Department of Plant Sciences, College of Agriculture and Bioresources, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
J Agric Food Chem. 2010 Apr 14;58(7):4193-201. doi: 10.1021/jf903861q.
An in vitro culture system is useful to study grain development under defined conditions to minimize confounding effects associated with whole plant studies and metabolite movement into the developing grains. The objective of this study was to monitor the expression patterns of carbohydrate metabolism genes during grain development in an in vitro wheat spike culture system. Immature spikes were cultured prior to anthesis, and grains were collected at various days postanthesis (DPA). Grains from cultured spikes showed maximum expression of starch metabolic genes by 10 DPA, with a rapid decline thereafter. The rapid increase and decrease in expression rate in the in vitro system was thought to be due to fructan exohydrolase (1-FEH and 6-FEH) or sucrose transporter 1 (SUT1) and sucrose synthase (SuSy) genes being highly expressed. SUT1 reached peak expression at 8 DPA, two days earlier than the other genes, and may account for the rapid early stage trigger in expression of the other genes. However, expression of 1-FEH and 6-FEH genes in in vitro-cultured spikes peaked at 12 DPA, two days later than the other genes, and could indicate that fructan catabolism was not a factor in the rapid accumulation of starch in the in vitro-cultured spikes. Accumulation of GBSSI polypeptides generally showed similar patterns in both systems, with the maximum amount in the in vitro system observed four days later than in the in planta spikes, reflecting different turnover controls of GBSSI transcripts. The in vitro system offers opportunities for further refinement and detailed grain development studies.
体外培养系统有助于在特定条件下研究谷物发育,以最大程度地减少与整体植物研究相关的混杂效应和代谢物向发育中的谷物的转移。本研究的目的是监测体外小麦穗培养系统中谷物发育过程中碳水化合物代谢基因的表达模式。在开花前对未成熟的穗进行培养,并在开花后不同天数(DPA)收集谷物。培养穗的谷物在 10 DPA 时表现出最大的淀粉代谢基因表达量,此后迅速下降。体外系统中表达率的快速增加和下降被认为是由于果聚糖外切酶(1-FEH 和 6-FEH)或蔗糖转运蛋白 1(SUT1)和蔗糖合酶(SuSy)基因的高度表达。SUT1 在 8 DPA 时达到表达高峰,比其他基因早两天,可能是其他基因早期快速表达的触发因素。然而,在体外培养的穗中,1-FEH 和 6-FEH 基因的表达在 12 DPA 时达到高峰,比其他基因晚两天,这表明果聚糖分解代谢不是体外培养的穗中淀粉快速积累的因素。GBSSI 多肽的积累在这两种系统中通常表现出相似的模式,体外系统中最大量的观察到比在体内穗中晚四天,反映了 GBSSI 转录物不同的周转率控制。体外系统为进一步细化和详细的谷物发育研究提供了机会。