普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育

[目的]为进一步利用簇毛麦2V染色体上的有益基因,为小麦育种提供新种质。[方法]通过普通小麦-簇毛麦2V（ZD）二体代换系（DS2V）与普通小麦农林26-离果山羊草3c染色体二体异附加系（DA3C）杂交,综合运用染色体C-分带、基因组原位杂交和分子标记分析,并结合性状调查。[结果]从杂种后代中选育出小麦-簇毛麦纯合易位系T6BS·6BL-2VS,性状调查发现该易位系植株护颖颖脊上有刚毛。[结论]该易位系为杀配子染色体诱发的小片段易位;簇毛麦护颖颖脊刚毛基因定位于2VS的中部至端部。     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育

[目的]为进一步利用簇毛麦2V染色体上的有益基因,为小麦育种提供新种质。[方法]通过普通小麦-簇毛麦2V(2D)二体代换系(DS2V)与普通小麦农林26-离果山羊草3C染色体二体异附加系(DA3C)杂交,综合运用染色体C-分带、基因组原位杂交和分子标记分析,并结合性状调查。[结果]从杂种后代中选育出小麦-簇毛麦纯合易位系T6BS.6BL-2VS,性状调查发现该易位系植株护颖颖脊上有刚毛。[结论]该易位系为杀配子染色体诱发的小片段易位;簇毛麦护颖颖脊刚毛基因定位于2VS的中部至端部。     

普通小麦-簇毛麦2V染色体端体异附加系的选育与鉴定

簇毛麦(Haynaldia villosa) 具有抗多种病害、耐旱、耐寒等优良性状,是可供小麦改良利用的优良遗传资源.本研究综合利用根尖细胞有丝分裂中期染色体Giemsa C-分带、花粉母细胞减数分裂中期Ⅰ染色体构型分析、荧光原位杂交(genomic in situ hybridization,GISH) 及分子标记等技术,从普通小麦-簇毛麦2V(2D) 异代换系与含有Ph抑制基因的中国春高配对材料(pairing homoeologous inhibitor,Ph1) CO4-13的杂交后代中选了出分别含有簇毛麦2V长臂和短臂的普通小麦-簇毛麦2V端体异附加系.含有簇毛麦2V短臂的附加系在护颖颖脊上有刚毛,而在含2V长臂的附加系的护颖颖脊上没有刚毛,可将控制护颖颖脊刚毛性状的基因进一步定位在簇毛麦2V染色体的短臂上.筛选出町以追踪2V染色体短臂的SSR标记wmc25.该分子标记和护颖颖脊刚毛形态标记可用来追踪导入小麦遗传背景中的簇毛麦2V染色体短臂.     

涉及簇毛麦6V染色体结构变异体的筛选与鉴定

簇毛麦是小麦的一个野生近缘种,抗病性、抗逆性强,蛋白质和赖氨酸含量高,是小麦遗传改良的重要基因源.为定位、转移和利用簇毛麦有益基因,南京农业大学细胞遗传所通过花粉辐射,获得了一批涉及簇毛麦不同染色体且不同区段的结构变异体.该研究从这些结构变异体中筛选鉴定涉及簇毛麦6V染色体的结构变异体,共筛选鉴定出13种涉及簇毛麦6V染色体的结构变异体,包括2个整臂易位、5个小片段易位、4个大片段易位、1个缺失和1个中间插入易位,这些结构变异体的筛选鉴定将为进一步定位和利用簇毛麦6V染色体上的优异基因奠定基础.     

普通小麦-簇毛麦易位系T4VS·6AL的选育

在小麦背景中离果山羊草3C染色体具有优先传递的作用.当离果山羊草3C染色体处于单体状态时会导致后代不含有杀配子染色体的配子中产生包括缺失和易位等染色体结构变异.簇毛麦4V染色体携有抗小麦眼斑病和全蚀病基因.为进一步利用簇毛麦4V染色体上的有益基因.利用染色体C-分带和基因组原位杂交分析,从普通小麦-簇毛麦4V染色体二体异附加系(DA4V)与普通小麦农林26-离果山羊草3C染色体二体异附加系(DA3C)杂种后代中选育出小麦-簇毛麦纯合易住系T4VS·6AL.该易位系为杀配子染色体诱发形成的非补偿型易位;易位系T4VS·6AL高抗梭条花叶病.是小麦抗病育种的新种质.     

普通小麦-簇毛麦T5VS·5DL易位染色体对小麦主要农艺性状、品质和白粉病抗性的遗传效应分析

【目的】分析普通小麦-簇毛麦T5VS·5DL易位染色体对主要农艺性状、品质性状及白粉病抗性的遗传效应,了解T5VS·5DL易位染色体通过雌雄配子的传递情况,筛选便于育种利用的共显性分子标记,进一步明确T5VS·5DL易位系的高代回交品系在育种改良中的利用价值。【方法】分别以扬麦13、扬麦15为轮回亲本的高代T5VS·5DL易位系回交品系(BC4F4)及其分离群体(BC5F2)为材料,利用GISH及5VS特异分子标记对这些材料进行了鉴定;在大棚及大田2种环境下调查了这些材料的株高、穗长、小穗数、穗粒数、千粒重等主要农艺性状,并对这些材料的水溶剂保持力、碳酸钠溶剂保持力、蔗糖溶剂保持力、乳酸溶剂保持力、蛋白和籽粒硬度等品质性状进行了分析;还利用白粉菌混合生理小种对这些材料的苗期(二叶期)和成株期(抽穗期)进行了接种鉴定。【结果】含有T5VS·5DL易位染色体高代回交品系的株高、穗长、小穗数、穗粒数、千粒重等主要农艺性状与其轮回亲本相比,2种环境下的差异均不显著,表明簇毛麦5VS染色体臂代替普通小麦5DS染色体臂后,对产量性状的补偿性较好,没有显著的不利影响。品质性状的分析结果表明,含有T5VS·5DL易位染色体高代回交品系的籽粒硬度值(SKCS)均显著低于其轮回亲本,表明簇毛麦Dina/Dinb基因型比普通小麦的Pina/Pinb基因型具有更软质胚乳特性;另外,含有T5VS·5DL易位染色体高代回交品系的水溶剂保持力与碳酸钠溶剂保持力也显著低于轮回亲本,而蔗糖溶剂保持力、乳酸溶剂保持力及蛋白质含量的差异不显著,表明T5VS·5DL易位染色体对弱筋小麦品质指标可能有一定的正向效应。白粉菌混合生理小种接种鉴定的结果表明,在二叶期,含有T5VS·5DL易位染色体高代回交品系及其轮回亲本均高感白粉病,但在成株期含有T5VS·5DL易位染色体高代回交品系高抗白粉病,而其轮回亲本仍高感白粉病,表明含有T5VS·5DL易位染色体高代回交品系携带白粉病成株抗性基因。对BC5F2群体单株的白粉病及T5VS·5DL易位染色体鉴定的结果表明,所有含有T5VS·5DL易位染色体的单株均高抗白粉病,无T5VS·5DL易位染色体的单株均高感白粉病,推测一个显性的白粉病成株抗性基因与T5VS·5DL易位染色体共分离。同时在分离群体中,纯合易位染色体单株数、杂合单株数及无易位染色体单株数之比,经χ2测验符合1﹕2﹕1分离比例,表明T5VS·5DL易位染色体在小麦背景中遗传传递正常。通过对5VS特异分子标记的扩增条件及带型分析发现,共显性EST分子标记5EST-237、Pinb-1,扩增条件简单,特异带型易于识别,可以利用这些标记对T5VS·5DL易位染色体进行分子标记辅助选择。【结论】鉴于T5VS·5DL易位系良好的品质及抗病特性,建议在弱筋小麦的育种项目中加以利用。     

普通小麦-簇毛麦易位系T4VS·6AL的选育

在小麦背景中离果山羊草3C染色体具有优先传递的作用。当离果山羊草3C染色体处于单体状态时会导致后代不含有杀配子染色体的配子中产生包括缺失和易位等染色体结构变异。簇毛麦4V染色体携有抗小麦眼斑病和全蚀病基因。为进一步利用簇毛麦4V染色体上的有益基因,利用染色体C-分带和基因组原位杂交分析,从普通小麦-簇毛麦4V染色体二体异附加系（DA4V）与普通小麦农林26-离果山羊草3C染色体二体异附加系（DA3C）杂种后代中选育出小麦-簇毛麦纯合易位系T4VS·6AL。该易住系为杀配子染色体诱发形成的非补偿型易位;易位系T4VS·6AL高抗梭条花叶病。是小麦抗病育种的新种质。     

小麦-簇毛麦易位系中贮藏蛋白的鉴定及分析

为探讨簇毛麦染色体对小麦品质的影响,利用A-PAGE、SDS-PAGE和高效毛细管电泳(High-performance capillary electrophoresis,HPCE)技术对12个易位系及7个附加系的贮藏蛋白进行鉴定。将簇毛麦的一条ω-醇溶蛋白基因定位于1VL。结果表明:易位系T1DL·1VS可用来改良小麦品质;定位于簇毛麦1VS染色体上的高分子谷蛋白在小麦中表达不稳定;其他材料醇溶蛋白组成发生改变较小,可能对面筋相关品质影响不大,在育种工作中不会引起面筋相关品质下降;易位系材料中的小麦醇溶蛋白的质量分数有所增加。     

簇毛麦抗白粉病基因向四川小麦转移的分子标记育种研究

利用小麦 6VS/6AL易位系的抗白粉病基因Pm2 1的SCAR标记 ,对不同簇毛麦来源的小麦抗白粉病材料及其杂交后代进行了分子鉴定和标记辅助选择研究 ,结果表明利用该标记可以鉴定小麦遗传背景下的簇毛麦 6VS染色体臂 ,而且引物对小麦背景中的多年生簇毛麦也同样得到扩增。对簇毛麦染色体易位系 6VS/6DL与四川小麦品种杂交的后代群体 ,可以利用SCAR标记进行辅助抗性选择。本文还对四川小麦分子标记辅助育种的策略进行了讨论     

组织培养诱导普通小麦--簇毛麦抗白粉病易位系研究

采用白粉病抗性鉴定、谷草转氨酶GOT-2同工酶及分子原位杂交相结合的方法, 从幼胚培养T240组合(普通小麦×小麦-簇毛麦6D/6V异代换系) 的32个SC2 代株系中,筛选出T240-7株系, 其所有的抗病单株均缺失簇毛麦6V染色体长臂的GOT-V2 位点, 而具有6V染色体短臂上的抗白粉病基因。细胞学观察表明, 该株系易位染色体与小麦染色体可正常配对。经原位杂交分析, T240-7为杂合的臂间易位。     

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa (2n =2X = 14, VV), a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetie Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge (a remarkable morphology) mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V(2D) and the hybrid F1 was then self-crossed. Chromosome C-banding, genomie in situ hybridization (GISH), and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids F2 and F3. To date, four translocations including one small segmental translocation T6BS.6BL-2VS, two whole arm translocations (preliminarily designed as T3DS·2VL and T2VS·7DL) and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one iso- chromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25-120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 (2BL) tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS·7DL, Mt2VS, 2VS·2VS as well as the parent DS2V(2D), whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C of Aegilops triuncialis could successfully induce chromosome 2V structural changes.     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育(英文)

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育(英文)

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.     

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.     

Development of EST-PCR Markers for the Chromosome 4V of Haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of Haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.     

小麦贵农775抗条锈病新基因YrGA的研究

小麦抗病种质贵农775具有抗条锈性,研究其抗条锈遗传,对揭示其抗病机制和培育持久抗病品种具有重要意义。以西农97148×贵农775的杂交群体为材料,利用RAPD,SCAR分子标记和荧光原位杂交技术研究其抗性基因来源及在染色体上的位置。贵农775中的YrGA基因来自于簇毛麦,特异标记与抗条锈病基因YrGA(暂时命名)遗传距离为(0.355+0.001)cM,荧光原位杂交结果显示,贵农775为小麦-簇毛麦新的易位系。由于抗条锈病基因Yr26来源于簇毛麦,位于6VS,而与YrGA连锁的特异片段位于染色体长臂,综合分子生物学试验结果,可以推断YrGA很可能是一个来自簇毛麦并与已知抗条锈病基因不同的新基因。     

Molecular Characterization of a Triticum durum-Haynaldia villosa Amphiploid and Its Derivatives for Resistance to Gaeumannomyces graminis var. tritici

Take-all is a serious disease found in wheat across the world. Haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid （2n =42, AABBVV） between Triticum durum and Haynaldia viUosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization （GISH） analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.