Kirch H H, van Berkel J, Glaczinski H, Salamini F, Gebhardt C
Max-Planck-Institut für Züchtungsforschung, Köln, Germany.
Plant Mol Biol. 1997 Mar;33(5):897-909. doi: 10.1023/a:1005759925962.
Cold storage of potato tubers at 4 degrees C is associated with the accumulation of several cold-induced transcripts. By using a previously characterized cDNA (CI7) as probe, we isolated and sequenced the corresponding ci7 gene. The putative promoter of ci7 contains sequence elements that have been shown to mediate expression of stress-responsive genes of Arabidopsis thaliana. CI7 transcripts were differentially induced in response to cold, drought, high salt or exogenous ABA treatment in potato tubers and leaves. Whereas accumulation of CI7 transcript during cold storage occurred within days, induction of CI7 transcript in response to drought, ABA and salt occurred rapidly within few hours. In tubers, accumulation of CI7 protein in response to abiotic stresses and ABA was small when compared to transcript levels. In leaves, the CI7 protein was undetectable after all treatments tested. 3 kb of the 5'-flanking ci7 promoter region were fused to the GUS reporter gene and introduced into S. tuberosum plants. The analysis of tubers of independent transgenic lines did not reveal significant induction of enzymatic GUS activity in response to low temperature. When RNA gel blotting was used to analyze the level of induction of the GUS gene driven by the ci7 promoter, the heterologous GUS fusion was, however, strongly responsive to low temperature. Nuclear run-on transcription studies of the ci7 gene, in comparison with RNA gel blot analyses of the transgenic plants, indicated that most of the temperature-regulated expression of the ci7 gene in tubers may be accounted for by post-transcriptional control mechanisms.
将马铃薯块茎在4℃下冷藏与几种冷诱导转录本的积累有关。通过使用先前鉴定的cDNA(CI7)作为探针,我们分离并测序了相应的ci7基因。ci7的推定启动子包含已被证明可介导拟南芥应激反应基因表达的序列元件。CI7转录本在马铃薯块茎和叶片中对冷、干旱、高盐或外源ABA处理有差异诱导。虽然冷藏期间CI7转录本的积累在数天内发生,但对干旱、ABA和盐的CI7转录本诱导在数小时内迅速发生。在块茎中,与转录本水平相比,CI7蛋白对非生物胁迫和ABA的积累较少。在叶片中,在所有测试处理后均未检测到CI7蛋白。将3 kb的5'-侧翼ci7启动子区域与GUS报告基因融合,并导入马铃薯植株。对独立转基因系块茎的分析未发现低温诱导酶促GUS活性有显著增加。然而,当使用RNA凝胶印迹分析由ci7启动子驱动的GUS基因的诱导水平时,异源GUS融合对低温有强烈反应。与转基因植物的RNA凝胶印迹分析相比,ci7基因的核运行转录研究表明,块茎中ci7基因的大多数温度调节表达可能由转录后控制机制解释。