Allen R D, Trelease R N, Thomas T L
Department of Biology, Texas A&M University, College Station, Texas 77843.
Plant Physiol. 1988 Feb;86(2):527-32. doi: 10.1104/pp.86.2.527.
A cDNA sequence that encodes a portion of sunflower (Helianthus annuus L.) seedling isocitrate lyase was selected from a lambda gt11 cDNA library derived from sunflower seedling cotyledon poly(A)(+) messenger RNA. The library was screened for bacteriophage recombinants that expressed antigens which reacted with antisera directed against cotton seed isocitrate lyase. The isolated cDNA hybridized with a 2 kilobase RNA species that was first detectable in maturing sunflower embryos 19 days after flowering and remained at a constant low level through seed desiccation. The prevalence of this transcript in sunflower cotyledons increased by about 10-fold within 2 days after inhibition in darkness, and transcript levels began to decrease by 5 days after imbibtion. During the first 2 days of germination and growth of sunflower seedlings in light, the rate of isocitrate lyase mRNA accumulation was greater than the rate observed during this period in dark-grown seedlings, giving peak levels about 2-fold higher than corresponding levels in dark-grown seedlings. Illumination of seedlings also promoted an earlier, and more rapid decline of isocitrate lyase transcripts. Peak levels of isocitrate lyase mRNA preceded a corresponding peak in immunologically detectable isocitrate lyase polypeptides by about 24 hours. Isocitrate lyase expression in sunflower cotyledons is developmentally regulated and is modulated, in seedlings, by exposure to light. Mechanisms that control these processes appear to function primarily at the level of mRNA accumulation and are likely to involve changes in transcription rates and/or mRNA stability.
从一个由向日葵幼苗子叶多聚腺苷酸(poly(A))信使核糖核酸构建的λgt11 cDNA文库中,挑选出了一个编码向日葵(Helianthus annuus L.)幼苗异柠檬酸裂解酶部分序列的cDNA。该文库用于筛选表达与抗棉籽异柠檬酸裂解酶抗血清发生反应的抗原的噬菌体重组体。分离得到的cDNA与一种2千碱基的RNA杂交,这种RNA在开花后19天的成熟向日葵胚中首次可检测到,并在种子干燥过程中保持在恒定的低水平。在黑暗中抑制2天内,这种转录本在向日葵子叶中的丰度增加了约10倍,而在吸胀后5天转录本水平开始下降。在向日葵幼苗在光照下发芽和生长的头2天里,异柠檬酸裂解酶mRNA积累的速率大于在此期间黑暗中生长的幼苗的观察速率,峰值水平比黑暗中生长的幼苗相应水平高约2倍。幼苗的光照也促进了异柠檬酸裂解酶转录本更早、更快的下降。异柠檬酸裂解酶mRNA的峰值水平比免疫可检测的异柠檬酸裂解酶多肽的相应峰值提前约24小时出现。向日葵子叶中异柠檬酸裂解酶的表达受发育调控,在幼苗中受光照影响。控制这些过程的机制似乎主要在mRNA积累水平起作用,并且可能涉及转录速率和/或mRNA稳定性的变化。