Bowman V B, Huang V, Huang A H
Biology Department, University of South Carolina, Columbia 29208.
J Biol Chem. 1988 Jan 25;263(3):1476-81.
The storage lipids of seeds are packaged into lipid bodies, simple organelles containing a triacylglycerol core surrounded by a layer of phospholipids and proteins. A cDNA encoding the major protein from lipid bodies of maize (L3) was used as a hybridization probe to analyze the regulation of lipid body biosynthesis during seed development and germination. Immunofluorescent microscopy demonstrates that L3 protein accumulation is tissue-specific, confined to the embryo (scutellum and embryonic axis) and the aleurone layer of developing seed. Northern analyses show that L3 mRNA also accumulates to high levels in the embryo and is not found in any nonseed tissue. The steady-state level of L3 mRNA is developmentally regulated, increasing during early seed development and peaking at about the midpoint of seed development. It then decreases slowly, to 20% of the peak level, in the embryo of the mature seed and declines rapidly to undetectable levels as the seed germinates. The high level of L3 gene expression during seed development is not due to amplification of the L3 gene, since Southern analyses of maize genomic DNA indicate that the L3 gene has the same relatively low copy number throughout development (1-4 copies/haploid) genome). Rather, the increase in the steady-state level of L3 mRNA during seed development is transcriptionally regulated. The developmental changes in L3 mRNA levels are paralleled by changes in the transcription of the L3 gene, as measured by run-off transcription in isolated nuclei. The role of the plant growth regulator abscisic acid in regulating L3 gene expression was investigated. When the mature seed is allowed to imbibe in the presence of exogenous abscisic acid, germination is retarded, and the period of L3 gene expression is extended. The extended period of L3 gene expression in the presence of exogenous abscisic acid is due, at least in part, to a dramatic and specific increase in L3 gene transcription.
种子的储存脂质被包装成脂质体,这是一种简单的细胞器,其含有一个三酰甘油核心,周围环绕着一层磷脂和蛋白质。编码玉米脂质体主要蛋白质(L3)的cDNA被用作杂交探针,以分析种子发育和萌发过程中脂质体生物合成的调控。免疫荧光显微镜显示,L3蛋白的积累具有组织特异性,局限于发育种子的胚(盾片和胚轴)和糊粉层。Northern分析表明,L3 mRNA在胚中也积累到高水平,而在任何非种子组织中均未发现。L3 mRNA的稳态水平受到发育调控,在种子发育早期增加,并在种子发育中期左右达到峰值。然后它缓慢下降,在成熟种子的胚中降至峰值水平的20%,随着种子萌发迅速下降至不可检测水平。种子发育过程中L3基因的高表达并非由于L3基因的扩增,因为对玉米基因组DNA的Southern分析表明,L3基因在整个发育过程中具有相同的相对低拷贝数(单倍体基因组中1 - 4个拷贝)。相反,种子发育过程中L3 mRNA稳态水平的增加是由转录调控的。通过分离细胞核中的 runoff 转录测量,L3 mRNA水平的发育变化与L3基因转录的变化平行。研究了植物生长调节剂脱落酸在调节L3基因表达中的作用。当成熟种子在存在外源脱落酸的情况下吸水时,萌发受到抑制,L3基因表达的时期延长。在存在外源脱落酸的情况下,L3基因表达时期的延长至少部分是由于L3基因转录的显著且特异性增加。