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马铃薯冷调节基因ci21A的表达模式及启动子活性

Expression patterns and promoter activity of the cold-regulated gene ci21A of potato.

作者信息

Schneider A, Salamini F, Gebhardt C

机构信息

Max-Planck-Institut für Züchtungsforschung, Cologne, Germany.

出版信息

Plant Physiol. 1997 Feb;113(2):335-45. doi: 10.1104/pp.113.2.335.

Abstract

Storage of potato (Solanum tuberosum) tubers at 4 degrees C is associated with the accumulation of several transcripts. DNA sequence analysis of cDNA clone CI21, which corresponds to one of the cold-induced transcripts, revealed high homology to transcripts of tomato (Lycopersicon esculentum) and wild potato (Solanum chacoense) induced by ripening and water stress. Two homologous, nonallelic genes, ci21A and ci21B, were isolated and sequenced. Northern blot analysis showed that CI21 transcripts were present at the highest levels in cold-stored tubers, at lower levels in stems and roots, and at the lowest levels in leaves and tubers stored at room temperature. Treatment with abscisic acid, heat, and a high concentration of salt had no marked effect on CI21 transcript levels in tubers and leaves. Drought was the only stress treatment that induced CI21 transcripts in leaves, but it did not do so in tubers. Western blot analysis detected CI21 protein only in tubers. Chimeric gene constructs between the putative ci21A promoter region and the uidA reporter gene were tested in transgenic potato plants for induction of beta-glucuronidase activity by low temperature. A 2-fold increase of beta-glucuronidase activity in response to tuber storage at 4 degrees C was observed for fragments between 380 and 2000 bp of the ci21A promoter region.

摘要

将马铃薯(Solanum tuberosum)块茎贮藏于4℃与几种转录本的积累有关。对与一种冷诱导转录本相对应的cDNA克隆CI21进行DNA序列分析,发现其与番茄(Lycopersicon esculentum)和野生马铃薯(Solanum chacoense)成熟及水分胁迫诱导的转录本具有高度同源性。分离并测序了两个同源的非等位基因ci21A和ci21B。Northern印迹分析表明,CI21转录本在冷藏块茎中含量最高,在茎和根中含量较低,在室温贮藏的叶片和块茎中含量最低。用脱落酸、热和高浓度盐处理对块茎和叶片中的CI21转录本水平没有显著影响。干旱是唯一能诱导叶片中CI21转录本的胁迫处理,但对块茎无效。Western印迹分析仅在块茎中检测到CI21蛋白。在转基因马铃薯植株中测试了推定的ci21A启动子区域与uidA报告基因之间的嵌合基因构建体,以检测低温对β-葡萄糖醛酸酶活性的诱导作用。对于ci21A启动子区域380至2000 bp之间的片段,观察到在4℃贮藏块茎时β-葡萄糖醛酸酶活性增加了2倍。

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