甜菜遗传连锁图谱初步构建

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王茂芊, 李博, 王华忠. 甜菜遗传连锁图谱初步构建. 作物学报, 2014, 40(2): 222-230[WANG Mao-Qian, LI Bo, WANG Hua-Zhong. Construction of Molecular Genetic Linkage Map of Sugarbeet. 作物学报, 2014, 40(2): 222-230] 复制到剪切板

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作物学报编辑部

甜菜遗传连锁图谱初步构建

王茂芊^1,^2,³, 李博¹, 王华忠^1,^2,^3,^*

¹黑龙江省普通高校甜菜遗传育种重点实验室 / 黑龙江大学农作物研究院, 黑龙江哈尔滨 150080

²中国农业科学院甜菜研究所, 黑龙江哈尔滨 150080

³中国农业科学院北方糖料作物资源与利用重点开放实验室, 黑龙江哈尔滨 150080

^* 通讯作者(Corresponding author): 王华忠, E-mail:wwhhzz0451@sohu.com, Tel: 0451-86609494

收稿日期:2013-09-06

基金:本研究由国家自然科学基金项目(31271780), 国家现代农业产业技术体系建设专项(CARS-210104)和黑龙江大学青年基金(QL201038)项目资助。

摘要

以甜菜高产低糖型JV34-2和低产高糖型2B023两材料杂交, 构建了200个单株的F₂作图群体, 利用所筛选出的56对SRAP引物组合和20对SSR引物, 对F₂作图群体进行PCR扩增和遗传连锁分析, 初步构建了一张包含9个连锁群、141个(123个SRAP和18个SSR)标记位点的甜菜遗传连锁图谱。该图谱覆盖长度为1399.88 cM, 平均图距9.92 cM。未进入连锁群的有4个标记。9个连锁群包含3~26个标记不等, 连锁群遗传距离15.69~237.21 cM。连锁群上有20.56%的标记出现偏分离, 主要集中在Chr.3连锁群上, 其余分散在Chr.1、Chr.2、Chr.8和Chr.9中。该图谱是我国甜菜领域利用SRAP和SSR相结合方法, 构建的第一个较精密的分子遗传图谱, 为重要性状的基因定位和优良基因的克隆奠定了基础。

关键词: 甜菜; SRAP; SSR; 遗传图谱

Construction of Molecular Genetic Linkage Map of Sugarbeet

WANG Mao-Qian^1,^2,³, LI Bo¹, WANG Hua-Zhong^1,^2,^3,^*

¹Key Laboratory of Sugar Beet Genetic Breeding/ Crop Academy of Heilongjiang University, Harbin 150080, China

²Sugar Beet Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150080, China

³Key Laboratory of North Sugar Crop Resource and Utilization, Chinese Academy of Agricultural Sciences, Harbin 150080, China

Abstract

Molecular genetic map is a fundamental tool for genomic research. In this study, a molecular genetic map was constructed using an F₂ population consisting of 200 individuals from a cross between sugarbeet JV34-2 (the high beet yield and low sugar content line) and sugarbeet 2B023 (the low beet yield and high sugar content line) and a total of 561 SRAP primers and 114 SSR primers. Approximately 56 out of 561 and 20 out of 114 primers, respectively, showed polymorphisms. Each of these polymorphic primers produced at least one scorable polymorphic DNA band which was visible enough for detection and scoring. The map consisted of nine linkage groups which included 141 (123 SRAP and 18 SSR) markers, and covered 1399.88 cM with an average distance of 9.92 cM. Three markers were still unlinked. All nine linkage groups consisting of 3-26 markers were 15.69-237.21 cM in length. Twenty point five six percent partially segregated markers distributed in the map, were mainly located on Chr.3 linkage groups, and the others were mapped on Chr.1, Chr.2, Chr.8, and Chr.9. This is the first gene map of sugarbeet, constructed by SRAP and SSR markers in China. It will provide the basis for gene mapping of important characters and cloning of excellent gene in sugarbeet.

Keyword: Sugarbeet; SRAP; SSR; Genetic linkage map

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我国甜菜育种技术相对落后, 近几年国外品种大量进入我国, 我国甜菜育种以及甜菜种子、甜菜糖业存在重大安全隐患。为防止国外品种垄断, 培育出国产高产、高糖、抗病的新品种, 进一步研究优良基因定位和克隆等, 有必要建立我国自主的甜菜分子标记遗传图谱。目前国内尚未建立一套精密的甜菜遗传图谱, 严重影响了我国甜菜分子育种的发展。国外虽早有甜菜遗传图谱的相关报道, 但应用亲本不同, 构建的群体不同, 所得到的遗传信息也不同。而且到目前为止, 已构建的甜菜分子遗传连锁图谱中主要应用的DNA标记是AFLP、RFLP、RAPD和SSR, 尚未见报道应用SRAP。如1996年Hallden等^{[ 1]}利用RFLP构建了包括431个标记, 覆盖620 cM的甜菜遗传图谱, 标记间平均间距1.5 cM; 1995年Barzen^{[ 2]}构建了包含248个RFLP和50个RAPD位点, 全长815 cM的遗传连锁图谱, 并对抗丛根病基因、下胚轴颜色基因和单粒型性状基因进行定位; Uphoff^{[ 3]}构建了包括85个RAPD标记位点的糖甜菜遗传图谱, 定位了下胚轴颜色基因、一年生基因及一个恢复基因; 1999年Nisson等^{[ 4]}用221个AFLP和46个RFLP标记对204个植株群体进行了甜菜叶斑病抗性的QTL分析; 2002年Schneider等^{[ 5]}对甜菜的含糖率、产量和品质进行了QTL分析; 2007年Grimmer和Trybush建立在AFLP、SNP、RAPD标记基础上的甜菜锚定连锁图和抗甜菜坏死黄脉病毒的QTL定位^{[ 6]}。本研究利用SRAP和SSR两种分子标记方法, 以甜菜材料JV34-2 (高产低糖品系)和2B023 (低产高糖品系)为亲本, 构建F₂代作图群体, 初步构建甜菜的遗传图谱, 为甜菜QTL定位、标记辅助选择育种等奠定基础。

1 材料与方法

1.1 作图群体的构建及田间性状调查

2010年春季以二倍体甜菜单胚品系JV34-2 (高产低糖型)和二倍体甜菜多胚品系2B023 (低产高糖型)为亲本, 杂交获得F₁种子, 同年8月初在山东高密地区南繁, 11月初收获F₁母根并运回北方放入地窖内储藏使之通过春化处理(2~4℃), 60 d后即2011年1月初栽植于温室进行F₁自交, 2011年5月初获得F₂代种子, 播种于呼兰校区试验田, 得到F₂作图群体, 从中选用200株作为试验样品。

1.2 DNA的提取

在苗期每次取50份供试材料的单株幼嫩叶片2片, 用冷冻真空抽干机处理(-50℃左右, 36~48 h), 后研磨成粉末, 用CTAB法提取基因组DNA^{[ 7]}。

1.3 SRAP-PCR和SSR-PCR

SRAP-PCR反应体系含2.5 mmol L^-1的dNTPs 1.2 μL, 上下引物各40 ng, 10×buffer (已含Mg²⁺) 2 μL, TaqDNA聚合酶1 U, DNA 40 ng, 以ddH₂O调整至20 μL。反应程序为94℃ 3 min; 94℃ 1 min, 35℃ 1 min, 72℃ 1 min, 5个循环; 94℃ 1 min, 50℃1 min, 72℃ 1 min 35个循环; 72℃ 10 min。20 μL SSR-PCR反应体系含2.5 mmol L^-1 dNTPs 0.6 μL, 正反引物各40 ng 10×buffer (已含Mg²⁺), TaqDNA聚合酶1 U, 模板DNA 60 ng。反应程序为94℃ 4 min; 94℃ 1 min, 50℃ 1 min, 72℃ 1.5 min, 30个循环; 72℃ 10 min。用8%非变性聚丙烯酰胺凝胶电泳检测PCR产物, 电泳缓冲液为0.5×TBE。硝酸银染色观察结果。试验中选用的SSR引物和SRAP引物均来源于已发表的参考文献^{[ 8, 9, 10]}, 并由北京华瑞生物技术公司合成, 引物序列和组合见表1、表2和表3。

1.4 数据处理

按点样顺序记录多态性条带: (1)共显性标记中同父本带型相同的记为“1”, 与母本带型相同的记为“0”, 杂合条带记为“2”, 缺失或带型不清的记为“-”(2)显性标记中同父本带型相同的记为“1”, 同母本带型相同的记为“0”, 缺失或带型不清的记为“-”。利用卡方检验检测多态性标记是否符合(3∶1显性, 或者1∶2∶1共显性)分离比例^{[ 11]}。用MapMaker3.0软件划分连锁群、标记各连锁群内的顺序和计算位置等, 用QTL Icimapping 3.1绘制连锁图谱(设置LOD=3.0, 最大距离37.2 cM), 可在http://download. csdn.net/网站下载Mapmaker 3.0, 在http://www.isbr eeding.net/网站下载QTL IciMapping。

1.5 图谱预期长度和框架图谱覆盖率的计算

按照Postlethwait^{[ 12]}方法估算遗传图谱长度 G_e1= G_of+2 s× n, 其中 G_of是框架图的总长, s是连锁图谱的标记平均间隔, n是连锁群数目。根据Chakravarti^{[ 13]}的方法4, G_e2= G_of( m+1)/( m-1), m是连锁群上的总标记数。遗传图谱的预期长度 G_e是 G_e1和 G_e2的平均值, 框架图谱覆盖率即框架图总长占图谱预期长度的百分比。

2 结果与分析

2.1 SRAP和SSR多态性分析

利用亲本DNA从561对SRAP引物组合和114对SSR引物中选出多态性好的SRAP引物组合56对和SSR引物20对, 再利用筛选出的引物对作图群体进行多态性分析。56对SRAP引物组合扩增出126个多态性标记, 平均每对引物组合能扩增出2.25个多态性标记, 标记数为1~5条不等; 20对SSR引物获得19个多态性标记, 有1对SSR引物ms0331在F₂群体中未出现多态性。

2.2 标记的偏分离分析

卡方检验表明, 145个分子标记中(126个SRAP标记, 19个SSR标记), 有31个出现偏分离, 占标记总数的21.38%, 其中SRAP标记28个, 占SRAP总标记的22.22%; SSR标记3个, 占SSR总标记的15.78%。连锁群上共有29个标记偏分离(26个SRAP和3个SSR), 占连锁群标记数的20.56%, 主要集中在Chr.3连锁群上, 有17个偏分离标记, 其余分别分散在Chr.1、Chr.2、Chr.8和Chr.9中, 标记数各为3个。通过对照亲本基因型, 这些偏分离标记中有23个偏向父本, 均为SRAP标记, 占连锁群偏分离标记的79.31%, 其中有20个( P<0.01)表现为极显著偏离父本, 占偏父本标记的86.96%; 有6个偏向母本, 占20.69%, 3个SSR标记和3个SRAP标记, 其中3个标记( P<0.01)表现为极显著偏离母本, 占偏母本标记的50%。偏离度最大的标记是SRAP标记m2e8-4, 卡方值为25.62 (表4和表5), 偏分离标记的分离程度可见图1, 纵坐标越大, 表明偏离度越大。未发生连锁的有4个标记, 其中偏分离标记2个。

表1 SRAP引物序列名称 Table 1 SRAP primers and their sequences

表2 56对SRAP引物组合名称 Table 2 Fifty-six pairs of SRAP primer combination

2.3 分子遗传图谱的构建

利用MapMaker 3.0和QTL IciMapping 3.1软件, 取LOD值为3.0, 构建了一张覆盖9个连锁群(用Ch加数字表示连锁群), 包括141个标记位点的分子连锁图谱(123个SRAP和18个SSR), 其中未连锁上的有4个标记位点(3个SRAP和1个SSR), 偏分离标记用*标记(图2)。图谱覆盖基因组总长度为1399.88 cM, 标记间平均间距9.92 cM, 平均标记数15.67个。连锁群的标记位点为3~26个, 长度是15.69~237.21 cM, 其中Chr.1最长, 为237.21 cM, Chr.9最短, 为15.69 cM。每个连锁群间标记的个数和密度有较大差别, Chr.3标记数目最多, 包含26个标记; Chr.9标记数目最少, 只含3个标记; Chr.2标记密度最大, 标记平均间距为4.10 cM; Chr.7标记密度最小, 标记平均间距为19.47 cM (表6)。通过遗传学公式计算得知甜菜遗传图谱的预期长度是1499.15 cM, 框架图谱覆盖率是93.38%。在31个偏分离的标记中有29个表现出连锁遗传关系, 分布于5个不同的连锁群上, 有的形成了偏分离区域(3个或3个以上偏分离标记紧密连锁的区域, SDR)。分布在Chr.3连锁群上的SDR最多, 为3个, 分别有5个、6个、4个标记。Chr.1、Chr.9连锁群上的只有1个SDR, 且较小, 只有3个标记。

表3 SSR引物序列名称 Table 3 SSR primers and their sequences

表4 SSR 偏分离标记卡方检验 Table 4 Chi-square test of SSR with segregation distortion

3 讨论

3.1 SRAP标记和SSR标记

SRAP属于显性标记, 具有操作简便、低成本、多位点以及基因组分布均匀等优点, 因而被广泛应用于植物遗传多样性分析以及遗传育种等研究中。现已成功应用在黄瓜、棉花、油菜、棉花、烟草等作物中^{[ 14, 15, 16, 17]}。甜菜分子标记方面, SRAP只应用到遗传多样性分析, 如王茂芊等^{[ 18]}利用SRAP分子标记对我国甜菜三大产区骨干材料的遗传多样性分析。但在甜菜图谱构建方面, 尚未见报道。本研究利用SRAP构建的连锁图只是初步的框架图, 目的是检验SRAP在甜菜作图中的可靠性。本试验表明SRAP在甜菜F₂群体中的多态性较好, 适合甜菜遗传图谱的构建。而且SRAP标记在不同物种之间的使用可以相互借鉴, 将成为甜菜研究领域的一种有力工具。

表5 SRAP偏分离标记卡方检验 Table 5 Chi-square test of SRAP with segregation distortion

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	图1 偏分离标记分离程度柱形图横坐标: 标记名称; 纵坐标: -lg P。Abscissa: marker name; Ordinate: -lg P.Fig. 1 Separation column chart of loci with segregation distortion

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	图2 甜菜F₂代分子遗传图谱Fig. 2 Molecular genetic linkage map of F₂ sugar beet

表6 分子标记在分子遗传图谱上的分布 Table 6 Distribution of molecular markers on genetic map

为提高图谱构建饱和度, 本试验还采用了共显性标记SSR。SSR具有数量丰富、多态性好、呈共显性、对DNA要求不高、操作简便、重复性以及可靠性好等优点^{[ 19]}, 成为目前构建遗传连锁图谱、品种鉴定及品种选育的理想工具。现已广泛应用于玉米、大豆、花生等作物的研究中^{[ 20, 21, 22]}, 在甜菜方面, 吴则东等^{[ 23]}经优化, 建立了适合甜菜的SSR反应体系; 王华忠等^{[ 24]}利用9对SSR引物组合分析了49份甜菜材料的遗传多样性。2010年, 沙红等^{[ 25]}采用RAPD和SSR分子标记技术, 构建了包含7个连锁群, 16个标记位点的甜菜遗传图谱, 覆盖418.9 cM, 定位了3个与甜菜高糖性状有关的QTL。本试验中20对SSR引物获得19个多态性标记, 有1对SSR引物ms0331在亲本间表现多态性, 在F₂群体中却不分离, 这可能与群体类型、群体大小、标记类型等有关。

3.2 遗传图谱

本研究利用SRAP和SSR两种标记相结合方法构建了一张包含141个标记, 覆盖9个连锁群的甜菜分子遗传图谱。图谱长度长的原因可能是由于虽然两种标记的多态性位点较多, 却不够密集, 如Chr.7长度为194.79 cM, 标记数只有10个, 平均间距 19.47 cM; 有些连锁群标记间距过大, 如Chr.3中标记16到17间距为56.2 cM, 第25和26标记间的间距为94.6 cM, 均表明所构建图谱密度有待于加密。本试验构建的遗传图谱包含了9个连锁群, 虽然尚未完全覆盖甜菜基因组, 但与甜菜实际连锁群个数一致, 且平均间距较小, 为9.92 cM, 是目前国内覆盖率最大最完善的一张甜菜连锁图谱, 对于我国甜菜的分子育种具有一定应用价值。目前我们正继续应用多种标记对F₂群体构图, 将尽快构建出一张分布更均匀和位点密度较高的甜菜遗传连锁图谱。

3.3 偏分离

偏分离遗传现象在自然界普遍存在, 甘蓝^{[ 26]}、大豆^{[ 27]}、高粱^{[ 28]}等的分子遗传连锁图谱构建中都有表现。Knox等认为环境因素、非同源重组、基因转换和转座因子等也可能是引起偏分离的重要因素^{[ 29]}。本研究F₂作图群体连锁群上共有29个标记偏分离(26个SRAP和3个SSR), 占连锁群标记数的20.56%, 偏分离稍高可能与我们采用的亲本遗传距离相对较远, 或配子体选择性较强有关。偏分离标记中SRAP标记占22.22%, SSR标记占15.78%, 主要是由于显性标记比共显性标记易发生偏分离^{[ 30]}。本试验中有的连锁群上形成了偏分离区域, 可能与配子选择关系有关, 从而导致偏分离的方向趋于一致, 如Chr.3中的SDR的标记都偏向父本, Chr.9和Chr.2上的偏分离标记都偏向母本, 表明偏分离方向相同的标记之间连锁性可能更高。偏分离标记可能会影响连锁的检验, 但偏分离的标记和正常的分离标记有相同的作图效率, 去掉偏分离标记的遗传连锁图可能会使该区域的基因不能定位在连锁图上^{[ 31]}。本试验中就出现类似的现象, 去除偏分离标记后连锁图谱个数与实际个数不符, 反之加上偏分离标记构建出了与实际连锁群个数相等的9个连锁图谱, 表明在多态性标记较少的情况下可以考虑将偏分离标记用于作图。

4 结论

本研究构建了一张包括9个连锁群, 141个标记位点的分子连锁图谱, 标记间平均间距9.07 cM。有4个标记未进入连锁群, 连锁群的标记位点数在3~26个之间, 长度为15.69~237.21 cM, 其中Chr.1最长, Chr.9最短。连锁群上有20.56%的标记出现偏分离, 主要集中在Chr.3连锁群上, 形成3个偏分离区域, 其余分散在Chr.1、Chr.2、Chr.8和Chr.9中。偏分离标记中, SRAP标记占22.22%, SSR标记占15.78%; 并有23个偏向父本, 占连锁群偏分离标记的79.31%, 6个偏向母本, 占20.69%。图谱覆盖基因组总长度为1399.88 cM, 甜菜遗传图谱的预期长度是1499.15 cM, 框架图谱覆盖率是93.38%。SRAP在甜菜F₂群体中的多态性较好, 将成为甜菜图谱构建的有力工具。

The authors have declared that no competing interests exist.

作者已声明无竞争性利益关系。The authors have declared that no competing interests exist.

参考文献

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1	Halldén C, Hjerdin A, Rading I M, Sall T, Fridlundh B, Johannisdottir G, Tuvesson S, Akesson C, Nilsson N O. A high density RFLP linkage map of sugar beet. Genome, 1996, 39: 634-645 [本文引用:1] [JCR: 1.668]
2	Barzen E, Mechelke W, Ritter E, Seitzer J F, Salamini F. RFLP markers for suger beet breeding: chromosomal linkage maps and location of major genes for rhizomania resistance, monogermy and hypocotil colour. Plant J, 1995, 2: 601-611 [本文引用:1] [JCR: 6.582]
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5	Schneider K. Mapping QTLs for sucrose content, yield and quality in sugar beet population fingerprinted by EST-related markers. Theor Appl Genet, 2002, 104: 1107-1113 [本文引用:1] [JCR: 3.658]
6	Grimmer M K, Trybush S, Hanley S, Francis S A, Karp A, Ashe M J C. An anchored linkage map for sugar beet based on AFLP, SNP and RAPD markers and QTL mapping of a new source of resistance to beet necrotic yellow vein virus. Theor Appl Genet, 2007, 114: 1151-1160 [本文引用:1] [JCR: 3.658]
7	陈昆松, 李方, 徐昌杰, 张上隆, 傅承新. 改良CTAB法用于多年生植物组织基因组DNA的大量提取. 遗传, 2004, 26(4): 529-531 Chen K S, Li F, Xu C J, Zhang S L, Fu C X. An efficient macro-method of genomic DNA isolation from Actinidia chinensis leaves. Hereditas (Beijing), 2004, 26(4): 529-531 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.928]
8	赵娟. 辣椒分子连锁图谱的构建及抗黄瓜花叶病毒QTL定位. 内蒙古农业大学硕士学位论文, 2009. pp24-25 Zhao J. Construction of Molecular Linkage Map and QTL Analysis of Cucumber Mosaic Virus Resistance in Pepper. MS Thesis of Inner Mongolia Agricultural University, 2009. pp24-25 (in Chinese with English abstract) [本文引用:1]
9	王华忠, 吴则东, 王晓武, 方智远. 利用SRAP与SSR标记分析不同类型甜菜的遗传多样性. 作物学报, 2008, 34: 37-46 Wang H Z, Wu Z D, Wang X W, Fang Z Y. Analysis of the genetic diversity in different types of sugar beets by SRAP and SSR markers. Acta Agron Sin, 2008, 34: 37-46 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
10	赵尚敏. 甜菜抗丛根病种质资源遗传多样性分析. 内蒙古农业大学硕士学位论文, 2009. pp31-34 Zhao S M. Study on the Genetic Diversity of Germplasm Resource of Rhizomania Resistance Sugar Beet. MS Thesis of Inner Mongolia Agricultural University, 2009. pp31-34 (in Chinese) [本文引用:1]
11	宋宪亮, 孙学振, 张大真. 偏分离及对植物遗传作图的影响. 农业生物技术学报, 2006, 14: 286-292 Song X L, Sun X Z, Zhang D Z. Segregation Distortion and Its Effect on Genetic Mapping in Plants. J Agric Biotechnol, 2006, 14: 286-292 (in Chinese with English abstract) [本文引用:1]
12	Postlethwait J H, Johnson S L, Midson C N, Talbot W S, Gates M, Ballinger E W, Africa D, Andrews R, Carl T, Eisen J S. A genetic linkage map for the zebrafish. Science, 1994, 264: 699-703 [本文引用:1]
13	Chakravarti A, Lasher L K, Reefer J E. A maximum likelihood for estimating genome length using genetic linkage data. Genetics, 1991, 128: 175-182 [本文引用:1] [JCR: 4.389]
14	唐玉海, 郭春芳, 张木清. 相关序列扩增多态性(SRAP)标记及其应用研究进展. 生物技术通报, 2006, (增刊-1): 237-241 Tang Y H, Guo C F, Zhang M Q. Sequence-related amplified polymorphism (SRAP) markers and applications. Biotechnol Bull, 2006, (suppl-1): 237-241 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.49]
15	林忠旭, 张献龙, 聂以春. 新型标记SRAP在棉花F₂分离群体及遗传多样性评价中的适用性分析. 遗传学报, 2004, 6: 622-626 Lin Z X, Zhang X L, Nie Y C. Evaluation of application of a new molecular marker SRAP on analysis of F₂ segregation population and genetic diversity in cotton. Acta Genet Sin, 2004, 6: 622-626 (in Chinese with English abstract) [本文引用:1]
16	李媛媛, 沈金雄, 王同华, 傅廷栋, 马朝芝. 利用SRAP、SSR和AFLP标记构建甘蓝型油菜遗传连锁图谱. 中国农业科学, 2007, 40: 1118-1126 Li Y Y, Shen J X, Wang T H, Fu T D, Ma C Z. Construction of a linkage map using SRAP, SSR and AFLP markers in Brassica napus L. Sci Agric Sin, 2007, 40: 1118-1126 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.889]
17	马红勃, 祁建民, 李延坤, 梁景霞, 王涛, 兰涛, 陈顺辉, 陶爱芬, 林荔辉, 吴建梅. 烟草SRAP和ISSR分子遗传连锁图谱构建. 作物学报, 2008, 34: 1958-1963 Ma H B, Qi J M, Li Y S, Liang H X, Wang T, Lan T, Chen S H, Tao A F, Lin Z H, Wu J M. Construction of a molecular genetic linkage map of tobacco based on SRAP and ISSR markers. Acta Agron Sin, 2008, 34: 1958-1963 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
18	王茂芊, 吴则东, 王华忠. 利用SRAP标记分析我国甜菜三大产区骨干材料的遗传多样性. 作物学报, 2011, 37: 811-819 Wang M Q, Wu Z D, Wang H Z. Genetic diversity of major sugar beet varieties from three regions of China with SRAP markers. Acta Agron Sin, 2011, 37: 811-819 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
19	唐荣华, 张君诚, 吴为人. SSR分子标记的开发技术研究进展. 西南农业学报, 2002, 15(4): 106-109 Tang R H, Zhang J C, Wu W R. Progress in the way to develop SSR molecular marker. Southwest China J Agric Sci, 2002, 15(4): 106-109 (in Chinese with English abstract) [本文引用:1]
20	向道权, 曹海河, 曹永国, 杨俊品, 黄烈健, 王守才, 戴景瑞. 玉米SSR遗传图谱的构建及产量性状基因定位. 遗传学报, 2001, 28: 778-784 Xiang D Q, Cao H H, Cao Y G, Yang J P, Huang L J, Wang S C, Dai J R. Construction of a genetic map and location of quantitative trait loci for yield component traits in maize by SSR markers. Acta Genet Sin, 2001, 28: 778-784 (in Chinese with English abstract) [本文引用:1]
21	陈庆山, 张忠臣, 刘春燕, 王伟权, 李文滨. 应用Charleston×东农594重组自交系群体构建SSR大豆遗传图谱. 中国农业科学, 2005, 38: 1312-1316 Chen Q S, Zhang Z C, Liu C Y, Wang W Q, Li W B. Construction and analysis of soybean genetic map using recombinant inbred line of Charleston × Dongnong 594. Sci Agric Sin, 2005, 38: 1312-1316 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.889]
22	洪彦彬, 梁炫强, 陈小平, 刘海燕, 周桂元, 李少雄, 温世杰. 花生栽培种SSR遗传图谱的构建. 作物学报, 2009, 35: 395-402 Hong Y B, Liang X Q, Chen X P, Liu H Y, Zhou G Y, Li S X, Wen S J. Construction of genetic linkage map in peanut (Arachis hypogaea L. ) cultivars. Acta Agron Sin, 2009, 35: 395-402 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.667]
23	吴则东, 王华忠, 韩英. 甜菜SSR-PCR反应体系的优化. 中国糖料, 2008, (1): 11-13 Wu Z D, Wang H Z, Han Y. Optimization of sugarbeet SSR-PCR reaction system. Sugar Crops China, 2008, (1): 11-13 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.5408]
24	史树德, 魏磊, 张子义, 邵金旺, 田自华. 甜菜EST-SSR引物的开发与应用. 中国糖料, 2011, (3): 1-5 Shi S D, Wei L, Zhang Z Y, Shao J W, Tian Z H. Development and Utilization of EST-SSR Marker in Sugarbeet (Beta vulgaris L. ). Sugar Crops China, 2011, (3): 1-5 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.5408]
25	沙红, 王燕飞, 高文伟, 曲延英, 张立明. 甜菜离子注入诱变高糖性状的QTL分析. 中国糖料, 2010, (3): 27-28 Sha H, Wang Y F, Gao W W, Qu Y Y, Zhang L M. QTL analysis of impregnatingion into high-sugar mutant sugarbeet. Sugar Crops China, 2010, (3): 27-28 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.5408]
26	陆光远, 杨光圣, 傅廷栋. 甘蓝型油菜分子标记连锁图谱的构建及显性细胞核雄性不育基因的图谱定位. 遗传学报, 2004, 31: 1309-1315 Lu G Y, Yang G S, Fu T D. Linkage map construction and mapping of a dominant genic male sterility gene (Ms) in Brassica napus. Acta Genet Sin, 2004, 31: 1309-1315 (in Chinese with English abstract) [本文引用:1]
27	吴晓雷, 贺超英, 王永军, 张志永, 东方阳, 张劲松, 陈受宜, 盖钧镒. 大豆遗传图谱的构建和分析. 遗传学报, 2001, 28: 1951-1961 Wu X L, He C Y, Wang Y J, Zhang Z Y, Dong F Y, Zhang J S, Chen S Y, Gai J Y. Construction and analysis of a genetic linkage map of soybean. Acta Genet Sin, 2001, 28: 1951-1961 (in Chinese with English abstract) [本文引用:1]
28	赵姝华, 李玥莹, 邹剑秋, Folkertsma R, Hash T C. 高粱分子遗传图谱的构建. 杂粮作物, 2005, 25(1): 11-13 Zhao S H, Li Y Y, Zou J Q, Folkertsma R, Hash T C. Construction of a molecular genetic map of sorghum. Rain Fed Crops, 2005, 25(1): 11-13 (in Chinese with English abstract) [本文引用:1] [CJCR: 0.5562]
29	Knox M R, Ellis T H N. Excess heterozygosity contributes togenetic map expansion in pea recombinant inbred populations. Genetics, 2002, 162: 861-873 [本文引用:1] [JCR: 4.389]
30	刘海燕, 崔金腾, 高用明. 遗传群体偏分离研究进展. 植物遗传资源学报, 2009, 10: 613-617 Liu H Y, Cui J T, Gao Y M. Progress of segregation distortion. J Plant Genet Resour, 2009, 10: 613-617 (in Chinese with English abstract) [本文引用:1] [CJCR: 1.1628]
31	宋立君. 萝卜遗传连锁图谱构建与主要品质性状QTLs分析. 南京农业大学硕士学位论文, 2009. pp34-35 Song L J. Construction of a genetic linkage map and QTLs analysis of main quality traits in radish (Raphanus sativus L. ). MS Thesis of Nanjing Agricultural University, 2009. pp34-35 (in Chinese with English abstract) [本文引用:1]

1996

1.668

0.0

... 如1996年Hallden等^[1]利用RFLP构建了包括431个标记, 覆盖620 cM的甜菜遗传图谱, 标记间平均间距1 ...

1995

6.582

0.0

... 1995年Barzen^[2]构建了包含248个RFLP和50个RAPD位点, 全长815 cM的遗传连锁图谱, 并对抗丛根病基因、下胚轴颜色基因和单粒型性状基因进行定位 ...

1995

1.175

0.0

... Uphoff^[3]构建了包括85个RAPD标记位点的糖甜菜遗传图谱, 定位了下胚轴颜色基因、一年生基因及一个恢复基因 ...

1999

1.175

0.0

... 1999年Nisson等^[4]用221个AFLP和46个RFLP标记对204个植株群体进行了甜菜叶斑病抗性的QTL分析 ...

2002

3.658

0.0

Theor Appl Genet. 2002, 104(6-7):1107 - 1113 DOI:10.1007/s00122-002-0890-8

Mapping QTLs for sucrose content, yield and quality in a sugar beet population fingerprinted by EST-related markers

Schneider

.Mapping QTLs for sucrose content, yield and quality in sugar beet population fingerprinted by EST-related markers.Theor Appl Genet,2002,104:1107-1113

1.Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné Weg 10, 50829 K?ln, Germany
2.KWS SAAT AG, Grimsehlstra?e 31, 37555 Einbeck, Germany

... 2002年Schneider等^[5]对甜菜的含糖率、产量和品质进行了QTL分析 ...

2007

3.658

0.0

Theor Appl Genet. 2007, 114(7):1151 - 1160 DOI:10.1007/s00122-007-0507-3

An anchored linkage map for sugar beet based on AFLP, SNP and RAPD markers and QTL mapping of a new source of resistance toBeet necrotic yellow vein virus

M. K. Grimmer(1) S. Trybush(2) S. Hanley(2) S. A. Francis(1) A. Karp(2) M. J. C. Asher(1)

1.Broom’s Barn Research Station, Higham, Bury St Edmunds, IP28 6NP UK
2.Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK

Rhizomania, caused byBeet necrotic yellow vein virus (BNYVV), is an important sugar-beet disease worldwide and can result in severe losses of root yield and sugar content. We have identified a major QTL for BNYVV resistance from a new source in a segregating population of 158 individuals. The QTL explained an estimated 78% of the observed phenotypic variation and the gene conferring the partial resistance is referred to asRz4. AFLP was used in combination with bulked segregant analysis (BSA) to develop markers linked to the resistance phenotype. AFLP marker analysis was extended to produce a linkage map that was resolved into nine linkage groups. These were anchored to the nine sugar-beet chromosomes using previously published SNP markers. This represents the first anchored sugar-beet linkage map to be published with non-anonymous markers. The final linkage map comprised 233 markers covering 497.2 cM, with an average interval between markers of 2.1 cM. TheRz4 QTL and anRz1 RAPD marker were mapped to chromosome III, the known location of the previously identified BNYVV resistance genesRz1,Rz2 andRz3. The availability to breeders of new resistance sources such asRz4 increases the potential for breeding durable disease resistance.

... 2007年Grimmer和Trybush建立在AFLP、SNP、RAPD标记基础上的甜菜锚定连锁图和抗甜菜坏死黄脉病毒的QTL定位^[6] ...

2004

0.0

0.928

Hereditas. 2004, 26(4):529 - 531

An efficient macro-method of genomic DNA isolation from Actinidia chinensis leaves

改良CTAB法用于多年生植物组织基因组DNA的大量提取*An Efficient Macro-method of Genomic DNA Isolation from Actinidia chinensis Leaves

(陈昆松),(李方),(徐昌杰),(张上隆),(傅承新).(改良CTAB法用于多年生植物组织基因组DNA的大量提取).(遗传),2004,26(4):529-531

陈昆松1 李方1 徐昌杰1 张上隆1 傅承新2 CHEN Kun-Song1; LI Fang1; XU Chang-Jie1; ZHANG Shang-Long1; FU Cheng-Xin2

根据多年生植物组织富含多酚、多糖的具体特性，对现有的DNA提取方法进行了改进。通过增加提取缓冲液中b-巯基乙醇用量，简化氯仿/异戊醇抽提液步骤，改用经-20℃预冷异丙醇沉淀DNA等，对CTAB法加以改进。改进后方法具有以下优点：（1）获得的DNA质量良好，提取过程无明显的DNA降解，基本上排除了多酚物质的干扰；（2）用获得的DNA进行Southern杂交，可得到理想的杂交信号，可满足相关的分子研究要求；（3）操作简便。Abstract It is a difficult problem to isolate high quality DNA from plants containing a high contents of polyphenolics and polysaccharose, such as Actinidia plant. The protocol described in this paper is a modified CTAB (hexadecyltrimethylammonium bromide) method. High quality genomic DNA can be isolated from Actinidia plant using the improved method. The DNA is good enough for Southern blot and other uses in DNA research. The protocol is also efficient for quick and macro-DNA extraction.

... 2 DNA的提取在苗期每次取50份供试材料的单株幼嫩叶片2片, 用冷冻真空抽干机处理(-50℃左右, 36~48 h), 后研磨成粉末, 用CTAB法提取基因组DNA^[7] ...

2009

0.0

... 试验中选用的SSR引物和SRAP引物均来源于已发表的参考文献^[8,9,10], 并由北京华瑞生物技术公司合成, 引物序列和组合见表1、表2和表3 ...

2008

0.0

1.667

Acta Agron Sin. 2008, 34(1):37 - 46

Analysis of the Genetic Diversity in Different Types of Sugar Beets by SRAP and SSR Markers

利用SRAP与SSR标记分析不同类型甜菜的遗传多样性

WANG Hua-Zhong¹²,WU Ze-Dong¹²,WANG Xiao-Wu²,FANG Zhi-Yuan^1*

王华忠^1,2;吴则东^1,2;王晓武³;方智远^3,*

1 The Key Laboratory of Sugar Beet Genetic Breeding, Colleges of Heilongjiang Province/ Heilongjiang University, Harbin 150080, Heilongjiang; 2 Sugar Beet Research Institute of Chinese Academy of Agricultural Sciences/ Crop Academy of Heilongjiang University, Harbin 150080, Heilong- jiang; 3 Institute of Vegetable and Flower of Chinese Academy of Agricultural Sciences, Beijing 100081, China

Sugar beet (Beta vulgaris L.) is one of the most important sugar crops in the world. However, the application of molecular markers in sugar beet greatly lags behind that in other field crops. In the present study SRAP and SSR markers were employed to investigate the genetic diversity and relationships of sugar beet in order to make full use of germplasm reasonably, to select parents for breeding programs accurately, to improve breeding efficiency and to identify target cultivars quickly. We analyzed 49 sugar beet accessions, including monogerm lines (including male sterilities and maintainors), polygerm tetraploids, polygerm diploids, F₁of monogerm cross combinations, and foreign varieties introduced. All these materials are conserved at Sugar Beet Research Institute, CAAS. In a preliminary experiment, four accessions markedly distinct in phenotype were used to screen 64 pairs of SRAP primers and 11 pairs of SSR primers. Finally, 11 pairs of SRAP primers and nine pairs of SSR primers were selected and used for the present study. A total of 199 SRAP bands (including 86 polymorphic bands) were detected and one primer pair produced 18.0 bands on average (including 7.8 polymorphic bands). The ratio of polymorphic SRAP bands was from 33.3% to 62.5%, with an average of 43.7%. The selected SSR primers yielded 35 bands, with an average of 3.9 per primer pair, and the ratio of polymorphism was as high as 100%. The average genetic distance in the 49 accessions was 0.3860 and the average genetic similarity was 0.6795. About 30% of accessions were significantly different in average genetic distance or genetic similarity. The highest genetic similarity occurred between polygerm tetraploid lines (0.7264), followed by that between F₁ of monogerm cross combinations (0.7243) and that between foreign varieties introduced (0.7060), while the lowest one was found between monogerm lines (0.6837). The 49 accessions were divided into A, B, C and D groups based on cluster analysis. About 40% of accessions belonged to the D group which was divided into 4 subgroups, including diploids, tetraploids, F₁ of monogerm cross combinations and foreign varieties introduced, which also showed high genetic diversity. Thus, the combination method with SRAP and SSR markers is economic, effective and reliable to assess genetic diversity and relationships of sugar beet. The results indicate that sugar beet has a high level of heterogeneity. There are a definite difference between foreign genetic background and native genetic background, and the closer relationships and narrower genetic basis. Introducing foreign germplasm in breeding has shorted the gap between China and other countries in sugar beet genetic basis among cultivars in production. The classification based on the identification of biological and economical properties in field experiments was generally in accord with that based on SRAP and SSR molecular markers. Therefore, the application of the technology of molecular markers in sugar beet will accelerate breeding process and improve the quality, the yield and the content of sugar.

为选育优质甜菜新品种, 指导种质资源引进和利用, 为进行分子标记辅助选择育种提供科学依据, 采用SRAP和SSR两种分子标记方法相结合, 对甜菜单胚雄性不育系及保持系等49份材料进行遗传多样性分析。利用4个表型差异显著的甜菜品系对SRAP的64对引物组合及SSR的11对引物组合进行扩增, 分别筛选出有效引物组合11对和9对。SRAP的11对引物组合共产生199条扩增带, 其中有86条多态性带, 多态性带的比率平均为43.7%。SSR的9对引物共产生35条扩增带, 多态性比率为100%。全部材料的平均遗传距离为0.3860, 平均遗传相似系数为0.6795, 大约30%的材料遗传距离或遗传相似系数具显著或极显著差异。遗传相似系数平均值比较, 多胚四倍体品系0.7264＞单胚杂交组合0.7243＞国外品种0.7060＞多胚二倍体品系0.6908＞单胚品系0.6837。在遗传距离0.20处, 将49个甜菜材料划分为A、B、C、D 4个类群, D类群又分为4个亚类, 较好地显示了甜菜材料丰富的遗传多样性。表明不同甜菜品种具有相当高的异质性, 国外与国内材料的遗传基础存在一定差异, 但生产应用的甜菜品种间存在亲缘关系较近、遗传基础较窄的倾向。

... 试验中选用的SSR引物和SRAP引物均来源于已发表的参考文献^[8,9,10], 并由北京华瑞生物技术公司合成, 引物序列和组合见表1、表2和表3 ...

2009

0.0

... 试验中选用的SSR引物和SRAP引物均来源于已发表的参考文献^[8,9,10], 并由北京华瑞生物技术公司合成, 引物序列和组合见表1、表2和表3 ...

2006

0.0

... 1共显性)分离比例^[11] ...

1994

0.0

... 5 图谱预期长度和框架图谱覆盖率的计算按照Postlethwait^[12]方法估算遗传图谱长度G_e1=G_of+2s#cod#x000d7 ...

1991

4.389

0.0

... 根据Chakravarti^[13]的方法4, G_e2=G_of(m+1)/(m-1), m是连锁群上的总标记数 ...

2006

0.0

0.49

... 现已成功应用在黄瓜、棉花、油菜、棉花、烟草等作物中^{[14,15,16,17]} ...

2004

0.0

Yi Chuan Xue Bao. 2004, 31(6):622-6

[Evaluation of application of a new molecular marker SRAP on analysis of F2 segregation population and genetic diversity in cotton].

Lin ZX, Zhang XL, Nie YC

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China.

SRAP (Sequence-related Amplified Polymorphism), a new marker system, was applied in cotton genome analysis. We developed an efficient PCR reaction system for detecting SRAP that showed reliable, effective and reproductive. SRAP marker was used to detect the polymorphisms between 'Pima90' (G. barbodense) and ' Handan208' (G. hirsutum) and showed comparatively high polymorphism. Among the 30 primer pairs, 29 pairs generated 149 polymorphic bands with an average of 5.14 polymorphic bands per primer pair. The primer pair showing most polymorphism was the combination of me3 and em2, which produced 13 polymorphic bands. The SRAP marker was also tested among 11 upland cotton cultivars, 15 of the tested 30 primer pairs showed polymorphism yielding 22 polymorphic genetic loci, which is relatively high among upland cotton cultivars. The results showed that SRAP marker could be widely used in molecular genetic map construction and germplasm evaluation of cotton.

PMID: 15490882

... 现已成功应用在黄瓜、棉花、油菜、棉花、烟草等作物中^{[14,15,16,17]} ...

2007

0.0

1.889

Sci Agric Sin. 2007, 40(6):1118 - 1126

Construction of a Brassica napus L. linkage map using SRAP, SSR and AFLP markers

利用SRAP、SSR和AFLP 标记构建甘蓝型油菜遗传连锁图谱

(李媛媛),(沈金雄),(王同华),(傅廷栋),(马朝芝).(利用SRAP、SSR和AFLP标记构建甘蓝型油菜遗传连锁图谱).(中国农业科学),2007,40:1118-1126

李媛媛,沈金雄,王同华,傅廷栋,马朝芝

华中农业大学作物遗传改良国家重点实验室

A genetic linkage map of Brassica napus was constructed using 143 SSR, 118 AFLP, and 137 SRAP markers based on an F2 population of 184 individuals, which was developed by crossing “SI-1300” and “Eagle”. These markers were aligned in 21 linkage groups (LGs), and the map covered 1949.8 cM with an average marker interval of 4.9 cM. The public microsatellites (SSR markers) contributed to establish the links between the map of Parkin et al. and ours. Our results showed that most of LGs were aligned with the standard map except LG 20 and LG 21，and the distribution of markers on all LGs were uniform except N7 based on the analysis of Kolmogorov-Smirnov. However, the distorted markers were mainly cluster in LGs, and eight segregation distortion regions on seven LGs were detected. In addition, the results showed that SRAP markers may be a better type of markers than AFLP in map construction. In conclusion, according to the analysis of the marker distribution on LG, the map coverage and the marker density, the linkage map constructed in this study was of enough quality .Threrefore, this map will be a fine platform for the location of QTLs, studies on heterosis, and using the information of others.

【目的】建立甘蓝型油菜的高质量遗传图谱，为在分子水平上研究复杂性状打下基础并提供保证。【方法】以甘蓝型油菜自交不亲和系“SI-1300” 及其恢复系“Eagle”组配得到的184个F2单株为群体，利用SRAP、SSR和AFLP标记技术构建甘蓝型油菜遗传图谱。【结果】该图谱共包含21个连锁群，涉及137个SRAP标记、143个SSR标记和118个AFLP标记。图谱总长1 949.8 cM，标记间平均图距为4.9 cM。以SSR标记为锚定标记，该图谱与国际标准图谱进行了初步对应。研究共发现了8个偏分离区域。【结论】根据对标记分布的均匀程度，图谱覆盖程度以及标记密度等方面的分析，本研究构建的甘蓝型油菜分子遗传图谱质量较高，并且SRAP标记可能是比AFLP标记更适合于图谱构建的标记体系。

... 现已成功应用在黄瓜、棉花、油菜、棉花、烟草等作物中^{[14,15,16,17]} ...

2008

0.0

1.667

Acta Agron Sin. 2008, 34(11):1958 - 1963 DOI:10.3724/SP.J.1006.2008.01958

Construction of A Molecular Genetic Linkage Map of Tobacco Based on SRAP and ISSR Markers

烟草SRAP和ISSR分子遗传连锁图谱构建

MA Hong-Bo¹,QI Jian-Min^1*,LI Yan-Kun¹,LIANG Jing-Xia¹,WANG Tao²,LAN Tao¹,CHEN Shun-Hui³,TAO Ai-Fen¹,LIN Li-Hui¹,WU Jian-Mei¹

马红勃¹;祁建民^1,*;李延坤¹;梁景霞¹;王涛²;兰涛¹;陈顺辉³;陶爱芬¹;林荔辉¹;吴建梅¹

1College of Crop or Life science, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian; 2Fujian Shunchang Tobacco Company, Shunchang 353200, Fujian; 3Fujian Tobacco Company, Fuzhou 350003, Fujian, China

High-density genetic linkage maps are useful tools for studying genome, phylogeny and evolution, and for mapping and Cloning genes of interest. In this study, a molecular genetic map was constructed with F₂population consisting of 187 individuals from a cross of flue-cured tobacco cultivar Taiyan 7 and burley tobacco cultivar Bailei 21 by using a total of 513 SRAP primers and 42 ISSR primers. Approximately 58 out of 513 and 10 out of 42 primers, respectively, showed polymorphisms. Each of this polymorphic primers produced at least one scorable polymorphic DNA band which was visible enough for detection and scoring. The map consisted of 26 linkage groups which included 112 (92 SRAP and 10 ISSR) markers, and covered 1 560.2 cM with an average distance of 18.1 cM. Sixteen markers were still unlinked. All 26 linkage groups consisted of 2–20 markers ranging in length from 0–291.0 cM. A total of 24.1% distorted markers distributed in the map, mainly concentrated in the LG1 and LG4 linkage groups, and the others were mapped on diverse linkage groups respectively. This map will provide the basis for gene mapping and marker-assisted selection of important agronomic traits in tobacco.

采用烤烟品种台烟7号与白肋烟品种白肋21杂交，构建了187个单株的F₂遗传作图群体，利用所筛选的68个能扩增出多态性条带的SRAP和ISSR引物，对F₂作图群体进行PCR扩增和遗传连锁分析，初步构建了一张包含26个连锁群、112个(92个SRAP和20个ISSR)标记位点的烟草遗传连锁图谱。该图谱覆盖长度为1 560.2 cM，平均图距18.1 cM。有16个标记未进入连锁群。26个连锁群包含2~20标记不等，连锁群遗传距离0~291.0 cM。连锁群上有24.1%的标记出现偏分离，主要集中在LG1和LG4连锁群上，其余分散在不同连锁群。该图谱为烟草重要农艺性状的基因定位、以及分子标记辅助选择等研究奠定基础。

... 现已成功应用在黄瓜、棉花、油菜、棉花、烟草等作物中^{[14,15,16,17]} ...

2011

0.0

1.667

Acta Agron Sin. 2011, 37(5):811 - 819 DOI:10.3724/SP.J.1006.2011.00811

Analysis of Genetic Diversity of Major Sugar Beet Varieties from Three Regions of China with SRAP Markers

利用SRAP标记分析我国甜菜三大产区骨干材料的遗传多样性

WANG Mao-Qian^1,2,3, WU Ze-Dong^1,2,3, WANG Hua-Zhong^1,2,3,*

王茂芊^1,2,3,吴则东^1,2,3,王华忠^1,2,3,*

1 The Key Laboratory of Sugar Beet Genetic Breeding, Colleges of Heilongjiang Province / Heilongjiang University, Harbin 150080, Heilongjiang; 2 The Key Laboratory of North Sugar Crop Resource and Utilization, Chinese Academy of Agricultural Sciences, Harbin 150080, Heilongjiang; 3 Sugar Beet Research Institute of Chinese Academy of Agricultural Sciences / Crop Academy of Heilongjiang University, Harbin 150080, Heilongjiang

Shortage ofsugar beet germlasm resources results in the lag in researches of breeding and molecular biology. It is necessary to analyze the major varieties from three major production regions of China. Eighty-eight primer pairs were used to amplify the genomic DNA from leaves of four types of sugar beet varieties with different economic traits, which contain high yield type; high yield, low sugar and Rhizomania resistant type; standard type; medium yield, high sugar and anti-brown spot type, and 33 of which were obtained to be with availability. Two hundred and forty-one varieties from three major regions of China and nine varieties from abroad were detected with 33 primer pairs of SRAP markers. A total of 719 unambiguous bands were obtained, 459 of which were polymorphic. The average ratio of polymorphic bands was 63.8%. Compute over-all mean showed that genetic distance was 0.4165, genetic similarity among varieties was 0.6593, the genetic similarities were 0.7528 among foreign varieties, 0.6945 among monogerm varieties, 0.6816 among polygerm tetraploid varieties, and 0.6612 among polygerm diploid varieties. A total of 250 varieties were divided into four cluster groups based on cluster analysis by MEGA3.1 (at intercept of 0.2). Each genepool from three major region showed the highest. The varieties from China and abroad were classified into two different groups by POPGEN32. This indicated definite difference in the genetic background between foreign and native varieties.s of sugar beet production in China showed high level of genetic diversity, of which the Northeast genepool

利用SRAP分子标记, 选用甜菜中SRAP的88对引物组合分别对4个经济性状差异较大的代表性品系(高产型、高产低糖抗丛根病型、标准型、中产高糖抗褐斑病型)进行扩增，筛选出有效引物组合33对。采用筛选的33对引物检测全国三大产区的241份甜菜材料及9份外国材料，扩增到719条带，其中多态性条带459条，多态性条带的比率平均为63.8%。利用MEGA3.1软件中的Compute over-all mean计算，组内品种间平均遗传距离为0.4165，平均遗传相似系数为0.6593。遗传相似系数平均值为外国品种(0.7528)>单胚品系(0.6945)>多胚四倍体品系(0.6816)>多胚二倍体品系(0.6612)。利用MEGA3.1软件，在遗传距离0.20处，将供试材料分为4大类群。结果表明，我国三大产区供试材料遗传多样性丰富，其中东北产区优于华北与西北产区。利用POPGEN32软件将供试材料与外国品种分为两类，表明我国材料与外国品种的遗传基础存在较大差异。

... 甜菜分子标记方面, SRAP只应用到遗传多样性分析, 如王茂芊等^[18]利用SRAP分子标记对我国甜菜三大产区骨干材料的遗传多样性分析 ...

2002

0.0

... SSR具有数量丰富、多态性好、呈共显性、对DNA要求不高、操作简便、重复性以及可靠性好等优点^[19], 成为目前构建遗传连锁图谱、品种鉴定及品种选育的理想工具 ...

2001

0.0

Yi Chuan Xue Bao. 2001, 28(8):778-84

[Construction of a genetic map and location of quantitative trait loci for yield component traits in maize by SSR markers].

Xiang DQ, Cao HH, Cao YG, Yang JP, Huang LJ, Wang SC, Dai JR

Dept. of Plant Genetics and Breeding, China Agricultural University, Beijing 100094, China. xiangdq@263.net

Using 230 F2:3 lines derived from the hybrid of ND3138, a SSR genetic linkage map of maize (Zea mays L.) was constructed on which 80 markers distribute among 10 linkage groups and spanning maize genome about 2033.4 cM with average distance between markers being 25.42 cM. The 230 F2:3 lines were grown in a random design of one-row plot with three replication, and evaluated for ear length, bared tip, ear diameter, row number, kernel number row-1, one thousand grains weight, ear weight and grain weight plant-1. With interval mapping procedure, 30 QTLs were revealed for all traits, which distributed on 9 of 10 chromosomes of maize except chromosome 6 and accounted for from 9.5% to 55.3% of the total phenotypic variation. Genetic effects and gene action were also determined.

PMID: 11554353

... 现已广泛应用于玉米、大豆、花生等作物的研究中^[20,21,22], 在甜菜方面, 吴则东等^[23]经优化, 建立了适合甜菜的SSR反应体系 ...

2005

0.0

1.889

Sci Agric Sin. 2005, 38(7):1312 - 1316

Construction and Analysis of Soybean Genetic Map Using Recombinant Inbred Line of Charleston × Dongnong 594

应用Charleston×东农594重组自交系群体构建SSR大豆遗传图谱

,,,,

陈庆山,李文滨,张忠臣,刘春燕,王伟权

东北农业大学大豆研究所

F2:10 RIL population with 154 lines, crossed by Charleston as female parent and Dongnong 594 as male parent were used in this experiment. One hundred and sixty-four SSR primers were screened with the two parents and amplified on the 154 lines. A new soybean molecular genetic map, named NEAUSRI-GMS, was constructed by Mapmaker. The total length of the soybean genetic map is 1913.5 cM, and the average distance among markers is 11.89 cM. The length of linkage group varied from 0.4 cM to 309.5 cM, and the markers on the linkage group varied from 2 to 28. The distribution of SSR markers on every linkage group is not even. High density region of markers existed on linkage group A1, C2, and D1a. Compared with 5 soybean genetic maps constructed at home and abroad, NEAUSRI-GMS has high homologous with the public genetic map abroad.

以美国半矮杆大豆品种Charleston为母本，东北农业大学高蛋白大豆品系东农594为父本及其F2:10代重组自交系的154个株系为试验材料，利用164个在亲本之间表现多态的SSR引物对群体进行了分析，构建了一张大豆遗传图谱。该大豆遗传图谱总长度1 913.5 cM，标记间平均距离为11.89 cM。每个连锁群长度变动在0.4～309.5 cM之间，连锁群上的标记数在2～28个之间。各连锁群上的SSR标记并不是均匀分布的，其中A1、C2、D1a三个连锁群存在标记密集区。与国内外已构建完成的5张大豆遗传图谱比较表明，该图谱与国外的大豆公共遗传图谱对应性较好。

... 现已广泛应用于玉米、大豆、花生等作物的研究中^[20,21,22], 在甜菜方面, 吴则东等^[23]经优化, 建立了适合甜菜的SSR反应体系 ...

2009

0.0

1.667

Acta Agron Sin. 2009, 35(3):395 - 402 DOI:10.3724/SP.J.1006.2009.00395

Construction of Genetic Linkage Map in Peanut(Arachis hypogaea L.) Cultivars

花生栽培种SSR遗传图谱的构建

HONG Yan-Bin;LIANG Xuan-Qiang*;CHEN Xiao-Ping;LIU Hai-Yan;ZHOU Gui-Yuan;LI Shao-Xiong;WEN Shi-Jie

洪彦彬;梁炫强*;陈小平;刘海燕;周桂元;李少雄;温世杰

Crops Research Institute, GuangdongAcademy of Agricultural Sciences,Guangzhou 510640,China

Molecular genetic map is a fundamental organizational tool for genomic research. However, a comprehensive genetic linkage map for peanut cultivars has not yet been developed due toextremely low levels of DNA polymorphisms in cultivated peanut.In this study, 184 recombinant inbred lines (RIL), derived from a cross between two Spanish type peanut cultivars (Yueyou 13 × Fu95-5), were used as mapping population. A total of 652 pairs of genomic-SSR primers and 392 pairs of EST-SSR primers were used to detect the polymorphisms between the parental cultivars. Of them, 121 SSR primer pairs amplified 123 segregating loci and were selected to analyze the RIL population. A genetic linkage map consisting of 108 SSR loci (102 genomic-SSR and 6 EST-SSR) in 20linkage groups and covers 568 cM with an average distance of 6.45 cM of intermaker was constructed. By comparing the SSR genetic map ofArachis (A. duranensis ×A. stenosperma) with AA genome, 11 linkage groups in the linkage map constructed in this study were confirmed to have homology with 6 linkage groups of wild peanut species.

花生栽培种品种间分子多态性相对缺乏,至今未构建出较完整的分子遗传图谱。本研究以粤油13和阜95-5为亲本,通过杂交构建包含184个F6重组自交系的遗传作图群体。采用652对genomic-SSR引物和392对EST-SSR引物对亲本进行多态性检测,从中筛选出121对多态性引物,在亲本中共检测到123个多态性位点。利用作图群体对多态性SSR位点进行遗传连锁分析,获得包含108个SSR标记(102个genomic-SSR标记和6个EST-SSR标记),涉及20个连锁群,总长568 cM,平均图距为6.45 cM的花生栽培种遗传图谱。与前人构建的花生野生种(A. duranensis ×A. stenosperma, AA genome)SSR遗传图谱比较,初步确定本研究构建的遗传图谱中有11个连锁群与野生种遗传图谱的6个连锁群存在同源关系。

... 现已广泛应用于玉米、大豆、花生等作物的研究中^[20,21,22], 在甜菜方面, 吴则东等^[23]经优化, 建立了适合甜菜的SSR反应体系 ...

2008

0.0

0.5408

Sugar Crop China. 2008, (1):11 - 13

Optimization of Sugarbeet SSR-PCR Reaction System

甜菜SSR-PCR反应体系的优化

WUZe-dong;WANGHua-zhong;HANYing

吴则东;王华忠;韩英

为了建立适宜甜菜的SSR反应体系,笔者以甜菜不育系TB005A为试验材料,研究了甜菜SSR技术中PCR反应体系的主要成分对SSR扩增结果的影响.对SSR反应体系中的DNA模板浓度、引物浓度、Taq聚合酶浓度、dNTPs浓度以及退火温度进行了探索,确立了适合甜菜的SSR反应体系为:在20μL反应体系中,模板DNA为60ng,引物(50ng/μL)1.2μL,Taq DNA聚合酶1.0U,dNTPs(2.5mM/L)0.6μL.并利用该反应体系对30个甜菜不同品系进行SSR反应,用6%的变性聚丙烯酰胺凝胶电泳检测,不同品系间DNA谱带多态性丰富,证实该体系稳定可靠.

... 现已广泛应用于玉米、大豆、花生等作物的研究中^[20,21,22], 在甜菜方面, 吴则东等^[23]经优化, 建立了适合甜菜的SSR反应体系 ...

2011

0.0

0.5408

Sugar Crop China. 2011, (3):1 - 5

Development and Utilization of EST-SSR Marker in Sugarbeet (Beta vulgaris L.)

甜菜EST-SSR引物的开发与应用

SHIShu-de^[1];WEILei^[1];ZHANGZi-yi^[2];SHAOJin-wang^[3];TIANZi-hua^[1]

史树德^[1];魏磊^[1];张子义^[2];邵金旺^[3];田自华^[1]

利用NCBI公共数据库现有的甜菜(Beta vulgaris L.)表达序列标签(expressed sequence tags,EST)数据信息,开发了甜菜EST-SSR标记.在所有的29830条甜菜EST序列中共确认得到20109条非冗余EST序列,总长为11287.6kb.在含有微卫星重复的6951条EST序列中按照SSR引物设计要求,最终获得了2845个EST-SSR,平均每3.96 kb含有1个SSR.EST-SSR的分布频率和特征分析表明,A/T单碱基重复最多,其次是AAG/CTT三核苷酸重复,AG/CT二核苷酸重复,ACCTCC/AGGTGG等六核苷酸重复最少.随机合成了100对SSR引物,并分别选用6个甜菜品种进行多态性检验,将其按遗传相似性分为两组,多态信息含量(polymorphism information content,PIC)平均值为0.47.本研究证实这种全新的开发甜菜SSR标记的方法具有高效、多态性较高的特点,在甜菜遗传多样性分析、功能基因定位、遗传图谱构建以及比较基因组等研究方面有广阔的利用前景.

... 王华忠等^[24]利用9对SSR引物组合分析了49份甜菜材料的遗传多样性 ...

2010

0.0

0.5408

Sugar Crop China. 2010, (3):27 - 28

QTL Analysis of Impregnating Ion into High-sugar Mutant Sugarbeet

甜菜离子注入诱变高糖性状的QTL分析

SHAHong^[1];WANGYan-fei^[1];GAOWen-wei^[2];QUYan-ying^[2];ZHANGLi-ming^[1]

沙红^[1];王燕飞^[1];高文伟^[2];曲延英^[2];张立明^[1]

利用离子注入诱变突变体高糖材料S10与中糖F104构建高糖F2群体,通过800条RAPD,20对SSR引物的筛选及F2群体63个单株田间性状的调查,运用Maprnaker/expversion3.0和QTL2.0软件进行高糖性状QTL分析,获得了包含16个标记位点的7个连锁群,覆盖甜菜染色体总长度的418.9cM,确定了3个与甜菜高糖性状相关的QTL位点

... 2010年, 沙红等^[25]采用RAPD和SSR分子标记技术, 构建了包含7个连锁群, 16个标记位点的甜菜遗传图谱, 覆盖418 ...

2004

0.0

Yi Chuan Xue Bao. 2004, 31(11):1309-15

[Linkage map construction and mapping of a dominant genic male sterility gene (Ms) in Brassica napus].

Lu GY, Yang GS, Fu TD

National Key Lab of Crop Genetic Improvement, National Sub-Center of Rapeseed Improvement Huazhong Agricultural University, Wuhan 430070, China.

A backcross population derived from a cross between the dominant genic male sterile line Rs1046A and the double-low cultivar 'Samourai' was used for linkage map construction in Brassica napus. A linkage map with a total length of 2 646 cM was developed using 138 AFLP markers, 83 SSR markers and one morphology trait, which were distributed over 18 major linkage groups, two triplets and one linkage pair. 11.7% of the mapped markers distorted from the expected 1:1 ratio. The dominant genic male sterility gene (Ms) was mapped in LG10 and surrounded by five closely linked AFLP markers. Meanwhile,two evident marker segregation distortion regions were observed in both LG8 and LG16. The map constructed in the present study and the mapping of Ms gene are highly valuable in designing marker-assisted breeding program for the genic male sterile two-type line in Brassica napus. They are also important to map-based cloning of the Ms gene and to better understanding of the mechanism of genic male sterility at molecular level.

PMID: 15651685

... 3 偏分离偏分离遗传现象在自然界普遍存在, 甘蓝^[26]、大豆^[27]、高粱^[28]等的分子遗传连锁图谱构建中都有表现 ...

2001

0.0

... 3 偏分离偏分离遗传现象在自然界普遍存在, 甘蓝^[26]、大豆^[27]、高粱^[28]等的分子遗传连锁图谱构建中都有表现 ...

2005

0.0

0.5562

... 3 偏分离偏分离遗传现象在自然界普遍存在, 甘蓝^[26]、大豆^[27]、高粱^[28]等的分子遗传连锁图谱构建中都有表现 ...

2002

4.389

0.0

Am. J. Respir. Crit. Care Med.. 2000, 162(3 Pt 1):861-4

Familial sarcoidosis is linked to the major histocompatibility complex region.

Schürmann M, Lympany PA, Reichel P, Müller-Myhsok B, Wurm K, Schlaak M, Müller-Quernheim J, du Bois RM, Schwinger E

Institute of Human Genetics, Lübeck University Medical School, Lübeck, Germany. schuerma@medinf.mu-luebeck.de

Sarcoidosis is a systemic granulomatous disorder associated with high CD4+ cell activity, but no pathogen is detectable. Clustering in families occurs, and the existence of a genetic predisposition to sarcoidosis is widely accepted. The major histocompatibility complex (MHC) is believed to contribute to this susceptibility. Many studies testing this hypothesis have produced conflicting results. We have genotyped 122 affected siblings from 55 families for seven DNA polymorphisms that flank and cover the MHC region on chromosome 6, and for HLA-DPB1, a candidate gene for granulomatous disorders. Multipoint nonparametric linkage (NPL) analysis showed linkage (NPL score > 2.5; p< 0.006) for the entire MHC region with a maximum NPL score of 3.2 (p = 0.0008) at marker locus D6S1666 in the Class III gene cluster. There was a significant excess of marker haplotype sharing among affected siblings. However, the frequency of HLA-DPB1 alleles on 104 shared chromosomes did not differ from that of a control group of founders from the family panel. Transmission disequilibrium was found for allele DPB1*0201, but only nine families contributed to this result. We conclude that genes of the MHC are involved in the genetic predisposition to sarcoidosis, but HLA-DPB1 alone does not sufficiently explain this fact.

PMID: 10988096

... Knox等认为环境因素、非同源重组、基因转换和转座因子等也可能是引起偏分离的重要因素^[29] ...

2009

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1.1628

... 78%, 主要是由于显性标记比共显性标记易发生偏分离^[30] ...

2009

0.0

... 偏分离标记可能会影响连锁的检验, 但偏分离的标记和正常的分离标记有相同的作图效率, 去掉偏分离标记的遗传连锁图可能会使该区域的基因不能定位在连锁图上^[31] ...