小麦突变体AS208中Glu-B1位点缺失对籽粒中蛋白体形成和储藏蛋白合成与加工相关基因表达的影响

发布时间：2018-05-29 00:14

本文选题：小麦 + 高分子量谷蛋白亚基　；参考：《作物学报》2017年05期

【摘要】：高分子量谷蛋白亚基(HMW-GS)是小麦籽粒贮藏蛋白的重要成分,其组成、表达和含量决定面团弹性和加工品质。本研究以Glu-B1位点上1Bx20和1By20双亚基沉默的小麦无性系变异体AS208为材料,以AS208的来源亲本轮选987作为对照,对AS208中Glu-B1位点的沉默机制和1Bx20、1By20亚基沉默对种子中蛋白体融合以及合成加工相关基因表达的影响进行了研究。Southern blot分析发现AS208比轮选987少2条特异带,染色体原位杂交(FISH)结果显示轮选987中有6条染色体出现杂交信号,而在AS208中有4条染色体出现杂交信号,表明AS208基因组中缺失Glu-B1位点。透射电镜观察发现,与轮选987相比,AS208的籽粒蛋白体形成过程,以及蛋白体大小和形状基本一致。qRT-PCR检测发现,在12个与籽粒蛋白质合成与加工相关基因中,有7个的表达模式两品种相似,有8个的表达量在AS208中高于在轮选987中,特别是Bip、PDI-1和PDI-5基因(高1.5~2.0倍)。本研究结果表明,小麦Glu-B1位点的基因对种子中蛋白体的形成没有明显影响,但一定程度上刺激了多数蛋白质合成和加工相关基因的表达,保证了种子内蛋白含量、蛋白体外形和大小基本不变,表现负反馈调节效应。
[Abstract]:High molecular weight glutenin subunit (HMW-GSH) is an important component of wheat grain storage protein. Its composition, expression and content determine dough elasticity and processing quality. In this study, wheat clone AS208, which was silenced by 1Bx20 and 1By20 subunits at Glu-B1 locus, was used as the material, and 987, the parent of AS208, was used as control. The silencing mechanism of Glu-B1 locus in AS208 and the effect of 1Bx20 1 By20 subunit silencing on protein body fusion and gene expression related to synthetic processing in seeds were studied. Southern blot analysis showed that AS208 had two less specific bands than that of rotation 987. The results of in situ hybridization (fish) showed that there were hybridization signals in 6 chromosomes and 4 chromosomes in AS208, indicating the deletion of Glu-B1 loci in the AS208 genome. Transmission electron microscopy (TEM) showed that the grain proteome formation process, protein body size and shape of AS208 were basically the same as those of rotation 987. QRT-PCR showed that there were 12 genes related to protein synthesis and processing. The expression patterns of 7 cultivars were similar, and 8 of them were higher in AS208 than in rotational selection 987, especially in Bip-PDI-1 and PDI-5 genes. The results showed that the Glu-B1 locus gene had no significant effect on the formation of protein bodies in wheat seeds, but to some extent stimulated the expression of most protein synthesis and process-related genes, and ensured the protein content in seeds. The shape and size of the protein remained unchanged, showing negative feedback regulation effect.
【作者单位】：中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程;首都师范大学生命科学学院;中国科学院遗传与发育生物学研究所/植物细胞与染色体工程国家重点实验室;
【基金】：国家自然科学基金项目(31371621)资助~~
【分类号】：S512.1

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