高羊茅MAPK基因的克隆与表达分析

丝裂原活化蛋白激酶(M APK)是生物体内信号转导的重要组分,与生长、发育和逆境胁迫反应密切相关.为了研究草坪草对非生物逆境胁迫反应的分子机理,利用同源基因克隆法从4℃低温诱导的草坪草高羊茅(F estu-ca arund inacea Schreb.)幼苗cDNA文库中分离得到一个M APK的cDNA即F aMAPK 1,F aMAPK 1编码369个氨基酸残基的蛋白激酶,该蛋白激酶具有TEY的磷酸化基序.据推测的氨基酸序列的BLA ST同源性分析表明,F aM APK 1蛋白与水稻O sM APK 4蛋白的一致性为91.1%.N orthern杂交检测F aMAPK 1基因对逆境胁迫反应的结果表明冷(4℃)处理对根中F aMAPK 1基因的表达没有明显影响,但诱导叶中F aMAPK 1上调表达.而且低温(4℃)、高盐(250 mm o l/L N aC l)、干旱和100μm o l/L ABA都诱导叶中F aMAPK 1上调表达,表明F aM APK 1蛋白可能在高羊茅对非生物逆境胁迫的反应中起重要作用.

Cloning and Expression of MAPK Gene in Festuca arundinacea

Mitogen-activated kinase (MAPK) is the important component of the signal transmission in living things and closely related to growth,development and responses to adverse environments. In order to explore the molecular mechanism of turf-grass response (Festuca arundinacea Schreb. ) to abiotic stress,homologous gene cloning was adopted to isolate cDNA of a MAPK, FaMAPK1, from the turf-grass seedlings induced at 4 C. FaMAPK1 encoded a protein kinase with 369 amino acid residuals,which had phosphorylation activation motif of TEY. The amino acid sequencing by BLAST showed that the amino acid sequence of FaMAPK1 was 91.1% identical with that of OsMAPK4 in rice. The responses of FaMAPK1 to adverse environmental stress were tested by Northern blotting to show that chilling (at 4 C )did not exert influence on FaMAPK1 gene expression in the roots but FaMAPK1 gene expression was upregulated in the induced leaves. Furthermore, low temperature (at 4C), high salt concentration (250 mmol/L NaC1),drought, 100 μmol/L ABA could led to a up-regulated expression of FaMAPK1,and this indicated that FaMAPK1 probably played an important role in the responses of F. arundinacea to abiotic stresses.