合成六倍体小麦与其亲本在合成后小麦A/B染色体组微卫星位点的比较

构建人工合成六倍体小麦是利用小麦近缘材料优异基因的很有效的方法。但是目前在人工合成异源六倍小麦的过程中对微卫星位点的影响研究尚不完善。本研究直接比较了亲本四倍体小麦PS5与4个不同粗山羊草进行远缘杂交并经染色体自然加倍后获得4个人工合成六倍体小麦前后,位于普通小麦A/B染色体组不同染色体臂上的104对引物的变化。结果表明,104对微卫星引物的扩增产物在4个合成六倍体小麦中具有与普通小麦相同的带型;但在22对引物的扩增产物上存在差异,其中Am4与其它3个六倍体小麦Am1,Am2,Am3在15对引物扩增的条带存在差异;另外发现有45对特异与AB染色体的引物能够在粗山羊草中扩增出产物,其中特异于B染色体组的引物47.54%的可以在粗山羊草中扩增出产物,而A染色体组的引物占38.24%。因此,基于普通小麦开发的微卫星引物可以用于合成六倍体小麦的研究,而Am4材料与其它3个合成二倍体小麦的差异尚需进一步研究,另外我们推测普通小麦的B染色体组与A染色体组相比与粗山羊草存在较近的亲缘关系。     

粗山羊草抗条锈病新基因YrY206遗传分析和微卫星 标记

从小麦野生近缘属——粗山羊草中挖掘小麦条锈病抗病基因, 拓展小麦抗病性的遗传基础。利用抗小麦条锈病与感小麦条锈病的粗山羊草间杂交, 从粗山羊草[Aegilops tauschii (Coss.) Schmal] Y206中鉴定出1个显性抗小麦条锈病基因, 暂定名为YrY206。应用分离群体分组法(Bulked segregant analysis, BSA)筛选到Wmc11a、Xgwm71c、Xgwm161和Xgwm183标记, 与该基因之间的遗传距离分别为4.0、3.3、1.5和9.3 cM。根据连锁标记所在小麦微卫星图谱的位置, YrY206被定位在3DS染色体上。分析基因所在染色体的位置、抗病性特征, 认为YrY206是一个新的抗小麦条锈病基因。     

小麦-簇毛麦新种质山农05078的鉴定(简报)

簇毛麦（Dasypyrum villosum,VV,2n=14）属禾本科小麦族小麦亚族簇毛麦属,分蘖力比较强,多花,籽粒蛋白质含量高,耐旱,抗寒性好,高抗锈病、全蚀病和白粉病。硬簇麦（AABBVV,2n=42）是利用硬粒小麦和簇毛麦杂交人工合成的双二倍体,保留了簇毛麦的优良性状和特点,是进行小麦遗传改良的优良中间材料。人工合成小麦Am3（AABBDD,2n=42）是利用波斯小麦与粗山羊草杂交后染色体加倍而形成的双二倍体,但它所具有的A、B、D染色体组可能与经过多年分化的普通小麦所含的A、B、D染色体组不同,     

人工合成小麦抗白粉病未知基因的SSR标记

YAV-2/TEZ//A.SQ(895)是硬粒小麦与粗山羊草杂交获得的抗白粉病人工合成小麦。本研究利用人工合成小麦YAV-2/TEZ//A.SQ(895)与感白粉病的普通小麦品系品资50098杂交和自交获得的F2代群体及F3家系,在温室条件下鉴定群体的白粉病抗性。遗传分析结果表明,该抗白粉病基因为显性单基因遗传。利用647对小麦SSR引物进行了白粉病抗性基因的分子标记分析,结果表明该白粉病抗性基因与2A染色体的6个SSR标记连锁,与标记Xcfa2086的遗传距离最近,为11.8cM。     

粗山羊草居群间遗传分化及多样性的SSR分析

选用分布在粗山羊草14条染色体上的32对SSR引物,对来自中国河南、陕西、新疆和中东地区共147份粗山羊草材料进行遗传分化及多样性分析,结果表明在26个多态性位点中,等位基因数平均为4.15,Ne i基因多样性指数(He)平均为0.243,多态性信息含量指数(PIC)平均为0.226;居群间遗传变异差异明显,中东粗山羊草居群具有丰富的遗传变异(He=0.607,PIC=0.551),而来自陕西和河南的粗山羊草资源遗传多样性较低(He=0.055,PIC=0.047)和(He=0.024,PIC=0.021)。AMOVA分子变异分析显示,居群间遗传变异占总变异的52%,达到显著水平;河南粗山羊草和陕西粗山羊草间发生了一定的遗传分化(Fst=0.210),为研究中国粗山羊草资源的起源与分化问题提供了有用的信息与证据。     

大麦与小麦属间杂种的形态学和细胞遗传学研究

通过幼胚培养技术分别获得了大麦(Hordeum vulgare L., 2n=2x=14)与普通小麦(Triticum aestivum L., 2n=6x=42)、硬粒小麦(T. durum d. sf., 2n=4x=28)以及栽培二粒小麦(T. dicoccum Schrank 2n=4x=28)之间的属间杂种,平均杂交结实率分别为3.0％、1.2％和0.8％。杂种在形态上偏向小麦,自交不孕,用父本作回交亲本获得了大麦×普通小麦杂种的回交一代。所有杂种均表现细胞学上的不稳定性。杂种减数分裂中期1 PMC染色体平均配对频率和交叉结频率分别为27.31Ⅰ 0.33Ⅱ 0.011Ⅲ,0.45;20.571 0.20Ⅱ 0.008Ⅲ,0.25和20.41Ⅰ 0.28Ⅱ 0.005Ⅲ,0.32,表明双亲染色体组间不存在同源关系。大麦×四倍体小麦杂种小孢子的染色体计数表明,杂种中未减数雄配子的频率在60％以上。     

几种异质小麦核型的比较研究

本文以普通小麦中国春为对照,对(节节麦)中国春、(卵圆山羊草)中国春和(无芒山羊草)中国春3种同核异质杂种小麦的核型进行了分析比较,结果表明:它们的核型都与对照的一致,其核型公式均为2n=6x=42=36m+6sm,均属1A核型,从而证明了异源细胞质对普通小麦核型无明显影响。     

普通小麦与粗山羊草属间杂种的染色体构型及其后代的育性特征

利用3个推广品种(莱州953、山农辐63、陕7859)分别与原产地不同的抗白粉病的6份粗山羊草[Aegilops tauschii(Coss.)Schmal.]杂交,得到63个无胚乳的种子,将56枚幼胚接种到N6 0.5mg/L IBA 0.2 mg/L NAA的培养基上进行褓姆培养,得到37个植株。其中莱州953与粗山羊草的杂交结实率和成苗率较高,分别平均为8.58%和4.82%。粗山羊草对白粉病的抗性基因在不同的杂交组合中受到不同程度的改变或抑制。以莱州953为父本,分别与不同组合的杂种F_1回交,大多数组合均得到回交种子,回交结实率平均为1.70%;以莱州953作母本,与莱州953/Y225 F _1回交得到2粒种子,说明普通小麦与粗山羊草的杂种F_1也能产生少量有授精能力的花粉。以山农辐63为父本与山农辐63/Y219 F_1回交亦得到回交种子。通过对普通小麦与粗山羊草6个杂交组合的杂种F_1PMCMI染色体构型的分析,一般多出现14个左右单价体和一定频率的多价体,并观察到可能为A、B组染色体形成的异形二价体;粗山羊草的D组染色体和普通小麦的D组染色体联会正常,可发生自由重组,从而为将粗山羊草的有益基因导入普通小麦提供了细胞学依据。     

基因从普通小麦向山羊草属植物漂移可能性研究

分别以山羊草属4个不同的种为母本,以普通小麦为父本杂交,将获得的杂种再进行回交和自交。结果表明,不同种与普通小麦的可交配性存在较大差异,在人工多次授粉并有激素处理的条件下,粗山羊草与普通小麦的杂交结实率最高,2个基因型Ae42和Y92的杂交结实率分别为46.49%和22.58%;其次为卵穗山羊草,2个基因型Ae23和Y100的杂交结实率分别为12.11%和14.76%;柱穗山羊草位列第3,2个基因型Ae7和Y145的杂交结实率分别为2.23%和8.50%;拟斯卑尔脱山羊草最低,基因型Ae48的杂交结实率只有0.19%。不同种的杂种胚产生愈伤组织率不同,柱穗山羊草/小麦表现较高的水平,卵穗山羊草/小麦次之,粗山羊草/小麦第3,拟斯卑尔脱山羊草/小麦最低。卵穗山羊草/小麦的杂种幼胚直接成苗率最高,其次为粗山羊草/小麦,柱穗山羊草/小麦居第3位。山羊草与普通小麦杂种的育性较低,在自然状态下,只有卵穗山羊草/小麦能够自交结实,但自交结实率仅为0.044%,其他杂种自交不能结实。在人工多次授粉并激素处理条件下,用父、母本回交的结实率:卵穗山羊草/普通小麦组合分别为4.36%和3.71%,柱穗山羊草/普通小麦组合分别为0...     

小伞山羊草高分子量麦谷蛋白亚基及其基因的鉴定

运用SDS_PAGE和分子克隆技术 ,对小伞山羊草 (Aegilopsumbellulata ,UU ,2n =2x =14)的高分子量麦谷蛋白亚基 (1Ux ,1Uy)及其编码基因进行了鉴定。SDS_PAGE分析表明小伞山羊草不同基因型中的 1Ux的电泳迁移率接近或慢于普通小麦 1Dx2 .2亚基的电泳迁移率 ,1Uy亚基的电泳迁移率一般接近或慢于普通小麦的 1Dy类亚基。采用PCR扩增技术获得了 1Ux和 1Uy亚基编码基因的全长编码区 ,并对一个 1Uy基因的全长编码区进行了全序列测定。对推导的氨基酸序列进行比较发现 1Ux和 1Uy亚基具有与来自于其他物种的高分子量麦谷蛋白亚基一致的一级结构 ,聚类分析显示 1Ux和 1Uy亚基与D基因组编码的高分子量麦谷蛋白亚基在起源和进化上具有较高的相似性。     

Development of a set of Triticum aestivum-Aegilops tauschii introgression lines

New wheat introgression lines were obtained which contain different segments of individual chromosomes of Aegilops tauschii in the Triticum aestivum cv. 'Chinese Spring' background. The introgression lines were developed to examine various subsets of alleles from the wild grass in the genetic background of common wheat. As starting point substitution lines of 'Chinese Spring' in which single chromosomes of the D genome had been replaced by homologous chromosomes of a synthetic wheat were used. Synthetic wheat had been obtained earlier from a cross between the tetraploid emmer (genomes AABB) and wild grass Aegilops tauschii (genome DD). The seven wheat chromosome substitution lines carrying different chromosomes of Ae. tauschii were crossed twice to T. aestivum cv. 'Chinese Spring' and 259 BC1-progeny plants were analysed. Phenotypic evaluation was carried out for different traits such as plant height, spikelet number, peduncle length, flowering time, spike length, tiller number, grain weight per ear, fertility and thousand kernel weight. Genotypic analysis was performed using a set of 65 microsatellite markers previously mapped on the chromosomes of the D genome of wheat. During this analysis recombinant lines carrying different segments of Ae. tauschii chromosomes were detected. Plants containing small introgressions of the alien genetic material were selfed to get homozygous lines and plants carrying large pieces of the donor chromosome were backcrossed again to get smaller introgressions. Further microsatellite analysis of selected BC1F2-progeny plants resulted in detection of a first set of 36 homozygous lines carrying different pieces of Ae. tauschii genome.     

Analysis of 5S rDNA changes in synthetic allopolyploids Triticum x Aegilops

By the example of three synthetic allopolyploids: Aegilops sharonensis x Ae. umbellulata (2n =28), Triticum urartu x Ae. tauschii (2n =28), T. dicoccoides x Ae. tauschii (2n =42) the 5S rDNA changes at the early stage of allopolyploidization were investigated. Using fluorescent in situ hybridization (FISH), the quantitative changes affecting the separate loci of one of the parental genomes were revealed in plants of S3 generation of each hybrid combination. Souther hybridization with genomic DNA of allopolyploid T. urartu x Ae. tauschii (TMU38 x TQ27) revealed lower intensity of the fragments from Ae. tauschii compared with the T. urartu fragments. It may be confirmation of the reduction of signal on 1D chromosome that was revealed in this hybrid using FISH. Both appearance of a new 5S rDNA fragments and full disappearance of fragments from parental species were not showed by Southern hybridization, as well as PCR-analysis of 5-15 plants of S2-S3 generations. The changes were not found under comparison of primary structure of nine 5S rDNA sequences of allopolyploid TMU38 x TQ27 with analogous sequences from parental species genomes. The observable similarity by FISH results of one of the studied synthetic allopolyploids with natural allopolyploid of similar genome composition indicates the early formation of unique for each allopolyploid 5S rDNA organization.     

Simultaneous painting of three genomes in hexaploid wheat by BAC-FISH.

Fluorescence in situ hybridization (FISH) is widely used in the physical mapping of genes and chromosome landmarks in plants and animals. Bacterial artificial chromosomes (BACs) contain large inserts, making them amenable for FISH mapping. In our BAC-FISH experiments, we selected 56 restriction fragment length polymorphism (RFLP)-locus-specific BAC clones from the libraries of Triticum monococcum and Aegilops tauschii, which are the A- and D-genome donors of wheat (Triticum aestivum, 2n = 6x = 42), respectively. The BAC clone 676D4 from the T. monococcum library contains a dispersed repeat that preferentially hybridizes to A-genome chromosomes, and two BAC clones, 9I10 and 9M13, from the Ae. tauschii library contain a dispersed repeat that preferentially hybridizes to the D-genome chromosomes. These repeats are useful in simultaneously discriminating the three different genomes in hexaploid wheat, and in identifying intergenomic translocations in wheat or between wheat and alien chromosomes. Sequencing results show that both of these repeats are transposable elements, indicating the importance of transposable elements, especially retrotransposons, in the genome evolution of wheat.     

远缘杂交不需幼胚培养的节节麦基因型

六倍体普通小麦(Triticum aestivum L.)是由四倍体小麦(T.turgidum L.)与二倍体节节麦(Aegilops tanschii Coss.)天然杂交然后通过染色体自然加倍形成的异源多倍体.这一起源过程是自然条件下天然发生的,它的发生需要具备一个条件:四倍体小麦与节节麦的天然杂交种子在自然条件(没有幼胚培养等)下能够正常发芽出苗.我们从22份节节麦中发现来自中东的节节麦AS60在不采用幼胚培养等人工辅助条件下,仍然很容易与四倍体小麦和普通小麦产生有生活力的杂种植株.AS60与四倍体小麦的杂交种子有50.0%(反交)及57.1%(正交)的种子,而AS60与六倍体普通小麦的杂交种子则有45.5%不需幼胚培养等措施能够正常发芽、生长.AS60的这一特征正是普通小麦起源过程需要的条件.最后探讨了这一发现对小麦遗传改良和对普通小麦起源演化研究的意义.     

普通小麦与粗山羊草正,反交的育性研究

普通小麦与粗山羊草杂交需借助幼胚培养方可获得杂种,取授粉１２～１６天的幼胚进行拯拯救,成苗率较高,效果较好。而不同杂交方式对共杂种成胚率、成苗率及思胚拯救率影响很大。以普通小麦为母本,粗山羊草为父杂交时,１５个杂交组合平均杂种成胚率,成苗率及幼胚拯率分别为５．３５％和５８．７３％,而在１２个反交组合中,其平均成胚率,成苗率及幼胚拯救率分别为９８．８８％、１７．１０％和２４．４４％。由此可见,普通小     

加拿大披碱草和圆柱披碱草BC1植株的细胞遗传学鉴定

以加拿大披碱草(2n=4x=28)和圆柱披碱草(2n=6x=42)为材料,对其BC1植株染色体数目、配对构型以及花粉育性和结实性等进行了鉴定。结果显示:BC1代85%以上细胞染色体数目为28条(2n=4x=28);BC1植株的花粉母细胞减数分裂中期Ⅰ染色体配对行为较规则,其平均染色体构型分别为0.04Ⅰ 13.98Ⅱ,且环状二价体多于棒状二价体;BC1植株的花粉可育率和自然结实率分别为84.83%和70.98%,说明BC1植株的育性已得到恢复,为其后代优良株系的选育奠定了基础。     

Cytological Studies on F1 Hybrids Between Hexaploid Aegilops and Triticum aestivum

Morphologcal and cytological studies of hybrids between hexaploid Aegilops crassa Boiss. (2n = 6x = 42, DDD2 D2Mcr Mcr), Ae. vavilovii (Zhuk.) Chen. (2n = 6x = 42, DDMcr McrSp Sp) and Triticum aestivum L. (2n= 6x = 42, AABBDD) were carried out. The results showed that most of the F1 hybrids morphologically resembled their Aegilops parental species. Four Fl hybrids of Ae. vavilovii × T. aestivum and one of Ae. crassa × T. aestivum produced seeds containing few endosperms. The percentage of seed obtained ranged from 0.1% to 6.5 %. These seeds were not vigorous and only a few of them germinated. A large number of univalents appeared at meiosis MI. The frequencies of bivalents were lower than those being theoriticaly estimated. These results indicated that the D genomes in Ae. Crassa and Ae. vavilovii may have been substantially modified. Trivalents were observed in all Fl hybrids. Quadrivalents and pentavalents were also observed in some PMCs during meiosis. The chiasmata frequencies in hybrids using Aegilops species as maternal parents were higher than those in their reciprocal ones. Chromosome segrigations were abnormal at A Ⅰ and A Ⅱ . Polyads and micronuclei appeared frequently at telophase tetrad stage. A plant with 21 chromosomes was obtained in Ae. vavilovii × Jimai 30, although the reason was not yet clear.     

Barley chromosome identification using genomic in situ hybridization in the genome of backcrossed progeny of barley-wheat amphiploids [H. geniculatum All. (2n = 28) x T. aestivum L. (2n = 42)] (2n = 70)

Genomic in situ hybridization (GISH) has been used to study characteristics of the formation of alloplasmic lines detected among self-pollinated backcrossed progeny (BC1F5-BC1F8) of barley--wheat amphiploids [Hordeum geniculatum All. (2n = 28) x Triticum aestivum L. (2n = 42)] (2n = 70). The chromosome material of the wild barley H. geniculatum has been shown to contribute to these lines. For example, fifth-generation plants (BC1F5) had genotypes (2n = 42w + 2g), (2n = 42w + 1g + 1tg), and (2n = 41w + 1g), where w is common wheat chromosomes, g is barley (H. geniculatum) chromosomes, and tg is the telocentric chromosome of wild barley. Beginning from the BC1F6 generation, alloplasmic telocentric addition lines (2n = 42 + 2tg) and (2n = 42 + 1tg) appear. This lines has been found cytogenetically unstable. The progeny of each of these cytological types include not only the (2n = 42 + 2tg) and (2n = 42 + 2tg) addition plants, but also plants with the monosomic (2n = 41 + 1tg) and the disomic (2n = 40 + 2tg) substitutions, as well as the (2n = 41 + 2tg) plants, which lack one wheat chromosome and have two telocentric barley chromosomes. It has been demonstrated that the selection for well-filled grains favors the segregation of telocentric addition lines (2n = 42 = 2tg) and (2n = = 42 + 1tg).     

Molecular analysis, cytogenetics and fertility of introgression lines from transgenic wheat to Aegilops cylindrica host

Natural hybridization and backcrossing between Aegilops cylindrica and Triticum aestivum can lead to introgression of wheat DNA into the wild species. Hybrids between Ae. cylindrica and wheat lines bearing herbicide resistance (bar), reporter (gus), fungal disease resistance (kp4), and increased insect tolerance (gna) transgenes were produced by pollination of emasculated Ae. cylindrica plants. F1 hybrids were backcrossed to Ae. cylindrica under open-pollination conditions, and first backcrosses were selfed using pollen bags. Female fertility of F1 ranged from 0.03 to 0.6%. Eighteen percent of the sown BC1s germinated and flowered. Chromosome numbers ranged from 30 to 84 and several of the plants bore wheat-specific sequence-characterized amplified regions (SCARs) and the bar gene. Self fertility in two BC1 plants was 0.16 and 5.21%, and the others were completely self-sterile. Among 19 BC1S1 individuals one plant was transgenic, had 43 chromosomes, contained the bar gene, and survived glufosinate treatments. The other BC1S1 plants had between 28 and 31 chromosomes, and several of them carried SCARs specific to wheat A and D genomes. Fertility of these plants was higher under open-pollination conditions than by selfing and did not necessarily correlate with even or euploid chromosome number. Some individuals having supernumerary wheat chromosomes recovered full fertility.