利用SSR分子标记构建四倍体杂交冰草的遗传连锁图谱

引用本文

姜志艳, 于肖夏, 于卓, 张志成, 石悦, 姜超. .利用SSR分子标记构建四倍体杂交冰草的遗传连锁图谱. 草业学报, 2016,25(6): 94-101
JIANG Zhi-Yan, YU Xiao-Xia, YU Zhuo, ZHANG Zhi-Cheng, SHI Yue, JIANG Chao. .Construction of a genetic linkage map for tetraploid hybrid wheatgrass using a SSR molecular marker. Acta Prataculturae Sinica,2016,25(6): 94-101 复制到剪切板

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利用SSR分子标记构建四倍体杂交冰草的遗传连锁图谱

姜志艳^1,², 于肖夏¹, 于卓^1*,, 张志成¹, 石悦¹, 姜超¹

1.内蒙古农业大学农学院,内蒙古呼和浩特 010019

2.内蒙古科技大学数理与生物工程学院,内蒙古包头 014010

*通信作者Corresponding author. E-mail:yuzhuo58@sina.com

作者简介:姜志艳(1976-),女,内蒙古包头人,讲师,博士。E-mail: L2359916@sina.com

收稿日期: 2015-09-06

基金项目:国家自然科学基金面上项目(31172254)资助

摘要

为构建四倍体杂交冰草分子遗传连锁图谱,对深入开展冰草产量、抗性等重要性状的QTL定位及分子标记辅助育种提供依据,以四倍体杂种F₂分离群体的347个单株及亲本蒙古冰草和航道冰草为材料,采用SSR分子标记技术和Joinmap 4.0软件进行了遗传作图研究。试验从256对SSR引物中筛选出条带清晰稳定、多态性丰富的适宜引物30对,PCR扩增得到224个SSR标记位点,平均每对引物扩增出7.47个位点,其中多态性标记位点185个,占82.6%。偏分离分析显示,在185个SSR多态性标记位点中有24个标记产生偏分离,占13.0%,符合植物遗传作图时通常偏分离标记比率<30%的要求,可用于遗传作图。构建了1张四倍体杂交冰草的分子遗传连锁框架图谱,该图谱包含14个连锁群、185个标记,其长度范围在123.0~202.6 cM之间,连锁群LG4最长、LG12最短,各连锁群的平均长度167.32 cM,覆盖基因组总长度2342.5 cM,标记间的平均距离12.66 cM。

关键词: 杂交冰草; 四倍体; SSR标记; 遗传连锁图谱

Construction of a genetic linkage map for tetraploid hybrid wheatgrass using a SSR molecular marker

JIANG Zhi-Yan^1,², YU Xiao-Xia¹, YU Zhuo^1,^*, ZHANG Zhi-Cheng¹, SHI Yue¹, JIANG Chao¹

1.Agronomy College,Inner Mongolia Agricultural University,Huhhot 010019,China

2.School of Mathematics,Physics and Biological Engineering,Inner Mongolia University of Science & Technology,Baotou 014010,China

Abstract

To establish a genetic linkage map in tetraploid hybrid wheatgrass genetic mapping was conducted using a simple sequence repeats (SSR) molecular marker technique with ‘Joinmap’ 4.0 software. 347 individuals from the F₂ segregating population and their parents were utilized, this helped lay the foundation for further study of marker-assisted breeding, and quantitative trait locus (QTL) location of important traits in wheatgrass, such as disease resistance and yield. Thirty optimal primers with clear, stable and high polymorphic bands were screened from 256 tested SSR primers. A total of 224 SSR loci were obtained from polymerase chain reaction (PCR) amplification with an average of 7.47 loci per primer, of which 185 were polymorphic loci, accounting for 82.6% of all loci. Segregation distortion analysis showed that a total of 24 loci were distorted, accounted for 13.0% of all (185) polymorphic loci, which met the requirement of <30% segregation distortion for plant genetic mapping. These polymorphic loci would be useful for genetic mapping. In this study, a molecular genetic linkage map of tetraploid hybrid wheatgrass was constructed which contained 185 loci and 14 linkage groups with length ranging from 123.0 to 202.6 cM. The longest and the shortest linkage group were LG4 and LG12, respectively. The average length of each linkage group was 167.32 cM, total length of the genome was 2342.5 cM and the average marker distance was 12.66 cM.

Keyword: hybrid wheatgrass; tetraploid; SSR marker; genetic linkage map

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冰草为多年生草本植物, 隶属于禾本科小麦族(Triticeae)。其抗逆性强、适应性广, 饲用和生态价值高, 可用于退化的天然草地改良、人工草地建植及公路和铁路护坡等水土保持^{[1, 2]}。四倍体杂交冰草(Agropyron mongolicum× A. cristatum cv. Fairway, 2n=4x=28)是课题组利用国产蒙古冰草(A. mongolicum, 2n=2x=14)与产自美国的航道冰草(A. cristatum cv. Fairway, 2n=2x=14)相组配, 人工去雄授粉杂交获得的二倍体杂种F₁, 经秋水仙碱溶液加倍染色体而育成的优良牧草, 其既有母本蒙古冰草耐寒抗旱、适应性强等优良特性, 又有父本航道冰草再生性和分蘖性强、耐倒伏、叶量大、营养价值和产量高的特点^{[3, 4, 5]}。

简单序列重复SSR(simple sequence repeat)是一项成熟的DNA标记技术, 因其具有重复性好、多态性丰富、标记呈共显性等优点, 在作物品种和杂种真实性鉴定、种质资源遗传差异分析、辅助选择育种、分子遗传连锁图谱建立等方面已得到应用^{[6, 7, 8, 9]}。由于大多数多年生禾本科或豆科牧草都具有多倍性和异花授粉等生物学特性^[10], 其复杂的遗传背景使得牧草的遗传作图、重要性状QTL定位及分子标记辅助育种研究工作相比作物进展较慢。1993年Brummer等^[11]最早利用RFLP分子标记构建了四倍体紫花苜蓿(Medicago sativa)的遗传连锁图, 之后在二倍体的羊茅(Festuca spp.)、黑麦草(Lolium spp.)、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12, 13, 14, 15]}, 而有关四倍体杂交冰草的遗传作图至今未见有研究报道。本试验以四倍体杂交冰草F₂群体为作图材料, 拟利用SSR分子标记技术构建四倍体杂交冰草分子遗传连锁图谱, 为深入开展四倍体冰草重要农艺性状的QTL定位、分子标记辅助育种及基因图位克隆等奠定基础。

1 材料与方法

1.1 供试材料

材料为四倍体杂交冰草F₂ 群体的单株及其亲本蒙古冰草和航道冰草, 种植于呼和浩特市赛罕区内蒙古农业大学农场的饲草材料圃内。试验于2014年5月— 2015年5月在田间和室内进行。

1.2 DNA的提取与纯度检测

从四倍体杂交冰草F₂群体内随机取347个单株及其父母本各10个单株的嫩叶, 用北京天根公司生产的植物基因组试剂盒提取DNA, 1.6%琼脂糖凝胶电泳检测其DNA纯度后稀释至50 ng/μ L, 置于-20℃冰箱中备用^[16]。

1.3 SSR-PCR扩增反应体系和程序

SSR-PCR扩增体系:反应液总体积为20 μ L, 其中含50 μ mol/L 的10× Buffer 2.0 μ L、2.5 mmol/L的 dNTP 2.0 μ L、25 mmol/L的MgCl₂ 1.8 μ L、50 μ mol/L的Forward primer和 Reverse primer各 1.0 μ L、5 U/μ L的Taq DNA聚合酶0.1 μ L、模板DNA 1.0 μ L、ddH₂O 11.1 μ L。SSR-PCR扩增程序:94℃ 5 min, 94℃ 30 s, 56℃ 30 s, 72℃ 1 min, 30个循环, 72℃延伸10 min^[17]。

1.4 PCR扩增产物检测及SSR引物来源

PCR反应结束后, 加入6.0 μ L变性剂, 于PCR仪上95℃变性5 min, 用7%的变性聚丙烯酰胺凝胶进行电泳, 功率70 W, 预电泳30 min, 电泳时间2 h。电泳胶板在乙醇-冰乙酸溶液中固定, AgNO₃溶液中染色, NaOH-甲醛溶液中显影, 风干后照相^[18]。

试验所用的256对SSR引物, 是从NCBI网站公布的禾本科作物小麦(Triticum aestivum)、玉米(Zea mays)和高粱(Sorghum bicolor)上开发的部分SSR引物中选取, 送至上海生物工程有限公司进行合成。

1.5 SSR多态性位点统计

按胶板点样顺序对每个位点上的多态性位点条带进行记录, 有带记为“ 1” , 无带记为“ 0” , 带型模糊不清或缺失的记为“ — ” , 生成“ 0” 和“ 1” 组成的原始矩阵进行统计分析。多态性标记位点百分率计算公式:P(%)=(K/N)× 100, 式中, K为多态性位点数目、N为观测位点总数^[19]。同一SSR引物PCR扩增2~3次, 统计电泳胶板条带清晰稳定后的多态性位点。

1.6 SSR标记偏分离统计

依据孟德尔分离规律, 四倍体杂交冰草F₂群体基因型理论分离比为3∶ 1, 将每个SSR标记的基因型数目进行卡方检验(χ ²测验), 凡P< 0.05的SSR标记均列入偏分离标记之内。

1.7 SSR分子遗传连锁图谱构建

将统计的SSR标记数据中的“ 0” 和“ 1” 原始矩阵分别转换成F₂群体所要求的数据格式, 应用计算机作图软件Joinmap 4.0, 构建以SSR分子标记为基础的四倍体杂交冰草的分子遗传连锁图谱。具体步骤:开New project命令创建一个新的文件夹; 选择Options, 设置参数即选择作图群体模式、群体数量和标记位点数; 导入数据, 选择LOD Groupings (tree) 窗口, 用Calculate命令进行标记连锁分组(LOD=3.0~10.0), 选用LOD≥ 3.0标记连锁组群, 运行Create Groups for Mapping命令得到用于作图的连锁群; 选择Calculate map命令构建连锁图谱; 用Map chart完成图谱的绘制。

2 结果与分析

2.1 各材料基因组DNA的纯度检测

用DNA试剂盒提取的四倍体杂交冰草F₂群体347个单株及其亲本的基因组DNA, 其电泳条带清晰稳定、无降解、无蛋白和RNA污染(图1), 表明各材料的DNA纯度高, 符合SSR分子标记试验的要求。

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	图1 F₂群体部分单株基因组DNA电泳纯度检测 P₁:蒙古冰草; P₂:航道冰草; 1-31:F₂群体部分单株。Fig.1 Genomic DNA electrophoresis purity test of partial plants of the F₂ population M:Marker DL 2000.P₁:A. mongolicum; P₂:A. cristatum cv. Fariway; 1-31:Partial plants of the F₂ population.

2.2 SSR引物筛选及多态性分析

用亲本蒙古冰草和航道冰草及其四倍体杂种F₂群体1个单株的基因组DNA为模板进行适宜引物筛选, 从256对SSR引物中选出条带清晰、重复性和多态性好的SSR适宜引物30对(表1, 图2)。

表1 适宜SSR引物碱基序列及其多态性标记位点 Table 1 Nucleotide sequence of SSR primers and polymorphic markers loci

引物 Primer	正向引物碱基序列 Forward primer nucleotide sequence	反向引物碱基序列 Reverse primer nucleotide sequence	扩增位点数 Amplified loci No.	多态位点数 Polymorphic loci No.	多态位点百分率 Polymorphic loci percentage (%)
AH4	5'-GCGAAACCAACAGCAAGG-3'	5'-CGAAGTCGGCGTTCATCTC-3'	7	5	71.4
AH15	5'-AGAGCGATCCCAGAAACA-3'	5'-GCAACGGGTGGTAGAAAT-3'	8	7	87.5
AH25	5'-TTGAACCGAAGAAAGGGAT-3'	5'-CAAAGCAGTAAGGGGAAA-3'	5	3	60.0
AH63	5'-CACCATCCTCCTCCTACCC-3'	5'-AGCAGCAGCCATCTCAAAA-3'	8	6	75.0
AH70	5'-TTGCACTACCGTGACTTT-3'	5'-AGAAACCACGCTCACATA-3'	6	5	83.3
AH71	5'-TGCCCAACCGAATCACAA-3'	5'-CTGGACGGAGCCACATCA-3'	10	10	100.0
AH85	5'-CAAGGCTGAGGTCAAGAA-3'	5'-GGAAGCACCATGAAACAC-3'	6	5	83.3
B3107	5'-CTTGTTCACCTTGCTTGCCTAT-3'	5'-ATGCGTTTCTGCTCTGTTCACT-3'	5	5	100.0
B3252	5'-CGCCGAGTCCCAGTAGTGTT-3'	5'-CGAGGGCAATCCAGAGTTCA-3'	10	8	80.0
Brac124	5'-TGCACCCCTTCCAAATCT-3'	5'-TGCGAGTCGTGTGGTTGT-3'	5	4	80.0
Barc1060	5'-GCGTCTATTTTTGCCATTTCCA-3'	5'-GCGATGTTCTGTAGTTCTTAGTGTT-3'	5	4	80.0
C12	5'-CGCCGAGTCCCAGTAGTGTT-3'	5'-CGAGGGCAATCCAGAGTTCA-3'	12	10	83.3
C2743	5'-CCTACTACTGGCGGATGAAG-3'	5'-CTACTCCTGGACAGCGAAAC-3'	9	7	77.8
C3269	5'-TTTCCACTCGGCTCTTG-3'	5'-CCAGTTTGGCAGTTTCG-3'	7	7	100.0
C5511	5'-GCAGCAACACCAATCCTA-3'	5'-GCAGCAACACCAATCCTA-3'	5	5	100.0
D965	5'-CTTTGAGAAGTGCCAGACG-3'	5'-GGGAATAGCAGCAACGAA-3'	9	8	88.9
D5750	5'-CGCCGAGTCCCAGTAGTGTT-3'	5'-CGAGGGCAATCCAGAGTTCA-3'	6	6	100.0
F3974	5'-ATGCCTACGCCTACAACAA-3'	5'-CAACGGACGAAACGAGAA-3'	11	9	81.8
F2359	5'-GGATTTCCAGCCCTACCA-3'	5'-CTTTCCACCGATTCACCC-3'	9	8	88.9
S68	5'-AACCGAACCAAACCAAGC-3'	5'-GCAGAGCAACGCACAGAA-3'	8	6	75.0
Tc68268	5'-CAGTGTTCTTGGAGCTGCTG-3'	5'-GCAAACGAATAAGCAGGGTC-3'	7	5	71.4
Tc69675	5'-ACACAACATAGCGCATGGAA-3'	5'-CCTGTGGGTTTTGCTGAGAT-3'	7	5	71.4
Tc85804	5'-CCTCCTCCGACTCCTTTTTC-3'	5'-GCTCTGGTCTGTTTCCCTGT-3'	7	6	85.7
Txp83	5'-CGCTAGTCAAAATTGAGGAG-3'	5'-CAAACCGATAAACAAGTATG-3'	8	6	75.0
Txp158	5'-GCCATCAGGGATTGAC-3'	5'-GAGCTTCGCTGAGTAAGG-3'	6	6	100.0
Txp160	5'-AGTCGGCTGGTAAAGTGG-3'	5'-GGATATGGGATGGTTTGG-3'	7	5	71.4
Txp353	5'-CTCCAACATGCTCTGCTG-3'	5'-GGATCAACTTATTGGGGATG-3'	6	5	83.3
Wmc506	5'-CACTTCCTCAACATGCCAGA-3'	5'-CTTTCAATGTGGAAGGCGAC-3'	9	7	77.8
Wmc617	5'-CCACTAGGAAGAAGGGGAAA-3'	5'-ATCTGGATTACTGGCCAACTGT-3'	7	5	71.4
Xgwm169	5'-GTGCTCTGCTCTAAGTGTGGG-3'	5'-ACCACTGCAGAGAACACATACG-3'	9	7	77.9
合计Total			224	185	82.6

表1 适宜SSR引物碱基序列及其多态性标记位点 Table 1 Nucleotide sequence of SSR primers and polymorphic markers loci

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	图2 用F₂的1个单株及双亲的DNA对部分SSR引物筛选结果 F₂:F₂ 的1个单株; P₁:蒙古冰草; P₂:航道冰草。Fig.2 SSR primer screening result with one plant in F₂ population and parents F₂:One plant of hybrid F₂; P₁:A. mongolicum; P₂:A. cristatum cv. Fariway.

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图3 引物D965(A)和F3974(B)对F₂分离群体部分单株及其亲本DNA的扩增结果
P₁:蒙古冰草; P₂:航道冰草; 1-55:F₂群体部分单株。Fig.3 DNA amplified results of partial plants of F₂ segregating population in hybrid wheatgrass and their parents using the primer D965(A)and F3974(B)
M:Marker DL 1000.P₁:A. mongolicum; P₂:A. cristatum cv. Fariway; 1-55:F₂ partial plants.

利用这30对引物对F₂作图群体347个单株及亲本的DNA进行PCR扩增, 共获得224个位点, 平均每对引物扩增出7.47个位点, 其中多态性标记位点185个, 平均多态性标记位点百分率为82.6%, 表明该四倍体杂交冰草F₂群体的SSR多态性丰富(图3, 表1)。

2.3 SSR作图标记的分离及遗传连锁图谱特征分析

SSR标记分离及连锁图谱的基本特征如表2和图4所示。经卡方检验表明, 当P< 0.05时, 在已获得的185个SSR多态性标记位点中, 有161个标记符合孟德尔分离比例, 其占标记总数的87.0%。另有24个SSR标记产生偏分离, 偏分离比例仅为13.0%, 符合植物遗传作图时通常偏分离标记比率< 30%的要求^[20], 这185个SSR分子标记均可用于遗传作图。

利用Joinmap 4.0软件对185个SSR标记进行连锁分析, 构建出1张四倍体杂交冰草的SSR分子遗传连锁框架图谱, 该图谱包含14个连锁群、185个标记, 14个连锁群的长度范围在123.0~202.6 cM之间, 连锁群LG12最短、LG4最长, 各连锁群的平均长度为167.32 cM, 总长度为2342.5 cM, 标记间的平均距离为12.66 cM。

表2 四倍体杂交冰草SSR标记分离及遗传连锁图谱的基本特征 Table 2 The segregation of SSR markers and the basic characteristics of genetic map in tetraploid hybrid wheatgrass

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	图4 四倍体杂交冰草SSR分子遗传连锁框架图谱Fig.4 SSR genetic linkage map of tetraploid hybrid wheatgrass

3 讨论

高密度的分子遗传连锁图谱是深入开展植物重要性状QTL定位及分子标记辅助育种等研究的基础^{[21, 22]}。与小麦、玉米、水稻(Oryza sativa)等农作物构建的遗传连锁图相比, 目前国内外已构建的牧草遗传连锁图谱的图幅均偏小, 且标记间空隙较大, 使得遗传图谱在牧草分子标记辅助育种及重要性状QTL定位等研究方面较薄弱。本课题组曾利用蒙古冰草× 航道冰草杂种F₂分离群体构建了含7个连锁群、175个分子标记(AFLP标记152个、RAPD标记23个)的二倍体杂交冰草分子遗传图谱, 其全长为416 cM, 平均标记间密度为2.47 cM^[15]; 本试验构建了含14个连锁群、185个SSR标记的四倍体杂交冰草分子遗传连锁图谱, 其全长为2342.5 cM, 远超过二倍体杂交冰草图谱的长度, 表明四倍体的基因组明显大于二倍体; 但标记间的密度(12.66 cM)小于二倍体, 而且在各连锁群上标记的分布密度不均匀, 如连锁群LG6上的标记数较多, 密集了40个SSR标记, 而连锁群LG8和LG9上SSR标记数均只有3个, 这说明该图谱是一张SSR框架图谱, 还有待于进一步扩大SSR标记数量或与其他分子标记如AFLP及SRAP等进行整合, 来增加图谱中的分子标记数目、加大图谱密度, 目前课题组正在开展四倍体杂交冰草遗传连锁图谱的加密研究。

遗传图谱精确度的高低很大程度取决于构建群体的大小, 理论上作图群体越大, 作图精度就越高, 但若作图群体过大, 就会使实验工作量和费用相应增加, 故选择合适的群体数目很重要^{[23, 24]}。从作图效率看, 作图群体的大小主要决定于3个方面:一是参照随机分离的结果计算出最大图距, 二是两个标记间能够测算出重组的最小图距, 三是所选用作图群体的类型。为了能够提高作图精度, 在构建植物分子标记连锁图时需考虑作图群体的类型, 一般认为作图所需的群体大小的顺序为F₂> Rl> BC和DH^[25]。目前在牧草上遗传图谱构建所用的作图群体在100~200个之间, 本试验选用四倍体杂交冰草F₂分离群体的347个单株进行遗传作图研究, 主要考虑多年生牧草的多倍性, 其数量性状分离复杂且表型受环境影响较大, 因此控制试验误差显得尤为重要。

4 结论

试验筛选出条带清晰、重复性和多态性好的SSR适宜引物30对。对四倍体杂交冰草F₂群体347个单株及亲本的基因组DNA进行PCR扩增得到185个SSR标记。构建了1张四倍体杂交冰草SSR分子遗传连锁框架图谱, 其14个连锁群的长度变幅123.0~202.6 cM, 覆盖基因组总长度2342.5 cM, 标记间的平均距离12.66 cM。

The authors have declared that no competing interests exist.

参考文献

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[1]	云锦凤, 李瑞芬, 米福贵. 冰草的远缘杂交及杂种分析. 草地学报, 1997, 5(4): 221-227. [本文引用:1]
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[4]	包美莲. 蒙古冰草与航道冰草正反交杂种染色体加倍植株F₂遗传性状分析[D]. 呼和浩特: 内蒙古农业大学, 2012. [本文引用:1]
[5]	于肖夏, 姜志艳, 于卓, 等. 四倍体杂交冰草新品种—蒙杂冰草1号的选育. 草业科学, 2015, 32(5): 738-744. [本文引用:1]
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[15]	李小雷. 几种披碱草种间杂种F₁遗传特性及冰草分子图谱构建研究[D]. 呼和浩特: 内蒙古农业大学, 2008. [本文引用:2]
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[17]	张自强, 于肖夏, 鞠天华, 等. 3个马铃薯杂种优良株系的核型及SSR分析. 西北植物学报, 2014, 34(7): 1318-1324. [本文引用:1]
[18]	于卓, 谢锐, 于肖夏, 等. 低氢氰酸含量高丹草新品系及其亲本的SSR分析. 草业学报, 2014, 23(1): 223-228. [本文引用:1]
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1997

0.0

. 1997, 5(4):221-227

冰草的远缘杂交及杂种分析

云锦凤, 李瑞芬, 米福贵

Inner Monglia Institute of Agricultural and Animal Husbandry, Huhhot 010018

Agropyron mongolicum keng., the narrow linear-spiked diploid species (2n = 14) from Inner Mongolia was hybridized with the broad pectinate-spiked diploid (2n = 14) A.cristatum cv.Fair-way from Montana state of America.Interspecific hybridization was conducted for combining their hard re-sistan gene from A.mongoliicum and superior quality gene from Fairway.the F 1 hybrid was diploid and morhpologically intermediate to to their parents.Chromosome pairing at metaphase I in the hybrid averaged2.09I, 4.86Ⅱ,0.48Ⅲ,0.24Ⅳand 0.19Ⅴ per cell,demonstrating that the two parental genomes are very similar.The F 1 hybrid was partially fertile so that pe opportunity was offered for gene exchange and transfer.

蒙古冰草隶属于冰草属的二倍体种(2n＝14),原产于内蒙古。航道冰草是从北美引进的栽培品种隶属于扁稳冰草种,亦为二倍体种(2n＝14)。为了将蒙古冰革的优良抗性基因和航道冰草的优良品质基因相结合,进行二倍体种间的远缘杂交。杂种F1全部为二倍体,其形态学特征介于双亲之间。杂种F1PMCMⅠ的染色体平均配对构型为:2.09Ⅰ、4.86Ⅱ、0.48Ⅲ、0.24Ⅳ及0.19Ⅴ。杂种F

... 其抗逆性强、适应性广,饲用和生态价值高,可用于退化的天然草地改良、人工草地建植及公路和铁路护坡等水土保持^[1,2] ...

2009

0.0

... 其抗逆性强、适应性广,饲用和生态价值高,可用于退化的天然草地改良、人工草地建植及公路和铁路护坡等水土保持^[1,2] ...

2006

0.0

. 2006, 14(1):14-19

蒙古冰草#cod#x000D7;航道冰草杂种F 4 代株系同工酶分析

权威, 于卓, 王月华

College of Agronomy, Inner Mongolia Agriculture University, Huhhot, Inner Mongolia Autonomous Region 010019, China

The genetic differences of 11 lines of F 4 hybrid between Agropyron Mongolicum and A.cristatum were studied using isozyme electrophoresis technique.The results showed that there were certain differences in the number,locus and intensity of the esterase（EST）,proxidase（POD） and superoxide dismutase（SOD） isozyme bands among the tested materials at different developing stages.The differences of isozyme zymograms phenotypes were important genetic markers in protein level to the 11 lines of F 4 and parents;the isozyme zymograms characteristics of tested plants at earing and tillering stages in the summer and the autumn could accurately reflect their genetic differences and enhance the preciseness of identified results.According to the genetic distance standard of 0.40,the 13 tested plants were divided into 6 groups:lines 6,7,8,9 and 10 were in the first group,lines 4 and 5 in the second group,lines 1,3 and 2 in the third group,lines 11 was in the forth group,the male parent A.cristatum in the fifth group,and the female parent Agropyron mongolicum in the sixth group.The results had important theoryetic significance for clustering and breeding new wheatgrass cultivars.

利用同工酶电泳技术研究蒙古冰草与航道冰草种间杂交F 4 代11个株系的遗传差异性。结果表明:在相同或不同生育阶段,供试材料的酯酶同工酶(EST)、过氧化物酶同工酶(POD)和超氧化物岐化酶(SOD)的位点、数目和强弱均存在差异,其表型差异是11个株系间及其与亲本间在蛋白质分子水平识别的重要遗传标记;选择抽穗期、分蘖期取样分析的同工酶酶谱特征,更能体现材料间酶谱表型的遗传差异性和提高鉴定结果的准确性;以遗传距离(GD)值0.40为基准,将13个材料聚类成6类:第1类株系6、7、9、8和10,第2类株系4和5,第3类株系1、3和2,第4类株系11,第5类父本航道冰草,第6类母本蒙古冰草;该研究结果对冰草杂交后代新品系的归类选育具有理论意义。

... Fairway,2n=2x=14)相组配,人工去雄授粉杂交获得的二倍体杂种F₁,经秋水仙碱溶液加倍染色体而育成的优良牧草,其既有母本蒙古冰草耐寒抗旱、适应性强等优良特性,又有父本航道冰草再生性和分蘖性强、耐倒伏、叶量大、营养价值和产量高的特点^[3,4,5] ...

2012

0.0

2015

0.0

. 2015, 32(5):738-744 DOI:10.11829\j.issn.1001-0629.2014-0378

四倍体杂交冰草新品种#cod#x02014;蒙杂冰草1号的选育

于肖夏, 姜志艳, 于卓

Agronomy College, Inner Mongolia Agriculture University, Huhhot010019, China

The superior tetraploid hybrid wheatgrass variety SZB02 was selected from colchicine induced chromosome doubling F1 hybrids after widely crossing of Agropyron cristatum cv. Fairway and Agropyron mongolicum. And then, the new variety of tetraploid hybrid wheatgrassMengza No.1 was obtained after a series of tests including varieties comparison, regional and production which lasted 13 years. Mengza No.1 is a perennial gramineous herb with fibrous root system, regular plant growth, lanceolate leaf length, and wide panicle type. The fresh grass yield, dry grass yield and seed yield of Mengza No.1 with twice mowing per year was 25 938.35 kg·hm-2, 9 552.33 kg·hm-2 and 802.37 kg·hm-2, respectively, which suggest that this variety has relatively high grass yield. Mengza No.1 has crude protein content of 16.13 % in anthesis, and is rich in nutritional composition such as crude fat, nitrogen free extract, calcium and phosphorus. It also has characteristics of soft texture, high leaf yield and good palatability which can be used for green fodder, graze and hay. It has strong resistance to low temperature in earlier development stage and wide ecological adaptability, which is suitable to be applied in the areas with the effective accumulated temperature more than 2 200 ℃·d (≥10 ℃).

将二倍体航道冰草（Agropyron cristatum cv. Fairway）与蒙古冰草（Agropyron mongolicum）进行种间远缘杂交获得的杂交种F1代种子，利用秋水仙碱诱导染色体加倍，通过选育得到四倍体杂交冰草优良新品系SZB02，经品比、区域和生产试验，历时13年育成四倍体蒙杂冰草1号新品种。该品种为多年生疏丛型禾本科草本植物，须根系、植株生长整齐、叶长披针形、穗型较宽大。蒙杂冰草1号产量较高，年刈割两次的鲜草总产量平均为25 938.35 kg·hm-2，干草产量平均为9 552.33 kg·hm-2，种子产量平均为802.37 kg·hm-2。其开花期粗蛋白质含量高达16.13%，并富含粗脂肪、无氮浸出物、钙和磷等营养成分，营养价值高；草质柔软、叶量大、适口性好，可青饲、放牧、调制干草等；发育前期耐低温能力强，生态适应性广，适宜于在≥10 ℃有效积温≥2 200 ℃·d的地区推广种植。

2005

0.0

. 2005, 112(1):158-166

Isolation, characterization and mapping of simple sequence repeat markers in Zoysiagrass (Zoysia spp.).

Cai H W , Inoue M , Yuyama N

Japan Grassland Agriculture and Forage Seed Association, Forage Crop Research Institute, 388-5 Higashiakada, Nasushiobara, Tochigi 329-2742, Japan. hcai@jfsass.or.jp

The genus Zoysia consists of 16 species that are naturally distributed on sea coasts and grasslands around the Pacific. Of these, Zoysia japonica, Zoysia matrella, and Zoysia tenuifolia are grown extensively as turfgrasses, and Z. japonica is also used as forage grass in Japan and other countries in East Asia. To develop simple sequence repeat (SSR) markers for zoysiagrass (Zoysia spp.), we used four SSR-enriched genomic libraries to isolate 1,163 unique SSR clones. All four libraries contained a high percentage of perfect clones, ranging from 67.1 to 96.0%, and compound clones occurred with higher frequencies in libraries A (28.6%) and D (11.6%). From these clones, we developed 1,044 SSR markers when we tested all 1,163 SSR primer pairs. Using all 1,044 SSR markers, we tested one screening panel consisting of eight Zoysia clones for testing PCR amplifications, from which five unrelated clones, among the eight, were used for polymorphism assessment, and found that the polymorphic information content ranged from 0 (monomorphic loci) to 0.88. Of the 1,044 SSR markers, 170 were segregated in our mapping population and we mapped 161 on existing amplified fragment length polymorphism-based linkage groups, using this mapping population. These SSR markers will provide an ideal marker system to assist with gene targeting, quantitative trait locus mapping, variety or species identification, and marker-assisted selection in Zoysia species.

PMID: 16235048

... 简单序列重复SSR(simple sequence repeat)是一项成熟的DNA标记技术,因其具有重复性好、多态性丰富、标记呈共显性等优点,在作物品种和杂种真实性鉴定、种质资源遗传差异分析、辅助选择育种、分子遗传连锁图谱建立等方面已得到应用^[6,7,8,9] ...

2003

0.0

2014

0.0

1998

0.0

2002

0.0

... 由于大多数多年生禾本科或豆科牧草都具有多倍性和异花授粉等生物学特性^[10],其复杂的遗传背景使得牧草的遗传作图、重要性状QTL定位及分子标记辅助育种研究工作相比作物进展较慢 ...

1993

0.0

... 1993年Brummer等^[11]最早利用RFLP分子标记构建了四倍体紫花苜蓿(Medicago sativa)的遗传连锁图,之后在二倍体的羊茅(Festuca spp ...

2003

0.0

. 2003, 108(1):25-40

A linkage map of meadow fescue (Festuca pratensis Huds.) and comparative mapping with other Poaceae species.

Alm V , Fang C , Busso C S

Department of Chemistry and Biotechnology, Agricultural University of Norway, PO. Box 5040, 1432, As, Norway.

A genetic linkage map has been constructed for meadow fescue ( Festuca pratensis Huds.) (2n=2 x=14) using a full-sib family of a cross between a genotype from a Norwegian population (HF2) and a genotype from a Yugoslavian cultivar (B14). The two-way pseudo-testcross procedure has been used to develop separate maps for each parent, as well as a combined map. A total number of 550 loci have been mapped using homologous and heterologous RFLPs, AFLPs, isozymes and SSRs. The combined map consists of 466 markers, has a total length of 658.8 cM with an average marker density of 1.4 cM/marker. A high degree of orthology and colinearity was observed between meadow fescue and the Triticeae genome(s) for all linkage groups, and the individual linkage groups were designated 1F-7F in accordance with the orthologous Triticeae chromosomes. As expected, the meadow fescue linkage groups were highly orthologous and co-linear with Lolium, and with oat, maize and sorghum, generally in the same manner as the Triticeae chromosomes. It was shown that the evolutionary 4AL/5AL translocation, which characterises some of the Triticeae species, is not present in the meadow fescue genome. A putative insertion of a segment orthologous to Triticeae 2 at the top of 6F, similar to the rearrangement found in the wheat B and the rye R genome, was also observed. In addition, chromosome 4F is completely orthologous to rice chromosome 3 in contrast to the Triticeae where this rice chromosome is distributed over homoeologous group 4 and 5 chromosomes. The meadow fescue genome thus has a more ancestral configuration than any of the Triticeae genomes. The extended meadow fescue map reported here provides the opportunity for beneficial cross-species transfer of genetic knowledge, particularly from the complete genome sequence of rice.

PMID: 12923626

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

2007

0.0

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

2005

0.0

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

2008

0.0

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

... 47 cM^[15] ...

2012

0.0

... L,置于-20℃冰箱中备用^[16] ...

2014

0.0

... SSR-PCR扩增程序:94℃ 5 min,94℃ 30 s,56℃ 30 s,72℃ 1 min,30个循环,72℃延伸10 min^[17] ...

2014

0.0

. 2014, 23(1):223-228 DOI:10.11686/cyxb20140127

低氢氰酸含量高丹草新品系及其亲本的SSR分析

于卓, 谢锐, 于肖夏

College of Agronomy,Inner Mongolia Agricultural University,Hohhot 010019,China

To identify the difference degree of four new strains of sorghum-sudangrass with low hydrocyanic acid content SLCN-11,SLCN-12,SLCN-13 and SLCN-14,and their parents at a DNA level,we conducted SSR analyses using the variety Mengnong Qingsi No.3 as a control. A total of 12 SSR primer pairs with high polymorphism and good reproducibility were screened from 200 developed sorghum SSR primer pairs. The 10 materials were analyzed using these SSR primers. A total of 509 polymorphic loci were amplified,and the percentage of polymorphic loci was up to 87.30%. Each SSR polymorphic primer pairs could amplify clear and stable bands,which can be used as molecular basis for identifying the four new strains of sorghum-sudangrass. Genetic distance (GD) among 10 materials ranged from 0.3165-0.6692,with an average of using 0.5359; Using the GD value of 0.50 as a threshold,the 10 materials can be clustered into five groups: Mengnong qingsi No.3,loose spike sorghum,SLCN-11,SLCN-12 and SLCN-13; SLCN-14 and brown hull sudangrass; white hull sudangrass; black hull sudangras; red hull sudangrass. This study laid the groundwork for the next step breeding,registration and utilization of new sorghum-sudangrass varieties with low hydrocyanic acid content.

为明确4个低氢氰酸含量的高丹草新品系SLCN-11、SLCN-12、SLCN-13、SLCN-14及其亲本散穗高粱、黑壳苏丹草、白壳苏丹草、红壳苏丹草和棕壳苏丹草在DNA水平上的差异程度,以育成登记的高丹草品种蒙农青饲3号为对照,对其进行了SSR分析。试验从200个高粱SSR引物中筛选出多态性丰富、重复性好的12对引物,利用这些引物对10个材料进行PCR扩增共得到509个多态性位点,多态性百分率达87.30%。每对引物扩增的SSR指纹图清晰、稳定,可作为鉴别4个高丹草新品系及其亲本的分子依据。10个供试材料间的遗传距离(GD)变幅在0.3165~0.6692之间,平均为0.5359;以GD值0.50为基准,将10个材料划分为5类:蒙农青饲3号、散穗高粱、SLCN-11、SLCN-12、SLCN-13为一类;SLCN-14、棕壳苏丹草为一类;白壳苏丹草、黑壳苏丹草、红壳苏丹草各单独为一类。该研究为下一步低氢氰酸含量高丹草新品种育成及登记利用奠定了基础。

... 电泳胶板在乙醇-冰乙酸溶液中固定,AgNO₃溶液中染色,NaOH-甲醛溶液中显影,风干后照相^[18] ...

1973

0.0

. 1973, 70(12):3321-3323

Analysis of gene diversity in subdivided populations.

Nei M.

A method is presented by which the gene diversity (heterozygosity) of a subdivided population can be analyzed into its components, i.e., the gene diversities within and between subpopulations. This method is applicable to any population without regard to the number of alleles per locus, the pattern of evolutionary forces such as mutation, selection, and migration, and the reproductive method of the organism used. Measures of the absolute and relative magnitudes of gene differentiation among subpopulations are also proposed.

PMID: 4519626

... 100,式中,K为多态性位点数目、N为观测位点总数^[19] ...

2006

0.0

. 2006, 14(2):286-292

偏分离及对植物遗传作图的影响

宋宪亮, 孙学振, 张天真

National Key Laboratory of Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China

Segregation distortion, which is identified as a strong evolutionary force, is common in genetic mapping. In plants, the percentage, degree, origin and genetic effects of segregation distortion vary significantly with species, population types, crosses and marker types. The exhibition and common features, causes, methods of mapping segregation distortion loci, effects of segregation distortion on map construction and corresponding mapping strategies in plants, mainly in crops, are reviewed.

偏分离是遗传作图中的普遍现象，是一种强有力的进化动力。在植物方面，标记偏分离的比例、程度、来源、遗传效应等因物种、群体类型、杂交组合、标记类型等的不同而有很大变化。从偏分离的表现和共同特征、偏分离产生的原因、偏分离基因位点的标记定位、偏分离对遗传作图的影响及克服方法等方面，对植物(主要是作物)中的偏分离研究进展进行了综合分析。

... 30%的要求^[20],这185个SSR分子标记均可用于遗传作图 ...

2014

0.0

. 2014, 36(10):1036-1042 DOI:10.3724/SP.J.1005.2014.1036

利用向日葵重组自交系构建遗传图谱

张永虎, 于海峰, 侯建华

1. Agricultural College, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Huhhot 010031, China

A genetic linkage map of sunflower was constructed by combined applying the SSR and AFLP markers using 187 F 5:6 individuals of recombinant inbred lines (RILs) which derived from the cross between Helianthus annuus K55 and Helianthus annuus K58 through single-seed descent (SSD). Using 78 pairs of SSR primers and 48 pairs of AFLP primer, 341 and 1119 bands were amplified, respectively. Among these 1460 bands, 557 bands (39.52%) were polymorphic, including 184 bands by SSR markers and 393 bands by AFLP markers. In the group of these polymorphic bands, 84 bands from SSR markers and 108 bands from AFLP markers showed the genetic distortion ( P = 0.05). A total of 192 segregation distortion markers were obtained in this study. By using the JoinMap 4.0 software to do the linkage analysis, a genetic linkage map was established with length of 2759.4 cM, consisted of 17 linkage groups, and comprised of 495 polymorphic molecular markers including 170 segregation distortion markers. The mean marker interval distance is 5.57 cM between markers. In addition, the number of markers in the linkage groups varied from 5 to 72, and the length of linkage groups were from 68.88 cM to 250.17 cM. The genetic map developed in the present study could be used for QTL mapping and gene cloning of sunflower important genes.

以向日葵自选系K55为母本、K58为父本杂交组合,通过单粒传得到的187个F 5：6 代重组自交系群体为作图材料,联合应用SSR和AFLP标记构建遗传连锁图谱。经过78对SSR引物和48对AFLP引物组合选择性扩增,分别得到341和1119条带,共1460条,分别获得多态性条带184条和393条,共577条多态性条带,占所有条带的39.52%。SSR和AFLP标记各有84个和108个多态性标记偏离孟德尔分离比例( P =0.05),共192个偏分离标记。采用JoinMap4.0软件进行连锁分析,构建了1张总长度为2759.4 cM、包含17个连锁群、连锁495个多态性标记的遗传图谱,其中偏分离标记170个,标记间的平均图距为5.57 cM。每个连锁群上分布有5~72个标记,长68.88~250.17 cM。本图谱为向日葵永久性图谱,为向日葵重要性状QTL定位和基因克隆奠定基础。

... 3 讨论高密度的分子遗传连锁图谱是深入开展植物重要性状QTL定位及分子标记辅助育种等研究的基础^[21,22] ...

2015

0.0

. 2015, 24(8):118-129 DOI:10.11686/cyxb2015071

不同水分环境条件下小麦粒重QTL定位及遗传分析

胡亮亮, 叶亚琼, 吕婷婷

1.Gansu Provincial Key Lab of Aridland Crop Science, College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2.College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China

To better understand the molecular quantitative genetic and QTL patterns affecting thousand-grain weight (TGW) in wheat ( Triticum aestivum ) in different water environments, QTL mapping and genetic analysis were performed for TGW using a mixed linear model approach. TGW was evaluated for recombinant inbred lines (RIL) with 120 progenies from a cross between Longjian 19 (drought tolerant) and Q9086 (drought sensitive) under different water regimes in three environments. Phenotypic expression of TGW in the RILs was highly sensitive to water status and showed wide variation and transgressive segregation. TGW was found to be subject to complex quantitative genetic regulation by minor-effect polygenes, which were easily affected by water environments. A total of 19 additive QTL (A-QTL) and 38 pairs of epistatic QTL (AA-QTLs) were detected for TGW in wheat, distributed on all chromosomes including 1A, 3B, 4D and 6A. Expression of these QTLs influenced the phenotypic variation of TGW resulting in both up- and down-regulation. The magnitude of these effects on TGW ranged from 1.24%-10.94% and 0.38%-2.89%, respectively. Three A-QTLs, Qtgw.acs -1 B .1, Qtgw.acs -2 A .1 and Qtgw.acs -4 A .1, were detected in multiple environments. In addition, four A-QTL hot-spot regions for TGW were also found at some specific locations, e.g., Xmag2064-Xbarc181 on chromosome 1B, Xwmc522-Xgwn122 on chromosome 2A, Xwmc446-Xgwm610 on chromosome 4A and Xwmc603-Xbarc195 on chromosome 7A. Most of the interaction effects of A-QTLs and AA-QTLs associated with drought-stressed environments were linked to down-regulation of the TGW variations. The additive and the additive×environment interaction effects may be the main genetic factors in TGW inheritance, and if so their expression would decrease TGW. The findings of this study should be useful for the genetic improvement of drought tolerance using molecular marker-assisted selection in wheat.

为探讨小麦千粒重(TGW)分子数量性状遗传,及QTL与水分环境互作关系,本文以抗旱性强的冬小麦品种陇鉴19与水地高产品种Q9086杂交创建的重组近交系(recombinant inbred lines,RIL)群体120个株系为供试材料,采用条件复合区间作图法对3个环境不同水分条件下TGW进行QTL定位和遗传分析。结果表明,小麦RIL群体TGW对水分环境反应敏感,群体中各株系呈现广泛变异和超亲分离,属于微效多基因控制的复杂数量性状,易受水分环境影响。共检测到19个和38对控制TGW的加性QTL(A-QTL)和上位性QTL(AA-QTL),分布在除1A、3B、4D和6A以外的其他17条染色体上。这些A-QTL和AA-QTL表达通过正向或负向调控影响TGW表型变异,贡献率分别在1.24%~10.94%和0.38%~2.89%。发现了3个多环境均能稳定表达的A-QTL( Qtgw.acs -1 B .1, Qtgw.acs -2 A .1和 Qtgw.acs -4 A .1),以及4个A-QTL热点区域 [Xmag2064-Xbarc181(1B),Xwmc522-Xgwn122(2A),Xwmc446-Xgwm610(4A)和Xwmc603-Xbarc195(7A)]。所检测到的A-QTL和AA-QTL与干旱胁迫环境互作普遍负向调控TGW表型。加性效应和加性与环境的互作效应是决定小麦TGW的主要遗传因子。在干旱胁迫条件下,这种遗传主效应均不同程度降低TGW表型。本研究结果可为小麦抗旱遗传改良和分子标记辅助选择育种奠定理论基础。

... 3 讨论高密度的分子遗传连锁图谱是深入开展植物重要性状QTL定位及分子标记辅助育种等研究的基础^[21,22] ...

1992

0.0

... 遗传图谱精确度的高低很大程度取决于构建群体的大小,理论上作图群体越大,作图精度就越高,但若作图群体过大,就会使实验工作量和费用相应增加,故选择合适的群体数目很重要^[23,24] ...

2010

0.0

. 2010, 121(6):1071-1082 DOI:10.1007/s00122-010-1372-z

Effects of missing marker and segregation distortion on QTL mapping in F₂ populations.

Zhang L Y , Wang S Q , Li H H

School of Mathematical Sciences, Beijing Normal University, Beijing, 100875, China.

Missing marker and segregation distortion are commonly encountered in actual quantitative trait locus (QTL) mapping populations. Our objective in this study was to investigate the impact of the two factors on QTL mapping through computer simulations. Results indicate that detection power decreases with increasing levels of missing markers, and the false discovery rate increases. Missing markers have greater effects on smaller effect QTL and smaller size populations. The effect of missing markers can be quantified by a population with a reduced size similar to the marker missing rate. As for segregation distortion, if the distorted marker is not closely linked with any QTL, it will not have significant impact on QTL mapping; otherwise, the impact of the distortion will depend on the degree of dominance of QTL, frequencies of the three marker types, the linkage distance between the distorted marker and QTL, and the mapping population size. Sometimes, the distortion can result in a higher genetic variance than that of non-distortion, and therefore benefits the detection of linked QTL. A formula of the ratio of genetic variance explained by QTL under distortion and non-distortion was given in this study, so as to easily determine whether the segregation distortion marker (SDM) increases or decreases the QTL detection power. The effect of SDM decreases rapidly as its linkage relationship with QTL becomes looser. In general, distorted markers will not have a great effect on the position and effect estimations of QTL, and their effects can be ignored in large-size mapping populations.

PMID: 20535442

2013

0.0

... BC和DH^[25] ...

1997

0.0

. 1997, 5(4):221-227

冰草的远缘杂交及杂种分析

云锦凤, 李瑞芬, 米福贵

Inner Monglia Institute of Agricultural and Animal Husbandry, Huhhot 010018

... 其抗逆性强、适应性广,饲用和生态价值高,可用于退化的天然草地改良、人工草地建植及公路和铁路护坡等水土保持^[1,2] ...

2009

0.0

... 其抗逆性强、适应性广,饲用和生态价值高,可用于退化的天然草地改良、人工草地建植及公路和铁路护坡等水土保持^[1,2] ...

2006

0.0

. 2006, 14(1):14-19

蒙古冰草#cod#x000D7;航道冰草杂种F 4 代株系同工酶分析

权威, 于卓, 王月华

College of Agronomy, Inner Mongolia Agriculture University, Huhhot, Inner Mongolia Autonomous Region 010019, China

2012

0.0

2015

0.0

. 2015, 32(5):738-744 DOI:10.11829\j.issn.1001-0629.2014-0378

四倍体杂交冰草新品种#cod#x02014;蒙杂冰草1号的选育

于肖夏, 姜志艳, 于卓

Agronomy College, Inner Mongolia Agriculture University, Huhhot010019, China

2003

0.0

2014

0.0

2005

0.0

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

2008

0.0

... )、高丹草(Sorghum-Sudangrass)及二倍体杂交冰草等牧草上构建了分子遗传连锁图谱^{[12,13,14,15]},而有关四倍体杂交冰草的遗传作图至今未见有研究报道 ...

... 47 cM^[15] ...

2012

0.0

... L,置于-20℃冰箱中备用^[16] ...

2014

0.0

... SSR-PCR扩增程序:94℃ 5 min,94℃ 30 s,56℃ 30 s,72℃ 1 min,30个循环,72℃延伸10 min^[17] ...

2014

0.0

. 2014, 23(1):223-228 DOI:10.11686/cyxb20140127

低氢氰酸含量高丹草新品系及其亲本的SSR分析

于卓, 谢锐, 于肖夏

College of Agronomy,Inner Mongolia Agricultural University,Hohhot 010019,China

... 电泳胶板在乙醇-冰乙酸溶液中固定,AgNO₃溶液中染色,NaOH-甲醛溶液中显影,风干后照相^[18] ...

2006

0.0

. 2006, 14(2):286-292

偏分离及对植物遗传作图的影响

宋宪亮, 孙学振, 张天真

National Key Laboratory of Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China

... 30%的要求^[20],这185个SSR分子标记均可用于遗传作图 ...

2014

0.0

. 2014, 36(10):1036-1042 DOI:10.3724/SP.J.1005.2014.1036

利用向日葵重组自交系构建遗传图谱

张永虎, 于海峰, 侯建华

1. Agricultural College, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Huhhot 010031, China

... 3 讨论高密度的分子遗传连锁图谱是深入开展植物重要性状QTL定位及分子标记辅助育种等研究的基础^[21,22] ...

2015

0.0

. 2015, 24(8):118-129 DOI:10.11686/cyxb2015071

不同水分环境条件下小麦粒重QTL定位及遗传分析

胡亮亮, 叶亚琼, 吕婷婷

... 3 讨论高密度的分子遗传连锁图谱是深入开展植物重要性状QTL定位及分子标记辅助育种等研究的基础^[21,22] ...

2013

0.0

... BC和DH^[25] ...