同亚基相比,迁移时间不同。对于这些新的HMW-GS,分别赋予了以下名称:用By18代替By18,用By20代替By20。所提出的CZE方法是一种有效的替代SDS-PAGE程序的方法,可用于早期筛选具有良好面包制作品质的有用小黑麦基因型。