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为了确定耐热性不同的小麦基因型是否在离体条件下热冲击蛋白(HSP)的合成(可翻译的HSPmRNA)及HSP mRNA的稳态水平的不同,本文对2个小麦品种Mustang(耐热)和Sturdy(热敏感)热冲击蛋白(HSP)基因的表达情况进行了分析。在37℃下对小麦幼苗进行不同时间的热胁迫处理,然后从其叶组织中分离出几组mRNA,再将这些mRNA与HSP cDNA或基因组探针(HSP17,26,70,98及泛有素)进行杂交。用离体翻译和双向凝胶电泳比较蛋白质的剖面图,胁迫处理后的Northern印迹资料表明,热冲击期间,耐热品种Mustang中低分子量(LMW)HSP mRNA的合成较热敏感品种Sturdy的发生得早,LMW HSP mRNA的含量也比Sturdy的高,位于叶绿体上的HSP尤为如此。双向凝胶电泳显示的蛋白质剖面图表明,2个小麦品系间不仅HSP的数量有差异,而且一些特异性HSP只出现于Mustang的HSP剖面图中。以HSP17为探针进行的Southern印迹杂交表明,两品系间RFLP的水平较高。这些资料为进一步对HSP基因在小麦耐热性方面的遗传分析提供了分子学基础。
In order to determine whether different wheat genotypes with different heat tolerance have different steady-state levels of heat shock protein (HSP) synthesis (translatable HSP mRNA) and HSP mRNA in vitro, two wheat cultivars Mustang ) And Sturdy (heat sensitive) heat shock protein (HSP) gene expression were analyzed. The wheat seedlings were subjected to heat stress at 37 ° C for different periods of time, and then several mRNAs were isolated from their leaf tissues and compared with HSP cDNA or genomic probes (HSP17, 26, 70, 98 and panopressin ). The cross-sectional images of proteins were compared by in vitro translation and two-dimensional gel electrophoresis. Northern blotting data after stress treatment showed that the synthesis of low molecular weight (LMW) HSP mRNA from the heat-tolerant variety Mustang occurred more frequently than the heat-sensitive variety Sturdy Early, LMW HSP mRNA levels are also higher than those of Sturdy, especially in chloroplasts. The cross-section of the protein revealed by two-dimensional gel electrophoresis showed that not only the number of HSPs differed between the two wheat lines, but some specific HSPs only appeared in Mustang’s HSP profiles. Southern blot hybridization with HSP17 probe showed that RFLP levels were higher between the two lines. These data provide the molecular basis for further genetic analysis of HSP gene in wheat heat tolerance.