转磷酸甘露糖变位酶基因提高水稻维生素C含量
收稿日期: 2015-08-21
修回日期: 2016-10-20
网络出版日期: 2016-07-10
基金资助
国家转基因生物新品种培育重大专项（2014ZX08001002）； 国家自然科学基金资助项目（31300999）； 湖北省教育厅项目（B2015229）； 武汉市科学技术局项目（2014072704011250）； 江汉大学科研启动项目（300306000043）。
Transgenosis of the Phosphomannomutase Transgene Increases Vitamin C Content in Rice
Received date: 2015-08-21
Revised date: 2016-10-20
Online published: 2016-07-10
维生素C (VC) 是人体健康所必需的营养元素。人类由于缺乏VC合成途径中的最后一种酶（L古洛糖酸内酯氧化酶），自身不能合成VC。水稻是重要的粮食作物，增加水稻种子中VC含量，能够提高其营养价值。磷酸甘露糖变位酶(PMM)是VC合成通路中一种重要的酶，催化甘露糖6磷酸到甘露糖1磷酸的转变。将水稻PMM基因(OsPMM)构建在双右边界双元载体pMNDRBBin6上，并用种子特异表达的启动子BX14驱动其表达。通过农杆菌介导的转化系统，OsPMM基因被转入粳型三系恢复系C418中。通过分子检测，在T2代筛选到了无选择标记的转基因植株。对OsPMM基因在转基因植株中的表达进行分析，发现OsPMM基因在转基因水稻种子内的表达水平明显提高，相应地，转基因系种子中的VC含量也提高了25%~50%。
关键词： 维生素C; 磷酸甘露糖变位酶; 双右边界双元载体系统; 无选择标记; 转基因系
高利芬1,2,夏志辉2,3,张继1,王道文2,翟文学2,* . 转磷酸甘露糖变位酶基因提高水稻维生素C含量[J]. 中国水稻科学, 2016 , 30(4) : 441 -446 . DOI: 10.16819/j.1001-7216.2016.5129
VC (vitamin C) is an essential nutrient to human health. Due to lack of Lgulonolactone oxidase, the last enzyme involved in VC synthesis pathway, human could not synthesize VC by themselves. Rice is an important food crop and its nutritional value could be greatly improved by increasing the VC content in rice seeds. Phosphomannomutase (PMM) is an important enzyme in VC synthesis pathway, catalyzing the interconversion from mannose 6phosphate from mannose 1phosphate. In this study, Oryza PMM gene (OsPMM) that was under the control of seedspecific expressed promoter Bx14 was transferred into ‘C418’, a restorer line of threeline japonica hybrid rice, using the double rightborder vector pMNDRBBin6 through an Agrobacterium tumefaciensmediated system. Molecular analysis revealed that OsPMM was integrated into the genome of transgenic ‘C418’, and the homozygous and markerfree transgenic line was obtained in the T2 generation. Gene expression analysis of transgenic lines showed the expression level of OsPMM was significantly increased in seeds, and accordingly, the VC content in the seeds of transgenic plant also increased by 25-50%.
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