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丝氨酸/苏氨酸磷酸酶PP1家族在植物生长发育和响应逆境胁迫中发挥重要的作用。探索澳洲坚果MiSTPP5基因在不同组织中和低温胁迫下的表达规律，可为深入研究PP1家族成员在澳洲坚果生长发育和抗寒分子机制提供理论基础。本研究采用RT-PCR技术克隆澳洲坚果MiSTPP5基因，利用生物信息学分析其编码蛋白，并通过实时荧光定量PCR检测其在澳洲坚果不同组织中及低温胁迫下的表达模式。结果显示，澳洲坚果MiSTPP5含有PP1家族的典型结构域MPP＿PP1＿PPKL,属于稳定的亲水性蛋白和非分泌跨膜蛋白。MiSTPP5蛋白以丝氨酸残基磷酸化为主，定位于细胞质的可能性最大(高达65%),其二级和三级结构主要以α螺旋、无规则卷曲和延伸链交错而成。MiSTPP5基因在澳洲坚果的根、茎、叶、花和小果等器官组织中都有表达，其中在叶片中的表达量最高，在根中则最低。在低温胁迫下，MiSTPP5基因在叶片中的表达量呈现“升-降-升-降”的表达模式，总体上受诱导上调表达。因此，澳洲坚果MiSTPP5基因在不同组织中都有表达，并受低温胁迫诱导上调表达。
Abstract：The PP1 family of serine/threonine phosphatase plays important roles in plant growth and development and response to stress. In order to provide a theoretical basis for further study on PP1 family genes in growth and development of Macadamia integrifolia and their molecular mechanism of cold resistance, the expression patterns of MiSTPP5 gene in cold stress and the different tissues of macadamia were analyzed. In this study, MiSTPP5 gene was cloned from macadamia by RT-PCR, and its coding protein was analyzed by bioinformatics. Expression patterns of MiSTPP5 gene in cold stress and the different tissues of macadamia were detected by real-time fluorescent quantitative PCR. The results showed that MiSTPP5 was a stable hydrophilic protein and non-secreted transmembrane protein, containing the typical domain MPP＿PP1＿PPKL of PP1 family. MiSTPP5 was mainly phosphorylated by serine residues, which may be located in the cytoplasm(up to 65%). The secondary and 3 D main components of MiSTPP5 were α-helix, random coil and extended strand. MiSTPP5 gene was expressed in the roots, stems, leaves, flowers and small fruits of macadamia, which had the highest expression in leaves and the lowest expression in roots. Expression level of MiSTPP5 gene in leaves was increased under cold stress, with an “up-down-up-down” expression pattern. MiSTPP5 gene was expressed in different tissues of macadamia, and was up-regulated by cold stress.
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基本信息:
DOI：10.13417/j.gab.041.000617
中图分类号:S664.9
引用信息:
[1]蔡元保,杨祥燕,黄强,等.澳洲坚果MiSTPP5基因在不同组织及低温胁迫的表达分析[J].基因组学与应用生物学,2022,41(03):617-624.DOI:10.13417/j.gab.041.000617.
基金信息:
国家自然科学基金项目(31860537); 广西科技计划项目(桂科AB19245008,桂科AD18281005); 广西农业科学院基本科研业务专项(桂农科2021YT154,桂农科2020YM56)共同资助
2022-03-25
2022-03-25