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生长素调控完整大豆下胚轴和切取下胚轴切片中的基因表达。

Auxin-regulated gene expression in intact soybean hypocotyl and excised hypocotyl sections.

机构信息

Department of Botany, University of Minnesota, 55108, St. Paul, MN, USA.

出版信息

Planta. 1984 Sep;162(2):147-53. doi: 10.1007/BF00410211.

Abstract

A library of complementary DNA (cDNA) clones has been prepared from polyadenylated RNA (poly(A)(+)RNA) from auxin (2,4-dichlorophenoxyacetic acid)-treated soybean (Glycine max (L.) Merr. cv. Wayne) seedlings. Using differential hybridization, four clones were selected as auxin-responsive, and characterized. The levels of the RNA sequences homologous to the cDNA clones were examined in the hypocotyl of the intact seedling and in excised hypocotyl sections before and after auxin treatment, using RNA blot hybridization analysis. RNA levels are rapidly increased (within 0.25-0.5 h) following auxin treatment and the response in the hypocotyl of the intact seedling is transient, reaching maximum RNA levels 2-4 h after auxin application. Increases in RNA levels were also observed with the auxins indole 3-acetic acid and 2,4,5-trichlorophenoxyacetic acid, but not with the ethylene-producing compound, Ethephon (2-chloroethylphosphonic acid). Hybridization analysis of in-vitro transcription products made in nuclei isolated from untreated and auxin-treated soybean primary leaves and excised hypocotyl sections indicates that, for the two cDNA clones analyzed, the increased RNA levels in auxin-treated organs are at least partially the result of increased transcriptional activity of specific DNA sequences.

摘要

已从生长素(2,4-二氯苯氧乙酸)处理的大豆(Glycine max(L.)Merr. cv.Wayne)幼苗的多聚腺苷酸化 RNA(poly(A)(+)RNA)中制备了互补 DNA(cDNA)克隆文库。使用差异杂交,选择了四个作为生长素反应的克隆,并进行了表征。使用 RNA 印迹杂交分析,在完整幼苗的下胚轴和生长素处理前后的切除下胚轴切片中,检查与 cDNA 克隆同源的 RNA 序列的水平。在生长素处理后,RNA 水平迅速增加(在 0.25-0.5 h 内),并且完整幼苗的下胚轴中的反应是短暂的,在生长素处理后 2-4 h 达到最大 RNA 水平。在吲哚 3-乙酸和 2,4,5-三氯苯氧乙酸等生长素中也观察到 RNA 水平的增加,但在产生乙烯的化合物乙膦(2-氯乙基膦酸)中没有观察到。未处理和生长素处理的大豆初生叶片和切除的下胚轴切片分离的核中进行的体外转录产物的杂交分析表明,对于分析的两个 cDNA 克隆,生长素处理的器官中 RNA 水平的增加至少部分是特定 DNA 序列转录活性增加的结果。

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