强筋小麦分子标记多重PCR体系的构建与应用

doi:10.3724/SP.J.1006.2011.01942

Abstract

Abstract: Wheat strong-gluten quality is closely correlated with combinations of high-molecular-weight glutenin subunits (HMW-GS) and low-molecular-weight glutenin subunits (LMW-GS). The multiplex PCR system is a rapid and efficient approach to evaluate wheat germplasm and its quality in wheat. In this study, we developed a multiplex PCR system confering molecular markers on Ax1/Ax2^*, Dx5, Glu-A3d, Glu-B3i genes, and Glu-B3 locus and validated it with 12 cultivars with known subunit at each locus. This multiplex PCR system was proved to be effective and stable to amplify target bands for these genes (locus), and used to evaluate the glutenin subunit genes (locus) accociated with strong-gluten in 62 major cultivars in wheat prodution in Shaanxi Province, China. The results showed that the frequencies of genes Ax1/Ax2^*, Dx5, Glu-A3d, Glu-B3i, and locus Glu-B3 in the 62 cultivars were 56.5%, 9.6%, 33.9%, 1.6%, and 64.4%, respectively, whereas gene Bx7^OE was not found. Most of the cultivars carried two-gene (locus) combinations with the frequency of 48.3%, a few cultivars carried a single gene or locus (33.9%). The frequency of cultivars carrying three or four-gene (locus) combinations was 11.3%. The remaining cultivars (6.5%) were free of above elite gene (locus). Therefore, the frequency of combination of multiple strong-gluten subunits gene (locus) was low in cultivars from Shaanxi Province, which could be promoted through germplasm introduction and traditional breeding aided by molecular marker selection. The multiplex PCR system developed in this study may serve as a rapid and efficient method to select materials pyrimiding multiple genes (loci) associated with strong-gluten in wheat breeding for quality.

Key words: Common wheat cultivars from Shaanxi Province, Molecular marker, Multiplex PCR system

LIANG Qiang, ZHANG Xiao-Ke, WEI Qian, WANG Xiao-Long, ZHANG Jing, SUN Dao-Jie, FU Xiao-Ji. Establishment and Application of Multiplex PCR System Based on Molecular Markers of Glutenin Subunit Genes (Loci) Related to Strong-gluten in Wheat[J].Acta Agron Sin, 2011, 37(11): 1942-1948.

References

[1]Kasarda D D. Glutenin polymers: the in vitro to in vivo transition. Cereal Foods World, 1999, 44: 566–571
[2]Gupta, R B, Shepherd K W. Two-step one-dimensional SDS-PAGE analysis of LMW subunits of glutenin: 1. Variation and genetic control of the subunits in hexaploid wheats. Theor Appl Genet, 1990, 80: 65–74
[3]Radovanovic N, Cloutier S. Gene-assisted selection for high molecular weight glutenin subunits in wheat doubled haploid breeding programs. Mol Breed, 2003, 12: 51–59
[4]Gupta R B, Bekes F, Wrigley C W. Prediction of physical dough properties from glutenin subunit composition in bread wheats. Cereal Chem, 1991, 68: 328–333
[5]Liu L, He Z H, Yan J, Zhang Y, Xia X C, Peña R J. Allelic variation at the Glu-1 and Glu-3 loci, presence of the 1B•1R translocation, and their effects on mixographic properties in Chinese bread wheats. Euphytica, 2005, 142: 197–204
[6]D’Ovidio R, Anderson O D. PCR analysis to distinguish between alleles of a member of a multigene family correlated with wheat bread-making quality. Theor Appl Genet, 1994, 88: 759–763
[7]Lafiandra D, Tucci G F, Pavoni A, Turchetta T, Margiotta B. PCR analysis of x- and y-type genes present at the complex Glu-A1 locus in durum and bread wheat. Theor Appl Genet, 1997, 94: 235–240
[8]van Campenhout S, Stappen J, Sagi L, Volckaert G. Locus-specific primers for LMW glutenin genes on each of the group 1 chromosomes of hexaploid wheat. Theor Appl Genet, 1995, 91: 313–319
[9]Ragupathy R, Naeem H A, Reimer E, Lukow O M, Sapirstein H D, Cloutier S. Evolutionary origin of the segmental duplication encompassing the wheat GLU-B1 locus encoding the over-expressed Bx7 (Bx7OE) high molecular weight glutenin subunit. Theor Appl Genet, 2008, 116: 283–296
[10]Wang L H, Li G Y, Peña R J, Xia X C, He Z H. Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.). J Cereal Sci, 2010, 51: 305–312
[11]Wang L H, Zhao X L, He Z H, Ma W, Appels R, Peña R J, Xia X C. Characterization of low molecular weight glutenin subunit Glu-B3 genes and development of STS markers in common wheat (Triticum aestivum L.). Theor Appl Genet, 2009, 118: 525–539
[12]Ahmad M. Molecular marker-assisted selection of HMW glutenin alleles related to wheat bread quality by PCR-generated DNA markers. Theor Appl Genet, 2000, 101: 892–896
[13]Ma W, Zhang W, Gale K R. Multiplex-PCR typing of high molecular weight glutenin alleles in wheat. Euphytica, 2003,134: 51–60
[14]Moczulski M, Salmanowicz B P. Multiplex PCR identification of wheat HMW glutenin subunit genes by allele-specific markers. J Appl Genet, 2003, 44: 459-471
[15]Zhang X K, Liu L, He Z H, Sun D J, He X Y, Xu Z H, Zhang P P, Chen F, Xia X C. Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat. Plant Breed, 2008, 127: 109–115
[16]Zhang X-K(张晓科), Xia X-C(夏先春), Wang Z-W(王忠伟), Wan Y-X(万映秀), Zhang P-Z(张平治), He X-Y(何心尧), Yang Y(杨燕), He Z-H(何中虎). Establishment of multiplex-PCR for quality traits in common wheat. Acta Agron Sin (作物学报), 2007, 33(10): 1703–1710 (in Chinese with English abstract)
[17]Wan Y-X(万映秀), Zhang X-K(张晓科), Xia X-C(夏先春), Zhang P-Z(张平治), He Z-H(何中虎). Development of multiplex PCR and identification of major quality genes in cultivars from Yellow and Huai River valley wheat region. Sci Agric Sin (中国农业科学), 2008, 41(3): 643–653 (in Chinese with English abstract)
[18]Zheng H(郑寒), Chen J(陈静), Ren Y(任妍), Yu M-Q(余懋群), Fu T-H(付体华). Establishment and application of multiplex-PCR for wheat glutenin subunits relative to superior end-use quality. Acta Agron Sin (作物学报), 2009, 35(10): 1831–1835 (in Chinese with English abstract)
[19]Sui X X, Wang L H, Xia X C, Wang Z L, He Z H. Development of an allele-specific marker for Glu-B3 alleles in common wheat and establishment of a multiplex PCR assay. Crop & Pasture Sci, 2010, 61: 978–987
[20]Liu L(刘丽). Identification of glutenin composition in common wheat by proteomics technology. PhD Dissertation of Chinese Academy of Agricultural Sciences, 2008 (in Chinese with English abstract)
[21]Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual, 3rd Edn. New York: Cold Spring Harbor Laboratory Press, 2001
[22]Liu L(刘丽), Zhou Y(周阳), Liu J-J(刘建军), He Z-H(何中虎), Yang J(杨金). Effect of allelic variation at the Glu-1 and Glu-3 loci and presence of 1BL/1RS translocation on pan bread and dry white Chinese noodle quality. Sci Agric Sin (中国农业科学), 2004, 37(9): 1265–1273 (in Chinese with English abstract)
[23]Shewry P R, Halford N G, Tatham A S. High-molecular weight subunits of wheat glutenin. J Cereal Sci, 1992, 15: 105–120
[24]Payne P I, Holt L M, Law C N. Structural and genetical studies on the high-molecular-weight subunits of wheat glutenin: Part 1: allelic variation in subunits amongst varieties of wheat (Triticum aestivum). Theor Appl Genet, 1981, 60: 229–236
[25]Zhang X-Y(张学勇), Pang B-S(庞斌双), You G-X(游光霞), Wang L-F(王兰芬), Jia J-Z(贾继增), Dong Y-C(董玉琛). Allelic variation and genetic diversity at Glu-1 loci in Chinese wheat (Triticum aestivum L.) germplasms. Sci Agric Sin (中国农业科学), 2002, 35(11): 1302–1310 (in Chinese with English abstract)
[26]Chen D-S(陈东升), Liu L(刘丽), Dong J-L(董建力), He Z-H(何中虎), Zhang Y(张艳), Liu J-J(刘建军), Wang D-S(王德森). Effect of HMW and LMW glutenin subunits and presence of 1BL/1RS translocation on quality traits in spring wheat. Acta Agron Sin (作物学报), 2005, 31(4): 414–419 (in Chinese with English abstract)
[27]Shewry P R, Tatham A S. Disulphide bonds in wheat glutein proteins. J Cereal Sci, 1997, 25: 207-227
[28]Zhao H-X(赵会贤), Hu S-W(胡胜武), Ji W-Q(吉万全), Mares D. The effects of allelic variation at glutein subunit loci Glu-1 and Glu-3 on the size distribution of polymeric protein. Sci Agric Sin (中国农业科学), 1998, 31(1): 69–75 (in Chinese with English abstract)
[29]Barro F, Rooke L, Békés F, Gras P, Tatham A S, Fido R, Lazzeri P A., Shewry P R, Barceló P. Transformation of wheat with high molecular weight subunit genes results in improved functional properties. Nat Biotechnol, 1997, 15: 1295–1299
[30]Yang J(杨金), Zhang Y(张艳), He Z-H(何中虎), Yan J(阎俊), Wang D-S(王德森), Liu J-J(刘建军), Wang M-F(王美芳). Association between wheat quality traits and performance of pan bread and dry white Chinese noodle. Acta Agron Sin (作物学报), 2004, 30(8): 739–744 (in Chinese with English abstract)
[31]Liu A-H(刘爱华), He Z-H(何中虎), Wang G-R(王光瑞), Wang D-S(王德森), Zhang Y(张艳), Zhou G-Y(周桂英). Investigation of wheat flour quality for northern style Chinese steamed bread. J Chin Cereal Oils Assoc (中国粮油学报), 2000, 15(2): 10–15 (in Chinese with English abstract)
[32]He Z H, Yang J, Zhang Y, Quail K J, Peña R J. Pan bread and dry white Chinese noodle quality in Chinese winter wheats. Euphytica, 2004, 139: 257–267

Related Articles 15

[1]	Zhai Sheng-Nan, Cao Xin-You, Li Hao-Sheng, Li Ji-Hu, Li Fa-Ji, Liu Jin-Dong, Xia Xian-Chun, Lyu Ying-Ying, Ma Rui-Feng, Wang Ying, Geng Hong-Wei, Liu Jian-Jun. Analysis of the genetic effects of allelic variation at the Pod-A1, Pod-D1, and Pod-2D loci on peroxidase activity in wheat grains [J]. Acta Agronomica Sinica, 2026, 52(6): 1593-1603.
[2]	Peng Jia-Luo, Li Ying, Li Dan-Dan, Yang Jun-Ning, Guo Xue-Feng, Zhang Wen-Jiao, Yu Xiao-Xue, Zhou Ya-Rong, Wang Zhen-Yu, Wang Cai-Xiang, Ma Xiong-Feng, Su Jun-Ji. Identification of class I LBD family members in upland cotton and function and haplotype analyses of GhLBD6 in regulating flowering period [J]. Acta Agronomica Sinica, 2026, 52(6): 1682-1697.
[3]	Zhang Xian-Feng, Guo Li-Jian, Li Kang-Chun, Kong Bin-Xue, Liu Yu-Fang, Che Zhuo, Yang De-Long. Identification of the ABHD6 gene family and development of functional markers for grain weight in wheat [J]. Acta Agronomica Sinica, 2026, 52(6): 1711-1727.
[4]	Wang Chu-Rui, Li Kai-Xiang, Zhao Zhi, Xiao Lu, Tang Guo-Yong, Zhao Zhi-Gang, Xu Liang, Du De-Zhi, Liu Hai-Dong. Development and application of KASP markers for functional sites of the early flowering gene BnCRY2 in Brassica napus L. spring rapeseed [J]. Acta Agronomica Sinica, 2026, 52(3): 708-721.
[5]	Wang Yue-Sheng, Ge Dong-Dong, Cheng Lan-Fei, Chen Chun-Huan, Wang Chang-You, Liu Xin-Lun, Li Ting-Dong, Deng Ping-Chuan, Ji Wan-Quan, Zhao Ji-Xin. Molecular cytogenetic and disease resistance characterization of the wheat- Psathyrostachys huashanica disomic substitution line 16DH25-7 [J]. Acta Agronomica Sinica, 2026, 52(2): 433-445.
[6]	ZHAN Zong-Bing, JIN Qi-Feng, LIU Di, LYU Ying-Chun, GUO Ying, ZHANG Xue-Ting, HU Meng-Xia, WANG Shang, YANG Fang-Ping. Molecular characterization and evaluation of important traits of landrace wheat Laomangmai in Gansu province, China [J]. Acta Agronomica Sinica, 2025, 51(3): 609-620.
[7]	XU Xiao-Wei, FENG Jing, WANG Feng-Tao, TONG Zhao-Yang, ZHANG Jian-Zhou, LI Chun-Ying, LIN Rui-Ming. QTL mapping of adult plant resistance to stripe rust in the Chinese wheat landrace Canlaomai [J]. Acta Agronomica Sinica, 2025, 51(11): 2933-2943.
[8]	TIAN Han-Zhao, FENG Long-Ting, YING Kai, MENG Tian-Qi, WU Jun, LIU Yu-Xiu. Composition and evaluation of wheat glutenin subunits in exotic wheat germplasm [J]. Acta Agronomica Sinica, 2025, 51(10): 2663-2680.
[9]	LIU Xin-Yuan, CHENG Yu-Kun, WANG Li-Li, ZHAN Shuai-Shuai, MA Meng-Yao, GUO Ling, GENG Hong-Wei. Allelic variation and distribution of peroxidase activity genes TaPod-A1, TaPod-A3, and TaPod-D1 of wheat in Xinjiang, China [J]. Acta Agronomica Sinica, 2025, 51(1): 68-78.
[10]	FAN Zi-Pei, LI Long, SHI Yu-Gang, SUN Dai-Zhen, LI Chao-Nan, JING Rui-Lian. Cloning of TabHLH112-2B gene and development of its functional marker associated with the number of spikelet per spike in wheat [J]. Acta Agronomica Sinica, 2024, 50(2): 403-413.
[11]	KE Hui-Feng, SU Hong-Mei, SUN Zheng-Wen, GU Qi-Shen, YANG Jun, WANG Guo-Ning, XU Dong-Yong, WANG Hong-Zhe, WU Li-Qiang, ZHANG Yan, ZHANG Gui-Yin, MA Zhi-Ying, WANG Xing-Fen. Identification for yield and fiber quality traits and evaluation of molecular markers in modern cotton varieties [J]. Acta Agronomica Sinica, 2024, 50(2): 280-293.
[12]	CHEN Tian, LI Yu-Ying, RONG Er-Hua, WU Yu-Xiang. Character identification and floral organ transcriptome analysis on artificial allotetraploids of Gossypium hirsutum L. [J]. Acta Agronomica Sinica, 2024, 50(2): 325-339.
[13]	TAN Dan, CHEN Jia-Ting, GAO Yu, ZHANG Xiao-Jun, LI Xin, YAN Gui-Yun, LI Rui, CHEN Fang, CHANG Li-Fang, ZHANG Shu-Wei, GUO Hui-Juan, CHANG Zhi-Jian, QIAO Lin-Yi. Discovery of auxin pathway genes involving spike type and association analysis between TaARF23-A and spikelet number in wheat [J]. Acta Agronomica Sinica, 2024, 50(2): 506-513.
[14]	ZHAO Yang, LI Long, YANG Jin-Wen, JING Rui-Lian, SUN Dai-Zhen, WANG Jing-Yi. An E3 ubiquitin ligase gene TaSINA-3A is associated with plant height and 1000-grain weight in various environments in wheat [J]. Acta Agronomica Sinica, 2024, 50(10): 2654-2664.
[15]	SU Zai-Xing, HUANG Zhong-Qin, GAO Run-Fei, ZHU Xue-Cheng, WANG Bo, CHANG Yong, LI Xiao-Shan, DING Zhen-Qian, YI Yuan. Identification of wheat dwarf mutant Xu1801 and analysis of its dwarfing effect [J]. Acta Agronomica Sinica, 2023, 49(8): 2133-2143.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Establishment and Application of Multiplex PCR System Based on Molecular Markers of Glutenin Subunit Genes (Loci) Related to Strong-gluten in Wheat

PDF