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我国南方春大豆种子发育过程中,常处于高温、高湿季节,加之种子本身富含蛋白(约40%)和脂肪(约20%),导致南方春大豆种子易劣变。本项目组前期差异蛋白质组学研究发现蔗糖结合蛋白在高温高湿胁迫168 h时在种子田间劣变抗性品种湘豆3号发育种子中呈下调表达。为进一步从分子水平了解Gm SBP基因表达以及响应高温高湿胁迫的特性,本研究利用RT-PCR技术从大豆扩增出两个Gm SBP基因(Gm SBP2和Gm SBPL)。两个基因编码的蛋白均为亲水性,不完整的膜蛋白。荧光定量PCR分析表明:在高温高湿条件下,种子田间劣变不抗品种宁镇1号和抗性品种湘豆3号发育种子中Gm SBP2和Gm SBPL基因表达量均受高温高湿胁迫影响,也会导致种子中蔗糖和可溶性糖含量变化。在籽粒发育过程中,Gm SBP2和Gm SBPL基因表达量在花后30 d左右达到最高,对应时期的蔗糖和可溶性糖含量也达到最大值。组织特异性显示Gm SBP和Gm SBPL基因在不同组织间存在差异表达。亚细胞定位结果表明Gm SBP2和Gm SBPL蛋白均定位在细胞膜和细胞质中。以上结果表明Gm SBP2和Gm SBPL基因可能参与了植物非生物胁迫的应答过程,这将从一个侧面丰富我们对大豆种子田间劣变性和劣变抗性的认识。
During the development of spring soybean in southern China, it is often in the high temperature and high humidity season. In addition, the seeds themselves are rich in protein (about 40%) and fat (about 20%), which leads to the easy change of seeds in southern spring. The preliminary differential proteomics study in this project group showed that sucrose-binding protein was down-regulated in seed development of Xiangdou No.3 under the condition of high temperature and high humidity 168 h. In order to further understand the characteristics of Gm SBP gene expression and response to high temperature and high humidity at the molecular level, two Gm SBP genes (Gm SBP2 and Gm SBPL) were amplified from soybean by RT-PCR. Both genes encode proteins that are hydrophilic and incomplete membrane proteins. Fluorescence quantitative PCR analysis showed that under the conditions of high temperature and high humidity, Gm SBP2 and Gm SBPL gene expression in non-resistant variety Ningzhen 1 and resistant variety Xiangdou 3 were all affected by high temperature and high humidity , But also lead to changes in the content of sucrose and soluble sugar in the seeds. During grain development, the expression levels of Gm SBP2 and Gm SBPL genes reached the highest level at about 30 days after anthesis, and the contents of sucrose and soluble sugar also reached the maximum during the corresponding period. Tissue specificity showed that Gm SBP and Gm SBPL genes were differentially expressed in different tissues. Subcellular localization results showed that both Gm SBP2 and Gm SBPL proteins were localized in the cell membrane and cytoplasm. The above results indicated that the Gm SBP2 and Gm SBPL genes may be involved in the plant abiotic stress response process, which will enrich our understanding of the bad and bad resistance of soybean seed in one aspect.