小麦-华山新麦草异代换系DH2322的分子细胞遗传学鉴定

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王秀娟, 陈新宏, 庞玉辉, 敬樊, 张军, 胡思远, 昝凯, 武军, 杨群慧, 赵继新. 小麦-华山新麦草异代换系DH2322的分子细胞遗传学鉴定. 41(2): 207-213
WANG Xiu-Juan, CHEN Xin-Hong, PANG Yu-Hui, JING Fan, ZHANG Jun, HU Si-Yuan, ZAN Kai, WU Jun, YANG Qun-Hui, ZHAO Ji-Xin. Molecular Cytogenetics Identification of a wheat Psathyrostachys huashanica Substitution Line DH2322 . Acta Agronomica Sinica, 41(2): 207-213 复制到剪切板

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小麦-华山新麦草异代换系DH2322的分子细胞遗传学鉴定

王秀娟, 陈新宏, 庞玉辉, 敬樊, 张军, 胡思远, 昝凯, 武军, 杨群慧, 赵继新^*

西北农林科技大学农学院, 陕西杨凌 712100

^*通讯作者(Corresponding author): 赵继新, E-mail: zhjx881@163.com

第一作者联系方式: E-mail: wangxiujuanluck@163.com

收稿日期:2014-12-19

基金:本研究由国家高技术研究发展计划(863计划)项目(2011AA100501), 陕西省自然科学基础研究计划重点项目(2013JZ007), 中央高校基本科研业务费(QN2011001)和西北农林科技大学唐仲英育种基金资助;

摘要

利用分子标记、细胞学、基因组原位杂交(GISH)等技术, 结合田间农艺性状调查, 对小麦-华山新麦草七倍体材料H8911与硬粒小麦D4286杂交F₄代分离群体中株系DH2322进行了综合鉴定。华山新麦草基因组特异SCAR标记鉴定表明, DH2322含有华山新麦草遗传物质; 细胞遗传学观察显示, DH2322染色体构型为2 n=42=21 II; 有丝分裂和花粉母细胞减数分裂中期I基因组原位杂交(GISH)鉴定表明, DH2322的染色体由40条小麦染色体和2条华山新麦草Ns染色体构成, 且2条Ns染色体能完全配对为一个二价体; SSR和STS分子标记分析表明, DH2322缺失小麦D基因组的2D染色体, 而含有华山新麦草的2Ns染色体; 农艺性状分析结果表明, DH2322具有双亲的形态学特征, 结实性好, 穗长和穗粒数显著大于亲本。

关键词: 普通小麦; 华山新麦草; 异代换系; SCAR; 基因组原位杂交(GISH); SSR; STS

Molecular Cytogenetics Identification of a wheat Psathyrostachys huashanica Substitution Line DH2322

WANG Xiu-Juan, CHEN Xin-Hong, PANG Yu-Hui, JING Fan, ZHANG Jun, HU Si-Yuan, ZAN Kai, WU Jun, YANG Qun-Hui, ZHAO Ji-Xin^*

College of Agronomy, Northwest A&F University, Yangling 712100, China

Abstract

A wheat- P. huashanica alien substitution line DH2322 was isolated from the F₄ progeny of heptaploid hybrid H8911 × Triticum durum cv. D4286 and identified by using SCAR markers, genomic in situ hybridization (GISH) , SSR markers, and STS markers. The results using SCAR marker RHS141 showed that DH2322 carrys chromosome derived from P. huashanica. Cytogenetics observation indicated that DH2322 has a chromosome karyotype of 2 n = 42 = 21 II. GISH showed that DH2322 is a line with 40 chromosomes from wheat and two Ns chromosomes from P. huashanica, which formed a ring bivalent at metaphase I in pollen mother cells (PMC). According to SSR and STS analysis, the wheat chromosome 2D in DH2322 was substituted by the chromosome 2Ns from P. huashanica. Evaluation of the agronomic traits showed that DH2322 presented favorable agronomic traits in spike length and kernels per spike. Development of this new germplasm will be useful for wheat breeding programs in the future.

Keyword: Triticum aestivum; Psathyrostachys huashanica; Alien substitution line; SCAR; GISH; SSR; STS

Show Figures

华山新麦草(Psathyrostachys huashanica Keng, 2n=14, NsNs)是禾本科(Gramineae)、小麦族(Triticeae)、大麦亚族(Hordeinae)、新麦草属(Psathyrostachys)多年生异花授粉植物, 具有抗寒、抗旱、耐瘠薄、早熟、优质、矮秆、抗病等优良特性^[1], 仅分布于我国秦岭山脉陕西华山段, 是我国特有种^[2], 也是小麦(Triticum aestivum, 2n=14, AABBDD)品种改良的优异种质资源。越来越多的研究者开始将研究目标瞄准华山新麦草的利用并取得了一些进展。Kang等^{[3, 4]}利用中国春ph2b基因突变体和华山新麦草杂交获得了含有ph2b基因的衍生后代、并利用面包小麦“ J-11” 和华山新麦草杂交获得一个八倍体属间杂种PHW-SA (2n=8x=56, AABBDDNsNs)。Wang等^[5]研究发现华山新麦草3Ns染色体上携带抗小麦条锈病的基因, Kang等^[6]将华山新麦草3Ns染色体导入小麦, 获得了抗条锈病的小麦-华山新麦草中间材料。另外, Liu等^[7]和Wang等^[8]利用简单重复序列标记(simple sequence repeats, SSR)技术对华山新麦草种群基因的多样性进行了分析。

本课题组自20世纪90年代初开始进行普通小麦品种7182与华山新麦草的杂交研究, 选育出普通小麦-华山新麦草七倍体材料H8911 (2n=49, AABBDDNs)^[1]。在此基础上, 侯文胜等^[9]、傅杰等^[10]、赵继新等^{[11, 12, 13]}、武军等^[14]利用细胞遗传学对小麦-华山新麦草异附加系、异代换系进行了分子细胞遗传学方面的研究; Du等^{[15, 16, 17, 18, 19, 20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系; 王秀娟等^[21]获得一个附加两条华山新麦草7Ns染色体的矮秆材料12DH25。

本研究以小麦-华山新麦草七倍体材料H8911与硬粒小麦品种D4286杂交的F₄分离群体中筛选的株系DH2322为材料, 从细胞遗传学、分子标记和农艺性状等方面对其进行研究, 以了解其遗传行为、染色体构成、所含华山新麦草染色体的同源群归属及形态特征等, 为小麦品种改良和染色体工程育种提供新的种质资源。

1 材料与方法

1.1 供试材料

普通小麦品种7182、华山新麦草(Psathyrostachys huashanica)、倍半二倍体H8911 (2n=49, AABBDDNs)及其与硬粒小麦(Triticum durum, 2n=28, AABB)品种D4286杂交并回交后选育的F₄代材料DH2322均来自陕西省植物遗传工程育种重点实验室。

1.2 农艺性状调查

2012年10月和2013年10月于西北农林科技大学试验田种植供试材料, 调查田间农艺性状和室内考种, 统计株高、旗叶面积、穗下茎节长度、穗长、小穗数、穗粒数和结实率。用SAS软件分析数据和检验差异显著性。

1.3 DNA提取

采用CTAB法^[22]提取普通小麦品种7182、华山新麦草、硬粒小麦品种D4286、普通小麦-华山新麦草七倍体材料H8911及DH2322的基因组DNA。

1.4 华山新麦草Ns基因组特异SCAR标记鉴定

为提高小麦背景中华山新麦草染色质的鉴定效率, 根据Du等^[23]开发的华山新麦草染色体特异SCAR标记RHS141, 以7182、D4286、DH2322、H8911和华山新麦草基因组DNA为模板进行扩增。在PTC-200扩增仪上进行PCR, 20 μ L总反应体系包括10× buffer (含Mg²⁺) 2.0 μ L、引物RHS141 (2.5 μ mol mL^-1) 2.0 μ L、模板DNA (200 μ g μ L^-1) 2.0 μ L、dNTP (2.5 μ mol mL^-1) 1.6 μ L、Taq DNA聚合酶(5 U μ L^-1) 0.2 μ L、ddH₂O 12.2 μ L。PCR程序为, 94℃预变性3 min; 94℃变性50 s, 60℃复性40 s, 72℃延伸 1 min, 35个循环; 72℃最后延伸10 min, 4℃保存。以1%琼脂糖凝胶135 V恒压条件下电泳30 min, 溴化乙锭(EB)染色, 显影后照相观察。

1.5 细胞遗传学分析

将种子于室内浸泡至露白, 转移到垫有湿润滤纸的培养皿内发芽。当根长至1~2 cm时, 用镊子夹取根尖放入0~4℃冰水中预处理24 h, 卡诺氏I固定液(无水乙醇∶ 冰醋酸= 3∶ 1)固定, -20℃冰箱保存。45%醋酸洋红染色压片, 用Olympus BX60显微镜观察根尖细胞染色体数, 并照相。

从田间取适龄幼穗(旗叶与倒二叶距离约为4 cm), 用卡诺氏II固定液(无水乙醇∶ 氯仿∶ 冰乙酸= 6∶ 3∶ 1)固定, 45%醋酸洋红染色压片, 用Olympus BX60显微镜观察花粉母细胞减数分裂中期I (PMC MI)的染色体构型, 并照相。

1.6 基因组原位杂交(GISH)

将DH2322的根尖和幼穗固定一周后用体积分数45%醋酸压片, 液氮揭片, 备用。

依照Dig-Nick-Translation Mix试剂盒(Roche, Germany)产品说明书对华山新麦草的全基因组DNA进行标记(4 μ L Dig-Nick-Translation Mix, 1 μ L华山新麦草DNA (1000 μ g μ L^-1), 加ddH₂O至20 μ L)。参照Han等^[24]描述的方法进行原位杂交。每张制片加40 μ L杂交液, 该液含探针DNA 100 ng、100%去离子甲酰胺20 μ L、50% (w/v)硫酸葡聚糖8 μ L、20× SSC 4 μ L、ssDNA (鲑鱼精DNA, 5 μ g μ L^-1) 1 μ L、10% (w/v)十二烷基磺酸钠1 μ L、加ddH₂O至40 μ L。在Hybrite原位杂交仪上进行杂交反应, 75℃ 6 min, 然后37℃ 16 h以上。用荧光素(FITC)检测杂交信号, 洗脱后用PI复染及H-1300 (抗退色剂)封片, OLYMPUS BX60荧光显微镜观察检测, Pixera Penguin 150CL CCD成像系统照相。

1.7 SSR标记和 STS标记分析

根据Rö der等^[25]和Pestsova等^[26]公布的小麦21条染色体特异SSR标记, 分别选取各条染色体长、短臂上各10对引物进行扩增。从网站http://wheat. pw.usda.gov/SNP/new/pcr_primers.shtm上选取小麦7个部分同源群上的STS标记, 分别选取各同源群6对标记进行扩增。所选引物由生工生物工程(上海)有限公司合成。PCR扩增反应体系和步骤参照本文SCAR标记鉴定, 退火温度依引物而定。扩增产物点样量为6 μ L, 经8%非变性聚丙烯酰胺凝胶电泳, 180 V恒压电泳2~3 h。硝酸银染色, 用数码相机照相。

2 结果与分析

2.1 农艺性状表现

表型调查结果显示, DH2322农艺性状稳定, 穗长较小麦亲本7182、D4286显著增长, 穗粒数也较D4286和七倍体材料H8911显著增大, 但与小麦亲本7182差异不大(表1)。整体上看, DH2322具有双亲的形态学特征, 结实性好, 籽粒饱满, 是较好的遗传材料。

表1 异代换系DH2322及其亲本农艺性状 Table 1 Agronomic traits of the substitution line DH2322 and its parents

2.2 华山新麦草Ns基因组特异SCAR标记鉴定

用华山新麦草Ns基因组特异SCAR标记RHS141对普通小麦品种7182、硬粒小麦品种D4286、DH2322、H8911及华山新麦草的基因组DNA进行PCR扩增表明, 华山新麦草、H8911和DH2322的DNA可以扩增出934 bp的华山新麦草特异条带, 而普通小麦品种7182、硬粒小麦品种D4286的DNA不能扩增出条带, 说明DH2322含有华山新麦草Ns基因组染色体(图1)。

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	图1 华山新麦草基因组特异SCAR标记分析Fig. 1 Amplification profile of the SCAR marker specific to P. huashanicaM: marker D2000; 1: 7182; 2: D4286; 3: DH2322; 4: H8911; 5: P. huashanica.

2.3 细胞遗传学鉴定

DH2322根尖细胞染色体制片观察, 发现根尖细胞染色体数目为2n=42 (图2-a); 对52个花粉母细胞镜检显示, 47/52 (90.4%)花粉母细胞减数分裂中期I染色体形成21个二价体(图2-b), 其中环状二价体占有很高比例, 平均每个细胞有20.91个(表2); 以华山新麦草总基因组DNA作为探针, 对DH2322根尖细胞和花粉母细胞基因组原位杂交鉴定显示, 其根尖细胞中有两条染色体为黄绿色(图3-a), 花粉母细胞减数分裂中期I细胞有1个完整的二价体显示荧光信号, 表明DH2322中的2条Ns基因组染色体可以完全配对为1对染色体(图3-b)。由此说明, DH2322是一个遗传稳定, 包含40条小麦染色体和1对华山新麦草Ns基因组染色体的小麦-华山新麦草异代换系。

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	图2 DH2322根尖细胞(a)和花粉母细胞(b)观察Fig. 2 Cytogenetic observation of root-tip cell (a) and pollen mother cell (b) of DH2322

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	图3 DH2322根尖细胞原位杂交(a)花粉母细胞原位杂交(b)Fig. 3 Genomic in situ hybridization (GISH) pictures for root-tip cell (a) and pollen mother cell (b) of DH2322

2.4 SSR和STS结果分析

用小麦21条染色体上共210对SSR引物对DH2322及其亲本进行分子标记分析表明, 2D染色体上的3对引物(Xgwm249、Xgwm296、Xgwm484)在小麦品种7182和H8911中扩增出154、182、153 bp的目标条带, 但在硬粒小麦D4286、华山新麦草和DH2322中未扩增出目标条带(图4)。推测DH2322缺失了小麦2D染色体。

表2 DH2322花粉母细胞减数分裂中期I (PMC MI)染色体配对 Table 2 Chromosome pairing at metaphase I in pollen mother cells of substitution line DH2322

选取定位于小麦7个部分同源群上的42对EST- STS标记, 对普通小麦品种7182、硬粒小麦品种D4286、DH2322、H8911及华山新麦草的基因组DNA进行PCR检测, 结果只有定位于小麦第2同源群的引物CD453246和BQ160526在DH2322中扩增出华山新麦草特异条带(图4)。推测DH2322可能含有华山新麦草2Ns染色体。

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	图4 SSR及STS引物扩增图谱Fig. 4 Amplification profile of SSR and STS markersM: marker D2000; 1: 7182; 2: D4286; 3: DH2322; 4: H8911; 5: P. huashanica.

3 讨论

实现华山新麦草有益基因向小麦的转移, 研究华山新麦草染色体的遗传效应以及获得小麦-华山新麦草中间材料, 如双二倍体、异附加系、异代换系是十分必要的。目前, 育种工作者已获得多个小麦-华山新麦草杂交后代, 它们表现型上多种多样, 如小麦-华山新麦草异附加系有矮株、分蘖数增多、抗条锈病、叶锈病等性状^{[15, 16, 17, 18, 19, 20, 21]}; Cao等^[27]鉴定出一个抗条锈病易位系材料。小麦-华山新麦草异代换系DH2322作为小麦与华山新麦草远缘杂交的新类型, 其穗长与穗粒数较亲本小麦品种显著增加, 是创制小麦-华山新麦草易位系的重要桥梁。华山新麦草染色体导入的同时, 必然与小麦遗传背景发生相互作用, 而导致穗长与穗粒数显著增大的原因, 尚需进一步研究。

外源基因的有效转移很大程度上依赖于对导入小麦背景中的外源染色体或染色体片段的快速准确鉴定^[28]。GISH可以直观地检测外源染色体或染色体片段在小麦遗传背景中的分布, 在鉴定远缘杂种染色体构型和结构变异方面有重要作用, 但其技术难度大、费时且成本较高, 在初步鉴定大量遗传材料时有一定的局限性。SCAR标记技术的发展为小大麦远缘杂交研究提供了极大方便。RHS141是Du等^[23]用RAPD技术筛选的华山新麦草基因组特异DNA片段, 在小麦族其他物种和华山新麦草及全套小麦-华山新麦草二体异附加系中验证发现, 仅含华山新麦草染色质的物种能扩增出长为938 bp的片段。RHS141可用于含华山新麦草特异染色质材料的检测。本研究中RHS141的应用为快速初步判定12DH22中含有华山新麦草遗传物质提供了方便。

SSR分子标记在近几年的小麦遗传作图、基因定位、系谱或亲缘关系分析、标记辅助选择等研究中得到广泛应用^[29]。Rö der等^[25]建立了小麦的第一张微卫星遗传图谱, 将230对引物扩增的279个位点整合到国际小麦簇作图计划(ITIM)的Opata85 × W7984作图群体遗传框架图上; Pestsova等^[26]对从粗山羊草分离的D基因组专化性微卫星标记(GDM)在小麦上进行了作图。国际小麦微卫星协会(Wheat Microsatellite Consortium, WMC)也将开发的66个WMC标记整合到已有的小麦遗传框架图上^[30]。本研究利用七倍体H8911与硬粒小麦D4286杂交选育DH2322的过程中, 由于来自H8911的A和B基因组与来自硬粒小麦的A和B基因组, 理论上可以很好地配对而得以完整遗传, 因此推测, 二体异代换系DH2322最大可能是由华山新麦草的1对Ns基因组染色体替换了1对小麦D基因组染色体。为了确定DH2322所缺失的小麦染色体, 本研究采用Pestsova等^[26]开发的来自粗山羊草专化性微卫星标记对小麦-华山新麦草异代换系DH2322进行分析, 结果显示, D基因组的2D染色体上的3对引物, 在DH2322中没有扩增出预期的目标条带, 而其他基因组染色体上的引物, 在DH2322中扩增出预期的目标条带。由此推断, DH2322缺失了小麦D基因组的2D染色体。

STS (Sequence-tagged site)即序列标签位点, 是基因组上定位明确, 作为界标并能通过PCR扩增被唯一操作的短的单拷贝DNA序列^[31], 由于可直接在DNA分子水平上指出小麦中外源染色体片段的组成和位置, 在外源染色体鉴定方面应用广泛。前人利用STS标记已成功鉴定出小麦背景中的簇毛麦、黑麦等小麦近缘种的染色体片段^{[32, 33, 34]}。Du等^[15]利用12对STS标记鉴定出3-6-4-1附加两条华山新麦草2Ns染色体, 其中的2对标记在华山新麦草和DH2322中扩增出与华山新麦草一致的特异条带, 说明DH2322含有华山新麦草Ns基因组的2Ns染色体。CD453246扩增出的H8911的条带本应与华山新麦草特异条带高度一致, 但H8911的条带却普遍高于后代DH2322的扩增条带, 原因尚不清楚。3-6-4-1和DH2322穗长较亲本7182都显著增长3~4 cm, 是否与2Ns染色体上携带的基因有关尚需进一步研究。

4 结论

DH2322是一个稳定的小麦-华山新麦草2D(2Ns)二体异代换系材料, 结实性好, 穗长和穗粒数等农艺性状上得到改善, 对进一步研究华山新麦草染色体的形态结构特征及遗传机制、挖掘利用华山新麦草优异基因、丰富小麦遗传种质资源具有重要价值。

The authors have declared that no competing interests exist.

作者已声明无竞争性利益关系。

参考文献

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1991

0.0

陈漱阳, 张安静, 傅杰. 普通小麦与华山新麦草的杂交. 遗传学报, 1991, 18: 508-512

Chen S

, Zhang A

, Fu

The hybridization between Triticum aestivum and Psathyrostachys huashanica.

Acta Genet Sin, 1991, 18: 508-512 (in Chinese with English abstract)

Psathyrosachys huashanica Keng is a special diploid species and distributed in Huashan branches of the Qinling mountain chain.It is very difficult to cross Trit icum aestivum with P.huashanica.The hybrids have been obtained by means of embry oculture.The percentage of seed set of cross was 0.19% and germination rate of embryo culture was 33.3%.The F 1 plant appeared to be intermediate between their parents in morphology.The chromosome number of F 1 plant root tip cells was 2n=28 .The 0.99Ⅱ and 26.0Ⅱ per cell of F 1 plant were observed at PMC’s MI and the po llen grain were abortive.The percentage of backcross seed set of F 1 hybrid polli nated with common wheat pollen was 2.5%.Root tip chromosome numbers of BC1 plant s were 2n=49 and most of the chromosome configuration at PMC’s MI of BC1 was 2Ⅲ +71.

Northwest Institute of Botany,Yangling Shaanxi 712100

华山新麦草是分布在秦岭山脉华山段的1个特有种，经细胞学鉴定为二倍体种(2n=14)，利用普通小麦与之杂交并通过幼胚培养获得了杂种，杂交结实率为0.19%，幼胚培养出苗率为33. 3%。杂种表现为双亲的中间型，杂种F 1 体细胞染色体数为2n=28，花粉母细胞减数分裂中期 Ⅰ每细胞平均0.99个二价体，26.01个单价体。杂种花粉粒败育，以小麦花粉与杂种回交时获得了种子，回交结实率为2.5%。回交一代体细胞染色体数为2n=49，花粉母细胞减数分裂中期Ⅰ染色体构型多数为21Ⅱ+7Ⅰ。

... 华山新麦草(Psathyrostachys huashanica Keng, 2n=14, NsNs)是禾本科(Gramineae)、小麦族(Triticeae)、大麦亚族(Hordeinae)、新麦草属(Psathyrostachys)多年生异花授粉植物, 具有抗寒、抗旱、耐瘠薄、早熟、优质、矮秆、抗病等优良特性^[1], 仅分布于我国秦岭山脉陕西华山段, 是我国特有种^[2], 也是小麦(Triticum aestivum, 2n=14, AABBDD)品种改良的优异种质资源 ...

... 本课题组自20世纪90年代初开始进行普通小麦品种7182与华山新麦草的杂交研究, 选育出普通小麦-华山新麦草七倍体材料H8911 (2n=49, AABBDDNs)^[1] ...

1991

0.595

0.0

Baden

A taxonomic revision of Psathyrostachys (Poaceae).

Nordic J Bot, 1991, 11: 3-26

Dept of Botany, Dendrology and Forests Genetics, The Royal Vet. and Agricult. Univ., Rolighedsvej 23, DK-1958 Frederiksberg C, Denmark

> The genus Psathyrostachys is revised. A total of 23 taxa had previously been described, now the following taxa are recognized: P. fragilis ssp. fragilis (2n = 14), P. fragilis ssp. secaliformis, P. fragilis ssp. villosus ssp. nov., P. caduca, P. huashanica (2n = 14), P. rupestris ssp. rupestris, P. rupestris ssp. daghestanica, P. lanuginosa (2n = 14 and 28), P. kronenburgii, P. juncea (2n = 14), and P. stoloniformis (2n = 14). A previous infrageneric division into sections cannot be recognized. A key, descriptions, nomenclature, illustrations, distribution maps, and cytogenetic data are presented.

2008

1.643

0.0

Kang H

, Zhang H

, Fan

, Zhou Y

Morphological and cytogenetic studies on the hybrid between bread wheat and Psathyrostachys huashanica Keng ex Kuo.

Euphytica, 2008, 162: 441-448

Psathyrostachys huashanica Keng ex Kuo (2 n = 2 x = 14, NsNs ), a source of wheat stripe rust, take - all fungus, and powdery mildew resistance with tolerance to salinity and drought, has been successfully hybridized as the pollen parent to bread wheat without using immature embryo rescuing culture for the first time. All of the CS ph2b × P. huashanica hybrid seeds germinate well. Backcross derivatives were successfully obtained. F 1 hybrids were verified as intergeneric hybrids on the basis of morphological observation, cytological and molecular analyses. The results obviously showed the phenotypes of the hybrid plants were intermediate between bread wheat and P. huashanica . Chromosome pairing at MI of PMCs in the F 1 hybrid plants was low, and the meiotic configuration was 26.80 I + 0.60 II (rod). Cytological analysis of the hybrid plants revealed the ineffectiveness of the ph2b gene on chromosome association between the parents. Eight RAPD-specific markers for Ns genome were selected for RAPD analysis, and the results indicated that F 1 hybrids contained the Ns genome of P. huashanica . Furthermore, the significance of the finding for bread wheat improvement was discussed.

1.Sichuan Agricultural University Triticeae Research Institute Dujiangyan 611830 Sichuan China 2.Ministry of Education, Sichuan Agricultural University Key Laboratory of Crop Genetic Resources and Improvement Yaan 625014 Sichuan China

... Kang等^[3,4]利用中国春ph2b基因突变体和华山新麦草杂交获得了含有ph2b基因的衍生后代、并利用面包小麦“ ...

2010

0.0

Kang H

, Chen

, Wang

, Zhong M

, Zhang H

, Zhou Y

Molecular cytogenetic characterization of the amphiploid between bread wheat and Psathyrostachys huashanica.

Genet Resour Crop Evol, 2010, 57: 111-118

A new wheat- Psathyrostachys huashanica amphiploid, PHW-SA, was characterized using molecular cytological tools to evaluate the potential utilization of P. huashanica for wheat improvement. PHW-SA pollen mother cells (PMCs) regularly revealed averagely 0.57 univalents, 24.51 ring bivalents, 3.19 rod bivalents and 0.01 trivalents per cell. Complete homologous chromosome pairing was seen in 81% of the PMCs, indicating a degree of cytological stability. P. huashanica C-banding karyotypes of PHW-SA displayed strong heterochromatin terminal bands at either or both ends. 24 P. huashanica chromosome arms showed telomeric bands. Using P. huashanica DNA as a probe and J-11 DNA as blocker, distinctive P. huashanica chromosomes, which were presented entirely greenish-yellow hybridization signals, were observed in a somatic metaphase of PHW-SA. GISH also revealed the chromosomal breakage and translocation occurring in the amphiploid PHW-SA, which may represent a centromeric fusion between P. huashanica and wheat chromosomes. Seeds storage proteins electrophoresis indicated that PHW-SA expressed some of P. huashanica specific gliadin and glutenin bands, and a few gliadin and glutenin bands of the parents disappeared and new bands appeared. The results indicated that the amphiploid PHW-SA could serve as novel germplasm sources for wheat improvement.

1.Sichuan Agricultural University Triticeae Research Institute 611130 Wenjiang, Chengdu Sichuan China 2.Ministry of Education, Sichuan Agricultural University Key Laboratory of Crop Genetic Resources and Improvement 625014 Yaan Sichuan China

... Kang等^[3,4]利用中国春ph2b基因突变体和华山新麦草杂交获得了含有ph2b基因的衍生后代、并利用面包小麦“ ...

2011

1.839

0.0

... Wang等^[5]研究发现华山新麦草3Ns染色体上携带抗小麦条锈病的基因, Kang等^[6]将华山新麦草3Ns染色体导入小麦, 获得了抗条锈病的小麦-华山新麦草中间材料 ...

2011

3.73

0.0

Kang H

, Wang

, Fedak

, Cao W

, Zhang H

, Fan

, Sha L

, Xu L

, Zheng Y

, Zhou Y

Introgression of chromosome 3Ns from Psathyrostachys huashanica into wheat specifying resistance to stripe rust.

PloS One, 2011, 6: e21802

Wheat stripe rust is a destructive disease in the cool and humid wheat-growing areas of the world. Finding diverse sources of stripe rust resistance is critical for increasing genetic diversity of resistance for wheat breeding programs. Stripe rust resistance was identified in the alien species Psathyrostachys huashanica , and a wheat- P. huashanica amphiploid line (PHW-SA) with stripe rust resistance was reported previously. In this study, a P. huashanica 3Ns monosomic addition line (PW11) with superior resistance to stripe rust was developed, which was derived from the cross between PHW-SA and wheat J-11. We evaluated the alien introgressions PW11-2, PW11-5 and PW11-8 which were derived from line PW11 for reaction to new Pst race CYR32, and used molecular and cytogenetic tools to characterize these lines. The introgressions were remarkably resistant to CYR32, suggesting that the resistance to stripe rust of the introgressions thus was controlled by gene(s) located on P. huashanica chromosome 3Ns. All derived lines were cytologically stable in term of meiotic chromosome behavior. Two 3Ns chromosomes of P. huashanica were detected in the disomic addition line PW11-2. Chromosomes 1B of substitution line PW11-5 had been replaced by a pair of P. huashanica 3Ns chromosomes. In PW11-8, a small terminal segment from P. huashanica chromosome arm 3NsS was translocated to the terminal region of wheat chromosomes 3BL. Thus, this translocated chromosome is designated T3BL-3NsS. These conclusions were further confirmed by SSR analyses. Two 3Ns-specific markers Xgwm181 and Xgwm161 will be useful to rapidly identify and trace the translocated fragments. These introgressions, which had significant characteristics of resistance to stripe rust, could be utilized as novel germplasms for wheat breeding.

2010

0.0

1.0251

Liu W

, Liu W

, Wu

, Gao A

, Li L

Analysis of genetic diversity in natural populations of Psathyrostachys huashanica Keng using microsatellite (SSR) markers.

Agric Sci China, 2010, 9: 463-471

Abstract Psathyrostachys huashanica Keng is endemic to China and only distributed in Huashan Mountain in Shaanxi Province, China. In this study, 15 P. huashanica populations consisting of 450 individuals sampled across their main distribution were investigated by using the simple sequence repeats (SSRs) markers. A total of 184 alleles were detected on 24 SSR loci, and the number of alleles on each locus ranged from 2 to15, with an average of 7.667. The total gene diversity ( H T = 0.683) and the coefficient of population differentiation ( G ST = 0.125) showed that P. huashanica had a relatively high level of genetic variation, and the genetic variation was mainly distributed within the populations. The gene flow among the populations of P. huashanica ( Nm = 1.750) was much less than that of the common anemophytes ( Nm = 5.24). Correlation analysis demonstrated that the number of alleles as well as genetic diversity of the five populations of Huangpu valley decreased along with the increase of altitudes, but the correlation was not significant. Implications of these results for future P. huashanica collection, evaluation and conservation were discussed.

... 另外, Liu等^[7]和Wang等^[8]利用简单重复序列标记(simple sequence repeats, SSR)技术对华山新麦草种群基因的多样性进行了分析 ...

2006

1.153

0.0

Wang

, Guo

, Zhao G

Genetic diversity of the endangered and endemic species Psathyrostachys huashanica natural populations using simple sequence repeats (SSRs) markers.

Biochem Syst Ecol, 2006, 34: 310-318

Abstract Psathyrostachys huashanica Keng., a species endemic to China, is only distributed in Huashan Mountain in Shaanxi Province. It has been listed as “national first-class protected rare species.” In this study, the microsatellites of barley were used to analyze genetic diversity of P. huashanica populations sampled from three valleys (Huangpu, Xian and Huashan Valleys) in Mt. Huashan. A total of 33 alleles of 11 loci were detected from 266 individuals. The observed average number of alleles ( A ) is 2.75; the effective number of alleles (Ae) is 1.67. The percentage of polymorphic loci (PPB) is 58.33% in Huangpu Valley, 75% in Xian Valley, 83.33% in Huashan Valley, and the total PPB is 83.33%. About 77.6% of ( F ST = 0.324) genetic diversity was observed within the subpopulations. Genetic differentiation within each subpopulations was higher than that among the subpopulations. Mean genetic distance is 0.17 (range: 0.010–0.401). Correlation analysis detected significant correlation between genetic distance and vertical distance of altitude in Huashan Valley. Differentiation mainly occurred between the higher altitude subpopulations and the lower altitude subpopulations, suggesting that altitude might be the major factor that restricted the gene flow between different altitude subpopulations and resulted in differentiation of subpopulations.

... 另外, Liu等^[7]和Wang等^[8]利用简单重复序列标记(simple sequence repeats, SSR)技术对华山新麦草种群基因的多样性进行了分析 ...

1997

0.0

1.321

... 在此基础上, 侯文胜等^[9]、傅杰等^[10]、赵继新等^[11,12,13]、武军等^[14]利用细胞遗传学对小麦-华山新麦草异附加系、异代换系进行了分子细胞遗传学方面的研究 ...

2003

0.0

1.321

傅杰, 王美南, 赵继新, 陈漱阳, 侯文胜, 杨群慧. 抗全蚀病小麦-华山新麦草中间材料H8911的细胞遗传学研究与利用. 西北植物学报, 2003, 12: 2157-2162

, Wang M

, Zhao J

, Chen S

, Hou W

, Yang Q

Studies on cytogenetic and utilization of wheat-Psathyrostachys huashanica medium material H8911 with resistance to wheat take-all fungus.

Acta Bot Boreali-Occident Sin. 2003, 12: 2157-2162 (in Chinese with English abstract)

抗小麦全蚀病中间材料H8911(BC1F1)是通过小麦与华山新麦草杂种幼胚培养及杂种F1(ABDN 2n=28)再与小麦回交后得到的.根尖细胞染色体数目49条,花粉母细胞减数分裂中期I,染色体构型为20.85(19～21)Ⅱ+7.30(7～11)I,21Ⅱ+7I的细胞占86.67%.BC1F2和BC1F3体细胞染色体数目范围分别为45～53和44～52,49条染色体的植株类型分别占30.19%和27.50%,华山新麦草染色体丢失率分别为11.85%和13.14%;花粉母细胞减数分裂中期I,染色体构型分别为20.62(18～22)Ⅱ+7.64(5～13)I+0.04(0～1)Ⅲ和20.53(17～22)Ⅱ+7.79(5～15)I+ 0.05(0～1)Ⅲ, 21Ⅱ+7I的细胞分别占77.24%和 69.42%.随着自交世代的延续,21Ⅱ+7I细胞的传递能力逐渐降低.利用H8911作供体,选育出小麦-华山新麦草抗全蚀病新种质13个,其中1个附加系表现近高度抗病性,6个附加系、3个代换系和3个易位系材料表现中度抗病性.

2004

0.0

赵继新, 陈新宏, 王小利, 武军, 傅杰, 何蓓如, 孙志刚. 普通小麦-华山新麦草异附加系的分子细胞遗传学研究. 西北农林科技大学学报(自然科学版), 2004, 32: 105-108

Zhao J

, Chen X

, Wang X

, Wu

, Fu

, He B

, Sun Z

Molecular cytogenetic study on the alien addition lines of Triticum-Psathyrostachys.

J Northwest Sci-Tech Univ Agric For (Nat Sci Edn), 2004, 32: 105-108 (in Chinese with English abstract)

The alien addition lines of Triticum-Psathyrostachys were detected by fluorescent in situ hybridization (FISH) and chromosomes C-banding.FISH shows the alien addition line H9015-17-1-9 and H9017-14-16-5 are all added with the two chromosomes of Psathyrostachys huashanica. The analysis using the chromosomes C-banding indicates that H9015-17-1-9 might be an alien addition line of N 5 h and H9017-14-16-5 might be an alien addition line of N 6 h .

利用荧光原位杂交和染色体C-分带技术，对普通小麦-华山新麦草的异附加系进行了研究。荧光原位杂交结果显示：异附加系H9015-17-1-9，H9017-14-16-5都附加有2条华山新麦草的染色体。对这2个材料和华山新麦草进行染色体C-分带带型比较，初步推断，H9015-17-1-9附加的是N 5 h 染色体，H9017-14-16-5附加的是N 6 h 染色体。

2004

0.0

1.321

赵继新, 陈新宏, 王小利, 武军, 傅杰, 何蓓如, 孙晓娟. 普通小麦-华山新麦草异代换系的分子细胞遗传学研究. 西北植物学报, 2004, 24: 2277-2281

Zhao J

, Chen X

, Wang X

, Wu

, Fu

, He B

, Sun X

Molecular cytogenetic study on the alien substitution lines of Triticum-Psathyrostachys.

Acta Bot Boreali-Occident Sin, 2004, 24: 2277-2281 (in Chinese with English abstract)

利用荧光原位杂交和染色体C-分带技术对普通小麦-华山新麦草的异代换系进行了研究.荧光原位杂交结果显示:异代换系H921-6-12和H924-3-4均含有2条华山新麦草的染色体.对这2个材料和华山新麦草进行染色体C-分带带型比较,结果认为:H921-6-12可能是普通小麦-华山新麦草的5A/N5h代换系,H924-3-4可能是3D/N4h代换系.

2010

0.0

Zhao J

, Ji W

, Wu

, Chen X

, Cheng X

, Wang J

, Pang Y

, Liu S

, Yang Q

Development and identification of a wheat-Psathyrostachys huashanica addition line carrying HMW-GS, LMW-GS and gliadin genes.

Genet Resour Crop Evol, 2010, 57: 387-394

Psathyrostachys huashanica Keng, in the family Poaceae, subtribe Triticeae is an endangered and endemic species of China. It is characterized as Shaanxi, drought tolerant and fungus and disease resistant. In this paper, a wheat ( Triticum aestivum L.)— P. huashanica disomic addition line “H9021-28-5” was developed using cytogenetics methods including SDS–PAGE, A-PAGE, and GISH. These investigations revealed that the chromosome number and configuration of “H9021-28-5” were 2 n = 44 = 22 II. The mitotic and meiotic GISH analysis indicated that “H9021-28-5” contained 42 wheat chromosomes and a pair of P. huashanica chromosomes. The SDS–PAGE and A-PAGE analysis showed that “H9021-28-5” carried the HMW-GS, LMW-GS and gliadin genes of P. huashanica . The results suggest that these storage protein genes of P. huashanica had been transferred into common wheat. Development of this new germplasm is significant for enriching and improving wheat resources of storage proteins genes, and for continuing to exploit the advantageous genes of P. huashanica .

1.Northwest A & F University College of Agronomy 712100 Yangling Shaanxi China 2.Northwest A & F University College of Life Sciences 712100 Yangling Shaanxi China

2010

0.0

2014

2.881

0.0

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

... 目前, 育种工作者已获得多个小麦-华山新麦草杂交后代, 它们表现型上多种多样, 如小麦-华山新麦草异附加系有矮株、分蘖数增多、抗条锈病、叶锈病等性状^{[15,16,17,18,19,20,21]} ...

... Du等^[15]利用12对STS标记鉴定出3-6-4-1附加两条华山新麦草2Ns染色体, 其中的2对标记在华山新麦草和DH2322中扩增出与华山新麦草一致的特异条带, 说明DH2322含有华山新麦草Ns基因组的2Ns染色体 ...

2014

1.668

0.0

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

2014

3.347

0.0

Du W

, Wang

, Lu

, Sun S

, Chen X

, Zhao J

, Yang Q

, Wu

Characterization of a wheat-Psathyrostachys huashanica Keng 4Ns disomic addition line for enhanced tiller numbers and stripe rust resistance.

Planta, 2014, 239: 97-105

1. Shaanxi Key Laboratory of Genetic Engineering for Plant Breeding, College of Agronomy, Northwest A&F University, Yangling, 712100, Shaanxi, China

Exploiting and utilizing excellent gene(s) from wild species has become an essential strategy for wheat improvement. In the disomic addition line 24-6-3, the 4Ns chromosomes from Psathyrostachys huashanica Keng (2 n = 2 x = 14, NsNs) carried valuable tiller and strip rust resistance gene(s), which was selected from the progeny of common wheat cv. 7182 and P. huashanica via embryo culture. Cytology, genomic in situ hybridization (GISH), and EST-STS analyses were used to detect the 4Ns chromosome in wheat background and its homoeologous relationship. Cytological studies demonstrated that 24-6-3 contained 44 chromosomes and formed 22 bivalents during meiotic metaphase I. GISH using P. huashanica genomic DNA as a probe indicated that a pair of Ns-chromosomes with strong hybridization signals had been introduced into 24-6-3. Ten EST-STS markers, i.e., BE404973 , BE442811 , BE446061 , BE446076 , BE497324 , BE591356 , BF473854 , BG274986 , BQ161513 and CD373484 , which were located on the homoeologous group 4 chromosomes of wheat, amplified bands unique to P. huashanica in 24-6-3. This indicated the presence of an introgressed P. huashanica Ns chromosome pair belonging to homoeologous group 4, which we designated the 4Ns disomic addition line. After it was inoculated using mixed races of stripe rust in the adult stages, 24-6-3 expressed high stripe rust resistance, which was possibly derived from its P. huashanica parent. Moreover, its increased number of tillers was probably controlled by gene(s) located in P. huashanica chromosome 4Ns. These high levels of disease resistance and excellent agronomic traits make the 24-6-3 line a promising germplasm for breeding in wheat.

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

2013

3.251

0.0

Du W

, Wang

, Lu

, Sun S

, Chen X

, Zhao J

, Yang Q

, Wu

Molecular cytogenetic identification of a wheat- Psathyrostachys huashanica Keng 5Ns disomic addition line with stripe rust resistance.

Mol Breed, 2013, 31: 879-888

1. Shaanxi Key Laboratory of Genetic Engineering for Plant Breeding, College of Agronomy, Northwest A&F University, Yangling, 712100, Shaanxi, China

We developed a new stripe rust resistant line of common wheat– Psathyrostachys huashanica Keng (2 n = 2 x = 14, NsNs) from a cross between wheat cv. 7182 and P. huashanica via embryo culture, and we refer to this line as 3-8-10-2. We characterized this new line by cytology, genomic in situ hybridization (GISH), EST-SSR, EST-STS, and disease resistance screening. GISH using P. huashanica genomic DNA as the probe indicated that a pair of Ns chromosomes with strong hybridization signals was introduced into 3-8-10-2. We screened 255 EST-SSR and EST-STS multiple-loci markers from seven wheat homoeologous groups in the parent lines. Of these, 90 markers were polymorphic with a polymorphism frequency of 40 %, while two EST-SSR markers and six EST-STS markers located on wheat chromosome group 5 produced specific bands in P. huashanica and 3-8-10-2, respectively. This suggested that the introduced Ns chromosome pair belonged to homoeologous group 5, which was identified using new genome-specific markers. After inoculation with stripe rust isolates, 3-8-10-2 exhibited stripe rust resistance that probably originated from its P. huashanica parent. 3-8-10-2 can be used as a donor source for introducing novel disease resistance genes into wheat during breeding programs with the assistance of molecular and cytogenetic markers. Moreover, 3-8-10-2 had improved agronomic characteristics compared with its parents. Therefore, the addition line could be exploited as an important bridge for wheat breeding and chromosome engineering.

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

2013

3.73

0.0

Du W

, Wang

, Pang Y

, Li Y

, Chen X

, Zhao J

, Yang Q

, Wu

Isolation and characterization of a Psathyrostachys huashanica Keng 6Ns chromosome addition in common wheat.

PloS One, 2013, 8: e53921

The development of alien addition lines is important for transferring useful genes from exotic species into common wheat. A hybrid of common wheat cv. 7182 (2 n = 6 x = 42, AABBDD ) and Psathyrostachys huashanica Keng (2 n = 2 x = 14, NsNs ) via embryo culture produced the novel intergeneric disomic addition line 59-11. The seed morphology of 59-11 resembled the parent 7182 and it exhibited extreme agronomic characteristics, i.e., twin stable spikelets, fertile florets, and multi-kernel clusters. Furthermore, 59-11 produced plump kernels with a high seed-setting percentage during the advanced maturation stage. The line was screened based on genomic in situ hybridization, EST-SSR, EST-STS, and gliadin to identify P. huashanica chromatin in the wheat background. The chromosome number and configuration of 59-11 was 2 n = 44 = 22 II and we confirmed the 6 Ns disomic chromosome additions based on A-PAGE analysis and molecular markers. The results suggested that the production of twin spikelets and multiple kernels per spike in the wheat- P. huashanica addition line was related to homologous group 6 in the wheat chromosome. This is the first report of the introduction of improved spike traits into common wheat from the alien species P. huashanica and it opens up the possibility of increasing the wheat yield based on this enlarged gene pool.

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

2013

3.73

0.0

Du W

, Wang

, Wang L

, Zhang

, Chen X

, Zhao J

, Yang Q

, Wu

Development and Characterization of a Psathyrostachys huashanica Keng 7Ns Chromosome Addition Line with Leaf Rust Resistance.

PloS One, 2013, 8: e70879

The aim of this study was to characterize a Triticum aestivum - Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs ) disomic addition line 2-1-6-3. Individual line 2-1-6-3 plants were analyzed using cytological, genomic in situ hybridization (GISH), EST-SSR, and EST-STS techniques. The alien addition line 2-1-6-3 was shown to have two P. huashanica chromosomes, with a meiotic configuration of 2 n = 44 = 22 II. We tested 55 EST-SSR and 336 EST-STS primer pairs that mapped onto seven different wheat chromosomes using DNA from parents and the P. huashanica addition line. One EST-SSR and nine EST-STS primer pairs indicated that the additional chromosome of P. huashanica belonged to homoeologous group 7, the diagnostic fragments of five EST-STS markers ( BE404955 , BE591127 , BE637663 , BF482781 and CD452422 ) were cloned, sequenced and compared. The results showed that the amplified polymorphic bands of P. huashanica and disomic addition line 2-1-6-3 shared 100% sequence identity, which was designated as the 7 Ns disomic addition line. Disomic addition line 2-1-6-3 was evaluated to test the leaf rust resistance of adult stages in the field. We found that one pair of the 7 Ns genome chromosomes carried new leaf rust resistance gene(s). Moreover, wheat line 2-1-6-3 had a superior numbers of florets and grains per spike, which were associated with the introgression of the paired P. huashanica chromosomes. These high levels of disease resistance and stable, excellent agronomic traits suggest that this line could be utilized as a novel donor in wheat breeding programs.

... Du等^{[15,16,17,18,19,20]}选育出一批附加华山新麦草2Ns-7Ns染色体的小麦-华山新麦草二体异附加系 ...

2014

0.0

1.007

王秀娟, 赵继新, 庞玉辉, 孙树贵, 张军, 鲁敏, 王亮明, 武军, 杨群慧, 陈新宏. 小麦-华山新麦草7Ns异附加系的分子细胞遗传学研究. 麦类作物学报, 2014, 34: 454-459

Wang X

, Zhao J

, Pang Y

, Sun S

, Zhang

, Lu

, Wang L

, Wu

, Yang Q

, Chen X

Molecular and cytogenetic characterization of a Triticum-Psathyrostachys 7Ns addition line.

J Triticeae Crops, 2014, 34: 454-459 (in Chinese with English abstract)

A wheat P. huashanica alien addition line 12DH25 was developed from the F 4 progeny of H8911 99 and Triticum durum cv. D4286. Analysis on morphology, cytology and SCAR markers were adopted to identify this alien addition line 12DH25. The results showed that 12DH25 presented favorable agronomic traits and its average plant height was 55.64 cm, which was very low. A SCAR marker was developed to distinguish the wheat lines carrying chromosomes derived from P. huashanica , with a specific band in the plant of 12DH25. The chromosome number of 12DH25 in root tip cells were 44 with 22 bivalents observed in most PMC at M I, and a pair of Ns chromosome with fluorescence signals appeared in genomic in situ hybridization (GISH) analysis using P. huashanica whole genomic DNA. STS markers BE591127 and BG274576 , which mapped on the wheat homocelogous group 7, produced the specific bands for P. huashanica , H8911 99 and 12DH25. It demonstrated that the chromosome from P. huashanica had a homoeologous relationship with the seventh group. Consequentially, 12DH25 was a dwarf material of a wheat P. huashanica disomic alien addition line, which could be exploited as an intermediate material in wheat genetic and breeding programs.

为了给有效利用华山新麦草基因提供新的种质材料，利用细胞遗传学、分子标记等技术，结合田间农艺性状考察，对从小麦华山新麦草七倍体材料H8911 99与硬粒小麦D4286杂交F 4 代分离群体中选育的杂交后代12DH25进行了鉴定。华山新麦草基因组特异SCAR标记鉴定表明，12DH25含有华山新麦草遗传物质；有丝分裂和花粉母细胞减数分裂中期I染色体数为2n=44=22Ⅱ，基因组原位杂交（GISH）出现两条杂交信号，且能配对，表明两条外源染色体是华山新麦草的一对同源染色体。选取定位于小麦7个部分同源群上的28对STS标记对12DH25及其亲本基因组DNA进行扩增，定位于小麦第7同源群上的STS标记 BE591127 [KG-*3]和 BG274576 [KG-*3]能在12DH25中扩增出华山新麦草特征条带，将12DH25附加的华山新麦草染色体归属于小麦第7部分同源群。由此确定矮秆材料12DH25是一个稳定的小麦华山新麦草7Ns二体异附加系。

... 王秀娟等^[21]获得一个附加两条华山新麦草7Ns染色体的矮秆材料12DH25 ...

2006

5.319

0.0

Cota-sánchez J

, Remarchuk

, Ubayasena

. Ready-to-use DNA extracted with a CTAB method adapted for herbarium specimens and mucilaginous plant tissue. Plant Mol Biol Rep, 2006, 24: 161-167

This report summarizes major changes in previously published protocols for DNA extraction to improve the quality of DNA extracted from plants. Here, we highlight the critical modifications in the original protocols. The efficiency of these changes results in high-quality DNA ready to use in a variety of phytogenetically distant plant families, in particular species with mucopolysaccharides. The DNA obtained can be used without further purification in various molecular biology assays, including direct sequencing and AFLP and RAPD (random-amplified polymorphic DNA) analyses. The effectiveness of this method is proven by the amplification and sequencing of PCR products of up to 1 kb with DNA extracted from herbarium tissue ≥60 years old. This versatility is not usually found in DNA extraction protocols. In addition, this method is quick, adaptable to standard laboratories, and most important, safer and more cost-effective.

1.University of Saskatchewan Biology Department 112 Science Place S7N 5E2 Saskatoon SK Canada

... 3 DNA提取采用CTAB法^[22]提取普通小麦品种7182、华山新麦草、硬粒小麦品种D4286、普通小麦-华山新麦草七倍体材料H8911及DH2322的基因组DNA ...

2013

0.994

0.0

Du W

, Wang

, Wang L

, Pang Y

, Wu

, Zhao J

, Yang Q

, Chen X

A novel SCAR marker for detecting Psathyrostachys huashanica Keng chromatin introduced in wheat.

Genet Mol Res, 2013, 12: 4797-4806

In this study, we cloned and sequenced a 938-base pair polymorphic band, pHs27, in the tightly linked random amplified polymorphic DNA marker OPU10 and converted it into a sequence-characterized amplified region (SCAR) marker referred to as RHS141, which was specific for the Ns genome of Psathyrostachys huashanica. A GenBank basic local alignment search tool search showed that the sequence of pHs27 had no primary sequence homology with known sequences, and Southern blotting confirmed this result. This SCAR marker was used to detect Ns genome chromatin in wheat, and it was successfully amplified in P. huashanica itself, a complete set of wheat-P. huashanica disomic addition lines (1Ns-7Ns), and undetermined homoeologous group addition lines. This SCAR marker will be a powerful tool for the marker-assisted selection of P. huashanica chromosome(s) in a wheat background, and it should also allow wheat breeders to screen for the excellent traits found in P. huashanica chromatin.

Du, W. L. 1 ;Wang, J. 1 ;Wang, L. M. 1 ;Pang, Y. H. 1 ;Wu, J. 1 ;Zhao, J. X. 1 ;Yang, Q. H. 1 ;Chen, X. H. 1 ;

... 4 华山新麦草Ns基因组特异SCAR标记鉴定为提高小麦背景中华山新麦草染色质的鉴定效率, 根据Du等^[23]开发的华山新麦草染色体特异SCAR标记RHS141, 以7182、D4286、DH2322、H8911和华山新麦草基因组DNA为模板进行扩增 ...

... RHS141是Du等^[23]用RAPD技术筛选的华山新麦草基因组特异DNA片段, 在小麦族其他物种和华山新麦草及全套小麦-华山新麦草二体异附加系中验证发现, 仅含华山新麦草染色质的物种能扩增出长为938 bp的片段 ...

2004

3.658

0.0

Han F

, Liu

, Fedak

, Liu

Genomic constitution and variation in five partial amphiploids of wheat-Thinopyrum intermedium as revealed by GISH, multicolor GISH and seed storage protein analysis.

Theor Appl Genet, 2004, 109: 1070-1076

Genomic in situ hybridization (GISH) and multicolor GISH (mcGISH) methodology were used to establish the cytogenetic constitution of five partial amphiploid lines obtained from wheat × Thinopyrum intermedium hybridizations. Line Zhong 1, 2 n =52, contained 14 chromosomes from each of the wheat genomes plus ten Th. intermedium chromosomes, with one pair of A-genome chromosomes having a Th. intermedium chromosomal segment translocated to the short arm. Line Zhong 2, 2 n =54, had intact ABD wheat genome chromosomes plus 12 Th. intermedium chromosomes. The multicolor GISH results, using different fluorochrome labeled Th. intermedium and the various diploid wheat genomic DNAs as probes, indicated that both Zhong 1 and Zhong 2 contained one pair of Th. intermedium chromosomes with a significant homology to the wheat D genome. High-molecular-weight (HMW) glutenin and gliadin analysis revealed that Zhong 1 and Zhong 2 had identical banding patterns that contained all of the wheat bands and a specific HMW band from Th. intermedium . Zhong 1 and Zhong 2 had good HMW subunits for wheat breeding. Zhong 3 and Zhong 5, both 2 n =56, possessed no gross chromosomal aberrations or translocations that were detectable at the GISH level. Zhong 4 also had a chromosome number of 2 n =56 and contained the complete wheat ABD-genome chromosomes plus 14 Th. intermedium chromosomes, with one pair of Th. intermedium chromosomes being markedly smaller. Multicolor GISH results indicated that Zhong 4 also contained two pairs of reciprocally translocated chromosomes involving the A and D genomes. Zhong 3, Zhong 4 and Zhong 5 contained a specific gliadin band from Th. intermedium . Based on the above data, it was concluded that inter-genomic transfer of chromosomal segments and/or sequence introgression had occurred in these newly synthesized partial amphiploids despite their diploid-like meiotic behavior and disomic inheritance.

1.Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave, Bldg 50 Ottawa ON Canada K1A 0C6 2.Division of Biological Sciences University of Missouri 117 Tucker Hall Columbia MO 65211-7400 USA 3.School of Life Sciences Northeast Normal University Changchun 130024 China 4.Department of Plant Pathology North Darkata University Fargo ND 58105 USA

... 参照Han等^[24]描述的方法进行原位杂交 ...

1998

4.389

0.0

... der等^[25]和Pestsova等^[26]公布的小麦21条染色体特异SSR标记, 分别选取各条染色体长、短臂上各10对引物进行扩增 ...

... der等^[25]建立了小麦的第一张微卫星遗传图谱, 将230对引物扩增的279个位点整合到国际小麦簇作图计划(ITIM)的Opata85 × ...

2000

1.668

0.0

... der等^[25]和Pestsova等^[26]公布的小麦21条染色体特异SSR标记, 分别选取各条染色体长、短臂上各10对引物进行扩增 ...

... Pestsova等^[26]对从粗山羊草分离的D基因组专化性微卫星标记(GDM)在小麦上进行了作图 ...

... 为了确定DH2322所缺失的小麦染色体, 本研究采用Pestsova等^[26]开发的来自粗山羊草专化性微卫星标记对小麦-华山新麦草异代换系DH2322进行分析, 结果显示, D基因组的2D染色体上的3对引物, 在DH2322中没有扩增出预期的目标条带, 而其他基因组染色体上的引物, 在DH2322中扩增出预期的目标条带 ...

2.922

0.0

Cao Z

, Deng Z

, Wang M

, Wang X

, Jing J

, Zhang X

, Shang H

, Li Z

Inheritance and molecular mapping of an alien stripe-rust resistance gene from a wheat-Psathyrostachys huashanica translocation line. Plant Sci, 174: 544-549

... Cao等^[27]鉴定出一个抗条锈病易位系材料 ...

1991

0.0

0.928

Zhang X

Production and utilization of wheat alien translocation lines.

Hereditas, 1991, 13: 39-44

... 外源基因的有效转移很大程度上依赖于对导入小麦背景中的外源染色体或染色体片段的快速准确鉴定^[28] ...

1998

3.658

0.0

Peil

, Korzun

, Schubert

, Schumann

, Weber W

, Röder M

The application of wheat microsatellites to identify disomic Triticum aestivum-Aegilops markgrafii addition lines.

Theor Appl Genet, 1998, 96: 138-146

A1. Institut für Pflanzenzüchtung und Pflanzenschutz, Martin-Luther-Universität Halle-Wittenberg, Berlinerstrasse 2, D-06188 Hohenthurm, Germany Fax: 034 602 154617 E-mail: peil@landw.uni-halle.de, DE A2. Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstrasse 3, D-06466 Gatersleben, Germany, DE

We describe the use of wheat microsatellites for the discrimination of Aegilops markgrafii chromosomes. Twenty out of eighty eight wheat microsatellites (WMS) tested were able to distinguish Triticum aestivum-Ae. markgrafii addition lines. Six, three, three, one and six of 18 WMS can be used as markers for single Ae. markgrafii chromosomes B, C, D, F and G, respectively. Addition line A is not available but additional bands, appearing only in Ae. markgrafii and the T. aestivum-Ae. markgrafii amphiploid and not in any of the available addition lines, indicate that three WMS detect markers for Ae. markgrafii chromosomes A. Addition line E could not be detected by any of the WMS markers applied, although the 20 WMS represented all the homologous groups of wheat. All three WMS located on the short arm of group-2 chromosomes were located on Ae. markgrafii chromosome B; three of four WMS, located on the long arm of wheat group-2 chromosomes, were specific to Ae. markgrafii chromosome G and three of four WMS, specific to group-5 chromosomes, were markers for Ae. markgrafii chromosome C, indicating the homoeology of these wheat chromosome arms with the respective Ae. markgrafii chromosomes.

... SSR分子标记在近几年的小麦遗传作图、基因定位、系谱或亲缘关系分析、标记辅助选择等研究中得到广泛应用^[29] ...

2002

3.658

0.0

Gupta P

, Balyan H

, Edwards K

, Isaac

, Korzun

, Röder

, Gautier M

, Joudrier

, Schlatter A

, Dubcovsky

, De la Pena

R C

, Khairallah

, Penner

, Hayden

M J

, Dubcovsky

Sharp P, Dubcovsky

Keller B, Wang

R C C

, Hardouin

J P

, Jack

, Leroy

. Genetic mapping of 66 new microsatellite (SSR) loci in bread wheat. Theor Appl Genet, 2002, 105: 413-422

A1. CCS University, Meerut, India, A2. Department of Biological Sciences, University of Bristol, UK, A3. Agrogene, Moissy-Cramayel, France, A4. Lochow-Petkus GmbH, Grimsehlstr. 31, 37574 Einbeck, Germany, A5. Institute for Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, 06466 Gatersleben, Germany, A6. INRA, Montpellier, France, A7. IRB-INTA, 1712 Castelar, Buenos Aires, Argentina, A8. University of California, Davis, USA., A9. Monsanto, USA, A10. CIMMYT, Mexico, A11. University of Sydney, Quality Wheat CRC and Plant Breeding Institute, Camden NSW 2570, Australia, A12. Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, 8008 Zurich, Switzerland, A13. USDA-ARS-FRRL, 695 N 1100 E, Logan, Utah 84322-6300, USA, A14. Benoist Ets, France, A15. UMR INRA-UBP ASP, 234 Avenue du Brézet, 63 039 Clermont-Ferrand Cedex 2., France,

In hexaploid bread wheat ( Triticum aestivum L. em. Thell), ten members of the IWMMN ( International Wheat Microsatellites Mapping Network ) collaborated in extending the microsatellite (SSR = simple sequence repeat) genetic map. Among a much larger number of microsatellite primer pairs developed as a part of the WMC ( Wheat Microsatellite Consortium ), 58 out of 176 primer pairs tested were found to be polymorphic between the parents of the ITMI ( International Triticeae Mapping Initiative ) mapping population W7984 × Opata 85 (ITMI pop ). This population was used earlier for the construction of RFLP ( Restriction Fragment Length Polymorphism ) maps in bread wheat (ITMI map ). Using the ITMI pop and a framework map (having 266 anchor markers) prepared for this purpose, a total of 66 microsatellite loci were mapped, which were distributed on 20 of the 21 chromosomes (no marker on chromosome 6D). These 66 mapped microsatellite (SSR) loci add to the existing 384 microsatellite loci earlier mapped in bread wheat.

... 国际小麦微卫星协会(Wheat Microsatellite Consortium, WMC)也将开发的66个WMC标记整合到已有的小麦遗传框架图上^[30] ...

1998

9.737

0.0

Gale M

, Devos K

Comparative genetics in the grasses.

Proc Natl Acad Sci USA, 1998, 95: 1971-1974

John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH,United Kingdom

Genetic mapping of wheat, maize, and rice and other grass specieswith common DNA probes has revealed remarkable conservation of genecontent and gene order over the 60 million years of radiation of Poaceae. The linear organization of genes in some ninedifferent genomes differing in basic chromosome number from 5 to 12 andnuclear DNA amount from 400 to 6,000 Mb, can be described in terms ofonly 25 “rice linkage blocks.” The extent to which thisintergenomic colinearity is confounded at the micro level by geneduplication and micro-rearrangements is still an open question.Nevertheless, it is clear that the elucidation of the organization ofthe economically important grasses with larger genomes, such as maize(2 n = 10, 4,500 Mb DNA), will, to a greater orlesser extent, be predicted from sequence analysis of smaller genomessuch as rice, with only 400 Mb, which in turn may be greatly aided byknowledge of the entire sequence of Arabidopsis , whichmay be available as soon as the turn of the century. Comparativegenetics will provide the key to unlock the genomic secrets of cropplants with bigger genomes than Homo sapiens.

... STS (Sequence-tagged site)即序列标签位点, 是基因组上定位明确, 作为界标并能通过PCR扩增被唯一操作的短的单拷贝DNA序列^[31], 由于可直接在DNA分子水平上指出小麦中外源染色体片段的组成和位置, 在外源染色体鉴定方面应用广泛 ...

1996

3.658

0.0

... 前人利用STS标记已成功鉴定出小麦背景中的簇毛麦、黑麦等小麦近缘种的染色体片段^[32,33,34] ...

1996

1.668

0.0

... 前人利用STS标记已成功鉴定出小麦背景中的簇毛麦、黑麦等小麦近缘种的染色体片段^[32,33,34] ...

1995

3.658

0.0

... 前人利用STS标记已成功鉴定出小麦背景中的簇毛麦、黑麦等小麦近缘种的染色体片段^[32,33,34] ...