欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2013, Vol. 39 ›› Issue (09): 1530-1537.doi: 10.3724/SP.J.1006.2013.01530

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

opaque-2玉米近等基因系的构建与赖氨酸含量快速检测

陈岩1宋丽雅2,周志强1,张德贵1,颜娜1,张世煌1,李新海1,郝转芳1,翁建峰1,白丽1,李明顺1,*   

  1. 1中国农业科学院作物科学研究所 / 农业部遗传育种重点实验室，北京100081；2北京工商大学 / 北京市植物资源研究开发重点实验室，北京100048
  • 收稿日期:2012-12-18 修回日期:2013-04-22 出版日期:2013-09-12 网络出版日期:2013-07-09
  • 通讯作者: 李明顺, E-mail: mshunli@aliyun.com
  • 作者简介:李明顺, E-mail: mshunli@aliyun.com
  • 基金资助:

    本研究由国家现代农业产业技术体系专项(CARS-02-01)，中央级公益性科研院所基本科研业务费专项和北京市自然科学基金项目(6112003)资助。

Construction of Maize opaque-2 Near-isogenic Lines and Rapid Detection of Lysine Content

CHEN Yan1,SONG Li-Ya2,ZHOU Zhi-Qiang1,ZHANG De-Gui1,YAN Na1,ZHANG Shi-Huang1,LI Xin-Hai1,HAO Zhuan-Fang1,WENG Jian-Feng1,BAI Li1,LI Ming-Shun1,*   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Beijing 100081, China; 2 Beijing Key Laboratory of Plant Resources Research and Development / Beijing Technology and Business University, Beijing 100048, China
  • Received:2012-12-18 Revised:2013-04-22 Published:2013-09-12 Published online:2013-07-09
  • Contact: 李明顺, E-mail: mshunli@aliyun.com
  • About author:李明顺, E-mail: mshunli@aliyun.com

RichHTML

PDF

1340

被引次数

3

摘要：

我国高赖氨酸玉米种质资源狭窄，opaque-2(o2)突变基因能大幅提高玉米赖氨酸含量，通过分子标记辅助选择构建o2玉米近等基因系并检测其赖氨酸含量具有重要意义。其中，要解决的两个关键问题是如何准确地将不同供体的o2突变基因导入受体系和如何快速地检测导入系的赖氨酸含量。本研究利用O2基因内紧密连锁的SSR共显性标记引物phi057检测玉米供体和受体自交系的多态性，利用其特异性和共显性构建o2近等基因系；参考已有研究，改进染料结合(DBL)法，测定18组构建成功的o2近等基因系的赖氨酸含量。结果表明，不仅在不同供体(CA339和山东2548)之间存在多态性，而且在不同受体系间也存在多态性，利用phi057能够成功地将不同供体的o2突变基因导入受体系，构建o2近等基因系；改进的DBL法分析表明，不同受体系赖氨酸含量变化较大，不同背景的受体系导入o2突变基因后赖氨酸含量增加的幅度差异也较大；普通玉米自交系间赖氨酸含量为0.223%~0.368%，构建成功的不同o2近等基因系间，赖氨酸含量为0.373%~0.527%，与受体亲本相比，赖氨酸增加幅度最低为13%，最高为74%。分析表明，phi057能准确筛选导入o2突变基因的受体系，结合改进的DBL法能快速地选择赖氨酸含量高的玉米。

关键词: 高赖氨酸玉米, 分子标记辅助选择(MAS), o2近等基因系, 染料结合法(DBL), 赖氨酸

Abstract:

The germplasm of high lysine maize in China is narrow, while the opaque-2 (o2) mutant gene can largely increase the lysine level in maize, so it is important to create opaque-2 near-isogenic lines (o2-NILs) by molecular marker assisted selection (MAS). There are two key problems in creating such lines, one is how to introgress the o2 gene into normal receptor lines correctly, and the other is the fast and convenient way to monitor the lysine content. In our study, the closely linked co-dominant SSR marker phi057 was used to detect the polymorphism between the maize donor and receptor lines, and the specific and co-dominant marker was used to construct o2-NILs. Meanwhile, we tested the lysine content of 18s group of o2-NILs by our own improved dye binding lysine (DBL) method. The results demonstrated that the normal maize inbred lines existed polymorphism, moreover, polymorphism also existed among QPM donors CA339 and Shandong2548, the marker phi057 was effective to construct o2-NILs, using different QPM donors. The lysine content varied in different genetic backgrounds, which was from 0.223% to 0.368% among normal maize inbred lines, and from 0.373% to 0.527% among o2-NILs. The increment of lysine content in o2-NILs ranged from 13% to 74%. We included that the SSR marker phi057 can select the mutant genotype of o2 gene correctly, and the improved DBL method is effective to be used in high lysine maize breeding.

Key words: High lysine maize, Molecular marker assisted selection (MAS), o2-NILs, Dye binding lysine (DBL), Lysine

[1]Vasal S. The quality protein maize story. Food Nutr Bull, 2000, 21: 445–450



[2]Kawakatsu T, Takaiwa F. Differences in transcriptional regulatory mechanisms functioning for free lysine content and seed storage protein accumulation in rice grain. Plant Cell Physiol, 2010, 51: 1964–1974



[3]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: 279–280



[4]Wenefrida I, Utomo H S, Blanche S B, Linscombe S D. Enhancing essential amino acids and health benefit components in grain crops for improved nutritional values. Recent Pat DNA Gene Seq, 2009, 3: 219–225



[5]Li M-S(李明顺), Zhang S-H(张世煌), Li X-H(李新海), Tian Q-Z(田清震), Bai L(白丽). Strategies of quality protein maize (QPM) development. J Maize Sci (玉米科学), 2004, 12(3): 36–39 (in Chinese with English abstract)



[6]Schmidt R J, Burr F A, Burr B. Transposon tagging and molecular analysis of the maize regulatory locus opaque-2. Science, 1987, 238: 960



[7]Maddaloni M, Di Fonzo N, 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



[8]Chin E, Shu H, Smith J, Senior M. Maize simple repetitive DNA sequences: abundance and allele variation. Genome, 1996, 39: 866–873



[9]Senior L. Polymorphism Rate and Utility of SSRs Using Agarose Gel. 39th Maize Genetics Conference Abstracts, 1997



[10]Tian Q-Z(田清震), Li X-H(李新海), Li M-S(李明顺), Jiang W(姜伟), Zhang S-H(张世煌). Molecular marker assisted selection to quality protein maize. J Maize Sci (玉米科学), 2004, 12(2): 108–110 (in Chinese with English abstract)



[11]Song M(宋敏), Tian Q-Z(田清震), Li X-H(李新海), Li M-S(李明顺), Zhang S-H(张世煌). Molecular markers assisted selection in quality protein maize breeding. Sci Agric Sin (中国农业科学), 2005, 38(9): 1748–1754 (in Chinese with English abstract)



[12]Lei K-R(雷开荣), Shi C-Y(石春焱), Li M-S(李明顺), Zhang S-H(张世煌), Li X-H(李新海), Xiao M-J(肖木楫), Zhang D-G(张德贵), Hao Z-F(郝转芳). SSR marker assisted selection for opaque-2 gene in early generations of NILs development in QPM. J Maize Sci (玉米科学), 2006, 14(6): 28–31 (in Chinese with English abstract)



[13]Zhao G(赵刚), Wu Z-K(吴子恺), Chen L(陈亮), Zhang D-G(张德贵), Zhang S-H(张世煌), Lu Z-Y(卢振宇), Bai L(白丽), Li M-S(李明顺). Comparison of several donor inbred lines on development of quality protein maize (QPM) near-isogenic lines. J Maize Sci (玉米科学), 2010, 18(6): 11–14 (in Chinese with English abstract)



[14]Bai P-F(白鹏飞), Zhang S-H(张世煌), Zhang D-G(张德贵), Li X-H(李新海), Zhao G(赵刚), Chen Y(陈岩), Li M-S(李明顺), Dong J-G(董金皋). Built QPM NILs using SSR molecular marker-assisted selection. J Maize Sci (玉米科学), 2011, 19(5): 32–38 (in Chinese with English abstract)



[15]Zhao G, Li M, Zhang D, Li X, Wu Z, Ci X, Xie C, Bai L, Lu Z, Chen L. Kernel lysine content does not increase in some maize opaque2 mutants. Planta, 2012, 235: 205–215



[16]Ciceri P, Castelli S, Lauria M, Lazzari B, Genga A, Bernard L, Sturaro M, Viotti A. Specific combinations of zein genes and genetic backgrounds influence the transcription of the heavy-chain zein genes in maize opaque-2 endosperms. Plant Physiol, 2000, 124: 451–460



[17]Moro G L, Habben J E, Hamaker B R, Larkins B A . Characterization of the variability in lysine content for normal and opaque2 maize endosperm. Crop Sci, 1996, 36: 1651–1659



[18]Jia H, Nettleton D, Peterson J M, Vazquez-Carrillo G, Jannink J L, Scott M P. Comparison of transcript profiles in wild-type and o2 maize endosperm in different genetic backgrounds. Crop Sci, 2007, 47: 45–59



[19]Lu Z-Y(卢振宇), Li M-S(李明顺), Xie C-X(谢传晓), Li X-H(李新海), Qu Y-Y(曲延英), Zhang S-H(张世煌). An improved study on rapid DNA extraction method from young leaves of maize. J Maize Sci (玉米科学), 2008, 16(2): 50–53 (in Chinese with English abstract)



[20]Zhao T-N(赵铁男), Chang Y-L(常瀛莲), Wang W-J(王维军). Seed Quality Analysis (种子品质分析). Harbin: Heilongjiang Science & Technology Press, 1982. pp 77–92(in Chinese)



[21]Zheng Y-L(郑云兰), Zhu C(朱才). The method for determining lysine content in soybean by using 721 type photometer. Biotechnology (生物技术), 1993, (3): 38–41 (in Chinese with English abstract)



[22]Yang W-P(杨文鹏), Zheng W-T(郑文婷), Wu J(吴菁), Feng R(冯睿), Zhang F-D(张方东), Zheng Y-L(郑用琏). Screening of the chemical methods applied to lysine content determination of the family seeds from segregating population in maize. Guizhou Agric Sci (贵州农业科学), 2007, 35(6): 19–23 (in Chinese with English abstract)



[23]Zhang W-L(张文龙), Yang W-P(杨文鹏), Chen Z-W(陈志伟), Wang M-C(王明春), Yang L-Q(杨留启), Cai Y-L(蔡一林). Molecular marker-assisted selection for corn o2 introgression lines with o16 gene. Acta Agron Sin (作物学报), 2010, 36(8): 1302–1309 (in Chinese with English abstract)



[24]Hopkins C G. Improvement in the chemical composition of the corn kernel. Ill Agric Exp Stn Bull, 1899, 55: 205–240



[25]Crow J F, Kermicle J. Oliver Nelson and quality protein maize. Genetics, 2002, 160: 819–821



[26]Wu Y R, Holding D R, Joachin M. γ-Zeins are essential for endosperm modification in quality protein maize. Proc Natl Acad Sci USA, 2010, 107: 12810–12815



[27]Rosales A, Galicia L, Oviedo E, Islas C, Palacios-Rojas N. Near-infrared reflectance spectroscopy (NIRS) for protein, tryptophan, and lysine evaluation in quality protein maize (QPM) breeding programs. J Agric Food Chem, 2011, 59: 10781–10786



[28]Xiao L(肖玲), Gong Y-S(龚月生). The determination of effective amino acid in soybean meal with dyestuff combination method. Feed Ind (饲料工业), 2001, 22(4): 33–34 (in Chinese with English abstract)
[1] 张晓星,朱慧,张东民,宋丽雅,张德贵,翁建峰,郝转芳,李明顺. 糯玉米opaque2基因近等基因系的创制[J]. 作物学报, 2017, 43(12): 1760-1766.
[2] 汤继华, 季洪强, 刘义宝, 张君, 谭晓军, 胡彦民, 刘宗华. 玉米籽粒赖氨酸含量的遗传及其与产量的关系分析[J]. 作物学报, 2011, 37(09): 1585-1591.
[3] 陈国林,吴建国,张莉,石春海. 花后不同时期油菜籽饼粕赖氨酸含量的遗传分析[J]. 作物学报, 2010, 36(4): 680-687.
[4] 张文龙,杨文鹏,陈志伟,王明春,杨留启,蔡一林. 玉米o16基因回交渗入o2系的分子标记辅助选择[J]. 作物学报, 2010, 36(08): 1302-1309.
[5] 胡洁云;杨文鹏;张祖新;郑用琏. 玉米Opaque-2基因内3个SSR位点的显性等位变异及其对赖氨酸含量的影响[J]. 作物学报, 2006, 32(09): 1265-1271.
[6] 唐俐;刘巧泉;邓晓湘;武小金;辛世文. 无抗性选择标记的转高赖氨酸蛋白(LRP)基因籼稻恢复系的获得[J]. 作物学报, 2006, 32(08): 1248-1251.
[7] 王为民;赵倩;于静娟;朱登云;敖光明. 水稻转高赖氨酸蛋白质基因(sb401)植株的获得及种子中蛋白质和氨基酸的含量分析[J]. 作物学报, 2005, 31(05): 603-607.
[8] 汪黎明;石德权;刘仁东;白丽. 高赖氨酸玉米主要品质性状的配合力分析[J]. 作物学报, 1994, 20(04): 446-452.
[9] 刘仁东. 玉米籽粒蛋白质、赖氨酸和油分含量的遗传成分的比较研究[J]. 作物学报, 1994, 20(01): 93-98.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2