Yabe N, Takahashi T, Komeda Y
Molecular Genetics Research Laboratory, University of Tokyo, Japan.
Plant Cell Physiol. 1994 Dec;35(8):1207-19. doi: 10.1093/oxfordjournals.pcp.a078715.
We have isolated three HSP90-family genes from Arabidopsis: HSP81-1 which is heat-inducible, and HSP81-2 and -3 which are highly expressed under normal growth temperatures. Northern blot analysis and RNase protection analysis, using gene specific probes, showed that HSP81-2 and -3 mRNA were present in all tissues and abundant in roots, floral bud clusters, and flowers at 22 degrees C. A small amount of HSP81-1 mRNA was detected only in roots. In situ hybridization and histochemical analysis using transgenic plants carrying chimeric gene fusions, with an HSP81 promoter region fused to a beta-glucuronidase (GUS) gene, confirmed these results. At 22 degrees C, high GUS activity was observed in the root apical meristems, pollen and tapeta in HSP81-2::GUS and HSP81-3::GUS transgenic plants, while only branches of the root in HSP81-1::GUS transgenic plants expressed high GUS activity. After 2 hours of 35 degrees C treatment, extensively high GUS activity was observed in all tissues in HSP81-1::GUS transgenic plants, while elevated but tissue specific expression was observed in HSP81-2 and -3 transgenic plants. Exogenous application of various chemicals such as ABA, GA3, kinetin, IAA, NaCl, and mannitol revealed that 10 mM IAA and 0.1 M NaCl significantly enhanced the accumulation of HSP81-2 and -3 transcripts. Only a slight response to IAA was observed in HSP81-1 mRNA accumulation at 22 degrees C; the increase was possibly caused by a novel pathway other than heat-shock-response pathway.
我们从拟南芥中分离出了三个热休克蛋白90(HSP90)家族基因:热诱导型的HSP81-1,以及在正常生长温度下高表达的HSP81-2和HSP81-3。使用基因特异性探针进行的Northern印迹分析和核糖核酸酶保护分析表明,HSP81-2和HSP81-3的信使核糖核酸(mRNA)存在于所有组织中,在22摄氏度时,根、花芽簇和花中含量丰富。仅在根中检测到少量的HSP81-1 mRNA。使用携带嵌合基因融合体(HSP81启动子区域与β-葡萄糖醛酸酶(GUS)基因融合)的转基因植物进行原位杂交和组织化学分析,证实了这些结果。在22摄氏度时,在HSP81-2::GUS和HSP81-3::GUS转基因植物的根尖分生组织、花粉和绒毡层中观察到高GUS活性,而在HSP81-1::GUS转基因植物中只有根的分支表达高GUS活性。在35摄氏度处理2小时后,在HSP81-1::GUS转基因植物的所有组织中均观察到广泛的高GUS活性,而在HSP81-2和HSP81-3转基因植物中观察到活性升高但具有组织特异性表达。外源施加各种化学物质,如脱落酸(ABA)、赤霉素(GA3)、激动素、吲哚乙酸(IAA)、氯化钠(NaCl)和甘露醇,结果显示10 mM IAA和0.1 M NaCl显著增强了HSP81-2和HSP81-3转录本的积累。在22摄氏度时,HSP81-1 mRNA积累对IAA仅表现出轻微反应;这种增加可能是由热休克反应途径以外的新途径引起的。