Isolation, identification and characterization of disomic and translocated barley chromosome addition lines of common wheat

Two disomic barley chromosome addition lines and five translocated chromosome addition lines of common wheat cultivar Shinchunaga were isolated. They were derived from a hybrid plant between Shinchunaga and cultivated barley Nyugoruden (New Golden) by backcrossing with wheat and self pollination. Barley chromosomes added to chromosome arms involved in the translocated chromosomes were identified by C-banding method and by crossing these lines with Chinese Spring/Betzes addition lines. Two disomic addition lines were identified to have chromosome 6 and 7 of barley, respectively. Two of the five translocated chromosome addition lines were clarified to have same chromosome constitution, 42 wheat chromosomes and a pair of translocated chromosomes constituted with a long arm of chromosome 5B of wheat and a short arm of chromosome 7 of barley. The other three lines could not be identified due to chromosome rearrangement. Performances of these seven lines on agronomic characters were examined. Addition of barley chromosome 7 induced early heading, and chromosome 6 showed lated heading. Almost all of the lines except that of chromosome 6 showed short culm length and all showed reduced number of tillers, spikelets and grains per ear, and low seed fertility. These lines would be useful for genetic analyses in wheat and barley and for induction of useful genes of barley into wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

Substituting Ability of Individual Barley Chromosomes for Wheat Chromosomes 1. Substitutions Involving Barley Chromosomes 1, 3 and 6

In order to determine the genetic relatedness of individual barley chromosomes to wheat chromosomes, ‘Betzes’ barley chromosomes 1, 3 and 6 were substituted for individual ‘Chinese Spring’ wheat chromosomes of homoeologous groups 7, 3 and 6, respectively. The substitution status of these lines has been confirmed using isozyme selective markers, chromosome pairing behaviour in F₁ hybrids between the substitution lines and the appropriate double ditelocentric stocks of wheat, and hybridization of cDNA probes to the genomic DNA digests of these substitution lines. Each of the three barley chromosomes provided genetic compensation for the wheat chromosomes they replaced in the substitution plants. From the basis of this compensation with respect to plant vigour and fertility, barley chromosomes 1, 3 and 6 have been designated 7H, 3H and 6H.     

The effect of temperature on seed set and embryo development in reciprocal crosses of wheat and barley

Summary The effect of temperature on seed set and embryo development in reciprocal crosses of barley and wheat was assessed in crosses involving two spring barley varieties (Betzes, Martonvásári 50) and one wheat variety (Chinese Spring). Detached tillers placed in nutrient solution were pollinated in controlled environments at constant day-night temperature regimes (12, 15, 18 and 21° C) with a light intensity of 30,000 lux and a relative humidity of 80%. When barley was used as the female, lower temperatures (12 and 15° C) produced the maximum seed set, whereas for the reciprocal cross, the highest temperature (21° C) produced the best seed set in the Chinese Spring × Betzes combination.Low temperature retarded the embryo development. The highest numbers of hybrid plants were produced at 18° C and 21° C in the barley × wheat cross and in the wheat × barley cross, respectively. Embryos of about 1.5 mm length in the barley × wheat cross, and of about 1.0 mm length in the wheat × barley cross germinated successfully. The smallest embryo giving rise to hybrid plants was 0.57 mm in the barley × wheat cross and 0.51 mm in the wheat × barley cross.     

Detection of an earliness per se quantitative trait locus in the proximal region of wheat chromosome 5AL

A homoeologous quantitative trait locus to that of eps5L on barley chromosome 5H was identified in a syntenic region of wheat chromosome 5A. Wheat single chromosome recombinant lines (SCRs) were developed from a cross between ‘Chinese Spring’(‘Cappelle-Desprez’ 5A) and ‘Chinese Spring’(Triticum spelta 5A), these were grown together with the parental controls under different vernalization and photoperiod regimes. The variation for ear emergence time accelerated heading induced by the T. spelta segment indicated an effect associated with the Xcdo412-Xbcd9 interval. Since no differences between the SCRs and controls in responses to vernalization and photoperiod treatments were detected, this effect was identified as an earliness per se gene, Q Eetocs-5 A.2, which may be homoeologous to the eps5L quantitative trait locus of barley. Xbcd926 has been found to be closely linked to the rice flowering time quantitative trait loci, QHd9a or FLTQ2, on chromosome 9, suggesting possible relationships among the quantitative trait loci across wheat, barley and rice genomes.     

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC₂F₂ progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

小麦EST-SSR标记的开发和染色体定位及其在追踪黑麦染色体中的应用

根据小麦盐胁迫诱导和茎秆组织相关EST序列开发了81对EST-SSR引物, 其中67、46、18和61对分别在小麦、黑麦、簇毛麦和大麦基因组中稳定扩增, 在不同小麦和大麦品种间具有多态性的引物分别有22和23对。利用小麦缺体-四体系共定位了43对引物的81个位点, 其中A、B和D染色体组上分别有29、30和22个位点, 涉及除4B、3D和6D外的18条染色体。此外30对引物在黑麦基因组中具有特异扩增, 其中8对分别在黑麦1R、4R、5R和R7染色体上具有特异扩增, 7对在多条黑麦染色体具有相同扩增。这些新标记可有效用于小麦及其近缘物种的遗传作图与比较遗传研究。     

Mapping of genes involved in glutathione, carbohydrate and COR14b cold induced protein accumulation during cold hardening in wheat

Using some of the chromosome substitution lines developed from thecrosses of the donor Cheyenne to Chinese Spring we showed that theaccumulation of water soluble carbohydrates during different stages ofhardening was time dependent. Moreover there was a significantcorrelation between the rate of carbohydrate accumulation and the frosttolerance. The expression and regulation of a wheat gene homologous tothe barley cold regulated cor14b gene was compared in frost sensitiveand frost tolerant wheat genotypes at different temperatures. Studies madewith chromosome substitution lines showed that the threshold inductiontemperature polymorphism of the cor14b wheat homologous genewas controlled by loci located on chromosome 5A of wheat, while cor14b gene was mapped, in Triticum monococcum, onto the longarm of chromosome 2A^m. Our study on the effect of cold hardeningon glutathione (GSH) metabolism showed that chromosome 5A of wheathas an influence on the GSH accumulation and on the ratio of reduced andoxidised glutathione as part of a complex regulatory function during coldhardening. In addition, the level of increase in GSH content duringhardening may indicate the degree of the frost tolerance of wheat.     

簇毛麦1V染色体的传递及品质效应分析

簇毛麦是小麦的一个野生近缘种,小麦-簇毛麦1V异附加系和异代换系的蛋白质含量和沉降值均高,将簇毛麦1V染色体的优质基因导入普通小麦,进一步创造小麦-簇毛麦1V染色体易位系是小麦品质改良的有效途径.以小麦-簇毛麦1V异染色体系材料为基础,用普通小麦连续回交,结合原位杂交和PCR标记鉴定方法,分析1V染色体以及1V结构变异...     

普通小麦背景中长穗偃麦草[Lophopyrum elongatum(Host) A. Lve]E染色体组的RGAP特异标记

利用已知植物抗病基因编码氨基酸保守区域NBS-LRR(核苷酸结合位点-富亮氨酸区域)设计了42个简并引物组合,运用抗病基因类似物多态性(resistance gene analog polymorphism,RGAP)分子标记技术,对中国春、中国春-长穗偃麦草双二倍体及其附加系和代换系基因组DNA进行PCR扩增。结果表明,共有38对引物组合获得扩增产物,其中35对在普通小麦中国春、中国春-长穗偃麦草双二倍体中能扩增出多态性,平均每个引物组合扩增出38.5个片段。在普通小麦背景下,共获得275条长穗偃麦草E基因组多态性谱带,占扩增总谱带数的17.44%,揭示出在普通小麦背景下E基因组和普通小麦A、B、D基因组间的高丰度遗传变异。另外,利用RGAP分子标记技术,构建了一套完整的长穗偃麦草1E~7E染色体的特异RGAP标记。为小麦背景中长穗偃麦草外源遗传物质的快速检测提供了新途径。     

Molecular and physical mapping of genes affecting awning in wheat

Quantitative trait loci (QTL) for three traits related to awning (awn length at the base, the middle and the top of the ear) in wheat were mapped in a doubled‐haploid line (DH) population derived from the cross between the cultivars ‘Courtot’ (awned) and ‘Chinese Spring’ (awnless) and grown in Clermont‐Ferrand, France, under natural field conditions. A molecular marker linkage map of this cross that was previously constructed based on 187 DH lines and 550 markers was used for the QTL mapping. The genome was well covered (more than 95%) and a set of anchor loci regularly spaced (one marker every 20.8 cM) was chosen for marker regression analysis. For each trait, only two consistent QTL were identified with individual effects ranging from 8.5 to 45.9% of the total phenotypic variation. These two QTL cosegregated with the genes Hd on chromosome 4A and B2 on chromosome 6B, which are known to inhibit awning. The results were confirmed using ‘Chinese Spring’ deletion lines of these two chromosomes, which have awned spikes, while ‘Chinese Spring’ is usually awnless. No quantitative trait locus was detected on chromosome 5A where the B1 awn‐inhibitor gene is located, suggesting that both ‘Courtot’ and ‘Chinese Spring’ have the same allelic constitution at this locus. The occurrence of awned speltoid spikes on the deletion lines of this chromosome suggests that ‘Chinese Spring’ and ‘Courtot’ have the dominant B1 allele, indicating that B1 alone has insufficient effect to induce complete awn inhibition.     

筛选一组用于鉴定小麦缺体-四体真实性的SSR分子标记

[目的]旨在筛选一组数量较大,可靠性较高的SSR分子标记,用于辅助鉴定小麦缺体-四体的真实性。[方法]方法是在小麦SSR分子标记图谱上,选取单一位点标记引物,并在中国春和已鉴定的缺体-四体DNA中扩增检测,筛选带型清晰,易识别的单扩增产物标记。[结果]结果表明,在小麦21个连锁群上共选择标记150个,长短臂均有分布,最终筛选出标记67个,这些标记在中国春和所有非连锁的缺体-四体中扩增为阳性,在对应的连锁的缺体-四体中扩增为阴性,扩增产物为单条带,每个连锁群上最少3个,最多5个。[结论]该套引物,带型清晰,易识别,单一位点,适用于所有中国春小麦缺体-四体及其他非整倍体的真实性鉴定。     

Locating supplementary RFLP markers on barley chromosome 7 and synteny with homoeologous wheat group 5

In order to develop QTL applications, eight new loci were mapped on barley chromosome 7 using 124 doubled haploid lines of the North American Barley Genome Mapping Project (NABGMP) progeny (‘Steptoe’בMorex’)- These loci involve six genomic DNA restriction fragment length polymorphisms (RFLPs) and two cDNA-RFLPs including a puroindoline gene. The distribution of these markers on barley chromosome 7 was compared with that of homoeologous wheat counterparts, i.e. wheat group 5. One locus on chromosome 7 was associated with a QTL for β-glucanase activity measured in green and finished barley malt.     

普通小麦背景中长穗偃麦草[Lophopyrum elongatum (Host) A. Löve]E染色体组的RGAP特异标记

利用已知植物抗病基因编码氨基酸保守区域NBS-LRR（核苷酸结合位点-富亮氨酸区域）设计了42个简并引物组合，运用抗病基因类似物多态性（resistance gene analog polymorphism，RGAP）分子标记技术，对中国春、中国春-长穗偃麦草双二倍体及其附加系和代换系基因组DNA进行PCR扩增。结果表明，共有38对引物组合获得扩增产物，其中35对在普通小麦中国春、中国春-长穗偃麦草双二倍体中能扩增出多态性，平均每个引物组合扩增出38.5个片段。在普通小麦背景下，共获得275条长穗偃麦草E基因组多态性谱带，占扩增总谱带数的17.44%，揭示出在普通小麦背景下E基因组和普通小麦A、B、D基因组间的高丰度遗传变异。另外，利用RGAP分子标记技术，构建了一套完整的长穗偃麦草1E~7E染色体的特异RGAP标记。为小麦背景中长穗偃麦草外源遗传物质的快速检测提供了新途径。     

小麦品种苏麦3号抗赤霉病基因的染色体定位研究

本研究以苏麦3号为染色体供体,一套“中国春”小麦单体系列分别作为受体和轮回母本,连续回交4次,并建立两套独立转育的重复系,对赤霉病抗性进行了染色体定位研究。结果表明,重复系Ⅰ中,苏麦3号染色体2B、3B和6B与赤霉病抗性有关;重复系Ⅱ中,染色体7A、2B、3B和6B与赤霉病抗性有关。由此推断,苏麦3号的抗性基因位于染色体2B、3B和6B上,染色体7A是否具有抗性基因,还有待于进一步证实。2D染色体载有促进赤霉病扩展的感病基因。     

利用SSR鉴定普通小麦-多枝赖草二体异附加系 Line24中外源染色体同源群的归属

用227对小麦微卫星引物进行PCR扩增，76对可在多枝赖草和耐黄矮病的普通小麦-多枝赖草二体异附加系Line24的小麦亲本中国春、丰抗13间检测到多态性。和多枝赖草相同而与Line24其他小麦亲本不同的扩增带。在这76对引物中，发现有4对引物能从Line24中扩增出进一步用Line24和普通小麦杂交得到的7个不同的单体异附加系进行验证，也得到同样的结果，说明这4对微卫星引物扩增出的特异带可以作为Line24中多枝赖草染色体的分子标记。根据这4对引物各自对应的微卫星标记位点在小麦染色体上的位置，说明Line24中附加的一对多枝赖草染色体是第3，5，6和7部分同源群多枝赖草染色体相互易位形成的。     

Intergeneric Transfer (Rye to Wheat) of a Gene(s) for Russian Wheat Aphid Resistance

An octoploid triticale was derived from the F, of a Russian wheat aphid-resistant rye, ‘Turkey 77’, and ‘Chinese Spring’ wheat. The alloploid was crossed to common wheat, and to ‘Imperial’ rye/‘Chinese Spring’ disomic addition lines. F₂, progeny from these crosses were tested for Russian wheat aphid resistance and C-banded. A resistance gene(s) was found to be associated with chromosome arm IRS of the ‘Turkey 77’ rye genome. A monotelosomic IRS (‘Turkey 77’) addition plant was then crossed with the wheat cultivar ‘Gamtoos’, which has the 1BL.1RS ‘Veery’ translocation. Unlike the IRS segment in ‘Gamtoos’, the ‘Turkey 77’-derived 1 RS telosome did not express the rust resistance genes Sr31 and Ar26, which could then be used as markers. From the F, a monotelosomic 1 RS addition plant that was also heterozygous for the 1BL. 1 RS translocation was selected and testerossed with an aphid-susceptible common wheat, ‘Inia 66’ Meiotic pairing between the rye arms resulted in the recovery of five euploid Russian-wheat-aphid-resistant plants. One recombinant also retained Sr31 and Lr26 and was selfed to produce translocation homozygotes.     

Involvement of Chromosomes 5A and 5D in Cold-Induced Abscisic Acid Accumulation in and Frost Tolerance of Wheat Calli

The relationship between frost tolerance and abscisic acid (ABA) accumulation was studied in callus cultures of three wheat cultivars differing in the level of frost-tolerance, and of the 5A and 5D chromosome substitution lines from the frost-tolerant variety ‘Cheyenne’ into frost-sensitive ‘Chinese Spring’. Following cold hardening, the increase in ABA level in the calli of the two frost tolerant cultivars was significantly higher than in those of the frost-susceptible cultivar. Similarly, in 5A and 5D substitution lines, significantly higher ABA levels were detected than in the recipient ‘Chinese Spring’. One week-long ABA treatment at 26 °C induced a significantly higher level of frost tolerance than that achieved by cold hardening, irrespective of the frost sensitivity of the examined genotypes.     

A physical map of chromosome 4Hch from H. chilense containing SSR, STS and EST-SSR molecular markers

The construction of a physical map of chromosome 4H^ch from Hordeum chilense containing molecular markers capable of detecting segments of this chromosome in a wheat background would be very useful for marker-assisted introgression of 4H^ch chromatin into both durum and common wheat. With this aim, the applicability of 106 barley chromosome 4H primers (62 SSRs and 44 STSs) to amplify markers showing polymorphism between H. chilense and both common or bread and durum wheat was investigated. Twenty-five SSR (40.3%) and six STS (13.6%) barley primer pairs consistently amplified H. chilense products. Eight SSR (12.9%) and four STS (9.1%) barley primers were polymorphic between H. chilense and both common and durum wheat, 10 of them (6 SSRs and 4 STSs) were located on chromosome 4H^ch using both the addition line of chromosome 4H^ch in Chinese Spring wheat and a tritordeum line (an amphiploid between H. chilense and T. turgidum) nullisomic for chromosome 4H^ch. Additionally, 18 EST-SSR barley markers previously located on chromosome 4H^ch were screened for polymorphism; 15 were polymorphic between H. chilense and both durum and common wheat. For physical mapping we used a ditelosomic tritordeum line for the short arm of chromosome 4H^ch and a tritordeum line homozygous for a 70% terminal deletion of the long arm of 4H^ch. A total of 25 markers (6 SSRs, 4 STSs and 15 EST-SSRs) were mapped to chromosome 4H^ch. Eight markers were allocated on the 4H^chS, eight were mapped in the 30% proximal region of 4H^chL and nine were on the 70% distal region of 4H^chL, respectively. Arm location on barley chromosome 4H was also carried out using both 4HS and 4HL ditelosomic addition lines in wheat. All markers mapped may have a role in marker-assisted introgression of chromatin segments of chromosome 4H^ch in both durum and common wheat backgrounds. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

QTL alleles from a winter feed type can improve malting quality in barley

Mapping quantitative trait loci (QTLs) responsible for malting quality traits in barley populations has been the main genetic approach to malting quality breeding. A ‘winter × spring’ doubled haploid barley population ‘Nure’ × ‘Tremois’, where such traits were segregating, has been recently developed. Our objective was to map QTLs for malting quality from 2 years of trials in two contrasting locations. QTLs were found on six chromosomes, with a main cluster on chromosome 1H. For wort viscosity and malt extract, favourable alleles at two loci on chromosome 5H were carried by the winter feeding parent ‘Nure’. Doubled‐haploids with higher quality than the spring malting cultivar ‘Tremois’ showed either a facultative or a winter growth habit and a level of frost tolerance comparable with that of the winter tolerant parent ‘Nure’. Markers and QTLs of quality traits were further validated on a separate set of DH lines, coming from the same cross, by means of marker‐assisted selection. This showed that, at least in the present cross combination, positive contributions to malting quality can be found in winter feed barley.     

Chromosome location of genes affecting polyphenol oxidase activity in seeds of common and durum wheat

This study used cytogenetic stocks to investigate the chromosomal location of genes responsible for polyphenol oxidase (PPO) activity in common and durum wheat seeds. Substitution lines of chromosome 2A of hexaploid varieties ‘Cheyenne’, ‘Thatcher’ and ‘Timstein’ in ‘Chinese Spring’ showed significantly higher PPO activity than all other substitution lines of the same variety, with the exception of substitutions of ‘Cheyenne’ chromosome 3A and ‘Thatcher’ chromosome 4B. Substitution lines of chromosome 2A of Triticum turgidum var. dicoccoides and of chromosome 2D of ‘Chinese Spring’ into the tetraploid variety ‘Langdon’ showed a significant increase in PPO activity relative to all other substitution lines in Langdon. The gene(s) responsible for high PPO activity in chromosome 2D from ‘Chinese Spring’ was mapped on the long arm within a deletion that represents 24% of the distal part of the arm. This study shows that genes located in homoeologous group 2 play a major role in the activity of PPO in wheat.