南方珍珠豆型花生籽仁蛋白质积累及亚基肽组分分析

Protein Accumulation in the Seed of Southern Spanish-type Peanut and the Analysis of Subunit Polypeptide Component

为研究南方珍珠豆型花生荚果发育过程中,蛋白质的积累及各类蛋白亚基的分布、可溶性蛋白质相对浓度变化规律,采用凯氏定氮法和SDS-PAGE电泳分析花生籽仁蛋白质及亚基肽组分,利用Bio-RadImagelabTM分析软件对花生蛋白质条带的相对浓度进行测算。结果表明,珍珠豆型花生籽仁蛋白质含量随生育进程,在开花40～50天急剧增加,后平缓上升。花生蛋白质合成积累一直伴随花生籽仁发育的全过程中,蛋白质高值出现在成熟期。因基因型的差异蛋白质合成积累的速度和强度有所不同,蛋白质增长曲线存在差异。花生蛋白种类存在较高的多态性,不同基因型花生,蛋白质的亚基构成不同,蛋白质积累过程的亚基条带数量均比成熟时丰富。不同基因型花生可溶性蛋白质相对浓度存在差异。可溶性总蛋白质相对浓度的峰值在70～80天,蛋白质亚基相对浓度分布在0.01～4.59,不同基因型存在差异。亚基相对浓度变化可分未缺型和缺失型2类亚基,缺失型亚基有前期缺失、中期缺失、后期缺失和前后期缺失亚型;前期缺失、中期缺失亚型亚基与未缺型亚基共同形成不同基因型花生的蛋白质构成。后期缺失亚型和前后期缺失亚型亚基在花生蛋白质积累的某个时期参与花生蛋白质的合成,终因后期被降解而得不到表达。2种基因型花生蛋白质亚基相对浓度变化除68kDa条带呈抛物线型积累外,其余条带均呈波动性积累。

In order to know the discipline of protein accumulation, distribution of protein subunits and variation of soluble protein relative concentration during the pod-setting of southern Spanish-type peanut, the Kjeldahl method and SDS-PAGE electrophoresis were used to analyze the peanut seed protein and subunits of peptide fractions. The relative concentration of peanut protein band had been estimated by Bio-Rad Image lab TM. The results showed that, Spanish-type peanut seed protein content increased dramatically after 40-50 d of the flower, then gentle rising. Protein accumulated with the seed development of whole process, and arrived the highest point in mature period. Protein growth curve was different, because unlike genotype lead to difference of protein accumulation speed and strength. The type of peanut protein species had the higher polymorphism, protein subunit composition of different between genotype. The number of bands of mature stage was the least.The soluble protein relative concentration was different in unlike genotypes. The total soluble protein relative concentration distribution in 0.01-4.59, the peak appeared in 70-80 d. Subunits relative concentration could be divided without missing and missing type, missing subunits have appeared early, medium, late and both early and late. The type of later missing and both early and late take part in peanut protein synthesis, then degrade and disappear in late stage. Two genotypes of peanut protein subunits relative to the concentration change parabola accumulation to 68 kDa strip, the remaining strip showed fluctuation accumulation.