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两种浊化剂在小麦谷蛋白大聚合体测定中的有效性

Comparison of Two Sorts of Turbidimetry Reagents in Measurement of Wheat Glutenin Macropolymer Content

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【作者】 王世杰; 康明辉; 尤明山; 李保云; 刘广田;

【Author】 WANG Shi-jie~(1,2),KANG Ming-hui~3,YOU Ming-shan~1,LI Bao-yun~1,LIU Guang-tian~1(1.China Agricultural University,Key Lab.of Crop Genomics & Genetic Improvement,Ministry of Agriculture,Beijing Key Lab.of Crop Genetic Improvement,Beijing 100094,China;2.Wheat Breeding Center,Henan Institute of Education,Zhengzhou,Henan450014,China;3.Institute of Bio.&Tech.,Henan Academy of Agricultural Sciences,Zhengzhou,Henan 450002,China)

【机构】 中国农业大学农业部作物基因组学与遗传改良重点开放实验室北京市作物遗传改良重点实验室; 河南省农科院生物技术研究所; 中国农业大学农业部作物基因组学与遗传改良重点开放实验室北京市作物遗传改良重点实验室 北京100094; 河南教育学院人口与生命科学系/小麦育种研究中心; 河南郑州450014; 河南郑州450002; 北京100094;

【摘要】 为比较两种浊化剂5-磺基水杨酸-无水硫酸钠(SSA-SS)和三氯乙酸(TCA)在比浊法测定小麦谷蛋白大聚合体(GMP)含量中的有效性,选用黄淮冬麦区有代表性的4个强筋小麦品种和4个中弱筋小麦品种组配的杂种F2代面粉为材料,采用1%DTT的50%正丙醇溶液提取GMP,分别用浊化剂SSA-SS和TCA浊化,再用酶标仪(波长540 nm)连续(10~90 min)测定不同基因型小麦的GMP提取液的吸光值。结果表明:(1)以SSA-SS为浊化剂时,牛血清蛋白浓度与吸光值的直线回归关系极其密切(r=0.9980**);以TCA为浊化剂时,牛血清蛋白浓度与吸光值的直线回归关系较差。(2)以SSA-SS为浊化剂时,各种基因型小麦GMP提取液的吸光值在30 min内具有较好的稳定性;以TCA为浊化剂时,各种基因型小麦GMP提取液的吸光值随时间不断增加。说明比浊法测定小麦GMP含量时,以SSA-SS为浊化剂,加样振荡后10~30min内测定的结果最为稳定。

【Abstract】 The flours of F2 derived from the crosses of four strong wheats and four medium or soft wheats,which were representative cultivars in the Huanghuai winter wheat region,were used to compare the efficiency of the turbidimetry reagents,5-sulfosalicylic acid and sodium sulfate(SSA-SS) and trichloroacetic acid(TCA),on the measurement of wheat Glutenin macropolymer(GMP).50% 1-propernol with 1% DTT was used to extract the wheat GMP,and the solutions of GMP were turbided using SSA-SS or TCA.The absorbance values of turbid solutions were obtained continuously(1090 min) by means of microplate reader at 540 nm of wavelength.The results were as follows:(1) It was by SSA-SS assay that the regression between the various concentrations of BSA and absorbance values is significantly linear(r=0.9980**),while by TCA assay,it was not.(2) It was with SSA-SS that the absorbance values of the GMP of solution are stable and timeless(1030 min),while with TCA,that those continuously grow up(1050 min).In conclusion,SSA-SS assay was available to measure the GMP content of wheat.

【基金】 国家自然科学基金项目(30270823,30471076);国家“863”计划项目(2002AA207003,2003AA207080)
  • 【文献出处】 麦类作物学报 ,Journal of Triticeae Crops , 编辑部邮箱 ,2006年06期
  • 【分类号】S512.1
  • 【被引频次】7
  • 【下载频次】72
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