Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important...Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.展开更多
Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourco...Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourcomponent of rice prolamines, 16KDa and10KDa contain more sulfate-riched amino acidresidues. So the total prolamine content andthe percentages of each component were im-portant to parameters for the rice storage pro-tein and its nutrient value. Prolamines of 36rice lines were studied with SDS-PAGE, andrelative quantities of 16 KDa, 13 a, 13 b, and展开更多
Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far...Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far, several sufur-rich seed protein genes have been isolated and the essential amino acid contents of seed proteins were increased in transgenic tobacco and Brassica napus. In this paper we report the isolation and sequencing of the 10kd prolamin gene from seeds. Poly(A) RNA were prepared from the immature endosperms of japonica rice, Sachiminori, 10 d after flowering. Complementary DNAs were synthesized according to Promega Instruction Manual. Two primers were synthesized and their sequences were Primer Ⅰ: CGTCTACACCATCTGGAATC, Primer Ⅱ: GTGTTTGCAGATAGTATGC. The amplified fragraents were inserted into the Sma I site of pGEM-7zf(+) and was used to transform E. coli JM101 after PCR reaction. DNA sequence were determined by Sanger’s Chain-termination method. Synthesis of cDNA. Using mRNAs as展开更多
The 5’ upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super...The 5’ upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super rice cultivar Zhonghua 8. Physical map and DNA sequence analysis showed strong homology with the 5’-flanking region of rice prolamin gene reported by Kim in 1988. No changes in the signal peptide coding sequence and a long leader sequence with several small ORFs were found. Chimeric gene containing 5’-flanking region of the prolamin gene has been transcriptionally fused with the β-glucuronidase reporter gene. The fusion junction was confirmed by both physical map and DNA sequence analysis. The resultant chimeric gene was used to transform the tabacco explants by Ti binary system of Agrobacterium tumefaciens LBA4404. With dot and Southern blotting hybridization, three transgenic tobacco plants with the copies of chimeric GUS genes as many as 20 were obtained. Histochemical analysis展开更多
酒糟是酿酒产业的最大副产物。酒糟富含醇溶蛋白,然而由于水溶性差,限制其进一步应用。采用醇碱法从清香型白酒糟中提取醇溶蛋白(prolamin from distiller's grains,PDG),用三聚磷酸钠对PDG进行磷酸化改性,通过溶解度、持水性、持...酒糟是酿酒产业的最大副产物。酒糟富含醇溶蛋白,然而由于水溶性差,限制其进一步应用。采用醇碱法从清香型白酒糟中提取醇溶蛋白(prolamin from distiller's grains,PDG),用三聚磷酸钠对PDG进行磷酸化改性,通过溶解度、持水性、持油性、乳化性等方面的分析,研究磷酸化改性对PDG功能性质的影响。结果表明,醇碱法提取的白酒糟PDG主要由4种不同分子质量的蛋白组成,原材料酒糟粉粉碎度影响PDG的提取率,粒度小时有利于提高蛋白提取率,以200目细粉为原料时PDG的提取率可达22.69%±0.26%。差示扫描量热实验结果显示PDG的热变性温度为123.47℃,PDG具有良好的热稳定性。傅里叶红外光谱结果表明与三聚磷酸钠反应后磷酸基团修饰到PDG上。磷酸化改性改善了PDG在水中的溶解度,30℃时磷酸化后PDG的溶解度较PDG提高了3.53倍,达到943.08±16.04μg/mL。由于引入磷酸基团,PDG的功能性质显著改善,磷酸化改性后PDG较PDG的持水性和持油性分别增加了44.9%和33.8%(30℃),乳化性及乳化稳定性分别提高了54.9%和3.6倍(pH10)。PDG功能性质的改善有利于扩大其应用领域,该研究为酒糟的高附加值利用奠定了一定基础。展开更多
利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5...利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5-12条谱带，平均8．5条，所有材料有1条共有带。红花种子醇溶蛋白可作为评价红花遗传多样性的工具之一。不同材料的遗传相似系数（GS）变异范围为0．375—1．000，平均值为0．752。聚类分析结果表明，在GS值为0．752的水平上供试材料聚为6大类，其亲缘关系远近与地理来源关系不大。其中，来自中国的7份材料分别被聚在了3个大类中，表明中国红花种子醇溶蛋白遗传多样性比较丰富。展开更多
文摘Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.
文摘Polamines are the major seed storage proteinsin most of the sereals. The rice prolaminescontain a high mole percentage of glutamineresidues and low levels of lysine, histidine,cystine, and methionine. Among the fourcomponent of rice prolamines, 16KDa and10KDa contain more sulfate-riched amino acidresidues. So the total prolamine content andthe percentages of each component were im-portant to parameters for the rice storage pro-tein and its nutrient value. Prolamines of 36rice lines were studied with SDS-PAGE, andrelative quantities of 16 KDa, 13 a, 13 b, and
文摘Nutritious value of seed storage protein is low due to deficiency in essential amino acid contents. Cereals are mainly deficient in lysine and legumes in sulfur-containing amino acids (methionine and cysteine). So far, several sufur-rich seed protein genes have been isolated and the essential amino acid contents of seed proteins were increased in transgenic tobacco and Brassica napus. In this paper we report the isolation and sequencing of the 10kd prolamin gene from seeds. Poly(A) RNA were prepared from the immature endosperms of japonica rice, Sachiminori, 10 d after flowering. Complementary DNAs were synthesized according to Promega Instruction Manual. Two primers were synthesized and their sequences were Primer Ⅰ: CGTCTACACCATCTGGAATC, Primer Ⅱ: GTGTTTGCAGATAGTATGC. The amplified fragraents were inserted into the Sma I site of pGEM-7zf(+) and was used to transform E. coli JM101 after PCR reaction. DNA sequence were determined by Sanger’s Chain-termination method. Synthesis of cDNA. Using mRNAs as
基金Project supported by Rockefeller Foundation for International Program on Rice Biotechnology.
文摘The 5’ upstream region (—680—+40) containing the complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super rice cultivar Zhonghua 8. Physical map and DNA sequence analysis showed strong homology with the 5’-flanking region of rice prolamin gene reported by Kim in 1988. No changes in the signal peptide coding sequence and a long leader sequence with several small ORFs were found. Chimeric gene containing 5’-flanking region of the prolamin gene has been transcriptionally fused with the β-glucuronidase reporter gene. The fusion junction was confirmed by both physical map and DNA sequence analysis. The resultant chimeric gene was used to transform the tabacco explants by Ti binary system of Agrobacterium tumefaciens LBA4404. With dot and Southern blotting hybridization, three transgenic tobacco plants with the copies of chimeric GUS genes as many as 20 were obtained. Histochemical analysis
文摘酒糟是酿酒产业的最大副产物。酒糟富含醇溶蛋白,然而由于水溶性差,限制其进一步应用。采用醇碱法从清香型白酒糟中提取醇溶蛋白(prolamin from distiller's grains,PDG),用三聚磷酸钠对PDG进行磷酸化改性,通过溶解度、持水性、持油性、乳化性等方面的分析,研究磷酸化改性对PDG功能性质的影响。结果表明,醇碱法提取的白酒糟PDG主要由4种不同分子质量的蛋白组成,原材料酒糟粉粉碎度影响PDG的提取率,粒度小时有利于提高蛋白提取率,以200目细粉为原料时PDG的提取率可达22.69%±0.26%。差示扫描量热实验结果显示PDG的热变性温度为123.47℃,PDG具有良好的热稳定性。傅里叶红外光谱结果表明与三聚磷酸钠反应后磷酸基团修饰到PDG上。磷酸化改性改善了PDG在水中的溶解度,30℃时磷酸化后PDG的溶解度较PDG提高了3.53倍,达到943.08±16.04μg/mL。由于引入磷酸基团,PDG的功能性质显著改善,磷酸化改性后PDG较PDG的持水性和持油性分别增加了44.9%和33.8%(30℃),乳化性及乳化稳定性分别提高了54.9%和3.6倍(pH10)。PDG功能性质的改善有利于扩大其应用领域,该研究为酒糟的高附加值利用奠定了一定基础。
文摘利用A—PAGE（Acid—polyacrylamide gel electrophoresis）技术对来源于32个国家的53份红花材料进行了种子醇溶蛋白检测。结果表明，这些红花材料具有丰富的醇溶蛋白等位变异，共分离出15条迁移率不同的谱带。其中，每份材料可电泳出5-12条谱带，平均8．5条，所有材料有1条共有带。红花种子醇溶蛋白可作为评价红花遗传多样性的工具之一。不同材料的遗传相似系数（GS）变异范围为0．375—1．000，平均值为0．752。聚类分析结果表明，在GS值为0．752的水平上供试材料聚为6大类，其亲缘关系远近与地理来源关系不大。其中，来自中国的7份材料分别被聚在了3个大类中，表明中国红花种子醇溶蛋白遗传多样性比较丰富。
