Characterization of QEet.ocs-5A.1, a quantitative trait locus for ear emergence time on wheat chromosome 5AL

QEet.ocs‐5A.1, a quantitative trait locus controlling ear emergence time, has been detected on wheat chromosome 5AL using single chromosome recombinant lines (SCRs) developed from a cross between ‘Chinese Spring’ (CS) (‘Cappelle‐Desprez’ 5A) and CS (Triticum spelta 5A). This locus has little influence on grain yield and its components, and thus has breeding potential for changing ear emergence time without yield reduction. To characterize the phenotypic expression of QEet.ocs.1 and to test its interaction with the Vrn‐A1 gene for vernalization response, six near‐isogenic SCRs differing for these two gene regions were grown together with the parental controls under different vernalization and photoperiod regimes. The T. spelta allele of QEet.ocs.1 accelerated heading time when vernalization and photoperiod were satisfied, demonstrating that the function of this QTL is earliness per se. There was no interaction between Vrn‐A1 and QEet.ocs.1.     

Microsatellite monitoring of recombination around the Vrn-B1 locus of wheat during early backcross breeding

The length of chromosomal segments retained around the Vrn‐B1 gene controlling sensitivity to vernalization in wheat (Triticum aestivum L.) was studied in the first and third backcrosses by using microsatellite markers. Eleven polymorphic markers located on chromosome 5B were used for microsatellite analysis. It was shown in the first backcross that plants with a donor segment around the gene of interest not longer than 50% of chromosome 5B could be selected. When selection is not molecular‐marker assisted, the length of the chromosomal donor segment with the target gene may reach 94% of chromosome 5B even in plants of the third backcross generation. The considerable length differences in the 5B microsatellite loci between the winter and spring lines of wheat studied indicate that these markers are promising in marker‐assisted backcrossing or marker‐assisted selection for the Vrn‐B1 gene using different combinations of Spring and Winter genotypes.     

Identifying AFLP and microsatellite markers for vernalization response gene Vrn-B1 in hexaploid wheat using reciprocal mapping populations

Marker‐assisted selection may be useful for combining specific vernalization response (Vrn) alleles into a single wheat genotype for yield enhancement; however, DNA markers are only available for two of the three genes identified to date. The objectives of this study were to investigate reciprocal effects on days to heading using F₂ populations generated by cross‐hybridizing near‐isogenic lines (NILs) carrying spring (Vrn‐B1; TDB) and winter (vrn‐B1; TDC) alleles, and to identify markers linked to Vrn‐B1 through genetic linkage analysis. Heading data were recorded for 91 and 89 progeny from reciprocal mapping populations TDB/TDC and TDC/TDB, respectively, and significant (P < 0.0001) reciprocal and dominance effects were detected. Among 207 amplified fragment length polymorphisms primer pairs and seven wheat microsatellite markers screened, two and one, respectively, were linked distally to Vrn‐B1 on wheat chromosome 5BL. Microsatellite Xgwm408 was most closely linked to Vrn‐B1 at 3.9 and 1.1 cM in the TDB/TDC and TDC/TDB map, respectively. Reciprocal differences in recombination distances emphasize the importance of female parent choice when generating mapping populations. Molecular markers are now available for three Vrn loci in wheat.     

Intrachromosomal mapping of genes for dwarfing (Rht12) and vernalization response (Vrn1) in wheat by using RFLP and microsatellite markers

For intrachromosomal mapping of the dominant GA-sensitive dwarfing gene Rht12 and the vernalization response gene Vrn1 on chromosome 5 A, an F₂ population was established using a wide (synthetic) wheat cross. In addition to restriction fragment length polymorphism (RFLP) probes four microsatellite markers were incorporated. Rht12 was mapped distally to four RFLP loci (Xmwg616, Xpsr164, Xwg114, Xpsr1201) and three microsatellite markers (Xgwm179, Xgwm410, Xgwm291), known to be located on the segment of chromosome SAL which was ancestrally translocated and is homoeologous to Triticeae 4 L. The map position of Rht12 suggests that it is homoeologous to the dominant GA-sensitive dwarfing gene Ddw1, present on chromosome 5RL. The vernalization response gene Vrn1 showed linkage to Xwg644, as might be expected from comparative maps.     

Validation of a major QTL for scab resistance with SSR markers and use of marker-assisted selection in wheat

The objectives of this study were to validate the major quantitative trait locus (QTL) for scab resistance on the short arm of chromosome 3B in bread wheat and to isolate near‐isogenic lines for this QTL using marker‐assisted selection (MAS). Two resistant by susceptible populations, both using ‘Ning7840’ as the source of resistance, were developed to examine the effect of the 3BS QTL in different genetic backgrounds. Data for scab resistance and simple sequence repeat (SSR) markers linked to the resistance QTL were analyzed in the F_2:3 lines of one population and in the F_3:4 lines of the other. Markers linked to the major QTL on chromosome 3BS in the original mapping population (‘Ning7840’/‘Clark’) were closely associated with scab resistance in both validation populations. Marker‐assisted selection for the QTL with the SSR markers combined with phenotypic selection was more effective than selection based solely on phenotypic evaluation in early generations. Marker‐assisted selection of the major QTL during the seedling stage plus phenotypic selection after flowering effectively identified scab resistant lines in this experiment. Near‐isogenic lines for this 3BS QTL were isolated from the F₆ generation of the cross ‘Ning7840’/‘IL89‐7978’ based on two flanking SSR markers, Xgwm389 and Xbarc147. Based on these results, MAS for the major scab resistance QTL can improve selection efficiency and may facilitate stacking of scab resistance genes from different sources.     

黄淮麦区Fhb1基因的育种应用

小麦赤霉病是由禾谷镰孢菌引起的一种世界性重要病害,严重威胁小麦生产安全。黄淮麦区作为我国小麦主产区,赤霉病危害日趋严重,因缺乏半冬性抗源,抗赤霉病育种进展缓慢。Fhb1基因是迄今发现的效应最大、抗性最稳定,也是被广泛应用于全球小麦赤霉病抗性育种的主效基因,但Fhb1基因在黄淮麦区尚未被广泛应用。本研究以感病品种矮抗58为轮回亲本, H35为Fhb1基因供体亲本,通过有限回交和分子标记辅助选择,同时利用双单倍体育种和传统系谱选育两种方法,培育出了一批综合性状较好、具有Fhb1基因的优良新品系,其中徐麦DH9和徐麦17252经多年鉴定均达到中抗水平。在以徐麦36和徐麦2023为杂交父本的后代品系中,含Fhb1基因的家系赤霉病平均抗性明显优于感病对照。Fhb1基因的导入显著提高了赤霉病抗性,但部分家系对赤霉病仍旧表现出高感水平,说明赤霉病抗性还受到Fhb1基因以外其他遗传因素的显著影响。本研究为Fhb1基因在黄淮麦区抗赤霉病小麦育种中的应用提供了成功的经验。     

小麦株高QTL Qph.nau-5B的效应评价

株高直接影响小麦的产量潜力,也是植株抗倒伏性的重要组成部分。目前虽有大量株高相关QTL被鉴定到,但大多QTL的遗传效应仍不清楚。本研究前期利用小麦品种群体,通过关联分析鉴定到一个小麦株高主效QTL Qph.nau-5B。为了评价该QTL的效应,通过分子标记辅助选择分别构建了以南大2419、吉春1016和郑麦9023为供体亲本,中优9507为背景的3种等位变异的近等基因系,背景回复率均高于93%。在7个独立的试验环境中,所有近等基因系的株高较轮回亲本均显著降低,平均降幅为11.1 cm(10.3%)。Qph.nau-5B不同等位变异效应强弱不同,其中来源于吉春1016和郑麦9023的等位变异平均降秆效应相似(12.4 cm),显著大于南大2419的等位变异(8.6 cm),但各等位变异相对降秆效应大小受环境影响。此外,Qph.nau-5B对单株穗数、穗长、千粒重等农艺性状无明显负效应。本研究结果表明Qph.nau-5B具有重要的育种价值,可为小麦的株型分子设计育种提供基因资源。     

Boron deficiency induced male sterility in wheat (Triticum aestivum L.) and implications for plant breeding

Boron (B) deficiency causes grain set in wheat to fail. A wide range of genotypic variation in the response to low B has been observed. Genotypes were screened in low B in soil and sand culture, and classified into five groups, namely, very sensitive, sensitive, moderately sensitive, moderately tolerant and tolerant. At very low levels of B, the very sensitive to sensitive genotypes were completely male sterile and set only a few or no grain, while the tolerant genotypes set grain normally. Natural outcrossing was detected in these male sterile plants when a tolerant genotype was growing nearby. Grain set by cross fertilisation was markedly enhanced by a B application directly on the ear of the male sterile plants. Three practical implications are suggested. Firstly, genotypes that are tolerant to low B can provide a solution for grain set failure caused by B deficiency. Secondly, the potential for outcrossing in male sterile B deficient wheat has to be taken into account in the maintenance of pure lines in low B soils even though wheat is normally self pollinated. Thirdly, a simple and novel method for hybridization is suggested, in which B deficiency is used as fertility selective medium and male sterile female parents and fertile male parents are provided by genotypic variation in the response to low B. This revised version was published online in July 2006 with corrections to the Cover Date.     

Distribution of the wheat–rye translocation 1RS.1BL among bread wheat varieties of Bulgaria

The distribution of the wheat–rye translocation 1RS.1BL was studied in 31 winter wheat varieties from Bulgaria. The presence of the translocation was verified in 17 varieties using chromosome N‐banding analysis, PAGE‐analysis of grain storage proteins and DNA‐marker analysis. The 1RS.1BL has been transmitted in 54% of varieties with a known source of the translocation in their pedigree.     

Identification of powdery mildew resistance genes in common wheat (Triticum aestivum L. em Thell.). IX. Cultivars, land races and breeding lines grown in China

The objective of the study was to provide information about the occurrence and distribution of resistance genes in wheat cultivars, including old cultivars, land races and advanced breeding lines grown in China. Ninety-four accessions were analysed with a set of 11 differential powdery mildew isolates. Forty-four cultivars did not possess any major mildew resistance genes. Thirty cultivars revealed the response pattern of individual resistance genes. The most frequently encountered gene was Pm8, which occurred singly in 11 cultivars, combined either with Pm4a in three cultivars or with Pm4b in another three cultivars. However, 12 cultivars possessing the wheat-rye translocated chromosome pair T1BL-1RS did not express Pm8. Gene Pm2 was found in four cultivars and in combination with Pm6 in one cultivar. Genes Pm4a and Pm4b were observed in four and five cultivars, respectively. Another six cultivars carried Pm5. A gene combination of Pm2+Pm4b+Pm6 was found in one cultivar. Twelve cultivars and breeding lines exhibited a response pattern that could not be assigned to resistance genes or gene combinations present in the differential cultivars. Five out of these 12 cultivars/lines showed resistance to all the isolates tested. There is an urgent need to search for novel sources of mildew resistance in order to sustain resistance to existing and emerging powdery mildew pathogens.     

Rapid screening of selected European winter wheat varieties and segregating populations for the Glu-D1d allele using PCR

Forty-two winter wheat varieties and 193 F₂ and BC₁F₂ seeds were screened for Glu-D1d allele encoding the HMW glutenin subunits 5 + 10 using polymerase chain reaction (PCR). The segregating populations originated from crosses involving wheat parents with good and poor bread-making quality. A clear PCR product of 450 bp, representing 1Dx5 of the Glu-D1d allele was identified in 24 varieties and 111 hybrid seeds. Four different Glu-D1 alleles: a (2 + 12), b (3 + 12), c (4 + 12) and d (5 + 10) were detected using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Only genotypes possessing Glu-D1d gave a positive PCR signal, hexaploid triticale and 4 × wheat lacking Glu-D1 locus gave a negative signal. The efficiency of PCR selection for bread-making quality in early generations using half seed is discussed.     

An assessment of the potential of 4DS.4DL-4s l L translocation lines as a means of eliminating tall off types in semi-dwarf wheat varieties

Summary Wheat varieties tend to be chromosomally unstable producing on average 2–3% of plants with abnormal chromosome numbers. A number of semi dwarf wheat varieties, carrying the gibberellic acid insensitive dwarfing genes Rht1 or Rht2, have been seen to produce distinct tall off types due to reduction in dosage of the chromosome carrying the dwarfing gene. The UK variety Brigand, carrying Rht2 on chromosome 4D, produced very distinct tall off types when this chromosome was reduced in dosage. The frequency of tall off types was sufficiently high to cause the variety to fail United Kingdom statutory uniformity tests. An attempt to prevent the loss of chromosome 4D was made by constructing translocation chromosomes involving the short arm of chromosome 4D, which carries Rht2, and the long arm of chromosome 4S ^l from Aegilops sharonensis, which carries a gene(s) conferring preferential transmission. The work in this paper describes the field evaluation of two lines carrying 4DS.4DL-4S ^l L translocations, and demonstrates their success in preventing spontaneously occurring monosomy of chromosome 4D in semi-dwarf wheats.     

A PCR-based DNA marker for detection of mutant and normal alleles of the Wx-D1 gene of wheat

To assist waxy wheat breeding a DNA marker was developed to discriminate mutant and normal alleles at the Wx‐D1 locus. This polymerase chain reaction‐based marker distinguishes the mutant from the normal allele by targeting the previously reported deletion basis of the mutant. The marker codominantly identifies the normal allele of the Wx‐D1 gene from the mutant allele originated from the Chinese landrace ‘Baihoumai’. However, attempts with a number of primer combinations targeting this deletion failed to amplify the corresponding fragment from an unrelated wheat line (NP150) that has a mutant null allele at the same locus. This indicates that NP150 has a different mutant allele from that of ‘Baihoumai’. This marker is a useful tool to identify wheat cultivars with mutant and normal alleles of the Wx‐D1 gene, and is used in marker‐assisted selection of the Wx‐D1 gene in our waxy wheat breeding programme.     

Efficient application of in vitro anther culture for different European winter wheat (Triticum aestivum L.) breeding programmes

The efficiency of our anther culture protocol was tested with high‐ and low‐responding genotypes, ‘Svilena’ and ‘Berengar’, and 93 F₁ winter wheat crosses in 2010 and 2011. Based on data for these genotypes, the effect of genotype influenced the number of embryo‐like structures, regenerated plantlets and green plantlets, while the number of albino plantlets was affected by genotype, year and environmental factors. Although genotype also influenced the production of green plantlets from breeding crosses, with green plantlets per 100 anthers ranging from 0.04 to 28.67, the average regeneration rate over all crosses was 5.3 green plantlets/100 anthers, which resulted in a total of 11 416 well‐rooted green plantlets. The survival rate of green plantlets following acclimatization was 97.21% in 2010 and 96.34% in 2011. In this study, the phenomenon of albinism and genotype dependency did not hinder the production of more than five thousand green plantlets each year. In our experiments, anther culture proved to be an efficient method in winter wheat breeding programmes with lower costs than alternative technologies.     

小麦脱落酸受体基因TaPYR1介导植株抵御干旱逆境功能研究

脱落酸(ABA)受体PYR/PYL/RCAR通过与渗透胁迫诱导的ABA结合,参与植株体内ABA介导的信号转导过程,在调控植株干旱逆境抵御过程中具有重要的生物学功能。本文研究了鉴定的1个干旱响应的PYR家族成员TaPYR1的分子特征、应答干旱表达模式及其介导植株抵御干旱逆境的功能。结果表明,TaPYR1与植物种属中部分PYR基因在氨基酸序列水平上高度同源,编码蛋白含有PYR家族成员保守结构域,翻译蛋白经内质网分选后定位于细胞质膜。TaPYR1基因在小麦根、叶中均呈明显的干旱诱导表达模式,在干旱胁迫48 h表达量达到峰值。与野生型对照(WT)相比,超表达TaPYR1烟草转化株系,干旱处理下植株长势增强,干鲜重增加。干旱处理下,转化株系较对照的光合能力增强,细胞保护酶活性提高,渗透调节物质脯氨酸和可溶性糖含量增加。研究表明, TaPYR1通过在转录水平上应答干旱逆境,改善干旱胁迫下的植株相关生理过程,在增强植株抵御干旱逆境能力中发挥重要作用。     

Potential use of doubled haploid lines for the screening of resistance to yellow rust (Puccinia striiformis) in hexaploid wheat

Doubled haploid lines derived from anther culture of two Iranian spring wheat genotypes‘Ghods’susceptible and‘9106’resistant to yellow rust in Iranian field conditions, and their F₁ hybrids were used in this study. Seedlings of 36 doubled haploid lines, selected out of 96 according to their agronomic traits and the two parental genotypes were inoculated with eight races of yellow rust. The parental genotypes (‘Ghods’and‘9106’) were segregating for some of the races but their doubled haploid lines were either resistant or susceptible to them.‘Ghods’was susceptible to three of the races studied but three doubled haploid lines derived from it were resistant to them. Five selected doubled haploids from the‘9106’genotype and six from F₁ hybrid plants were resistant to all eight races tested. After further investigations in Iranian field conditions it was found that some of these lines can be used as donor genotypes for resistance to yellow rust in wheat breeding programmes. Use of these genotypes should be possible if the French yellow rust races used for selection also represent the dominant races in Iran. It can be concluded that anther culture provides an efficient method for fixing genes of resistance to yellow rust and desirable doubled haploids from F₁ plants can be derived.     

Cytogenetical studies in wheat. XVIII. Gene Yr24 for resistance to stripe rust

A new gene, Yr24, for resistance to stripe rust was transferred from a durum accession to common wheat via an amphiploid (synthetic wheat) with Aegilops tauschii. Yr24 was located in chromosome 1B by monosomic analysis. Its genetic linkage of 4 cM with Yr15 indicated its localization to the short arm.     

Genetic Diversity for Adult Plant Resistance to Leaf Rust (Puccinia recondita) in Near-Isogenic Lines and in Indian Wheats

Variation for adult plant resistance in near-isogenic wheat lines carrying Lrl4b, Lrl4ab and Lr30 in a ‘Thatcher’ background indicated the possible presence of novel adult plant resistance genes effective against the Indian leaf rust population. Sixty-one wheats released for cultivation in India were grown in isolated nurseries. Each nursery was separately inoculated with one of four leaf rust pathotypes which had been selected to aid identification of resistance effective only in the adult plant stage. Seven distinct response groups were recognised and a minimum of six sources of adult plant resistance were postulated. In a group of 14 wheats, resistance was explained on the basis of the seedling response genes that were identified. Similar results for two years with pathotype 77-1 gave support to the reliability of field tests. Adult plant resistance (APR) sources were either race-specific or effective against all pathotypes used. Seedlings of cultivars with APR showed susceptible reactions. The possible presence of Lr34 in Indian wheats and its role in durable leaf rust resistance are discussed.     

Segregation for isozymes and HMW glutenins in a doubled-haploid cross of winter wheat carrying 1BL.1RS and 5DL.5RS translocations from rye

The doubled haploid (DH) wheat line ‘dh 5841’ carrying two translocations from rye, 5DL.5RS and 1BL.1RS, has been crossed to the subline of wheat cultivar ‘Amadeus 7143’ with a 1BL.1RS translocation. The resulting F₁ hybrid IJ 98 with a heterozygous 5DL.5DS‐5DL.5RS chromosome pair has been used to produce doubled haploids. A total of 57 DH lines were obtained from plantlets regenerated in anther culture after successful colchicine treatment and seed set. These lines were identified regarding the constitution of chromosome 5D (5DL.5DS or 5DL.5RS) by means of isoenzyme marker analysis. Thirty DH lines possessed the 5DL.5DS chromosome, while the remaining 27 lines carried the 5DL.5RS translocation. For some of these lines, the 5DL.5RS chromosome was cytologically confirmed by C‐banding. Furthermore, the DH lines were evaluated for their high molecular weight glutenin subunit composition. All possible combinations for the four independent loci —Skdh, Glu‐Al, Glu‐B1 and Glu‐D1— were detected in only 57 DH lines and no segregation distortion was observed.     

Comparison of a modified assay method for the endopeptidase marker Ep-D1b with the Sequence Tag Site marker XustSSR2001–7DL for strawbreaker foot rot resistance in wheat

The endopeptidase marker Ep‐D1b and Sequence Tag Site (STS) marker XustSSR2001–7DL were reported to be closely associated with the most effective resistance gene (Pch1) in wheat (Triticum aestivum L.) for strawbreaker foot rot [Pseudocercosporella herpotrichoides (Fron) Deighton]. Our objectives were to: (i) develop an efficient assay method for Ep‐D1b in wheat; (ii) correlate endopeptidase zymograms to strawbreaker foot rot reactions of various wheat genotypes; and (iii) compare the utility of Ep‐D1b and XustSSR2001–7DL for predicting disease response. An improved method of assaying for the Ep‐D1b marker using roots from a single seedling was developed, which is a 2.5‐fold improvement over the previous method. Thirty‐eight wheat genotypes with known reactions to strawbreaker foot rot were analysed for Ep‐D1b and the STS marker. Six distinct endopeptidase zymograms were identified among these 38 genotypes tested, and three of these patterns were novel. The endopeptidase marker was 100% accurate for predicting strawbreaker foot rot disease response, whereas the STS marker predicted the correct phenotype with approximately 90% accuracy. The endopeptidase marker Ep‐D1b was more effective and was more economical for use in marker‐assisted selection strategies for Pch1 in our laboratory compared with the STS marker.