卵穗山羊草1My亚基特异性分子标记开发

doi:10.16036/j.issn.1000-2650.2018.06.001

摘要/Abstract

摘要： 【目的】开发高分子量谷蛋白亚基1My的特异性分子标记,方便对其追踪。【方法】通过分析1M°染色体上的高分子量谷蛋白亚基1My编码基因与1A、1B、1D、1U°、1M°染色体上的高分子量谷蛋白亚基编码基因之间的单核苷酸多态性（SNPs）,开发了一个基于PCR扩增技术的1My特异性分子标记。【结果】该分子标记在1M°附加系及亲本卵穗山羊草中扩增出清晰的PCR条带,而其他对照材料无扩增产物。【结论】测序及序列比对分析表明该标记扩增片段与1My编码基因的部分序列完全一致,表明该标记能有效区分卵穗山羊草1My和普通小麦HMW-GS的编码基因。由于谷蛋白x和y型亚基基因紧密连锁,该标记也为筛选含有1Mx的小麦育种材料提供了方便。

关键词: 卵穗山羊草, 高分子量谷蛋白, 分子标记

Abstract: 【Objective】 Development of specific molecular marker to facilitate the tracking of high molecular weight glutenin subunit 1My. 【Method】 We developed 1My subunit-specific PCR marker based on the single nucleotide polymorphisms (SNPs) among 1My and other HMW-GS encoding genes from 1A, 1B, 1D, 1U°, and 1M°. 【Result】 The specific molecular marker amplified a clear band in the monosomic addition line of 1M° and Ae. ovata, but absent in the non-1M° sister lines and other bread wheat parents. 【Conclusion】 Sequence alignment conformed that the amplified fragment is target region of 1My-encoding gene, indicating the effectiveness of the marker for distinguishing 1My of Ae. ovata from bread wheat HMW-GS genes. The marker is also useful for tracing the 1Mx in molecualr breeding due to the closely link between the gluten genes of x and y types.

Key words: Aegilops ovata, HMW-GS, molecular markers

中图分类号:

S512.1

郑柯, 易滢瑾, 赵来宾, 刘冬梅, 许凯, 郝明, 张连全, 刘登才, 甯顺腙. 卵穗山羊草1My亚基特异性分子标记开发[J]. 四川农业大学学报, 2018, 36(06): 717-721.

ZHENG Ke, YI Yingjin, ZHAO Laibin, LIU Dongmei, XU Kai, HAO Ming, ZHANG Lianquan, LIU Dengcai, NING Shunzong. Development of PCR Marker Specific for Subunit 1My of Aegilops ovata[J]. Journal of Sichuan Agricultural University, 2018, 36(06): 717-721.

参考文献 25

[1]	RASHEED A, XIA X, YAN Y, et al.Wheat seed storage proteins: Advances in molecular genetics, diversity and breeding applications[J]. Journal of Cereal Science, 2014, 60(1): 11-24.
[2]	PAYNE P I, NIGHTINGALE M A, KRATTIGER A F, et al.The relationship between HMW glutenin subunit composition and the bread-making quality of British-grown wheat varieties[J]. Journal of the Science of Food & Agriculture, 1987, 40(1): 51-65.
[3]	HARBERD N P, BARTELS D, THOMPSON R D.DNA restriction-fragment variation in the gene family encoding high molecular weight (HMW) glutenin subunits of wheat[J]. Biochemical Genetics, 1986, 24(7): 579-596.
[4]	陈文杰. 节节麦高分子量麦谷蛋白基因的分子鉴定与育种利用[D]. 雅安: 四川农业大学, 2012.
[5]	翁跃进. 山羊草属基因库的开拓利用[J]. 中国种业, 1995(2): 16-18.
[6]	MONIKA G, HISASHI T, KUMAR G R, et al.Chromosome specific substitution lines of Aegilops geniculata alter parameters of bread making quality of wheat[J]. Plos One, 2016, 11(10): e0162350.
[7]	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[J]. Theoretical & Applied Genetics, 1994, 88(6/7): 759-763.
[8]	MA W, ZHANG W, GALE K R.Multiplex-PCR typing of high molecular weight glutenin alleles in wheat[J]. Euphytica, 2003, 134(1): 51-60.
[9]	AHMAD M.Molecular marker-assisted selection of HMW glutenin alleles related to wheat bread quality by PCR-generated DNA markers[J]. Theoretical & Applied Genetics, 2000, 101(5/6): 892-896.
[10]	XU Q, XU J, LIU C L, et al.PCR-based markers for identification of HMW-GS at Glu-B1x loci in common wheat[J]. Journal of Cereal Science, 2008, 47(3): 394-398.
[11]	LIANG X, ZHEN S, HAN C, et al.Molecular characterization and marker development for hexaploid wheat-specific HMW glutenin subunit 1By18 gene[J]. Molecular Breeding, 2015, 35(12): 221.
[12]	WEI L, WANG Q, ZHANG L L, et al.Genetic analyses of Glu-1Ssh in wheat/Aegilops sharonensis hybrid progenies and development of alien HMW-GSs gene-specific markers[J]. Molecular Breeding, 2015, 35(12): 230.
[13]	YAN Z, WAN Y, LIU K, et al.Identification of a novel HMW glutenin subunit and comparison of its amino acid sequence with those of homologous subunits[J]. Science Bulletin, 2002, 47(3): 222-226.
[14]	PAYNE P I.Catalogue of alleles for the complex gene loci, GluA1, Glu-B1 and Glu-D1 which code for high-molecular-weight subunits of glutenin in hexaploid wheat[J]. Cereal Research Communications, 1983, 11(1): 29-35.
[15]	DOYLE J.Isolation of plant DNA from fresh tissue[J]. Focus, 1990(12): 13-15.
[16]	BUTOW B J, MA W, GALE K R, et al.Molecular discrimination of Bx7 alleles demonstrates that a highly expressed high-molecular-weight glutenin allele has a major impact on wheat flour dough strength[J]. Theoretical & Applied Genetics, 2003, 107(8): 1524-1532.
[17]	BEDO Z, KARPATI M, KRAMARIK KISSIMON J, et al.Good breadmaking quality wheat (Triticum aestivum L.) genotypes with 2+12 subunit composition at the Glu-D1 locus[J]. Cereal Research Communications: hungary, 1995, 23(3): 283-289.
[18]	许自成, 牛本永. 应用灰色关联分析法综合评估小麦高分子量麦谷蛋白亚基的研究[J]. 粮食储藏, 1993, 22(1): 18-24.
[19]	GARG M, TANAKA H, ISHIKAWA N, et al.A novel pair of HMW glutenin subunits from Aegilops searsii improves quality of hexaploid wheat[J]. Cereal Chemistry, 2009, 86(1): 26-32.
[20]	WANG S, YU Z, CAO M, et al.Molecular mechanisms of hmw glutenin subunits from 1sl genome of Aegilops longissima positively affecting wheat breadmaking quality[J]. Plos One, 2013, 8(4): e58947.
[21]	LI G R, LIU C, LI C H, et al.Introgression of a novel Thinopyrum intermedium St-chromosome-specific HMW-GS gene into wheat[J]. Molecular Breeding, 2013, 31(4): 843-853.
[22]	XU Q, XU J, LIU C L, et al.PCR-based markers for identification of HMW-GS at Glu-B1x loci in common wheat[J]. Journal of Cereal Science, 2008, 47(3): 394-398.
[23]	SHEWRY P R, HALFORD N G, TATHAM A S.High molecular weight subunits of wheat glutenin[J]. Journal of Cereal Science, 1992, 15(2): 105-120.
[24]	RAFALSKI A.Applications of single nucleotide polymorphisms in crop genetics[J]. Current Opinion in Plant Biology, 2002, 5(2): 94-100.
[25]	刘会云, 刘畅, 王坤杨, 等. 小麦高分子量麦谷蛋白亚基鉴定及其品质效应研究进展[J]. 植物遗传资源学报, 2016, 17(4): 701-709.