Roach Melissa J, Deyholos Michael K
Department of Biological Sciences, University of Alberta, Edmonton, Canada.
Ann Bot. 2008 Sep;102(3):317-30. doi: 10.1093/aob/mcn110. Epub 2008 Jun 30.
Hypocotyls are a commonly used model to study primary growth in plants, since post-germinative hypocotyls increase in size by cell elongation rather than cell division. Flax hypocotyls produce phloem fibres in bundles one to two cell layers thick, parallel to the protoxylem poles of the stele. Cell wall deposition within these cells occurs rapidly at a well-defined stage of development. The aim was to identify transcripts associated with distinct stages of hypocotyl and phloem fibre development.
Stages of flax hypocotyl development were defined by analysing hypocotyl length in relation to fibre secondary wall deposition. Selected stages of development were used in microarray analyses to identify transcripts involved in the transition from elongation to secondary cell wall deposition in fibres. Expression of specific genes was confirmed by qRT-PCR and by enzymatic assays.
Genes enriched in the elongation phase included transcripts related to cell-wall modification or primary-wall deposition. Transcripts specifically enriched at the transition between elongation and secondary wall deposition included beta-galactosidase and arabinogalactan proteins. Later stages of wall development showed an increase in secondary metabolism-related transcripts, chitinases and glycosyl hydrolases including KORRIGAN. Microarray analysis also identified groups of transcription factors enriched at one or more stages of fibre development. Subsequent analysis of a differentially expressed beta-galactosidase confirmed that the post-elongation increase in beta-galactosidase enzyme activity was localized to phloem fibres.
Transcripts were identified associated with specific stages of hypocotyl development, in which phloem fibre cells were undergoing thickening of secondary walls. Temporal and spatial regulation of beta-galactosidase activity suggests a role for this enzyme in remodelling of flax bast fibre cell walls during secondary cell wall deposition.
下胚轴是研究植物初生生长常用的模型,因为萌发后的下胚轴通过细胞伸长而非细胞分裂来增大尺寸。亚麻下胚轴产生的韧皮纤维束有一到两个细胞层厚,与中柱的原生木质部极平行。这些细胞内的细胞壁沉积在发育的特定阶段迅速发生。目的是鉴定与下胚轴和韧皮纤维发育不同阶段相关的转录本。
通过分析下胚轴长度与纤维次生壁沉积的关系来定义亚麻下胚轴发育阶段。选择发育的特定阶段用于微阵列分析,以鉴定参与纤维从伸长到次生细胞壁沉积转变过程的转录本。通过qRT-PCR和酶促分析证实特定基因的表达。
在伸长阶段富集的基因包括与细胞壁修饰或初生壁沉积相关的转录本。在伸长和次生壁沉积转变阶段特异性富集的转录本包括β-半乳糖苷酶和阿拉伯半乳聚糖蛋白。壁发育的后期阶段显示与次生代谢相关的转录本、几丁质酶和包括KORRIGAN在内的糖基水解酶增加。微阵列分析还鉴定了在纤维发育的一个或多个阶段富集的转录因子组。对差异表达的β-半乳糖苷酶的后续分析证实,伸长后β-半乳糖苷酶活性的增加定位于韧皮纤维。
鉴定出了与下胚轴发育特定阶段相关的转录本,其中韧皮纤维细胞正在经历次生壁增厚。β-半乳糖苷酶活性的时空调节表明该酶在次生细胞壁沉积过程中对亚麻韧皮纤维细胞壁重塑中起作用。