Melan M. A., Enriquez ALD., Peterman T. K.
Department of Biological Sciences, Wellesley College, Wellesley, Massachusetts 02181.
Plant Physiol. 1994 May;105(1):385-393. doi: 10.1104/pp.105.1.385.
We examined the temporal and spatial expression patterns of the LOX1 gene during the development of Arabidopsis thaliana seedlings. Measurements of steady-state LOX1 mRNA levels indicated that this gene is transiently expressed during germination. LOX1 mRNA was not detected in seed that had imbibed (T0) but reached a maximum level by 1 d in both light- and dark-grown seedlings. The induction of the LOX1 gene was not light dependent; however, mRNA levels were 4-fold greater in light-grown seedlings. Immunoblot analysis of lipoxygenase protein levels and measurements of enzyme activity suggested that the induction of the LOX1 gene resulted in the production of functional lipoxygenase enzyme. Lipoxygenase protein was not present in dry seed or seed that had imbibed, but was first detected by immunoblot analysis after 1 and 2 d of growth in the light and dark, respectively. In both cases, lipoxygenase protein levels remained high for 2 d and then declined. Lipoxygenase activity paralleled the changes in protein levels. In situ hybridization studies revealed that the LOX1 gene is transiently expressed in the epidermis and the aleurone layer during germination. LOX1 mRNA levels were particularly high in the epidermis of the radicle and the adaxial side of the cotyledons. These results suggest that the LOX1 gene product is produced specifically during early germination and plays a role in the functioning of the epidermis.
我们研究了拟南芥幼苗发育过程中LOX1基因的时空表达模式。稳态LOX1 mRNA水平的测量表明,该基因在萌发过程中短暂表达。在已吸胀的种子(T0)中未检测到LOX1 mRNA,但在光照和黑暗培养的幼苗中,到第1天时均达到最高水平。LOX1基因的诱导不依赖光照;然而,光照培养的幼苗中mRNA水平高4倍。脂氧合酶蛋白水平的免疫印迹分析和酶活性的测量表明,LOX1基因的诱导导致了功能性脂氧合酶的产生。脂氧合酶蛋白在干种子或已吸胀的种子中不存在,但分别在光照和黑暗中生长1天和2天后通过免疫印迹分析首次检测到。在这两种情况下,脂氧合酶蛋白水平在2天内保持较高,然后下降。脂氧合酶活性与蛋白水平的变化平行。原位杂交研究表明,LOX1基因在萌发过程中在表皮和糊粉层中短暂表达。LOX1 mRNA水平在胚根表皮和子叶近轴侧特别高。这些结果表明,LOX1基因产物在早期萌发过程中特异性产生,并在表皮功能中发挥作用。