Jacobsen S E, Olszewski N E
Department of Plant Biology, University of Minnesota, St. Paul 55108, USA.
Planta. 1996;198(1):78-86. doi: 10.1007/BF00197589.
In an effort to understand the molecular mechanism of gibberellin (GA) action, we have cloned and performed an initial characterization of three cDNAs (GAD1, 2, and 3) which correspond to RNAs that become less abundant by 2 h after treatment of tomato (Lycopersicon esculentum Mill.) shoot tissue with gibberellic acid (GA3). Treatment with either auxin or ethephon also decreases the abundance of all three of the GAD RNAs. The tomato ethylene-insensitive mutant, Nr, and the GA-deficient mutant, gib1, were used to show that GA or auxin regulation of GAD RNA abundance is not dependent on ethylene sensitivity, and that ethylene or auxin regulation is not dependent on normal levels of gibberellin biosynthesis. Treatment with abscisic acid (ABA) antagonizes the GA-induced suppression of the GAD1 and GAD2 RNAs. GAD1 is similar to type-II wound-inducible plant proteinase inhibitors. Like the well-characterized proteinase inhibitor II (pin II) of tomato, the GAD1 and GAD2 RNAs are wound inducible. Induction of pin II and GAD1 RNA in gib1 was found to require less-severe wounding than was required using wild-type plants or plants doubly mutant for gib1 and sit (the sit mutation causes ABA deficiency). The predicted GAD2 protein sequence is similar to 2-oxoglutarate-dependent dioxygenases while the predicted GAD3 protein sequence is similar to proteins belonging to the nonmetallo-short-chain alcohol-dehydrogenase family, especially the T ASSELSEED2 (TS2) gene of maize and bacterial hydroxysteroid dehydrogenases.
为了了解赤霉素(GA)作用的分子机制,我们克隆了三个cDNA(GAD1、2和3)并进行了初步表征,这三个cDNA对应的RNAs在用赤霉酸(GA3)处理番茄(Lycopersicon esculentum Mill.)茎尖组织2小时后丰度降低。用生长素或乙烯利处理也会降低所有三种GAD RNAs的丰度。番茄乙烯不敏感突变体Nr和GA缺陷突变体gib1被用于证明GA或生长素对GAD RNA丰度的调节不依赖于乙烯敏感性,并且乙烯或生长素的调节不依赖于正常水平的赤霉素生物合成。用脱落酸(ABA)处理可拮抗GA诱导的GAD1和GAD2 RNAs的抑制作用。GAD1与II型创伤诱导型植物蛋白酶抑制剂相似。与番茄中特征明确的蛋白酶抑制剂II(pin II)一样,GAD1和GAD2 RNAs是创伤诱导型的。发现在gib1中诱导pin II和GAD1 RNA所需的创伤程度低于使用野生型植物或gib1和sit双突变体植物(sit突变导致ABA缺乏)所需的创伤程度。预测的GAD2蛋白序列与2-氧代戊二酸依赖性双加氧酶相似,而预测的GAD3蛋白序列与属于非金属短链醇脱氢酶家族的蛋白相似,尤其是玉米的T ASSELSEED2(TS2)基因和细菌羟基类固醇脱氢酶。