油用型红花(Carthamus tinctoris L.)种子醇溶蛋白遗传多样性分析

利用A-PAGE（Acid-polyacrylamide gel electrophoresis）技术对来源于不同国家的79份油用型红花材料醇溶蛋白位点进行检测。结果表明，油用型红花醇溶蛋白位点存在丰富的变异类型，共分离出15条迁移率不同的谱带，每份材料具有3—12条不等，平均8．5条。材料间平均遗传相似系数（GS）为0．5303，变幅为0．1333—1．000，且各洲间遗传多样性大于洲内遗传多样性。在GS值为0．512的水平上，供试材料聚为六大类，聚类结果表明，红花醇溶蛋白图谱类型与其地理分布有一定的相关，但不显著。     

Allelic variation of the HMW glutenin subunits in Aegilops tauschii accessions detected by sodium dodecyl sulphate (SDS-PAGE), acid polyacrylamide gel (A-PAGE) and capillary electrophoresis

Variability of high molecular weight glutenin subunits (HMW-GS) was studied in198 accessions of Ae. Tauschii (2n=2x=14, DD) by sodium dodecyl sulphate(SDS-PAGE) and acid polyacrylamide gel electrophoresis (A-PAGE) and capillary electrophoresis (CE). A high allelic variation of HMW-GS, including some novel x- and y-type subunits and variable subunit combinations were observed. One accession(TD159) showed a x-type null form. The results by A-PAGE analysis revealed that the subunits Dx5 ^t and Dy10 ^t encoded by Glu-D ^t 1 locus in Ae. tauschii were different in relative mobilities in comparison with the subunits Dx5 and Dy10 found in bread wheats, whereas they had the same mobilities, respectively, when separated by SDS-PAGE. The higher resolution of Ae. tauschii HMW-GS separated by CE method showed two clear peaks in accordance with x- and y-type subunits, respectively,except the accession TD151 which possessed only subunit Dy12.1^*t. The electro elution time of the x-type and y-type subunits were about 13–14 and 7–8minutes, respectively. Characterization of wheat HMW-GS was facilitated by using CE which provides high resolution and increases the speed of analysis in conjunction with the traditional gel electrophoretic methods. A total of 42HMW-GS alleles were identified, among which were several alleles not presently detected in bread wheats. Hence Ae. tauschii is potentially a valuable genetic resource for quality improvement of bread wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

Unique gliadin patterns in Chinese winter wheat cultivars

Cultivated Chinese wheat germplasm has been a valuable genetic resource in international plant breeding. Patterns of gliadin among cultivated Chinese accessions are unknown, despite the proven value and potential novelty. The objective of this work was to analyse the diversity within improved Chinese wheat germplasm. The electrophoretic banding patterns of gliadin in winter wheat cultivars and advanced lines were determined by acid-polyacrylamide gel electrophoresis. For 148 leading commercial cultivars and promising advanced lines used in this study, 48 patterns were identified, 29 corresponding to ω -gliadin, nine to γ -gliadin, five to β -gliadin and five to α -gliadin. The most frequent patterns were A6 in ω ; B in γ ; B in β and A in the region of α . A total of 116 band types appeared in the 148 samples: 94 accessions had unique gliadin types, and 22 gliadin, types while not unique, were found in 54 accessions. The gliadin patterns of Chinese wheat cultivars and lines greatly differed from the patterns of wheat lines from other countries. Three patterns, E, J, H, M, N and O in the ω -zone had not been reported previously. Three wheat zones of China, the Northern Winter Wheat Region, the Yellow and Huai Valley River valleys Winter Wheat Region and the Southwestern Winter Wheat Region, showed different frequencies in their gliadin patterns. This information can be used to monitor genetic diversity with Chinese wheat germplasm.     

Effect of allelic variation at the Glu-3/Gli-1 loci on breadmaking quality parameters in hexaploid wheat (Triticum aestivum L.)

Low molecular weight glutenin subunits (LMW-GS) encoded by the Glu-3 loci are known to contribute to wheat breadmaking quality. However, the specific effect of individual Glu-3 alleles is not well understood due to their complex protein banding patterns in SDS-PAGE and tight linkage with gliadins at the Gli-1 locus. Using DNA markers and a backcross program, we developed a set of nine near isogenic lines (NILs) including different Glu-A3/Gli-A1 or Glu-B3/Gli-B1 alleles in the genetic background of the Argentine variety ProINTA Imperial. The nine NILs and the control were evaluated in three different field trials in Argentina. Significant genotype-by-environment interactions were detected for most quality parameters indicating that the effects of the Glu-3/Gli-1 alleles are modulated by environmental differences. None of the NILs showed differences in total flour protein content, but relative changes in the abundance of particular classes of proteins cannot be ruled out. On average, the Glu-A3f, Glu-B3b, Glu-B3g and Glu-B3i_Man alleles were associated with the highest values in gluten strength-related parameters, while Glu-A3e, Glu-B3a and Glu-B3i_Chu were consistently associated with weak gluten and low quality values. The value of different Glu-3/Gli-1 allele combinations to improve breadmaking quality is discussed.     

栽培荞麦种子醇溶蛋白遗传多样性分析

利用A-PAGE（Acid-polyacrylamide gel electrophoresis）对来源于7个国家的76份栽培荞麦（苦荞54份,甜荞22份）醇溶蛋白遗传多样性进行评价。结果表明,荞麦醇溶蛋白位点存在丰富的等位变异,共分离出18条迁移率不同的谱带,每份材料具有6~12条不等,平均9.5条,多态性带占88.89%。材料间平均遗传相似系数（GS）为0.777,变幅为0.389~1.000。在GS为0.63的水平上供试材料可聚为苦荞和甜荞两大类,绝大部分来自于相同或相似生态地理环境的材料聚成一类,表明荞麦醇溶蛋白所揭示的遗传关系与地理来源有较高的相关性。     

冬小麦种质矮孟牛及其衍生后代1BL/1RS的分子和生化标记鉴定

本研究以小麦骨干亲本矮孟牛及其33个衍生品种(系)为材料,利用低分子量(LMW)麦谷蛋白Glu-B3位点的STS-PCR标记、醇溶蛋白Gli-B1位点的SSR标记和黑麦碱SEC-1b位点的STS-PCR标记进行复合PCR,检测1BL/IRS易位.结果表明,矮孟牛Ⅱ、Ⅳ、Ⅴ、Ⅵ和Ⅶ型含有1BL/1RS染色体,矮孟牛Ⅰ和Ⅲ型不含1BL/1RS;在矮孟牛的33个衍生后代中,25个含1BL/1RS,其余8个则不含1BL/1RS.利用A-PAGE技术对上述材料进行了黑麦碱蛋白的检测,结果与复合PCR一致,两种方法相结合能准确的检测1BL/1RS.     

小麦春溶蛋白酸性聚丙烯酰胺凝胶电泳（A-PAGE）方法的优化与改良及其应用

依据国际种子检验协会（ISTA）1986年公布的标准小麦醇溶蛋白酸性聚丙烯酰胺凝胶电泳（A-PAGE）方法,结合国内实验室的实际情况,摸索出了适合我国试剂和仪器的小麦醇溶蛋白酸性聚丙烯酰胺凝胶电泳（A-PAGE）的改良方法。该方法克服了标准方法中的胶韧性差、剥胶困难、分辨率不高、谱带识别繁琐、实验效率较低的限制,适合我国小麦品种纯度的快速鉴定、血缘关系分析等。     

适用于小麦品质育种的一种A-PAGE方法

为育种需要大批量地分析5+10亚基龙麦19中对小麦品质有较大影响的醇溶蛋白块品系,利用双板夹芯式垂直槽,凝胶面积为182 mm(宽)×95 mm(高),分离胶浓度为6%,以甲酸为缓冲体系的不连续的酸性聚丙烯酰胺凝胶电泳(A-PAGE)来鉴定该醇溶蛋白块。结果表明:该系统可以较好地鉴定该醇溶蛋白块,具有简便、经济、高效、重复性好等特点,是适合于小麦育种过程中快速大量鉴定需要的一种A-PAGE电泳方法。     

利用两种谷蛋白电泳体系分析杂交油菜品种间差异

以8个杂交油菜品种（系）种子为试材,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和酸性-聚丙烯酰胺凝胶电泳（A-PAGE）体系,分析杂交油菜种子谷蛋白电泳图谱的差异。结果表明,SDS-PAGE谷蛋白电泳图谱多态性较强,图谱清晰度较高,能有效鉴别杂交油菜品种。     

应用A-PAGE电泳方法鉴定豇豆品种

分别用3种提取液提取种子醇溶蛋白，利用改进的ISTA小麦和大麦醇渣蛋白酸性聚丙烯酰胺凝胶电泳（A-PAGE）方法对豇豆不同品种种子进行了鉴定。结果表明，以20％二氯乙醇＋1％2．巯基乙醇＋18％的尿素为提取液。且加样量为10μl时，电泳图谱条带清晰，可以对豇豆品种进行有效地鉴定。利用超声波处理样品可以缩短样品提取时间并得到同样的提取效果。