Determination of Amylose Content and Its Relationship with RVA Profile Within Genetically Similar Cultivars of Rice （Oryza safiva L. ssp. japonica）

Anther culture pure lines with antisense Wx gene were generated by Agrobacterium tumefaciens-mediated co-transformation followed by anther culture in transgenic rice （Oryza sativa L. ssp. japonica）. The antisense Wx transgenic pure lines were used to analyze the differences and the relationship between RVA eigenvalues among the lines with different amylose contents. 77 pure lines of anther culture having antisense Wx gene were studied for the amylose content of its offspring cultivar, Wuyunjing 7. It was found that levels ofamylose content were similar to the control in 19 lines （16.0%）; 1-5% lower than control in 50 lines, of which, within 11.0-13.9% in 40 lines; and 3.1-4.0% in 8 lines. The effect of the amylose content on RVA eigenvalue was analyzed through the RVA profile graphic comparison and test of significance for RVA eigenvalues. The analysis of RVA profiles with different amylose contents indicated that there were two kinds of RVA profiles within genetically similar cultivars. The lines with 3.1-4.0% of amylose content were distinctly different from other lines and conventional glutinous rice. The comparison of the similarity curve of RVA profile of the lines with different amylose contents showed that the lines with lower amylose content had a distinguished RVA profile with the curve increasing gradually, but not exceeding the first apex. The test of significance for eigenvalues of the RVA profile in the lines with different amylose contents indicated that there were five eigenvalues remarkably different among the lines with 3.1-4.0% of amylose content compared with other two groups. Further, there were three eigenvalues remarkably different among the lines with 11.0-13.5% of amylose content compared with control group. An optimized mathematical model, which characterizes the relationships between the amylose content and RVA eigenvalue, has been established in this study. From this model, it was clearly indicated that the parameters PeT and SBV were more related to amylose content. It was concluded that the differences in amylose content not only affected the eigenvalues of RVA but also caused different RVA profiles within genetically similar cultivars. The introduction of antisense Wx gene could lead to reduce the amylose content and lay theoretical foundation for quality improvement in rice breeding programs.     

转反义Waxy基因改良龙特甫雄性不育系品质及其育种应用

通过转反义Wx基因育成了直链淀粉含量(AC)降低的籼稻龙特甫新保持系B(wxLB1,wxLB2),并转育成了相应的新不育系新龙保A1(wxLA1)和新龙保A2(wxLA2)。考察和测定了由新不育系和原不育系与恢复系配制的杂种F1代的农艺性状、直链淀粉含量、胶稠度及RVA谱。结果表明:由wxLA1,A2配制的杂种后代直链淀粉含量和粒重极显著地低于各自对照(CK1:LA/YH559,CK2:LA/MH63),胶稠度(GC)和崩解值(BDV)则明显高于对照,并达到极显著水平。而冷胶黏度(CPV)、消减值(SBV)、峰值时间(PeT)、起浆温度(PaT)和回复值(CSV)则显著或极显著低于对照。杂种后代淀粉胶稠度和RVA特征值变异来源分析显示,GC和RVA谱特征值主要受母本影响,且都达到极显著水平。综合结果表明,选择改良籼稻龙特甫雄性不育系品质是改善杂交稻米淀粉理化和蒸煮品质的有效途径。     

缺失不同Wx蛋白对普通小麦直链淀粉含量及淀粉特性的影响

为了明确普通小麦缺失不同W x蛋白对直链淀粉含量的影响以及部分糯小麦品系的淀粉特性,采用单向改良SDS-PAGE方法对1个非糯×糯杂交组合F2分离群体的196个单籽粒进行了W x蛋白筛选,测定了这批籽粒的直链淀粉含量,同时测定了1个部分糯小麦品系(缺失W x-B 1和W x-D 1)和2个正常型小麦品系粗面粉的直链淀粉含量、膨胀势和RVA粘度。结果表明,缺失1对W x蛋白的直链淀粉含量比正常型降低3.29~7.15个百分点,缺失2对W x蛋白的比正常型降低9.45~13.82个百分点,缺失3对W x蛋白的直链淀粉含量几乎为0;缺失W x-B 1和W x-D 1的部分糯小麦品系粗面粉的直链淀粉含量比正常型降低5.62~7.49个百分点,膨胀势则增加4.51~4.72,RVA高峰粘度显著高于正常型。     

Decrease of Waxy (Wx) Protein in Two Common Wheat Cultivars with Low Amylose Content

Two cultivars with a low amylose content, Kanto107 (K107) and K79, were discovered in Japanese common wheat (Triticum aestivum L.). The amount of Wx protein, identified as a single major starch granule-bound protein of about 61 kD by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), was greatly decreased in those two cultivars. Analysis of their Wx protein with a modified SDS-PAGE and two dimensional-PAGE showed that two of the three Wx proteins, produced by Wx-A1 and Wx-B1 loci, were not detected. It was thus concluded that only one locus, Wx-D1, was active in the two low amylose cultivars. These mutants were termed “partial waxy mutants” and considered to be very useful material for breeding waxy wheat.     

利用Wx基因分子标记辅助选择技术培育中等直链淀粉含量的恢复系

为培育中等直链淀粉含量的水稻恢复系,以HC086为供体亲本(其Wx基因来自美国水稻品种Francis),以优良的水稻恢复系R898、R476、R838、R6547为受体亲本,利用PCR-Accl分子标记检测技术在回交世代进行辅助选择,将供体材料控制直链淀粉的Wx基因导入到不同恢复系中.对后代株系的基因型及其直链淀粉含量的分析结果表明,直链淀粉含量普遍有了显著提高,受体亲本的直链淀粉含量由10.6%～14.4%提高到了18.0%～21.4%.改良株系与广占63S和Y58S的配组实验表明,F1代杂交种的直链淀粉含量较原组合有显著提高,并达到适中直链淀粉含量水平,且主要农艺性状没有发生显著改变.说明分子标记辅助选择是改良水稻品种直链淀粉含量的快速有效的方法.     

小麦胚乳Wx蛋白缺失对淀粉含量和淀粉特性的影响

为给面条小麦亲本材料的筛选提供可靠依据，用SDS-PAGE和STS-PCR结合的方法鉴定了24个品种的Wx蛋白亚基组成，测定了这些品种的淀粉含量、膨胀势和RVA参数。结果表明，与正常材料相比，Wx蛋白亚基缺失材料的胚乳中直链淀粉含量减少，支链淀粉和总淀粉含量变化不明显，Wx蛋白亚基缺失的数目越多，直链淀粉含量减少得就越多，糯麦的直链淀粉含量最少。在本研究参试材料特定遗传背景干扰的情况下，在Wx蛋白的三个亚基中Wx-A1蛋白亚基对直链淀粉含量影响最大。Wx蛋白亚基的缺失还使面粉的膨胀势、RVA参数的高峰粘度、崩解值增大，峰值时间延长，而使最终粘度、回生值和糊化温度降低，Wx蛋白亚基缺失的数目越多，这些特性变化的幅度越大。在缺失单个亚基的情况下，Wx-B1亚基的缺失使小麦粉的淀粉特性表现最好。相关分析表明，直链淀粉合成数量的减少是导致上述淀粉特性发生变化的主要原因。     

Wx蛋白缺失对水稻颖果和剑叶生理活性的影响

以转反义Wx基因粳稻和籼稻为材料的研究结果表明，直链淀粉含量降低后的转基因水稻颖果的光合和呼吸活性在发育前期显著高于亲本，但下降较快，到了发育中期(花后15~18 d)以后，就低于同期的亲本。但其SOD和CAT活性比其亲本下降得慢，从花后9 d开始一直保持着比亲本高的水平。花后转基因水稻剑叶的叶绿素含量、光合速率和SOD     

不同Wx蛋白缺失类型小麦穗发芽特性的初步研究

以11份Wx蛋白全部缺失类型（糯性），11份Wx蛋白部分缺失类型（非糯性）和大面积推广品种扬麦158、扬麦12（非糯性）为试验材料，以白皮品种豫麦47为穗发芽对照，研究了它们的穗发芽特性、α-淀粉酶活性和籽粒可溶性糖含量。结果表明，糯小麦的穗发芽率和籽粒可溶性糖含量均极显著高于非糯小麦，而α-淀粉酶活性与非糯小麦差异不显著。α-淀粉酶活性高是非糯小麦穗发芽的原因之一，两者呈指数关系，y=0.4145e^0.4864x(R²=0.3513*)；对于糯小麦，籽粒可溶性糖含量高可能是其穗发芽严重的诱导因素，两者亦呈指数关系，y=0.0001e^2.5599x(R²=0.5338**)。