Chinese Journal OF Rice Science >
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
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%.
GAO Lifen1, 2, XIA Zhihui2, 3, ZHANG Ji1, WANG Daowen2, ZHAI Wenxue2,* . Transgenosis of the Phosphomannomutase Transgene Increases Vitamin C Content in Rice[J]. Chinese Journal OF Rice Science, 2016 , 30(4) : 441 -446 . DOI: 10.16819/j.1001-7216.2016.5129
[1]Wheeler G L, Jones M A, Smirnoff N. The biosynthetic pathway of vitamin C in higher plants. Nature, 1998, 393(6683): 365369.
[2]Smirnoff N, Wheeler G L. Ascorbic acid in plants: Biosynthesis and function. Crit Rev Biochem Mol Biol, 2000, 35(4): 291314.
[3]Smirnoff N, Conklin P L, Loewus F A. Biosynthesis of ascorbic acid in plants:A renaissance. Annu Rev Plant Physiol Plant Mol Biol, 2001, 52: 437467.
[4]Wolucka B A,van Montagu M. GDPmannose 3′,5′epimerase forms GDPLgulose, a putative intermediate for the de novo biosynthesis of vitamin C in plants. J Biol Chem, 2003, 278(48): 4748347490.
[5]Lorence A, Chevone B I, Mendes P, et al. Myoinositol oxygenase offers a possible entry point into plant ascorbate biosynthesis. Plant Physiol, 2004, 134(3): 12001205.
[6]Agius F, GonzalezLamothe R, Caballero J L, et al. Engineering increased vitamin C levels in plants by overexpression of a Dgalacturonic acid reductase. Nat Biotechnol, 2003, 21(2): 177181.
[7]Keller R, Renz F S, Kossmann J. Antisense inhibition of the GDPmannose pyrophosphorylase reduces the ascorbate content in transgenic plants leading to developmental changes during senescence. Plant J, 1999, 19(2): 131141.
[8]Gatzek S, Wheeler G L, Smirnoff N. Antisense suppression of Lgalactose dehydrogenase in Arabidopsis thaliana provides evidence for its role in ascorbate synthesis and reveals light modulated Lgalactose synthesis. Plant J, 2002, 30(5): 541553.
[9]Tabata K, Oba K, Suzuki K, et al. Generation and properties of ascorbic aciddeficient transgenic tobacco cells expressing antisense RNA for Lgalactono1,4lactone dehydrogenase. Plant J, 2001， 27(2): 139148.
[10]Tokunaga T, Miyahara K, Tabata K, et al. Generation and properties of ascorbic acidoverproducing transgenic tobacco cells expressing sense RNA for Lgalactono1,4lactone dehydrogenase. Planta, 2005, 220(6): 854863.
[11]Jain A K, Nessler C L. Metabolic engineering of an alternative pathway for ascorbic acid biosynthesis in plants. Mol Breed, 2000, 6(1): 7378.
[12]李坤岚. 转小鼠古洛糖酸内酯氧化酶基因拟南芥的评估. 上海： 复旦大学, 2010.
Li K L. Evaluation of mouse gulonolactone oxidase gene transformed Arabidopsis thaliana. Shanghai: Fudan University, 2010.
[13]Chen Z, Young T E, Ling J, et al. Increasing vitamin C content of plants through enhanced ascorbate recycling. Proc Natl Acad Sci U S A, 2003, 100(6): 35253530.
[14]Qian W, Yu C, Qin H, et al. Molecular and functional analysis of phosphomannomutase (PMM) from higher plants and genetic evidence for the involvement of PMM in ascorbic acid biosynthesis in Arabidopsis and Nicotiana benthamiana. Plant J, 2007, 49(3): 399413.
[15]Badejo A A, Eltelib H A, Fukunaga K, et al. Increase in ascorbate content of transgenic tobacco plants overexpressing the acerola (Malpighia glabra) phosphomannomutase gene. Plant Cell Physiol, 2009, 50(2): 423428.
[16]Lu H J, Zhou X R, Gong Z, X, et al. Generation of selectable markerfree transgenic rice using double rightborder (DRB) binary vectors. Aust J Plant Physiol, 2001, 28(3): 241248.
[17]杨振玉, 张宗旭, 魏耀林, 等. 粳型特异亲和恢复系 C418 的选育及其特性. 杂交水稻, 1998, 13(3): 3132.
Yang Z Y, Zhang Z X, Wei Y L, et al. Breeding and characteristics of japonica type wide compatibility line C418. Hybrid Rice, 1998, 13(3): 3132.
[18]Zhai W X, Li X B, Tian W, et al. Introduction of a rice blight resistance gene,Xa21, into five Chinese rice varieties through an Agrobacteriummediated system. China C:Life Sci, 2000, 43(4): 361368.
[19]Livak K J, Schmittgen T D. Analysis of relative gene expression data using rea Ltime quantitative PCR and the 2ΔΔCT method. Methods, 2001, 25(4): 402408.
[20]Gillespie K M, Ainsworth E A. Measurement of reduced, oxidized and total ascorbate content in plants. Nat Protocols, 2007, 2(4): 871874.
[21]Noctor G, Foyer C H. Ascorbate and glutathione: Keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol, 1998, 49: 249279.
[22]Badejo A A, Eltelib H A, Fukunaga K, et al. Increase in ascorbate content of transgenic tobacco plants overexpressing the acerola (Malpighia glabra) phosphomannomutase gene. Plant&Cell Physiol, 2009, 50(2): 423428.
[23]Gao L, Xia Z, Jiang G, et al. Generation of markerfree, bacterial blightresistant transgenic sterile line and hybrid rice with Xa21. Plant Breed, 2011, 130(4): 438443.
[24]Gao L, Cao Y, Xia Z, et al. Do transgenesis and markerassisted backcross breeding produce substantially equivalent plants? A comparative study of transgenic and backcross rice carrying bacterial blight resistant gene Xa21. BMC Genom, 2013, 14: 738750.
[25]Baudo M M, Lyons R, Powers S, et al. Transgenesis has less impact on the transcriptome of wheat grain than conventional breeding. Plant Biotechnol J, 2006, 4(4): 369380.
[26]Coll A, Nadal A, Collado R, et al. Gene expression profiles of MON810 and comparable nonGM maize varieties cultured in the field are more similar than are those of conventional lines. Transgenic Res, 2009, 18(5): 801808.
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