A high-density genetic linkage map of Aegilops tauschii, the D-genome progenitor of bread wheat

 Aegilops tauschii is the diploid D-genome progenitor of bread wheat (Triticum aestivum L. em Thell, 2n=6x=42, AABBDD). A genetic linkage map of the Ae. tauschii genome was constructed, composed of 546 loci. One hundred and thirty two loci (24%) gave distorted segregation ratios. Sixty nine probes (13%) detected multiple copies in the genome. One hundred and twenty three of the 157 markers shared between the Ae. tauschii genetic and T. aestivum physical maps were colinear. The discrepancy in the order of five markers on the Ae. tauschii 3DS genetic map versus the T. aestivum 3D physical map indicated a possible inversion. Further work is needed to verify the discrepancies in the order of markers on the 4D, 5D and 7D Ae. tauschii genetic maps versus the physical and genetic maps of T. aestivum. Using common markers, 164 agronomically important genes were assigned to specific regions on Ae. tauschii linkage, and T. aestivum physical, maps. This information may be useful for map-based cloning and marker-assisted plant breeding. Received: 23 March 1998 / Accepted: 27 October 1998     

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.     

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。     

Identification of molecular markers linked to Pm13, an Aegilops longissima gene conferring resistance to powdery mildew in wheat

 RFLP, RAPD, STS and DDRT-PCR techniques were applied to find molecular markers linked to Pm13, an Aegilops longissima gene conferring resistance to powdery mildew in wheat. The experimental strategy was based on the differential comparison of DNAs from common wheat and from common wheat/Ae. longissima recombinant lines carrying short segments of the 3S ^l S chromosome arm containing the Pm13 gene. Sixteen RFLP clones that detect loci previously located in the short arms of group-3 wheat chromosomes were screened for their ability to hybridise to Ae. longissima restriction fragments derived from the 3S ^l S segments introgressed into the recombinant lines. Eight RFLP clones and one STS marker detected 3S ^l S-specific fragments whose location relative to the wheat-alien chromatin breakage point of the recombinant lines was determined. Four amplification products were identified through the screening of about 200 RAPD primers. Their polymorphism was associated with the introgression of the alien DNA. One of the differential fragments was derived from the 3S ^l S DNA segment, while the remaining three corresponded to the replaced 3DS DNA. Further analyses carried out using 40 combinations of DDRT-PCR primers detected an additional reproducible polymorphism associated with the presence of 3S ^l S DNA. In view of their possible utilisation in Pm13 marker-assisted selection, differentially amplified RAPD and DDRT-PCR fragments were cloned, transformed into RFLP markers and converted into STS markers. Received: 23 March 1998 / Accepted: 5 August 1998     

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。     

节节麦细胞不同分裂时期和阶段的G—带核型及其变动性分析

采用改良的ASG法获得了中期和3个染色体凝缩程度不同的早中期阶段（分别称为早中期Ⅰ、Ⅱ、Ⅲ）染色体的G-带,并进行了G-带核型和变动性分析。所分析的分裂时期和阶段,每条染色体的全长显示出了密切邻近的多重的带纹,带纹细窄、大小较相近,带间区小,带纹分布较密集而均匀。随着有丝分裂进程推进,染色体的带纹数目减少,早中期Ⅰ、Ⅱ、Ⅲ于中期单倍染色体组的G-带带纹总数分别减少41%、36%、28%,而染色体组的绝对长度分别缩短43%、37%、27%,带数减少幅度与染色体长度缩短的幅度几乎相等。早中期Ⅰ至早中期Ⅱ、Ⅲ和早中期Ⅱ至早中期Ⅲ的带纹减少幅度与染色体长度缩短幅度也基本一致。染色体组中各染色体之间带纹减少和染色体缩短的比例不尽相同,有一定的变幅。早中期Ⅰ、Ⅱ、Ⅲ和中期染色体组中每单位绝对长度的带数（带/μm）分别为2.19、2.22、2.32和2.29,差异不大。对节节麦G-带的特性等问题进行了讨论。     

粗山羊草(Aegilops tauschii)中Pinb基因的克隆和表达分析

puroindoline a(Pina)和puroindoline b(Pinb)是控制小麦籽粒硬度的主效基因。根据已报道的小麦Pinb基因的保守序列,设计合成了一对特异性引物,对粗山羊草Aegilops tauschii(DD)的基因组DNA进行Pinb基因扩增、克隆和序列分析,发现了一个新型Pinb等位基因。该基因长447 bp,编码148个氨基酸残基,具有麦类作物PinB蛋白所特有的WPTKWWK色氨酸结构域和10个半胱氨酸所形成的5个二硫键结构。与软粒小麦cv.Capitole的Pinb-D1a相比较,该基因含有14个氨基酸变异位点,其中包括一个紧邻色氨酸结构域的变异位点(Val66Phe),其核苷酸和氨基酸同源性分别为93.3%和90.5%。RT-PCR和Western Blot证实了Pinb基因在籽粒胚乳中的表达。Southern Blot分析结果表明,粗山羊草中Pinb基因为单拷贝。研究结果表明,粗山羊草中包含着与小麦差异较大的籽粒硬度控制基因,对此基因的进一步研究将加深对小麦籽粒硬度形成分子机制的了解。     

Expression of high zinc efficiency of Aegilops tauschii and Triticum monococcum in synthetic hexaploid wheats

The effect of varied zinc (Zn) supply on shoot and root dry matter production, severity of Zn deficiency symptoms and Zn tissue concentrations was studied in two Triticum turgidum (BBAA) genotypes and three synthetic hexaploid wheat genotypes by growing plants in a Zn-deficient calcareous soil under greenhouse conditions with (+Zn=5 mg kg^-1 soil) and without (−Zn) Zn supply. Two synthetic wheats (BBAADD) were derived from two different Aegilops tauschii (DD) accessions using same Triticum turgidum (BBAA), while one synthetic wheat (BBAAAA) was derived from Triticum turgidum (BBAA) and Triticum monococcum (AA). Visible symptoms of Zn deficiency, such as occurrence of necrotic patches on leaves and reduction in shoot elongation developed more rapidly and severely in tetraploid wheats than in synthetic hexaploid wheats. Correspondingly, decreases in shoot and root dry matter production due to Zn deficiency were higher in tetraploid wheats than in synthetic hexaploid wheats. Transfer of the DD genome from Aegilops tauschii or the AA genome from Triticum monococcum to tetraploid wheat greatly improved root and particularly shoot growth under Zn-deficient, but not under Zn-sufficient conditions. Better growth and lesser Zn deficiency symptoms in synthetic hexaploid wheats than in tetraploid wheats were not accompanied by increases in Zn concentration per unit dry weight, but related more to the total amount of Zn per shoot, especially in the case of synthetic wheats derived from Aegilops tauschii. This result indicates higher Zn uptake capacity of synthetic wheats. The results demonstrated that the genes for high Zn efficiency from Aegilops tauschii (DD) and Triticum monococcum (AA) are expressed in the synthetic hexaploid wheats. These wheat relatives can be used as valuable sources of genes for improvement of Zn efficiency in wheat. This revised version was published online in June 2006 with corrections to the Cover Date.     

Sequence evidence for sporadic intergeneric DNA introgression from wheat into a wild Aegilops species

Introgressive hybridization has played a crucial role in the evolution of many plant species, especially polyploids. The duplicated genetic material and wide geographical distribution facilitate hybridization and introgression among polyploid species having either homologous or homoeologous genomes. Such introgression may lead to the production of recombinant genomes that are more difficult to form at the diploid level. Crop genes that have introgressed into wild relatives can increase the capability of the wild relatives to adapt to agricultural environments and compete with crops or to compete with other wild species. Although the transfer of genes from crops into their conspecific immediate wild progenitors has been reported, little is known about spontaneous gene movement from crops to more distantly related species. We describe recent spontaneous DNA introgression from domesticated polyploid wheat into distantly related, wild tetraploid Aegilops peregrina (syn. Aegilops variabilis) and the stabilization of this sequence in wild populations despite not having homologous chromosomes. Our results show that DNA can spontaneously introgress between homoeologous genomes of species of the tribe Triticeae and, in the case of crop-wild relatives, possibly enrich the wild population. These results also emphasize the need for fail-safe mechanisms in transgenic crops to prevent gene flow where there may be ecological risks.     

节节麦的酯酶同工酶分析

对 30份不同来源的节节麦进行 4个时期的酯酶同工酶分析。结果表明 :不同来源节节麦的酯酶同工酶存在较大差异 ,共分成 1 5种基本类型。我国黄河流域的 1 0份节节麦被划分为 2个基本类型 ,但二者关系极为相近 ;新疆节节麦与之有一定差异 ,但在相似系数≤ 0 .82 0时可视为一类。所有材料在 4个时期之间没有出现一个完全相同的酶带类型 ,说明酯酶同工酶随发育时期而不断变化。     

Production of aneuhaploid and euhaploid sporocytes by meiotic restitution in fertile hybrids between durum wheat Langdon chromosome substitution lines and Aegilops tauschii

Fertile F1 hybrids were obtained between durum wheat (Triticum durum Desf.) Langdon (LDN) and its 10 disomic substitution (LDN DS) lines with Aegilops tauschii accession AS60 without embryo rescue. Selfed seedset rates for hybrids of LDN with AS60 were 36.87% and 49,45% in 2005 and 2006, respectively. Similar or higher selfed seedset rates were observed in the hybrids of ID (1A), 1D (1B), 3D (3A), 4D (4B), 7D (TA), and 2D (2B) with AS60, while lower in hybrids of 3D (3B) + 3BL, 5D (5A) + 5AL, 5D (5B) + 5B and 6D (6B) + 6BS with AS60 compared with the hybrids of LDN with AS60. Observation of male gametogenesis showed that meiotic restitution, both first-division restitution (FDR) and single-division meiosis (SDM) resulted in the formation of functional unreduced gametes, which in turn produced seeds. Both euhaploid and aneuhaploid gametes were produced in Fi hybrids. This suggested a strategy to simultaneously transfer and locate major genes from the ancestral species T. turgidum or Ae. tauschii. Moreover, there was no significant difference in the aneuhaploid rates between the F1 hybrids of LDN and LDN DS lines with AS60, suggesting that meiotic pairing between the two D chromosomes in the hybrids of LDN DS lines with AS60 did not promote the formation of aneuhaploid gametes.     

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。     

Results of long-term field studies into the effect of strobilurin containing fungicides on the production of mycotoxins in several winter wheat varieties

In total, 23 field trials have been performed during the past 5 years to assess the effect of strobilurin containing fungicides on mycotoxin production in winter wheat. In 85% of the plots, the treatment with strobilurins at growth stages before blossom increased the content of deoxynivalenol in comparison with untreated controls. This effect was intensified by treatments at BBCH stage 45–49 and could be demonstrated in a great number of various cultivars. Application of strobilurin containing fungicides at early growth stages seems to be a risk factor for mycotoxin formation in wheat, especially under favourable conditions forFusarium infections. Presented at the 26^th Mykotoxin-Workshop in Herrsching. Germany, May 17–19, 2004     

Microsatellite analysis of Aegilops tauschii germplasm

The highly polymorphic diploid grass Aegilops tauschii isthe D-genome donor to hexaploid wheat and represents a potential source for bread wheat improvement. In the present study microsatellite markers were used for germplasm analysis and estimation of the genetic relationship between 113 accessions of Ae. tauschii from the gene bank collection at IPK, Gatersleben. Eighteen microsatellite markers, developed from Triticum aestivum and Ae. tauschii sequences, were selected for the analysis. All microsatellite markers showed a high level of polymorphism. The number of alleles per microsatellite marker varied from 11 to 25 and a total of 338 alleles were detected. The number of alleles per locus in cultivated bread wheat germplasm had previously been found to be significantly lower. The highest levels of genetic diversity for microsatellite markers were found in accessions from the Caucasian countries (Georgia, Armenia and the Daghestan region of Russia) and the lowest in accessions from the Central Asian countries (Uzbekistan and Turkmenistan). Genetic dissimilarity values between accessions were used to produce a dendrogram of the relationships among the accessions. The result showed that all of the accessions could be distinguished and clustered into two large groups in accordance with their subspecies taxonomic classification. The pattern of clustering of the Ae. tauschii accessions is according to their geographic distribution. The data suggest that a relatively small number of microsatellites can be used to estimate genetic diversity in the germplasm of Ae. tauschii and confirm the good suitability of microsatellite markers for the analysis of germplasm collections. Received: 8 September 1999 / Accepted: 7 October 1999     

小麦赤霉病镰孢菌毒素的生物合成及其分子调控

禾谷镰刀菌是小麦赤霉病的主要致病菌,其真菌次生代谢产生的单端孢霉烯类B型毒素,如雪腐镰刀菌烯醇(nivalenol,NIV)、脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)和其它乙酰化衍生物等污染小麦籽粒后对人畜健康构成威胁。综述了近年来国内外对小麦赤霉病镰孢菌单端孢霉烯类B型毒素生物合成的主要途径及分子调控研究进展,对毒素合成过程中的重要调控基因如TRI5、TRI7和TRI13在农业中的应用进行了阐述。     

Retroelement insertional polymorphisms, diversity and phylogeography within diploid, D-genome Aegilops tauschii (Triticeae, Poaceae) sub-taxa in Iran

Background and Aims: The diploid goat grass Aegilops tauschii (2n = 2x = 14) is nativeto the Middle East and is the D-genome donor to hexaploid breadwheat. The aim of this study was to measure the diversity ofdifferent subspecies and varieties of wild Ae. tauschii collectedacross the major areas where it grows in Iran and to examinepatterns of diversity related to the taxa and geography. Methods: Inter-retroelement amplified polymorphism (IRAP) markers wereused to analyse the biodiversity of DNA from 57 accessions ofAe. tauschii from northern and central Iran, and two hexaploidwheats. Key Results: Eight IRAP primer combinations amplified a total of 171 distinctDNA fragments between 180 and 3200 bp long from the accessions,of which 169 were polymorphic. On average, about eight fragmentswere amplified with each primer combination, with more bandsbeing amplified from accessions from the north-west of the countrythan from other accessions. Conclusions: The IRAP markers showed high levels of genetic diversity. Analysisof all accessions together did not allow the allocation of individualsto taxa based on morphology, but showed a tendency to put accessionsfrom the north-west apart from others regions. It is speculatedthat this could be due to different activity of retroelementsin the different regions. Within the two taxa with most accessions,there was a range of IRAP genotypes that could be correlatedclosely with geographical origin. This supports suggestionsthat the centre of origin of the species is towards the south-eastof the Caspian Sea. IRAP is an appropriate marker system toevaluate genetic diversity and evolutionary relationships withinthe taxa, but it is too variable to define the taxa themselves,where more slowly evolving morphological, DNA sequence or chromosomalmakers may be more appropriate.