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西藏白菜型油菜油体特征与营养品质及油体提取物的SDS-PAGE分析
侯维海, 王建林, 旦 巴,等
西藏农牧学院 高原作物分子育种实验室,植物科学学院
摘要:
【目的】探索白菜型油菜种子油体、油体蛋白表型与含油量和蛋白质、脂肪酸含量之间的关系,分析油体提取物蛋白亚基的多态性,为西藏白菜型油菜高油新品种的选育奠定基础。【方法】测定36份来自西藏不同地区的白菜型油菜农家种、野生近缘种种子的含油量和蛋白质、脂肪酸含量;利用Leica DM5000生物显微镜观察所提取油体的大小及油体蛋白的表型特征;利用SDS-PAGE对供试材料油体提取物进行凝胶电泳,分析供试材料的遗传多态性和进化关系。【结果】36份供试材料芥酸、油酸、亚油酸和烯酸含量分别为27.39~132.33,3.22~143.22,31.22~53.62和14.27~111.48 mg/g,其他脂肪酸含量相对较低;36份材料的含油量为57.20~151.37 mg/g,蛋白质含量为38.68~73.87 mg/g。相关分析显示,含油量与蛋白质含量、棕榈酸、烯酸含量呈极显著负相关。根据含油量将供试材料划分为高油组(含油量≥138 mg/g,HO)、中油组(110 mg/g≤含油量<138 mg/g,MO)、 低油组(含油量<110 mg/g,LO)3个类别,不同含油组材料油体和油体蛋白形状均以近圆形为主,HO油体平均粒径2.12 μm,油体蛋白平均粒径5.24 μm;MO油体平均粒径1.82 μm,油体蛋白平均粒径4.58 μm;LO油体平均粒径1.44 μm,油体蛋白平均粒径3.81 μm。相关分析表明,油体平均粒径与含油量呈极显著正相关,与蛋白质含量呈极显著负相关,而与脂肪酸含量相关性不大。油体粗提液SDS PAGE电泳共分离出迁移率不同的蛋白谱带32 条,出现频率为5.6%~100%;蛋白谱带的分子质量为25.5~128.7 ku,且分布较集中区域在87.8~27.4 ku;蛋白谱带N J聚类分析表明,供试材料遗传相似系数为0.610~0.980,在遗传相似系数0.835处可将36份材料聚为9 个类群。【结论】西藏白菜型油菜遗传变异丰富,在西藏高油白菜型油菜品种选育时可将油体粒径大且数量多、油体蛋白少且粒径大作为筛选亲本的条件。
关键词:  白菜型油菜  油体  SDS-PAGE  遗传多样性
DOI:
分类号:
基金项目:国家自然科学基金项目(31560362);西藏自治区重点项目(XZXTCX-2016);西藏自治区高校青年教师创新支持计划项目(QC2015-35);国家重点研发计划项目(2016YFC0502005)
Oil body characteristics,nutritional quality and SDS-PAGE analysis of oil body extracts of Brassica campestris form Tibet
HOU Weihai, WANG Jianlin, DAN Ba,et al
Abstract:
【Objective】This study explored the relationship of oil body and oleosin phenotype of Brassica campestris L.seeds with contents of oil,protein and fatty acid, and analyzed the polymorphism of protein subunit of oil body extracts to provide basis for developing new varieties of Tibetan B. campestris.【Method】This study utilized Leica DM5000 biological microscope to obtain phenotypic characteristics of oil bodies and oleosin,and measured oil, protein and fatty acid contents of seeds of 36 local cultivars and wild relative species of B. campestris from different areas in Tibet.The protein electrophoresis of oil extracts was carried out by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) to analyze the genetic relationship between subunit polymorphism and the tested materials.【Result】Erucic acid,oleic acid,linoleic acid and arachidonic acid contents of 36 sample materials were 27.39-132.33 mg/g,3.22-143.22 mg/g,31.22-53.62 mg/g and 14.27-111.48 mg/g,while contents of other fatty acids were relatively low.The oil content was 57.20 to 151.37 mg/g and the protein content was 38.68 to 73.87 mg/g.The correlation analysis showed that there was a significant negative correlation between the content of oil and the contents of protein,palmitic acid and olefine acid.The tested materials were divided into high oil content group (oil content≥138 mg/g,HO),middle oil content group (110 mg/g≤oil content<138 mg/g,MO) and low oil content group (oil content<110 mg/g,LO) according to oil content.The shape of oil body and oleosin in tested materials were nearly round.The average diameters of oil body and oleosin of the HO group were 2.12 μm and 5.24 μm,respectively.The average diameters of oil body and oleosin of the MO group were 1.82 μm and 4.58 μm,respectively.The average diameters of the oil body and oleosin of the LO group were 1.44 μm and 3.81 μm,respectively.Correlation analysis showed that the average diameter of oil was extremely significant positive correlated with oil content and extremely negative correlated with protein content,but the correlation with fatty acid content was insignificant.A total of 32 protein bands with different mobility ratios were isolated by SDS PAGE electrophoresis for the oil body extracts,and the occurrence frequencies for all protein bands were 5.6%-100%.The relative molecular mass of the protein band was 25.5-128.7 ku and the denser area of the distribution was 87.8-27.4 ku.N J (Neighbor Joining) clustering of the protein bands showed that the genetic similarity coefficient of the tested materials was 0.610-0.980,and 36 materials could be clustered into 9 groups by similarity coefficient of 0.835.【Conclusion】Tibetan B. campestris have abundant genetic diversity.In future,large diameter and quantity for oil body as well as less quantity and large diameter for oleosin can be used to screen parental species for breeding new B. campestris varieties.
Key words:  Brassica campestris L  oil body  SDS-PAGE  genetic diversity