楸树花粉壁蛋白质提取及特异性  

Pollen-wall protein extraction and characteristics of Catalpa bungei

作　　者：王改萍[1] 徐涛[1] 樊莉丽[1] 彭方仁[1] 吕昕[1] 陈琳月 

机构地区：[1]南京林业大学林学院,江苏南京210037

出　　处：《浙江农林大学学报》2016年第1期65-70,共6页Journal of Zhejiang A&F University

基　　金："十二五"国家科技支撑计划子项目(2012BAD21B03);江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：为了进一步研究楸树Catalpa bungei自交不亲和特性,以楸树不同产地植株云台山楸树(CB^(-1)),老山楸树(CB-2)和滇楸Catalpa fargesi f.duclouxii(CF)的花粉为对象,利用普通研磨法及超声波破碎法,研究了适合于花粉壁蛋白质提取的方法。同时采用考马斯亮蓝染色法及聚丙烯酰胺凝胶电泳(SDS-PAGE)比较了不同树种花粉蛋白质质量分数及其主要蛋白质组分。结果表明:超声波破碎法适于花粉壁蛋白质的提取,最佳超声参数为功率400 W,超声时间3 s·次^(-1),间歇时间6 s·次^(-1),3个回合,工作120次,相隔5 min·回合^(-1)。蛋白质提取液三羟甲基氨基甲烷(Tris-HCl)的pH 7.8最佳。不同树种的花粉壁蛋白质质量分数表现为滇楸最高,云台山楸树和老山楸树较低。蛋白质组分研究表明:各树种花粉共有蛋白质有26个,特异蛋白质为连楸53.8 k D,35.0 k D,23.4 k D,26.5 k D;老楸53.8 k D,38.5 k D,23.4 k D;滇楸38.5 k D,26.5 k D。The research was conducted to further reveal the self-incompatibility that exists extensively t, atazpa bungei and to evaluate the best way to extract pollen-wall protein of Catalpa, including the Catalpa bungei plants CB-1 from Yuntaishan National Forest Park of Lianyungang and CB-2 from Laoshan Forest of Nanjing and Catalpafargesii f. duclouxii （CF） using incorporation and uhrasonication. Also, the content and component of pollen-wall protein from different varieties was compared by the Coomassie Blue Staining Method and SDS （sodium dodecyl sulfate）-polyacrylamide gel electrophoresis （PAGE）. Results showed that the relative opti- mization method to extract pollen-wall protein of C. bungei was ultrasonication with the optimal parameters be- ing： ultrasonication at 400 W power for 3 s every 6 s for 3 rounds taking 120 duplications at an interval of 5 min. The optimal pH value to extract pollen-wall protein was pH 7.8 （F〉F0c~）. For pollen-wall protein content, CF was higher （F〉F00J） than both CB-1 and CB-2. From the pollen of these three catalpas, 26 proteins were found. Molecular weight of the specific proteins for CB-1 were 53.8 kDa, 35.0 kDa, 23.4 kDa, and 26.5 kDa; for CB-2 they were 53.8 kDa, 38.5 kDa, and 23.4 kDa; and for CF they were 8.5 kDa and 26.5 kDa. [Ch, 6 fig. 1 tab. 18 ref.]

关 键 词：植物学 楸树 超声波破碎法 蛋白质 聚丙烯酰胺凝胶电泳(SDS-PAGE) 

分 类 号：S718.4[农业科学—林学]