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细胞壁转化酶及高粱(双色高粱)种子发育过程中其他几个糖代谢相关基因的表达

Expression of cell wall invertase and several other genes of sugar metabolism in relation to seed development in sorghum (Sorghum bicolor).

作者信息

Jain Mukesh, Chourey Prem S, Li Qin-Bao, Pring Daryl R

机构信息

Department of Plant Pathology, University of Florida, Gainesville, FL 32611-0680, USA.

出版信息

J Plant Physiol. 2008;165(3):331-44. doi: 10.1016/j.jplph.2006.12.003. Epub 2007 Feb 12.

Abstract

We report expression profiles of several genes of carbohydrate metabolism, cell wall invertase (CWI) in particular, to better understand sugar transport and its utilization in developing caryopses of grain sorghum [Sorghum bicolor (L.) Moench]. Gene expression analyses for CWI using RNA gel blot and real-time quantitative PCR approaches on developing caryopses, including the glumes (maternal tissue appended to the seeds), showed expression of SbIncw (ZmIncw2 ortholog) primarily in the basal sugar unloading zone of endosperm. The expression of ZmIncw1 ortholog was significantly less abundant and restricted to the glumes. The protein and enzyme activity data corroborated the temporal transcript expression profile that showed maximal CWI protein (INCW) expression preceding the starch-filling phase of endosperm development, i.e. 6-12d-after-pollination (DAP). Protein gel blot analysis using polyclonal maize INCW1 antibodies showed a single polypeptide of 72kDa. The highest level of enzyme activity was unique to the basal part of the endosperm, in particular the basal endosperm transfer cell (BETC) layer and the maternal pedicel region that were highly enriched for the INCW protein, as seen by immunolocalization. High hexose-to-sucrose ratio in 6-12 DAP seeds, and negligible starch deposition in glumes corroborated the CWI activity data. Additionally, we report transcription profiles of several other genes related to sugar-to-starch metabolism in developing sorghum endosperm. As in maize, the INCW-mediated apoplastic cleavage of sucrose in the BETC and pedicel during the early developmental stages of caryopses is essential for the normal development of filial tissues. The unique cell-specificity of the INCW protein to both proximal and distal ends of placental sac shown here for the first time is likely to greatly increase uptakes of both hexose sugars and water through turgor sensing into developing seed. This trait is unique to sorghum among cereals and may facilitate its survival in drought environment.

摘要

我们报告了几种碳水化合物代谢基因的表达谱,特别是细胞壁转化酶(CWI),以便更好地了解糖转运及其在谷物高粱 [高粱(L.)Moench] 发育中的颖果中的利用情况。使用RNA凝胶印迹和实时定量PCR方法对发育中的颖果(包括颖片,即附着在种子上的母体组织)进行CWI基因表达分析,结果显示SbIncw(ZmIncw2直系同源物)主要在胚乳的基部糖卸载区表达。ZmIncw1直系同源物的表达量明显较低,且仅限于颖片。蛋白质和酶活性数据证实了瞬时转录表达谱,该谱显示CWI蛋白(INCW)在胚乳发育的淀粉填充阶段之前,即授粉后6 - 12天(DAP)表达量最高。使用多克隆玉米INCW1抗体进行的蛋白质凝胶印迹分析显示有一条72kDa的单一条带。最高水平的酶活性是胚乳基部所特有的,特别是基部胚乳转移细胞(BETC)层和母体花梗区域,通过免疫定位可见这些区域富含INCW蛋白。6 - 12 DAP种子中高己糖与蔗糖比率以及颖片中可忽略不计的淀粉沉积证实了CWI活性数据。此外,我们报告了发育中的高粱胚乳中其他几个与糖 - 淀粉代谢相关基因的转录谱。与玉米一样,在颖果发育早期阶段,BETC和花梗中INCW介导的蔗糖质外体裂解对于子代组织的正常发育至关重要。此处首次展示的INCW蛋白对胎盘囊近端和远端的独特细胞特异性,可能会通过膨压感知极大地增加己糖和水进入发育中种子的吸收。这种特性在谷物中是高粱所特有的,可能有助于其在干旱环境中生存。

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