Glu-B1位点亚基色谱鉴定及7OE对面团强度的影响

准确鉴定高分子量谷蛋白亚基(HMW-GS)及探讨其对面团特性的影响是小麦品质研究的重要内容。用聚丙烯酰胺凝胶电泳(SDS-PAGE)和反相高效液相色谱(RP-HPLC)对62份品种(系)的HMW-GS组成进行鉴定。结果表明,结合SDS-PAGE和RP-HPLC两种方法可以对Glu-B1位点,尤其是Glu-B1(7 8)进行有效鉴定。选用13份姊妹系为材料,使用RP-HPLC和凝胶色谱(SE-HPLC)方法,研究了Glu-B1al(7OE 8*)以及贮藏蛋白组分含量对面团强度的影响。结果表明,Glu-B1al(7OE 8*)可显著提高HWM-GS总量和面团强度,7OE可作为优质亚基用于强筋小麦育种。相关性分析表明,谷蛋白总量、HWM-GS、LMW-GS以及x-HMW含量与最大抗阻力呈1%显著正相关,相关系数分别为0.76、0.76、0.77和0.72。SDS-不溶性谷蛋白大聚体(UPP)占谷蛋白聚合体总量的百分比与揉面仪峰值时间、粉质仪形成时间和稳定时间以及最大抗阻呈1%或0.1%显著正相关,相关系数分别为0.77、0.90、0.89和0.87。醇溶蛋白与谷蛋白含量的比值与揉面仪峰值时间呈1%显著负相关,相关系数为-0.69;与粉质仪稳定时间和最大抗阻呈5%显著负相关,相关系数分别为-0.58和-0.64。HPLC可有效分离和量化HWM-GS,并作为小麦品质育种有效的辅助手段。

Identification of HWM-GS in Glu-B1 Loci by HPLC and the Effects of 7OE on Wheat Dough Strength

Correct identification of high molecular weight glutenin subunits(HMW-GS),and clarify their effects on wheat dough properties are very important in wheat(Triticum aestivum L.) quality study.Sixty-two cultivars or lines were used in present study to identify the composition of HMW-GS by SDS-PAGE and reversed-phase high-performance liquid chromatography(RP-HPLC).The results showed that combination of SDS-PAGE and RP-HPLC can effectively identify the composition of HWM-GS in Glu-B1 loci,especially for Glu-B1(7 8).Thirteen sib lines were used to study the effect of Glu-B1al(7OE 8) and protein fractions on wheat quality properties related to dough strength by RP-HPLC and size-exclusion high-performance liquid chromatography(SE-HPLC).The results showed that sib lines with Glu-B1al(7OE 8) significantly increased the quantity of HMW-GS,thus significantly improved dough strength,which could be used as high quality subunits in improving wheat gluten strength.Correlation analysis showed that the quantity of total glutenin,HMW-GS,LMW-GS and x-HMW were highly and positively correlated with extensograph maximum resistance(Rmax) with r of 0.76,0.76,0.77,and 0.72(P<0.01),respectively.The relative content of SDS-unextractable glutenin polymeric protein(UPP percentage) were highly and positively correlated with mixograph development time(MDT),farinograph development time(DT) and stability(ST),and Rmax with r of 0.77(P<0.01),0.90(P<0.001),0.89(P<0.001) and 0.87(P<0.001),respectively.The ratio of gliadin to glutenin in quantity was negatively and significantly correlated with MDT,ST and Rmax with r of-0.69(P<0.01),-0.58(P<0.05) and-0.64(P<0.05),respectively.HPLC is a useful tool for wheat quality breeding.