Instituto de Agrobiotecnología, Universidad Pública de Navarra/Consejo Superior de Investigaciones Científicas/Gobierno de Navarra, Mutiloako etorbidea zenbaki gabe, 31192 Mutiloabeti, Nafarroa, Spain.
Plant Cell Physiol. 2013 Feb;54(2):282-94. doi: 10.1093/pcp/pcs180. Epub 2013 Jan 3.
Sucrose synthase (SuSy) is a highly regulated cytosolic enzyme that catalyzes the conversion of sucrose and a nucleoside diphosphate into the corresponding nucleoside diphosphate glucose and fructose. In cereal endosperms, it is widely assumed that the stepwise reactions of SuSy, UDPglucose pyrophosphorylase and ADPglucose (ADPG) pyrophosphorylase (AGP) take place in the cytosol to convert sucrose into ADPG necessary for starch biosynthesis, although it has also been suggested that SuSy may participate in the direct conversion of sucrose into ADPG. In this study, the levels of the major primary carbon metabolites, and the activities of starch metabolism-related enzymes were assessed in endosperms of transgenic maize plants ectopically expressing StSUS4, which encodes a potato SuSy isoform. A total of 29 fertile lines transformed with StSUS4 were obtained, five of them containing a single copy of the transgene that was still functional after five generations. The number of seeds per ear of the five transgenic lines containing a single StSUS4 copy was comparable with that of wild-type (WT) control seeds. However, transgenic seeds accumulated 10-15% more starch at the mature stage, and contained a higher amylose/amylopectin balance than WT seeds. Endosperms of developing StSUS4-expressing seeds exhibited a significant increase in SuSy activity, and in starch and ADPG contents when compared with WT endosperms. No significant changes could be detected in the transgenic seeds in the content of soluble sugars, and in activities of starch metabolism-related enzymes when compared with WT seeds. A suggested metabolic model is presented wherein both AGP and SuSy are involved in the production of ADPG linked to starch biosynthesis in maize endosperm cells.
蔗糖合酶(SuSy)是一种高度调控的细胞质酶,它催化蔗糖和核苷二磷酸转化为相应的核苷二磷酸葡萄糖和果糖。在谷类胚乳中,人们普遍认为 SuSy、UDP 葡萄糖焦磷酸化酶和 ADP 葡萄糖(ADPG)焦磷酸化酶(AGP)的逐步反应发生在细胞质中,将蔗糖转化为淀粉生物合成所需的 ADPG,尽管也有人提出 SuSy 可能参与蔗糖向 ADPG 的直接转化。在这项研究中,评估了转 StSUS4 基因的玉米胚乳中主要初级碳代谢物的水平以及淀粉代谢相关酶的活性,StSUS4 编码一种马铃薯 SuSy 同工型。共获得了 29 个可育的 StSUS4 转化株系,其中 5 个含有一个单拷贝的转基因,该拷贝在经过五代后仍具有功能。含有单个 StSUS4 拷贝的 5 个转基因株系的每穗种子数与野生型(WT)对照种子相当。然而,转基因种子在成熟阶段积累的淀粉多 10-15%,并且直链淀粉/支链淀粉的平衡比 WT 种子高。与 WT 胚乳相比,表达 StSUS4 的种子发育中的胚乳的 SuSy 活性、淀粉和 ADPG 含量显著增加。与 WT 种子相比,转基因种子中可溶性糖的含量以及淀粉代谢相关酶的活性没有明显变化。提出了一个代谢模型,其中 AGP 和 SuSy 都参与了与玉米胚乳细胞中淀粉生物合成相关的 ADPG 的产生。