桃PpSnRK1βλ1基因的克隆及在拟南芥中异源转基因的功能分析

发布时间：2018-07-10 20:11

  本文选题：桃树 + PpSnRKβλ　； 参考：《北方园艺》2017年11期

【摘要】：以实生毛桃为试材,采用PCR和实时荧光定量技术,克隆桃树SnRK1蛋白激酶(蔗糖非发酵-1-型相关蛋白激酶-1)的调节亚基βλ,并分析其组织表达特性。构建p35S::PpSnRK1βγ1重组载体,获得超表达PpSnRK1βλ1基因拟南芥植株用于分析其生物功能。结果表明:克隆获得桃树SnRK1βλ1,命名为PpSnRK1βλ1。该cDNA全长为1 476bp,编码492个氨基酸。序列分析表明,其包含CBM和4个CBS结构域;PpSnRK1βλ1与果梅的SnRK1βλ蛋白亲缘关系最近;PpSnRK1βλ1基因在实生毛桃的根、茎、叶均有表达,其中在茎中表达量最低。超表达PpSnRK1βγ1基因拟南芥株系A-βγ1的花期比野生型晚2.19d,单株莲座叶数量比野生型多1.17片;叶片的叶绿素、可溶性糖和可溶性蛋白质含量均显著高于野生型拟南芥,分别比野生型提高了13.7%、12.9%和23.3%,但淀粉含量却没有显著性差异。在氧化胁迫条件下,转基因植株与野生型的生长均受到抑制,但前者具有更好的萌发率和主根长度,以保证植株正常生长。因此,PpSnRK1βλ1基因参与调控植株的碳氮代谢,并影响植物的花期,以及在防御氧化胁迫过程中起重要作用。
[Abstract]:The regulation subunit 尾 位 of SnRK1 protein kinase (sucrose non-fermentation-associated protein kinase 1) was cloned by PCR and real-time fluorescence quantitative technique, and its tissue expression characteristics were analyzed. P35 S: PpSnRK1 尾 纬 1 recombinant vector was constructed, and the overexpression of PpSnRK1 尾 位 1 gene was obtained from Arabidopsis thaliana to analyze its biological function. The results showed that SnRK1 尾 位 1 was cloned and named PpSnRK1 尾 位 1. The cDNA is 1476 BP and encodes 492 amino acids. Sequence analysis showed that PpSnRK1 尾 位 1 containing CBM and four CBS domains was closely related to the SnRK1 尾 位 1 protein of fruit plum. PpSnRK1 尾 位 1 gene was expressed in roots, stems and leaves of Phyllostachys pubescens, and the lowest expression was found in stems. In Arabidopsis thaliana strain A- 尾 纬 1 with overexpression of PpSnRK1 尾 纬 1 gene, the flowering time of A- 尾 纬 1 strain was 2.19 days later than that of wild type, the number of rosette leaves per plant was 1.17 more than that of wild type, and the contents of chlorophyll, soluble sugar and soluble protein in leaves of Arabidopsis thaliana were significantly higher than those of wild type Arabidopsis thaliana. Compared with wild type, the content of starch increased by 13.9% and 23.3%, respectively, but there was no significant difference in starch content. Under the condition of oxidative stress, the growth of transgenic plants and wild-type plants were inhibited, but the former had better germination rate and main root length to ensure the normal growth of the plants. Therefore, PpSnRK1 尾 位 1 gene plays an important role in regulating the metabolism of carbon and nitrogen in plants, affecting the flowering period of plants, and in the process of defense against oxidative stress.
【作者单位】： 山东农业大学园艺科学与工程学院作物生物学国家重点实验室;
【基金】：国家现代农业产业技术体系建设专项资金资助项目(CARS-31)
【分类号】：S662.1
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本文编号：2114559