【作者】 郭世华；

【导师】 王洪刚； 何中虎；

【作者基本信息】 山东农业大学 ， 作物学， 2003， 博士

【摘要】 硬度是小麦重要的品质性状之一，主要影响磨粉品质和食品加工品质。选用我国主要麦区351个品种（系），于2001-02年度种植在北京和安阳，55份春麦区品种（系）于2002年种植于新疆和巴盟，用异丙醇/氯化钠提取Friabilin 蛋白，按Puroindoline a和b及其突变序列设计引物，研究籽粒硬度、Friabilin 蛋白表达量和Puroindoline等位基因的变异类型及其分布特点。65份品种（系）于2001-02年度种植于北方冬麦区11个地点，43份品种种植于南方冬麦区4个地点，研究硬度等性状在不同环境下的变异。主要结果如下：1.冬麦区品种的SKCS籽粒硬度分布范围为12～104，各种硬度类型皆有，但不同麦区差异较大。北部和黄淮麦区品种的硬度显著高于长江中下游和西南麦区，前者以硬质为主，后者分别以软质和混合类型为主。春麦区品种籽粒硬度分布范围为11～84，中等偏硬的品种占70.9%，极端类型较少。东北春麦区、西北春麦区、青藏高原冬春麦区和新疆冬春麦区品种以硬质类型为主，北部春麦区以软质和混合类型为主。2.Friabilin谱带相对表达量与籽粒硬度呈显著负相关，籽粒硬度低，Friabilin谱带强，籽粒硬度数值高，Friabilin谱带弱。用Friabilin蛋白表达量判断冬小麦、春小麦和中优9507穗系籽粒软硬的准确率分别为84.4% 、81.8%和85.3%，其相关系数分别为-0.40、-0.37和-0.68，均达到0.01显著水平。因此，改进的Friabilin蛋白SDS-PAGE技术可用于小麦籽粒硬度生化标记辅助选择。3.STS标记表明，中国小麦的Puroindoline亚基存在丰富变异，包括已报道的2种野生型和5种突变类型、新变异类型、双突变和多位点突变。冬麦区Pina-DIa/Pinb-DIb类型比例最高，达46.8%；Pina-DIa/Pinb-DIa（野生型）和Pina-DIa/Pinb-DIe 的频率也较高，分别为25.4%和10.3%；Pina-DIa/Pinb-DId 和Pina-DIa/Pinb-DIf的频率分别为5.2%和4.8%；4份Pina-DIa/Pinb-DIg类型品种以双突变或多突变出现，占1.6%。Pina-DIb/Pinb-DIa基因品种包括中优9701、原冬107、原冬971和85中33。双突变有15份品种，分别是中优9507-60、中优9507-69、中优9701、原冬107、原冬971、85中33、晋麦60、鲁94（6）006、山农2013、济南17、CA8686、CA9532、京农97-86、晋农218和云麦42，农大3395和晋农207为多位点突变品种；濮优9175、农大3213、扬麦5号和藁城8901为新变异类型。其中濮优9175 的 Ser-103 突变为 Leu-103（TCA to TTA），暂命名为Pina-DIa/ Pinb-DIh；农大3213富含色氨酸区域缺失碱基A，暂命名为Pina-DIa/ Pinb-DIi；扬麦5号Pin b的 442（A）和443（G）之间插入碱基C，暂命名为Pina-DIa/ Pinb-DIj；藁城8901有2个位点碱基突变，即Gln-96—His-96 （CAG to CAT）和 Ser-114——Asn-114（AGT to AAT），暂命名为Pina-DIa/ Pinb-DIk。不同类型间硬度存在差异，其中Pina-DIb/Pinb-DIa和Pina-DIa/Pinb-DIb品种的硬度值高于Pina-DIa/Pinb-DId、 Pina-DIa/Pinb-DIe 和<WP=7>Pina-DIa/Pinb-DIf基因的品种，差异达到0.05显著水平；4.春麦品种Puroindoline变异类型也较多，不同类型分布频率有较大差异，Pina-DIa/Pinb-DIb和Pina-DIa/Pinb-DId在5个麦区均有不同程度的分布，但前者频率（52.7%）大于后者（18.2%）；Pina-DIa/Pinb-DIa（野生型）频率也较高，为12.7%；麦区之间相比，东北春麦区、西北春麦区、青藏高原冬春麦区和新疆冬春麦区Pina-DIa/Pinb-DIb类型比例高，北部春麦区Pina-DIa/Pinb-DIa的频率较高，为40.0%。青藏高原冬春麦区类型最多，有Pina-DIa/Pinb-DIc、Pina-DIa/Pinb-DIe和Pina-DIb/Pinb-DIa ，其对应品种为青春566、97-100、高原2000鉴1和95-1218；东北麦区包括克旱16、沈免90和沈免99121三份双突变品种。Pina-DIb/Pinb-DIa基因品种只有2份，为95R498和95-1218；除野生型外，变异类型间硬度无显著性差异。5.籽粒硬度的品种、地点和品种×地点互作平方和分别占处理总变异平方和的89.53%、1.11%和9.36%，硬度变异主要由品种引起。北片各点籽粒硬度和千粒重明显高于南片，籽粒硬度在各试点的变异程度较大。硬质麦籽粒硬度和SDS沉淀值间存在极显著正相关（r = 0.38），软质麦二者呈显著负相关（r = -0.58）；硬质麦籽粒硬度和千粒重呈极显著负相关（r = -0.53），软质麦二者无显著相关性。更多还原

【Abstract】 Endosperm hardness is one of the most important quality characteristics of cultivated wheat (Triticum aestivum L.) and has a profound effect on milling，baking and the quality of the end products. Three hundred and fifty one wheat varieties and lines from major wheat zones were grown in Beijing and Anyang in 2001 and 2002 seasons,and 55 varieties and lines from spring wheat zones were grown in Xinjiang and Bameng in 2002 seasons. Friabilin was extracted by isopropyl alcohol and NaCl, PCR primers were designed from puroindoline mutation allele sequences. Relationship between their grain hardness and Friabilin level in SDS-PAGE, and distribution of puroindoline hardness mutations (allele) was evaluated. Each variety was assessed for Kernel texture using the Single Kernel Characterization System（SKCS），and the puroindoline hardness allele was determined by STS markers. Sixty five winter wheat varieties were grown at eleven sites in north winter wheat region and fourty three varieties were grown at four sites in south winter wheat region in 2001 and 2002 seasons, wheat quality traits variantions at different sites were analyzed. The major results were presented bellow:1.Grain hardness index of winter wheat varieties was different from 12 to 104. Hard, soft and mixed wheats were distributed among four winter wheat regions at different levels. The hardness of winter wheats from the North China Winter Wheat Zone and Huang Huai Facultative Wheat Zone was obviously higher than that of the Middle and Lower Yangtze Valley Winter-sown Wheat Zone and the Southwestern Winter-sown Wheat Zone, most of varieties in the former were hard, most of varieties in the latter were mixed and soft. Grain hardness index of spring wheat varieties was different from 11 to 84, varieties with harder than mixed shared 70.91%, and extreme type was fewer. Most of varieties from the Northeastern Spring-sown Spring Wheat Zone, the Northwestern Spring Wheat Zones, the Qinghai-Tibet Spring and Winter Wheat Zone and the Xinjiang winter and Spring Wheat Zones were hard , and most of varieties from the Northern Spring-sown Spring Wheat Zone were soft and mixed. 2.Grian hardness was negative correlation with level of friabilin protein. The results showed that high level of friabilin protein in soft wheat and relatively low level in hard wheat, with correlation coefficience -0.40, -0.37 and -0.68 for winter wheat,spring wheat and Zhongyou 9507 seleced lines,respectively, at 0.01 significant level.3.STS molecular markers showed eight kinds of Pin-DI allele were associated with Chinese wheat, Of the winter wheat, frequency of varieties possessed Pina-DIa /Pinb-DIb allele was 46.8%，which was higher than other types. Frequency of varieties with Pina-DIa /Pinb-DIa and Pina-DIa /Pinb-DIe alleles ,which was relatively higher than the other Pin-DI allele wheat,was 25.4% and 10.3%, respectively.For varieties possessed Pina-DIa /Pinb-DId and Pina-DIa /Pinb-DIf allele,the frequency was 5.2% and 4.8%,respectively. Hardness was different among puroindoline mutation alleles, Varieties possessed Pina-DIa /Pinb-DIb and Pina-DIb/ Pinb-DIa allele had high hardness than varieties possessed<WP=9>Pina-DIa /Pinb-DId、Pina-DIa /Pinb-DIe and Pina-DIa /Pinb-DIf alleles and got to a 0.05 significant level.Varieties possessed double-mutation were Zhongyou 9507-60, Zhongyou 9507-69, Zhongyou 9701, Yuandong 107, Yuandong 971 and 85zhong33, Jinmai 60, Lu 94（6）006, Shannong 2013, Jinan 17, Linhan 205, CA 8686, CA9532, Jingnong 97-86, Jinnong 218, Yunmai 42 and four above varieties. Varieties possessed multi-mutation were Nongda 3395 and Jinnong 217, varieties associated with novle mutation type were Nongda 3213, Puyou 9175, Gaocheng 8901 and Yangmai 5. Of the new mutation alleles, Nongda 3213 with one deletion of A in Pinb gene encoding tryptophan-rich domain, causes a frame-shift mutation. Gaocheng 8901 assiciated with Gln-96 to His-96 and Ser-114 to Asn-114. Puyou 9175 had the mutation of Ser-103 to Leu-103 in puroindoline b and Yangmai 5 with insertion “C” bewee更多还原