摘要
玉米(Zea mays L.)是世界上重要的粮食兼饲料作物,产量居农作物之首,也是重要的遗传模式植物。我国是世界上仅次于美国的第二大玉米生产国,玉米生产在国民经济中占有非常重要的地位。因此,玉米的基础研究和遗传改良具有重大的理论和现实意义。
遗传图谱的构建是基因组研究中的重要环节,是基因定位与克隆乃至基因组结构与功能研究的基础；构建玉米遗传图谱及寻找与目标性状紧密连锁的分子标记是进行分子标记辅助选择育种的前提,对数量性状基因QTL的定位尤为重要。随着分子标记技术的飞速发展,为高精度的遗传图谱的构建提供了可能,许多作物高饱和的分子连锁图谱得以构建,使得在分子水平上进行数量性状的研究更加深入。
本研究以玉米孤雌生殖诱导的DH群体(240个株系)为供试材料,构建了一张微卫星或简单序列重复(simple sequence repeats, SSR)标记连锁图。具体的研究结果如下：
1.从已发表的玉米微卫星引物中挑选出730对覆盖玉米10条染色体的引物,进行多态性检测,共检测到130个多态性位点对DH群体进行基因型分析。卡方检验结果表明,65个位点表现基因偏分离,占50%。以1,3和8染色体表现突出。
2.为了提高利用玉米DH群体进行遗传连锁图谱构建的效率,降低其成本,采用高通量SSR检测方法对“先玉335”DH群体进行多个引物复合电泳的策略,提出了针对特定群体的SSR标记检测方案。为提高玉米分子育种中的实验效率提供了新的思路。
3.将130个多态性位点引物用Mapmaker/Exp3.0 (Lincoln等,1992)软件构建SSR连锁图谱,最终拟合到图谱上的位点共119个。将119个多态性位点分成10个连锁群,图谱总长度857.5 cM,平均间距8.15 cM。标记数最多的是第1号染色体,为20个标记位点；最少的是第6号染色体,为8个标记位点。该遗传图谱谱能有效地进行数量性状QTL定位研究。
Maize (Zea mays L.) is an important crop and feed in the world, which ranks first in all crops production. It is also most tractable genetic system among the cereals.Maize yield in China is second following the United States, which plays an important role in national economy. Thus, maize basical research and genetic improvement have great theoretical and practical significance.
Construction of genetic map is the important elements of gene mapping and it is the basis of gene mapping, cloning and genome structure and function; Construction of maize genetic map and looking for molecular marker which is tightly linked to the target trait are the basis of molecular marker-assisted selection breeding. It is particularly important for QTL. New technology on molecular marker technique makes high-precision construction of genetic maps possible. Many crops highly saturated molecular linkage maps built help the further and deeper study of quantitative traits on the molecular level.
DH population of maize (240 DH lines) were researched. It constructed a microsatellite or simple sequence repeats (simple sequence repeats, SSR) marker linkage map. Specific results are as follows:
1.730 pairs of published PCR primers for amplifying microsatellite or simple sequence repeat (SSR) loci were used for the study.130 of them displayed polymorphism between the two parents. Chi-square test indicates that sixty-five markers (50% of all) skew from the expected gene segregation. Chromosomes 1,3, and 8 had outstanding performance.
2. In order to improve the efficiency of Mapping Population of maize and to reduce the cost, high throughput measurement SSR electrophoresis technique was applied "Xian Yu 335" DH population. It process to multiple composite primers. This method was discussed in the maize targeted groups of SSR analysis.and provided a new way to improve the efficiency in maize molecular breeding.
3.130 polymorphic loci are used to construct a genetic linkage map by the software Mapmaker/Exp3.0. Eventually a total of 119 sites which fit into the map were divided into 10 linkage groups. It covered a total length of 857.5cM with an average distance of 8.15cM between adjacent markers. The most one is Chromosomes 1 with 20 markers, and the least one is Chromosomes 6 with 8 markers. This mapping can be used to QTL research efficiently.
引文
[1]陈刚,陈丽,高洪敏.丹玉自交系在我国玉米育种和生产中的地位与作用[J].沈阳农业大学学报,2000,31(5)：522-525.
[2]毛传澡,程式华.水稻农艺性状QTL定位精确性及其影响因素的分析[J].农业生物技术学报,1999,7(4)：386-394.
[3]黎裕,贾继增,王天宇.分子标记的种类及其发展[J].生物技术通报,1999,4：19-22.
[4]李平,朱立煌等.利用分子标记和水稻籼粳交双单倍体群体进行遗传作图研究[J].植物学报,1996,38(11)：881-886.
[5]张耿,祝丽英,陈景堂.玉米产量相关性状QTL通用图谱的构建及功候选基因的开发[J].华北农学报,2008,23(6)：20-27.
[6]王凤格.玉米矮花叶病抗性QTL的初步分析[D].硕士论文.北京：中国农业科学院,2001.
[7]贾继增.分子标记种质资源鉴定和分子标记育种[J].中国农业科学,1996,29(4)： 1-10.
[8]王倩,王斌.DNA分子标记在果树遗传学研究上的应用[J].遗传,2000,22(5)：339-344.
[9]盖钧锰,章元明,王建康.QTL混合遗传模型扩展至2对主基因+多基因时的多世代联合分析[J].作物学报,2000,26(4)：355-359.
[10]ottaviano E, Gorla M S, Frova P E.Moleuclar mkarers(RFLPsandHSPs) of the genetic dissection of the thermotolerance in maize[J].Theor Appl Genet, 1991,81:713-719.
[11]刘峰,陈受宜等.微卫星标记在大豆遗传作图中的应用[J],高技术通讯,1999,6：8-11.
[12]李平,周开达等.利用分子标记定位水稻野败型核质互作雄性不育恢复基因[J].遗传学报,1996,23(5)：357-362.
[13]李建雄.一个优良杂交组合的杂种优势遗传成因的分子标记分析[D].博士学位论文.武汉：华中农业大学.1998.
[14]冯英.水稻DH群体的构建与微卫星标记分析[D].博士学位论文.浙江.浙江大学.
[15]Botstein DR, White RL, Skolnick M, Davis RW. Construction of a genetics linkage map in man using restriction fragment length polymorphisms. Ameriean Journal of Human Genetics,1980,32:314-331.
[16]Williams JGK, Kubelik AR, Livak KJ. DNA polymorphisms amplified by arbitrary primers are useful genetic markers [J]. Nucleic Acids Res,1990,18: 6531-6537
[17]Ronald P C, Albano B, Tabien R, Abenes L, Wu K, McCouch S R, Tanksley S D. Genetic and physical analysis of the rice bacterial blight disease resistance locus, Xa21. Molecular Gene Genetics,1992,236:113-120.
[18]Song WY,Wang GL,Chen LL,Kim HS,Holsten T,Wang B,Zhai WX Zhu LH,Fauquet C,Ronald P.The rice disease resistance gene,xa2i,encodes rcceptor kinase-like protein. Science,1995,270:1804-1806.
[19]Vos P,Hogers R,Bleeker M,Reijans M,Lee VD,Homes M,Fritjers A,Pot J,PelemanJ,Kuiper M,Zabeau M. AFLP:a new concept for DNA fingerprinting Nucleic Acids Research,1995,23:4407-4414.
[20]Wang D G, Fan J B, Sao C J, et al. Large-scale identification mapping, and genotyping of Single-nucleotide polymorphisms in the human genome[J]. Science,1998,280:1077-1082.
[21]Rafalski JA. NoVel genetic mapping tools in plants:SNPs and LD-based approaches. Plant Scienee,2002,162:329-333.
[22]毛传澡,程式华.水稻农艺性状QTL定位精确性及其影响因素的分析[J].农业生物技术学报,1999,7(4)：386-394.
[23]严建兵,汤华,黄益勤,郑用琏,李建生.玉米F2群体分子标记偏分离的遗传分析[J].遗传学报,2003,30(10)：913-918.
[24]Eshed Y, Zamir D. Introgression from Lycopersicon pennellii can improve the soluble-solids yield of tomato hybrids. Theor Appl Genet,1994,88:891-897.
[25]Yamamoto T, Liu HX, Sashki T and Y ano M. Fine mapping of quantitative trait lociHd-1, Hd-2 and Hd-3, controlling heading data of rice, as single Mendelian factors. Theor Appl Genct,1998,97:37-44.
[26]Yamamoto T. Liu HX, Sasaki T and Y ano M. Identification of heading data of quantitative trait locus Hd6 and characterization of its epistatic interaction with Hd2 in rice using advanced back-crosses progeny.Genetics,2000,154:855-891.
[27]于永涛,宋燕春,黎裕,王天宇.玉米种质资源H21和M017抗亚洲玉米螟的QTL分析[J].植物遗传资源学报,2003,4(2)：94-98.
[28]Doebley J, Stec A, Gustus C. Teostine branchedle and the origin of maize: evidence for epistasis and the evolution of dominance. Geneties,1995,141: 333-346.
[29]Tanksley S D, Nelson J C. Advanced backcross QTL analysis:a method for the simultaneous discovery and transfer of valuable QTLs from unadapted germplasm into elite breeding lines. Theor. Appl. Genet.1996,92:191-203.
[30]盖钧锚等.利用回交或F2：3世代鉴定主基因和多基因的混合遗传模型[J].作物学报,1998,24(4)：402-409.
[31]Eshed Y, Abu-Abied M, Saranga Y, Zamir D. Lycopersicon esculentum lines containing small overlapping introgressions from L. pennellii. Theoretical and Applied Genetics,1992,83:1027-1034.Genetics,1992,83:1027-1034.
[32]胡中立,张志红,张学富.双单倍体群体中区间分子标记定位(QTL)的相关方法[J].生物数学学报,1998,13(3)：366-371.
[33]章元明,黄方,喻德跃,等.在DH或RIL群体中两对重叠基因控制性状的定位[J].遗传学报,2001,25(1)：57-59.
[34]汪军妹,沈秋泉,杨建明.大麦加倍单倍体(DH群体)的建立及其在遗传育种中的应用[J].大麦科学,2001,4：10-13.
[35]朱惠琴,薛庆中,张宪银.烟草DH群体的构建及其性状表现[J].青海大学学报(自然科学版),2004,(6)：52-56.
[36]赵佐宁,谷明光.化学药剂诱导玉米孤雌生殖植株的细胞遗传研究[J].遗传学1988,15(2)：89-94.
[37]刘志增,宋同明.玉米单倍体雌雄育性的自然恢复以及染色体的化学加倍[J].作物学报,2000,26(6)：947-952.
[38]魏俊杰,池书敏,刘志增.关于玉米单倍体人工加倍方法及花粉活力测定的初步研究[J].玉米科学,2001,9(3)：12-13.
[39]宋炜.玉米DH群体的遗传分析及QTL定位初探[D].硕士论文,河北农业大学,2005,06.
[40]文科.玉米单倍体诱导和加倍方法及其SSR和ISSR标记分离研究[D].中国农业大学硕学位论文,2003.
[41]Griffing B.Efficiency changes due to use of doubled-haploids in recurrent selection methods [J].Theor Appl Genet,1975,46:367-386.
[42]A.Gallais.Quantitative genetics of doubled haploid populations and application to the theory of line development [J].Geneties,1990,124:199-206.
[43]Lander E Skbostein D. Mapping mendelian factors underlying quantitative traits using RFLP linkage maps [J]. Genetics 1989,121:185-199.
[44]Burr B, Burr F A, Thompson K H, Albertson M C, Stuber C W. Gene mapping with recombinant inbreds in maize.Genetics,1988,118:519-526.
[45]Ragot M, Sisco P H, Hoisington D A.etal Molecular-makre-mediated Characteri-zation of favorable exotic alleles at quantitative trait loci in maize [J]. Crop Seience,1995,35:1306-1315.
[46]方宣钧,吴为人,唐纪良.作物DNA标记辅助育种[J].北京,科学出版社,2001.
[47]Gardiner J M,Coe E H,Melia-Hancock S,et al.Development of a core RFLP map in maize using an immortalized F2 population. Genetics,1993, 134(7):917-917.
[48]Paterson A H, Lander E S, Hewitt J D, Peterson S, Lincoln S E, Tanksley S D. Resolution of quantitative traits intoMendelian factors, using a complete linkage map of restriction fragment Length polymorphisms. Nature,1988,12: 185-199.
[49]Helentjaris T, Slocum M, wright S, et al. Construction of gene tic linkage maps in maize and tomato using restriction fragment length polymorphisms [J].Theor. Appl.Genet,1986,72:761-769.
[50]程须珍,王素华,吴绍宇,等.绿豆抗豆象育种后代F2群体遗传变异分析[J]植物遗传资源学报,2004,5(4)：364-368.
[51]向道权,曹海河,曹永国,等.玉米SSR遗传图谱的构建及产量性状基因定位[J].遗传学报,2001,28：775-784.
[52]王凤格.玉米矮花叶病抗性QTL的初步分析[D].硕士论文.北京：中国农业科学院,2001.
[53]顾晓红.中国玉米种质资源品质性状的分析与评价[J].玉米科学,1997,6(1)： 14-16.
[54]杨华,杨俊品,荣廷昭,等.玉米抗纹枯病QTL分子标记定位[J].科学通报,2005,50(8)：772-776.
[55]陈志军,汤在祥,宋雯,徐辰武.基于遗传位置的水稻与玉米重要农艺性状QTL比较研究[J].中国水稻科学,2009,23(3)：229-236.
[56]王凤格,赵久然,等.一种改进的玉米SSR标记的PAGE快速银染检测新方法[J].农业生物学报,2004,12(5)：606-607.
[57]孙立娜.基于玉米DH群体的遗传连锁图谱初步构建[D].硕士论文.北京：北京农科院,2009,05.
[58]Lincoln S, Daly M, Lander E. Constructing genetic maps with Mapmaker/Exp. 3.0White head institute Technical Report.3rd edition.1992.
[59]Lincoln S E, Daly M J, Lander E S. Constructing Genetic Linkage Maps with Mapmaker/Exp Version 3.0:A Tutorial and Reference Manual. A Whitehead Institute for Biomedical Research Technical Report, Third Edition, Jan.,1993.
[60]何祯祥,林国忠,施季森.遗传作图软件应用及辅助软件的研制[J].南京林业大学学报,1999,23(3)：l-5.
[61]方宣钧,吴为人,唐纪良编.作物DNA标记辅助育种[M].北京：科学出版社,2001.
[62]Hyne V, Kearsey M J, Pike D J.et al. QTL analysis:unreliability and bias in estimation Procedures [J]. Molecular Breeding,1995,1:271-282.
[63]Griffing B. Efficiency changes due to use of doubled-haploids in recurrent selection methods [J]. Theor Appl Genet,1975,46:367-386.
[64]Bouehez A, Gallais A. Efficiency of the Use of Doubled-HaPloids in Recurrent Selection for Combining Ability[J].Crop Sei.,2000,40:23-29.
[65]Bordes J G.,Charmet R,Dumas de Vaulx, et al. Doubled haploid versus S1 family recurrent selection for test cross Performance in a maize Population [J].TheorAPPIGenet.2006,112:1063-1072.
[66]ChooT M.Reinbergs E. Analysis of skeweness and kurtosis for detecting gene interaction in a doubled haploid population [J].Crop Scien,1982,22:231-235.
[67]胡中立,张志红,张学富.双单倍体群体中区间分子标记定位(QTL)的相关方法[J].生物数学学报,1998,13(3)：366-371.
[68]章元明,黄方,喻德跃,等.在DH或RIL群体中两对重叠基因控制性状的定位[J].遗传,2001,25(1)：57-59.
[69]Lyttle T W.Segregation distorters. Annual Review of Genetics,991,25: 511-557.
[70]Kinshita T. Report of the committee on gene symbolization, nomenclature and linkage group [J].Rice Genet Newl,1993,8:7-39.
[71]Knox M R, Ellis T H N.Excess heterozygosity contributes to genetic map expansion in pea recombinant inbred populations [J].Genetics,2002,162: 861-873.
[72]Beaumont V H. Mapping the anther culture response genes in maize (Zea mays L.) [J]. Genome,1995,38:968-975.
[73]Beaumout VH, Mantet J, Rocheford T R, Widholm J M. ComParison of RAPD and RFLP markers for mapping F2 generations in maize(Zea mays L.). Theor. Appl. Genet.,1996,93:606-612.
[74]李平,朱立煌,等.利用分子标记和水稻籼粳交双单倍体群体进行遗传作图研究[J].植物学报,1996,38(11)：881-886.
[75]喻婷,张玲,胡中立,等.水稻釉粳交DH群体花器性状的遗传分析[J].武汉植物学研究,2003,21(6)：459-463.
[76]Chang M.T Stock6 indueed double haploidy is random [J].MaizeGenet. CooP. NwesLett.,1992,67:98-99.
[77]Lashermes P, Beckert M.Genetic contol of maternal haploidy in maize (zea mays L.and selec-tion of haploid-indueing lines [J].Theor.AppI.Genet.1988, 76:405-410.
[78]Chalky S, Baumann A, Daniel G. et al. Aneuploidy as a possible cause of haploid-induction in maize [J]. Maize Genet. Crop. NewsLett.2003,29-30.
[79]沈利爽,何平,徐云碧,谭震波,陆朝福,朱立煌.水稻DH群体的分子连锁图谱及基因组分析[J].植物学报,1998,40(2)：1115-1122.
[80]Gardinex J M,Coe E H,Chao S,et al.Development of a core RFLP map in maize using an immortalized F2 population [J]. Genetics,1993,134:917-930.
[81]严建兵,汤华,黄益勤,等.玉米F2群体分子标记偏分离的遗传分析[J].遗传学报,2003,30(10)：913-918.
[82]刘宗华,汤继华,王春丽,田国伟,卫晓轶,胡彦民,崔党群.氮胁迫与非胁迫条件下玉米不同时期株高的动态QTL定位[J].作物学报,2007,33(5)：782-789.
[83]Veldboom LR, Lee M Woodman WL. Molecular marke-facilitated studies in an elite Maize Population. I.linkage analysis and determination of QTL of morphological traits [J].Theor Appi Genet,1994,88:7-16.
[84]徐吉臣,朱立煌,等.用双单倍体群体构建水稻的分子连锁图[J].遗传学报1994,21(3)：205-14.
[85]曹永国,王国英,王守才等.玉米RFLP遗传图谱的构建及矮生基因定位[J].科学通报,1999,44(20)：2178-2181.
[86]杨俊品,荣廷昭,黄烈健等.玉米分子遗传框架图谱构建[J].作物学报,2004,30(1)：82-87.
[87]易红梅,王凤格,赵久然等.玉米品种SSR标记毛细管电泳荧光检测法与变性PAGE银染检测法的比较研究[J].华北农学报,2006,21(5)：64-67.
[88]Wang Fengge, Zhao Jiuran, Dai Jingrui, Yi Hongmei, Kuang Meng, Sun Yanmei, Yu Xinyan Guo Jinglun, Wang Lu,2007. Selection anddevelopment of represent-tative simple sequence repeat primers and fluorescent multiplex SSR sets for high throughput automated genotyping in maize.Chinese Science Bulletin,52(2):215-223.
[89]郝晨阳,王兰芬,贾继增等.SSR荧光标记和银染技术的比较分析[J]. 作物学报,2005(2)：144-149.
[90]M.D.Krakowsky,M.Lee,M.L.Woodman-Clikeman,M.J.Long,N.SharoPova.QTL Mapping of Resistance to Stalk Tunneling by the EuroPean Corn Borer in RILs of Maize PoPulation[J].Crop Sci.,2004,44:274-282.
[91]Pinar Bor, Johnny Hindkjar, Steen Kolvraa.A new approach for screening for Y microdeletions:capillary electrophoresis combined with fluorescent multiplex PCR [J] Journal of Assisted Reproduction and Genetics,2003,20(1):46-51.
