糯玉米opaque2基因近等基因系的创制
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摘要
opaque2基因能够提高糯玉米赖氨酸、色氨酸等必需氨基酸的含量。用2种优质蛋白玉米(QPM)CA339和鲁2548自交系作为opaque2基因供体,25个品质较好的糯玉米自交系作为受体,利用回交技术和SSR分子标记辅助选择育种,创制糯玉米opaque2近等基因系。分子标记结果表明,不同受体以及两种供体间都存在多态性。5套创制成功的opaque2近等基因系的赖氨酸含量比其轮回亲本分别提高了59.0%、52.7%、48.5%、46.3%和61.9%,分别由0.308%、0.313%、0.309%、0.341%、0.323%提高到0.489%、0.478%、0.458%、0.498%、0.522%。本研究表明,可以利用此方法通过向不同遗传背景的多种受体导入opaque2,选取赖氨酸含量提高较大、透明表型的近等基因系,提高糯玉米的营养价值和经济价值。
In waxy maize,the opaque2 gene can increase the content of essential amino acids,such as lysine and tryptophan.Two high quality protein maize(QPM) inbred lines,CA339 and Lu 2548,were used as opaque2 gene donors.Twenty-five waxy maize with advantageous traits were used as receptors.Application of backcrossing and SSR marker-assisted selection in breeding can create waxy maize opaque2 near-isogenic lines.The results of molecular marker indicated that there were polymorphisms in different receptors and between the two donors.Lysine content of five sets of successfully created opaque2 near-isogenic lines was tested,showing that the content of lysine in o2 w-NILs was 59.0%,52.7%,48.5%,46.3%,and 61.9% higher than that in their recurrent parents,which was increased from 0.308%,0.313%,0.309%,0.341%,0.323% to 0.489%,0.478%,0.458%,0.498%,and 0.522%,respectively.Using this method,opaque2 could be introgressed into multiple receptors with different genetic backgrounds to select near-isogenic lines with high lysine content and transparent phenotype,so as to improve nutritional and economic values of waxy maize.
In waxy maize,the opaque2 gene can increase the content of essential amino acids,such as lysine and tryptophan.Two high quality protein maize(QPM) inbred lines,CA339 and Lu 2548,were used as opaque2 gene donors.Twenty-five waxy maize with advantageous traits were used as receptors.Application of backcrossing and SSR marker-assisted selection in breeding can create waxy maize opaque2 near-isogenic lines.The results of molecular marker indicated that there were polymorphisms in different receptors and between the two donors.Lysine content of five sets of successfully created opaque2 near-isogenic lines was tested,showing that the content of lysine in o2 w-NILs was 59.0%,52.7%,48.5%,46.3%,and 61.9% higher than that in their recurrent parents,which was increased from 0.308%,0.313%,0.309%,0.341%,0.323% to 0.489%,0.478%,0.458%,0.498%,and 0.522%,respectively.Using this method,opaque2 could be introgressed into multiple receptors with different genetic backgrounds to select near-isogenic lines with high lysine content and transparent phenotype,so as to improve nutritional and economic values of waxy maize.
引文
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[10]Schmidt R J,Ketudat M,Aukerman M J,Hoschek G.Opaque-2 is a transcriptional activator that recognizes a specific target site in22-k D zein genes.Plant Cell,1992,4:689–700
[11]Schmitz D,Lohmer S,Salamini F,Thompson R D.The activation domain of the maize transcription factor Opaque-2 resides in a single acidic region.Nucl Acids Res,1997,25:756–763
[12]Hunter B G,Beatty M K,Singletary G W,Hamaker B R,Dilkes B P,Larkins B A,Junq R.Maize opaque endosperm mutations create extensive changes in patterns of gene expression.Plant Cell,2002,14:2591–2612
[13]Viotti A,Sala E,Marotta R,Alberi P,Balducci C,Soave C.Genes and m RNAs coding for zein polypeptides in Zea mays.Eur J Biochem,1979,102:211–222
[14]Crow J F,Kermicle J.Oliver Nelson and quality protein maize.Genetics,2002,160:819–821
[15]Vasal S K.High quality protein corn.In:Hallawer A R ed.Specialty Corns.Boca Raton:CRC Press,2001.pp 85–121
[16]宋敏,张世煌.优质蛋白玉米(QPM)育种研究进展.新疆农业科学,2007,44(增刊3):30–35Song M,Zhang S H.Comprehensive report on study advance of breeding of high-quality protein maize(QPM).Xinjiang Agric Sci,2007,44(suppl-3):30–35(in Chinese)
[17]Zhang W,Yang W,Wang M,Wang W,Zeng G,Chen Z,Cai Y.Increasing lysine content of waxy maize through introgression of Opaque-2 and Opaque-16 genes using molecular assisted and biochemical development.PLo S One,2013,8:e56227
[18]昂沃,郝小琴,吴子恺.Opaque2基因导入各类隐性纯合体玉米早代籽粒赖氨酸含量的效应研究.玉米科学,2012,20(2):56–58Ang W,Hao X Q,Wu Z K.Analysis of Opaque-2 gene effects on kernel lysine content into some early generations’homozygote recessive maize.J Maize Sci,2012,20(2):56–58(in Chinese with English abstract)
[19]卢振宇,李明顺,谢传晓,李新海,曲延英,张世煌.玉米叶片DNA快速提取方法改进研究.玉米科学,2008,16(2):50–53Lu Z Y,Li M S,Xie C X,Li X H,Qu Y Y,Zhang S H.An improved study on rapid DNA extraction method from young leaves of maize.J Maize Sci,2008,16(2):50–53(in Chinese with English abstract)
[20]Vasal S K.The quality protein maize story.Food Nutr Bull,2000,21:445–450
[21]Chin E C,Senior M L,Shu H,Smith J S.Maize simple repetitive DNA sequences:abundance and allele variation.Genome,1996,39:866–873
[22]Chen Y,Zhou Z,Zhao G,Li X,Song L,Yan N,Weng J,Hao Z,Zhang D,Li M,Zhang S.Transposable element rbg induces the differential expression of opaque-2 mutant gene in two maize o2 NILs derived from the same inbred line.PLo S One,2014,9:e85159
[23]赵刚,吴子恺,陈亮,张德贵,张世煌,卢振宇,白丽,李明顺.几个供体对优质蛋白玉米(QPM)近等基因系构建效果的比较.玉米科学,2010,18(6):11–14Zhao G,Wu Z K,Chen L,Zhang D G,Zhang S H,Lu Z Y,Bai L,Li M S.Comparison of several donor inbred lines on development of quality protein maize(QPM)near-isogenic lines.J Maize Sci,2010,18(6):11–14(in Chinese with English abstract)
[2]Wessler S R,Tarpley A,Purugganan M,Spell M,Okagaki R.Filler DNA is associated with spontaneous deletions in maize.Proc Natl Acad Sci USA,1990,87:8731–8735
[3]彭泽斌,田志国.我国糯玉米产业现状与发展战略.玉米科学,2004,12(3):116–118Peng Z B,Tian Z G.Industrialization present status and strategies for development of waxy corn in China.J Maize Sci,2004,12(3):116–118(in Chinese with English abstract)
[4]杨引福,郭强,陈婧,郑小亚,蔺崇明.中国温带糯玉米自交系遗传品质性状分析.西北植物学报,2009,29:2213–2220Yang Y F,Guo Q,Chen J,Zheng X Y,Lin C M.Analysis of genetic and quality traits of waxy corn inbred lines in China temperate zone.Acta Bot Boreali-Occident Sin,2009,29:2213–2220(in Chinese with English abstract)
[5]田清震,李新海,李明顺,姜伟,张世煌.优质蛋白玉米的分子标记辅助选择.玉米科学,2004,12(2):108–110Tian Q Z,Li X H,Li M S,Jiang W,Zhang S H.Molecular markers assisted selection to quality protein maize.J Maize Sci,2004,12(2):108–110(in Chinese with English abstract)
[6]Mertz E T,Bates L S,Nelson O E.Mutant gene that changes protein composition and increases lysine content of maize endosperm.Science,1964,145:270–280
[7]Hartings H,Maddaloni M,Lazzaroni N,Fonzo N D,Motto M,Salamini F,Thompson R.The O2 gene which regulates zein deposition in maize endosperm encodes a protein with structural homologies to transcriptional activators.EMBO J,1989,8:2795–2801
[8]Maddaloni M,Fonzo N D,Hartings H,Lazzaroni N,Salamini F,Thompson R,Motto M.The sequence of the zein regulatory gene opaque-2(O2)of Zea mays.Nucl Acids Res,1989,17:7532
[9]Schmidt R J,Burr F A,Aukerman M J,Burr B.Maize regulatory gene opaque-2 encodes a protein with a“leucine-zipper”motif that binds to zein DNA.Proc Natl Acad Sci USA,1990,87:46–50
[10]Schmidt R J,Ketudat M,Aukerman M J,Hoschek G.Opaque-2 is a transcriptional activator that recognizes a specific target site in22-k D zein genes.Plant Cell,1992,4:689–700
[11]Schmitz D,Lohmer S,Salamini F,Thompson R D.The activation domain of the maize transcription factor Opaque-2 resides in a single acidic region.Nucl Acids Res,1997,25:756–763
[12]Hunter B G,Beatty M K,Singletary G W,Hamaker B R,Dilkes B P,Larkins B A,Junq R.Maize opaque endosperm mutations create extensive changes in patterns of gene expression.Plant Cell,2002,14:2591–2612
[13]Viotti A,Sala E,Marotta R,Alberi P,Balducci C,Soave C.Genes and m RNAs coding for zein polypeptides in Zea mays.Eur J Biochem,1979,102:211–222
[14]Crow J F,Kermicle J.Oliver Nelson and quality protein maize.Genetics,2002,160:819–821
[15]Vasal S K.High quality protein corn.In:Hallawer A R ed.Specialty Corns.Boca Raton:CRC Press,2001.pp 85–121
[16]宋敏,张世煌.优质蛋白玉米(QPM)育种研究进展.新疆农业科学,2007,44(增刊3):30–35Song M,Zhang S H.Comprehensive report on study advance of breeding of high-quality protein maize(QPM).Xinjiang Agric Sci,2007,44(suppl-3):30–35(in Chinese)
[17]Zhang W,Yang W,Wang M,Wang W,Zeng G,Chen Z,Cai Y.Increasing lysine content of waxy maize through introgression of Opaque-2 and Opaque-16 genes using molecular assisted and biochemical development.PLo S One,2013,8:e56227
[18]昂沃,郝小琴,吴子恺.Opaque2基因导入各类隐性纯合体玉米早代籽粒赖氨酸含量的效应研究.玉米科学,2012,20(2):56–58Ang W,Hao X Q,Wu Z K.Analysis of Opaque-2 gene effects on kernel lysine content into some early generations’homozygote recessive maize.J Maize Sci,2012,20(2):56–58(in Chinese with English abstract)
[19]卢振宇,李明顺,谢传晓,李新海,曲延英,张世煌.玉米叶片DNA快速提取方法改进研究.玉米科学,2008,16(2):50–53Lu Z Y,Li M S,Xie C X,Li X H,Qu Y Y,Zhang S H.An improved study on rapid DNA extraction method from young leaves of maize.J Maize Sci,2008,16(2):50–53(in Chinese with English abstract)
[20]Vasal S K.The quality protein maize story.Food Nutr Bull,2000,21:445–450
[21]Chin E C,Senior M L,Shu H,Smith J S.Maize simple repetitive DNA sequences:abundance and allele variation.Genome,1996,39:866–873
[22]Chen Y,Zhou Z,Zhao G,Li X,Song L,Yan N,Weng J,Hao Z,Zhang D,Li M,Zhang S.Transposable element rbg induces the differential expression of opaque-2 mutant gene in two maize o2 NILs derived from the same inbred line.PLo S One,2014,9:e85159
[23]赵刚,吴子恺,陈亮,张德贵,张世煌,卢振宇,白丽,李明顺.几个供体对优质蛋白玉米(QPM)近等基因系构建效果的比较.玉米科学,2010,18(6):11–14Zhao G,Wu Z K,Chen L,Zhang D G,Zhang S H,Lu Z Y,Bai L,Li M S.Comparison of several donor inbred lines on development of quality protein maize(QPM)near-isogenic lines.J Maize Sci,2010,18(6):11–14(in Chinese with English abstract)