黑龙江省小麦品种HMW-GS的RP-HPLC分析及其方法研究  

Study on RP-HPLC Analysis on HMW-GS Wheat Cultivars in Heilongjiang Province and Its Method

作　　者：张欣欣[1,2] 张延滨[1,2] 赵海滨[2] 李集临[1] 金海涛[3] 宋维富[2] 宋庆杰[2] 张春利[2] 辛文利[2] 肖志敏[2] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院,黑龙江哈尔滨150025 [2]黑龙江省农业科学院作物育种研究所,黑龙江哈尔滨150086 [3]农业部谷物及制品质量监督检验测试中心,黑龙江哈尔滨150086

出　　处：《黑龙江农业科学》2013年第5期1-5,共5页Heilongjiang Agricultural Sciences

基　　金：黑龙江省自然科学基金资助项目(C2007-35)

摘　　要：为了分析黑龙江省小麦主要栽培品种是否含有超量表达的优质亚基,利用SDS-PAGE和RP-HPLC在2009年和2011年分析了48份黑龙江省小麦主要栽培品种(系)。研究中共使用了同一厂家生产的同一型号不同批次的3个色谱柱,由于批次不同它们在分离8*亚基时所需柱温是不一致的。其中一个色谱柱在60℃时可以将各HMW-GS有效分离,另两个在60℃时8*亚基淹没在2或5亚基的峰中,只有在柱温提高到70℃时各HMW-GS才取得了较好的分离效果。因此,在应用RP-HPLC对小麦贮藏蛋白分析时,要充分考虑色谱柱性能指标的差异来确定最适柱温才能取得较好的分离效果。结果表明:黑龙江省小麦品种(系)中超量表达的亚基仅限于来自加拿大超强筋小麦Glenlea的Bx7oe亚基。在具有5个HMW-GS的不同品系中,该亚基相对含量(单个亚基RP-HPLC曲线峰面积占全部HMW-GS曲线峰面积的百分比)的平均值为46%(变化范围为43%～49%),其它Glu-B1x位点亚基相对含量的平均值为31%(变化范围为26%～35%)。因此在强筋小麦育种及生产中利用Bx7oe亚基可以有效地改善小麦品质。To investigate the HMW-GS over-expression of main wheat cultivars and advanced lines in the Heilongjiang province, 48 wheat cultivars/lines were analyzed by SDS-PAGE and Reversed-phase high-performance liquid chromatography（RP-HPLC）in 2009 and 2011. Three HPLC columns from the same manufacturer and same Part No but different packing lots were used. The columns from different lots required inconsistent temperatures in separation of 8＊ subunit. Each HMW-GS could be effectively separated at a column temperature of 600C for one column,but 8＊ subunit submerged in the peak of 2 or 5 subunits at a column temperature of 60℃ for other two columns, and could only be effectively separated at a column temperature of 70℃. Therefore, in the analysis of wheat storage protein in the application of RP-HPLC, full account should be taken on the differences in column performance to determine the optimum column temperature in order to achieve better separation effect. The results indicated that over-expression subunit was limited to Bx7oc subunit in wheat cultivars/ lines developed in Heilongjiang province and the Bx7oc subunit were from Canada Western Extra strong wheat cultivar Glenlea. In the different cultivars/lines with five HMW-GSs, the mean of relative content（single subunit peak area/total HMW-GS peak area）〈 100）of Bx7oc subunit was 46% （range 43 % -49 %）, other GlwBlx subunit was 31% （range 26 % - 35 ;）. Therefore, Bx7oc subunit was useful in wheat breeding and production for strong gluten or extra strong gluten.

关 键 词：小麦 反相高效液相色谱 Bx7oe亚基 遗传育种 

分 类 号：S512.1[农业科学—作物学]