Theologis A, Huynh T V, Davis R W
J Mol Biol. 1985 May 5;183(1):53-68. doi: 10.1016/0022-2836(85)90280-3.
DNA sequences complementary to three indoleacetic acid (IAA)-inducible mRNAs in pea epicotyl tissue were isolated by differential plaque filter hybridization of cDNA libraries constructed in the vector lambda gt10. Clone pIAA6 hybridized to an mRNA encoding the previously identified translational product polypeptide 6 (Mr 22,000), and clone pIAA4/5 hybridized to one or two mRNAs, encoding polypeptides 4 and 5 (Mr 23,000 and 25,000, respectively). The cDNA clones were subsequently used to characterize the hormonally mediated mRNA accumulation. The induction of the mRNAs was rapid, within 15 minutes of exposure to the IAA, and specific to auxins. Anaerobiosis, heat and cold stress did not induce the mRNAs. Other plant hormones, such as gibberellic acid, kinetin, abscisic acid and ethylene were also unable to cause or interfere with the IAA-induced mRNA accumulation. The hormonally regulated mRNAs were induced at least 50 to 100-fold above control levels after two hours of treatment with IAA and the accumulation was (1) independent of protein synthesis, (2) completely abolished by alpha-amanitin, (3) not due to polyadenylylation of pre-existing RNAs, and (4) independent of IAA and fusicoccin-induced H+ secretion. The IAA-induced mRNAs returned to control levels within three hours after removal of IAA, and the hormonally regulated genes were primarily expressed in the third and second internode of the seven-day-old etiolated pea seedling. The data indicate that IAA increases the amount of specific mRNAs rather than alters the translatability of pre-existing mRNAs. Auxin-induced H+ secretion appears not to have a potential role in mediating the induction and perhaps is a consequence of the enhanced biosynthetic activity induced by the hormone. The IAA-mediated mRNA induction is the fastest known for any plant growth regulator and may represent a primary hormonal response to auxin.
通过对构建于λgt10载体中的cDNA文库进行差异噬菌斑滤膜杂交,分离出了与豌豆上胚轴组织中三种吲哚乙酸(IAA)诱导型mRNA互补的DNA序列。克隆pIAA6与编码先前鉴定的翻译产物多肽6(分子量22,000)的mRNA杂交,克隆pIAA4/5与一种或两种mRNA杂交,分别编码多肽4和5(分子量分别为23,000和25,000)。这些cDNA克隆随后被用于表征激素介导的mRNA积累。mRNA的诱导很快,在暴露于IAA的15分钟内发生,并且对生长素具有特异性。厌氧、热和冷胁迫均未诱导这些mRNA。其他植物激素,如赤霉素、激动素、脱落酸和乙烯也不能引起或干扰IAA诱导的mRNA积累。用IAA处理两小时后,激素调节的mRNA诱导水平比对照水平至少高50至100倍,并且这种积累(1)与蛋白质合成无关,(2)被α-鹅膏蕈碱完全消除,(3)不是由于预先存在的RNA的多聚腺苷酸化,以及(4)与IAA和腐草霉素诱导的H⁺分泌无关。去除IAA后三小时内,IAA诱导的mRNA恢复到对照水平,并且激素调节的基因主要在七天大的黄化豌豆幼苗的第三节和第二节间表达。数据表明,IAA增加了特定mRNA的量,而不是改变预先存在的mRNA的可翻译性。生长素诱导的H⁺分泌似乎在介导诱导过程中没有潜在作用,也许是激素诱导的增强生物合成活性的结果。IAA介导的mRNA诱导是已知的任何植物生长调节剂中最快的,可能代表了对生长素的主要激素反应。