Identification of glycinin and β-conglycinin subunits that contribute to the increased protein content of high-protein soybean lines
Identification of glycinin and β-conglycinin subunits that contribute to the increased protein content of high-protein soybean lines
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DOI：
10.1021/jf062497n
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发表时间：
2007-03-07
影响因子：
6.1
通讯作者：
Nelson, Randall L.
中科院分区：
文献类型：
--
作者：
Krishnan, Hari B.;Natarajan, Savithiry S.;Nelson, Randall L.
根据基因型和位置，以干重计算的商业大豆品种的种子蛋白质浓度范围为约37 - 42%。目前正在进行协调一致的研究，以进一步提高蛋白质浓度。已知几种大豆植物引种（PI）含有大于50%的蛋白质。育种者利用这些PI将高蛋白质性状纳入商业北美品种。目前，关于调节高蛋白的生化和遗传机制的信息有限。本研究对LG 00 -13260及其亲本高蛋白系PI 427138和BARC-6的种子蛋白进行了蛋白质组学和分子生物学分析。十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析表明，高蛋白线积累的β-伴大豆球蛋白和大豆球蛋白的量增加，当与威廉姆斯82。利用pH 4-7和pH 6-11两性电解质的高分辨率双向电泳能够提高大豆种子蛋白的分辨率。基质辅助激光解吸电离飞行时间质谱法共鉴定了38个蛋白质点，分别代表β-伴大豆球蛋白和大豆球蛋白的不同亚基。高蛋白与代表β-伴大豆球蛋白和大豆球蛋白的大多数亚基积累的增加相关。高蛋白大豆品系氨基酸谱的比较表明，含硫氨基酸的浓度，反映蛋白质的质量，不受蛋白质浓度的影响。Southern印迹分析显示，PI 427138和BARC-6之间编码组1大豆球蛋白、β-伴大豆球蛋白、Bowman-Birk抑制剂（BBI）和Kunitz胰蛋白酶抑制剂（KTI）的基因在DNA水平上存在基因型变异。LG 00 -13260遗传了亲本系PI 427138的大豆球蛋白、β-伴大豆球蛋白和KTI的等位基因变体，而BBI遗传自亲本系BARC-6。我们的研究结果表明，高种子蛋白质浓度是由于更大的积累β-伴大豆球蛋白和大豆球蛋白亚基的特定成分，推测这些基因在种子发育过程中的优先表达介导。
Seed protein concentration of commercial soybean cultivars calculated on a dry weight basis ranges from approximately 37 to 42% depending on genotype and location. A concerted research effort is ongoing to further increase protein concentration. Several soybean plant introductions (PI) are known to contain greater than 50% protein. These PIs are exploited by breeders to incorporate the high-protein trait into commercial North American cultivars. Currently, limited information is available on the biochemical and genetic mechanisms that regulate high-proteins. In this study, we have carried out proteomic and molecular analysis of seed proteins of LG00-13260 and its parental high-protein lines PI 427138 and BARC-6. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed that the high-protein lines accumulated increased amounts of beta-conglycinin and glycinins, when compared with Williams 82. High-resolution two-dimensional electrophoresis utilizing pH 4-7 and pH 6-11 ampholytes enabled improved resolution of soybean seed proteins. A total of 38 protein spots, representing the different subunits of beta-conglycinin and glycinin, were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. High-protein was correlated with an increase in the accumulation of most of the subunits representing beta-conglycinin and glycinin. Comparisons of the amino acid profiles of high-protein soybean lines revealed that the concentration of sulfur amino acids, a reflection of protein quality, was not influenced by the protein concentration. Southern blot analysis showed the presence of genotypic variation at the DNA level between PI 427138 and BARC-6 for the genes encoding group1 glycinin, beta-conglycinin, Bowman-Birk inhibitor (BBI), and the Kunitz trypsin inhibitor (KTI). LG00-13260 inherited the allelic variants of the parental line PI 427138 for glycinin, beta-conglycinin, and KTI, while BBI was inherited from the parental line BARC-6. The results of our study indicate that high-seed protein concentration is attributed to greater accumulation of specific components of beta-conglycinin and glycinin subunits presumably mediated by preferential expression of these genes during seed development.