Terada R, Nakayama T, Iwabuchi M, Shimamoto K
Plantech Research Institute, Yokohama, Japan.
Plant J. 1993 Feb;3(2):241-52. doi: 10.1046/j.1365-313x.1993.t01-16-00999.x.
To investigate developmental regulation of wheat histone H3 gene expression, the H3 promoter, which has its upstream sequence to -1711 (relative to the cap site as +1), was fused to the coding region of the gus A gene (-1711H3/GUS) and introduced into a monocot plant, rice. Detailed histochemical analysis revealed two distinct types of GUS expression in transgenic rice plants; one is cell division-dependent found in the apical meristem of shoots and roots and in young leaves, and another is cell division-independent detected in flower tissues including the anther wall and the pistil. In this study, replication-dependent expression occurring in non-dividing cells which undergo endoreduplication could not be discriminated from strict replication-independent expression. The observed expression pattern in different parts of roots suggested that the level of the H3/GUS gene expression is well correlated with activity of cell division in roots. To identify 5' sequences of the H3 promoter necessary for an accurate regulation of the GUS expression, two constructs containing truncated promoters, -908H3/GUS and -185H3/GUS, were analyzed in transiently expressed protoplasts, stably transformed calli and transgenic plants. The results indicated that the region from -909 to -1711 contains the positive cis-acting element(s) and that the proximal promoter region (up to -185) containing the conserved hexamer, octamer and nonamer motifs is sufficient to direct both cell division-dependent and -independent expression. The use of the meristem of roots regenerated from transformed calli for the analysis of cell division-dependent expression of plant genes is discussed.
为了研究小麦组蛋白H3基因表达的发育调控,将其上游序列至-1711(相对于转录起始位点为+1)的H3启动子与gus A基因的编码区融合(-1711H3/GUS),并导入单子叶植物水稻中。详细的组织化学分析揭示了转基因水稻植株中两种不同类型的GUS表达;一种是依赖细胞分裂的,见于茎尖分生组织、根尖分生组织和幼叶中,另一种是不依赖细胞分裂的,见于包括花药壁和雌蕊在内的花组织中。在本研究中,发生在进行核内复制的非分裂细胞中的依赖复制的表达无法与严格的不依赖复制的表达区分开来。在根的不同部位观察到的表达模式表明,H3/GUS基因的表达水平与根中的细胞分裂活性密切相关。为了鉴定准确调控GUS表达所需的H3启动子的5'序列,在瞬时表达的原生质体、稳定转化的愈伤组织和转基因植株中分析了两个含有截短启动子的构建体,即-908H3/GUS和-185H3/GUS。结果表明,-909至-1711区域含有正向顺式作用元件,而包含保守六聚体、八聚体和九聚体基序的近端启动子区域(至-185)足以指导依赖细胞分裂和不依赖细胞分裂的表达。本文还讨论了利用转化愈伤组织再生的根分生组织来分析植物基因的依赖细胞分裂的表达。