将Thinopyrum bessarabicum和Thinopyrum elongatum的种质导？…

２个双倍体即Ｃ．Ｓ－Ｔｈｉｎｏｐｙｒｕｍ ｂｅｓｓａｒａｂｉｃｕｍ（ＡＡＢＢＤＤＪＪ ２ｎ＝８ｘ＝５６）和ＧＨＫ－Ｔｈｉｎｏｐｙｒｕｍ ｅｌｏｎｇａｔｕｍ（ＡＡＢＢＤＤＥＥ ２ｎ＝８ｘ＝５６）与普通小麦“中国春”杂交,获得了２个七倍体杂种。对２３个双倍体、中国春、杂种Ｆ１和部分Ｆ２进行麦谷蛋白ＳＤＳ－Ｐａｇｅ电泳,及结合染色体分带技术和田间赤霉病抗性鉴定筛选出由Ｔｈ．ｂｅｓｓａｒａｂｉｃｕｍ和Ｔｈ     

Identification of genetic loci affecting amylose content and agronomic traits on chromosome 4A of wheat

 Chromosome 4A of wheat carries the Wx-B1 gene encoding the granule-bound starch synthase involved in amylose synthesis in the endosperm. To determine the pleiotropic effects of this locus and effects of independent QTLs on agronomic traits, genetical analysis of chromosome 4A was conducted using 98 single-chromosome recombinant substitution lines derived from a cross of Chinese Spring and Chinese Spring (Kanto107 4A) with a low amylose content due to the null Wx-B1b allele. For amylose content, most of the genetic variation was explained by the allelic difference at the Wx-B1 locus. An additional QTL of minor effect was mapped in the 6.2-cM Xbcd1738/Xcdo1387 interval on the short arm, where the allele from Kanto107 led to an increase in amylose content. Field trials over two seasons revealed a pleiotropic effect of Wx-B1, or else the effect of a closely linked QTL, on ear emergence time. A QTL linked to Wx-B1 was detected for plant height. For plant yield and its components, there was no evidence for significant main effects associated with Wx-B1 or adjacent regions. One plant-yield QTL was identified by RFLP markers on the short arm and this was identical to QTLs controlling spikelet number/ear and grain weight/ear. At these QTLs for agronomic traits, alleles from Kanto107 contributed to an earlier emergence time, a height reduction and an yield increase. Received: 10 August 1998 / Accepted: 3 November 1998     

Chromosome markers in the tetraploid wheat Aegilops ventricosa analysed by in situ hybridization

 Three lines of the tetraploid wheat Aegilops ventricosa Tausch (2n=4x=28), which contains good resistance to eyespot, were analysed using fluorescent in situ hybridization. Probes used included rDNA, cloned repeated sequences from wheat and rye, simple-sequence repeats (SSRs) and total genomic DNA. The banding patterns produced could be used to distinguish most chromosome arms and will aid in the identification of Ae. ventricosa chromosomes or chromosome segments in breeding programmes. All lines had a single major 18S-25S rDNA site, the nucleolar organizing region (NOR) in chromosome 5N and several minor sites of 18S-25S rDNA and 5S rDNA. A 1NL.3DL, 1NS.3DS translocation was identified, and other minor differences were found between the lines. Received: 11 August 1998 / Accepted: 28 November 1998     

Characterization and genetic mapping of simple repeat sequences in the tomato genome

Tomato genomic libraries were screened for the presence of simple sequence repeats (SSRs) with seventeen synthetic oligonucleotide probes, consisting of 2- to 5-basepair motifs repeated in tandem. GAn and GTn sequences were found to occur most frequently in the tomato genome (every 1.2 Mb), followed by ATTn and GCCn (every 1.4 Mb and 1.5 Mb, respectively). In contrast, only ATn and GAn microsatellites (n > 7) were found to be frequent in the GenBank database, suggesting that other motifs may be preferentially located away from genes. Polymorphism of microsatellites was measured by PCR amplification of individual loci or by Southern hybridization, using a set of ten tomato cultivars. Surprisingly, only two of the nine microsatellite clones surveyed (five GTn, three GAn and one ATTn), showed length variation among these accessions. Polymorphism was also very limited betweenLycopersicon esculentum andL. pennelli, two distant species. Southern analysis using the seventeen oligonucleotide probes identified GATAn and GAAAn as useful motifs for the detection of multiple polymorphic fragments among tomato cultivars. To determine the structure of microsatellite loci, a GAn probe was used for hybridization at low stringency on a small insert genomic library, and randomly selected clones were analyzed. GAn based motifs of increasing complexity were found, indicating that simple dinucleotide sequences may have evolved into larger tandem repeats such as minisatellites as a result of basepair substitution, replication slippage, and possibly unequal crossing-over. Finally, we genetically mapped loci corresponding to two amplified microsatellites, as well as nine large hypervariable fragments detected by Southern hybridization with a GATA8 probe. All loci are located around putative tomato centromeres. This may contribute to understanding of the structure of centromeric regions in tomato.     

In situ hybridization in Actinidia using repeat DNA and genomic probes

 In situ hybridization has been used to probe chromosome spreads of hexaploid Actinidia deliciosa (kiwifruit; 2n=6x=174) and tetraploid A. chinensis (2n=4x=116). When a species-specific repeat sequence, pKIWI516, was used, six hybridization sites were observed in some accessions of tetraploid A. chinensis and all of A. deliciosa. Southern analysis with the pKIWI516 probe revealed that there are two types of tetraploid A. chinensis. Genomic probes from diploid A. chinensis (2n=2x=58) did not differentiate the genomes of hexaploid A. deliciosa and tetraploid A. chinensis, irrespective of the presence or absence of blocking DNA. The results indicate that the genomes of polyploid Actinidia species are similar but not identical. The origin of A. deliciosa is discussed. Received: 29 June 1996 / Accepted: 5 July 1996     

Identification of simple sequence repeats (SSRs) in olive ( Olea europaea L.)

A small insert genomic library of Olea europaea L., highly enriched in (GA/CT)n repeats, was obtained using the procedure of Kandpal et al. (1994). The sequencing of 103 clones randomly extracted from this library allowed the identification of 56 unique genomic inserts containing simple sequence repeat regions made by at least three single repeats. A sample of 20 primer pairs out of the 42 available were tested for functionality using the six olive varieties whose DNA served for library construction. All primer pairs succeeded in amplifying at least one product from the six DNA samples, and ten pairs detecting more than one allele were used for the genetic characterisation of a panel of 20 olive accessions belonging to 16 distinct varieties. A total of 57 alleles were detected among the 20 genotypes at the ten polymorphic SSR loci. The remaining primer pair allowed the amplification of a single SSR allele for all accessions plus a longer fragment for some genotypes. Considering the simple sequence repeat polymorphism, 5.7 alleles were scored on average for each of the ten SSR loci. A genetic dissimilarity matrix, based on the proportion of shared alleles among all the pair-wise combinations of genotypes, was constructed and used to disentangle the genetic relationships among varieties by means of the UPGMA clustering algorithm. Graphical representation of the results showed the presence of two distinct clusters of varieties. The first cluster grouped the varieties cultivated on the Ionian Sea coasts. The second cluster showed two subdivisions: the first sub-cluster agglomerated the varieties from some inland areas of Calabria; the second grouped the remaining varieties from Basilicata and Apulia cultivated in nearby areas. Results of cluster analysis showed a significant relationship between the multilocus genetic similarities and the geographic origin of the cultivars. Received: 2 February 2001 / Accepted: 1 June 2001     

Automated sizing of fluorescent-labeled simple sequence repeat (SSR) markers to assay genetic variation in soybean

 Simple Sequence Repeat (SSR) allele sizing provides a useful tool for genotype identification, pedigree analysis, and for estimating genetic distance between organisms. Soybean [Glycine max (L.) Merr.] cultivars are identified for Plant Variety Protection (PVP) purposes by standard pigmentation and morphological traits. However, many commercial soybeans arise from a limited number of elite lines and are often indistinguishable based on these traits. A system based on SSR markers would provide unique DNA profiles of cultivars. Fluorescent labeling of alleles combined with automated sizing with internal size standards in each gel lane was used as an alternative to standard [³²P] labeling to assess genetic variability in soybean. Allelic frequencies at 20 SSR loci were determined in 35 soybean genotypes that account for greater than 95% of the alleles in North American soybean cultivars based upon pedigree analysis. An average of 10.1 alleles per locus (range: 5–17), with a mean gene diversity of 0.80 (range: 0.50 to 0.87) were observed at the 20 SSR loci. The 20 loci successfully distinguished modern soybean cultivars that are identical for morphological and pigmentation traits, as well as 7 soybean genotypes reported to be indistinguishable using 17 RFLP probes. Pedigrees of 7 cultivars were studied to estimate stability of SSRs in soybean across generations. Of the 7 pedigrees 6 had one locus in the progeny with an allele(s) that was not present in either parent. These new alleles are most likely the result of mutation. The mutation rate of SSR alleles in soybean was similar to that reported in humans. To avoid difficulty associated with mutation, DNA fingerprint data should be determined from the bulk of 30-50 plants of a cultivar. Received: 24 March 1997 / Accepted: 4 April 1997     

Inheritance of resistance to leaf and glume blotch caused by Septoria nodorum Berk. in winter wheat

Sixteen crosses between eight winter wheat cultivars were screened for resistance to Septoria nodorum leaf and glume blotch in the F₁ and F₄ generations using artificial inoculation in the field. The F₁ of most crosses showed dominance for susceptibility on both ear and leaf. The effects of general combining ability were of similar magnitude as the effects for specific combining ability. On the basis of the phenotypic difference of the parents, no prediction was possible about the amount and the direction of genetic variance in the segregating populations. The variation observed in this study both within and among the segregating populations suggests a quantitative inheritance pattern influencing the expression of the two traits. The components of variance between F₂ families within a population were as high as (for S. nodorum blotch on the ear) or higher (for S. nodorum blotch on the leaf) than those between populations. Therefore, strong selection within a few populations may be as effective to obtain new resistant genotypes as selection in a large number of populations. In almost all crosses, progenies were found that were more resistant than the better parent. Thus transgression breeding may be a tool to breed for higher levels of resistance to S. nodorum blotch. Highly resistant genotypes were found even in combination with two susceptible parents. The genetic source for Septoria resistance is probably broader than is generally assumed and could be used to improve S. nodorum resistance by combination breeding followed by strong selection in large populations. Received: 18 January / Accepted: 30 April 1999     

Comparative RFLP mapping of the chlorotoluron resistance gene (Su1) in cultivated wheat (Triticum aestivum) and wild wheat (Triticum dicoccoides)

Chlorotoluron is a selective phenylurea herbicide widely used for broad-leaved and annual grass weed control in cereals. Variation in the response to chlorotoluron (CT) was found in both hexaploid bread wheat (Triticum aestivum L.) and wild tetraploid wheat (Triticum dicoccoides KöRN.). Here, we describe the comparative mapping of the CT resistance gene (Su1) on chromosome 6B in bread and wild wheat using RFLP markers. In bread wheat, mapping was based on 58 F₄ single-seed descent (SSD) plants of the cross between a genotype sensitive to chlorotoluron, ‘Chinese Spring’ (CS), and a resistant derivative, the single chromosome substitution line, CS (‘Cappele-Desprez’ 6B) [CS (CAP6B). In T dicoccoides, mapping was based on 37 F₂ plants obtained from the cross between the CT-susceptible accession B-7 and the resistant accession B-35. Nine RFLP probes spanning the centromere were chosen for mapping. In bread wheat Su1 was found to be linked to α-Amy-1 (9.84 cM) and Xpsr371 (5.2 cM), both on the long arm of 6B, and Nor2 (2.74 cM) on the short arm. In wild wheat the most probable linkage map was Nor2-Xpsr312-Su1-Pgk2, and the genetic distances between the genes were 24.8cM, 5.3cM, and 6.8cM, respectively. These results along with other published map data indicate that the linear order of the genes is similar to that found in T. aestivum. The results of this study also show that the Su1 gene for differential response to chlorotoluron has evolved prior to the domestication of cultivated wheat and not in response to the development and use of chemicals.     

Identification of new low-molecular-weight glutenin subunit genes in wheat

To clarify the composition of low-molecular-weight glutenin subunits (LMW-GSs) in a soft wheat cultivar, we cloned and characterized LMW-GS genes from a cDNA library and genomic DNA in Norin 61. Based on alignment of the conserved N- and C- terminal domains of the deduced amino-acid sequences, these genes are classified into 12 groups. One of these groups (group 5), the corresponding gene of which has not been reported previously, contains two additional hydrophobic amino-acid clusters interrupting the N-terminal repetitive domain. Other groups (groups 11 and 12), which were not identified in other cultivars as a protein product, showed all eight cysteines in the C-terminal conserved domain. With specific primer sets for these groups it was revealed that Glu-D3 and Glu-A3 encoded the former and the latter, respectively. Both groups of genes were expressed in immature seeds. The presence of these groups of LMW-GSs may affect the dough strength of soft wheat. Received: 26 March 2001 / Accepted: 16 July 2001     

In vitro screening for resistance against Septoria nodorum blotch in wheat

This study was carried out to develop an in vitro test for the identification of genotypes resistant to Septoria nodorum blotch. The basis for this project was a previous study in which a crude extract of S. nodorum was used as a selective agent (Keller et al. 1994). It was possible to distinguish resistant and susceptible cultivars in an in vitro test with zygotic embryos. In our project we wanted to test whether this in vitro test can also be used to detect resistant and susceptible genotypes in early segregating populations. Specific crosses between eight winter wheat lines showing contrasting resistance reaction for S. nodorum blotch on leaves and ears were made. The resistance level of both leaf and ear was evaluated after artificial inoculation in the field for the parental lines, the F₁ progenies, as well as for segregating F₃ and F₄ populations. In addition, this plant material was tested in vitro using methods similar to those described by Keller et al. (1994), i.e. culturing immature zygotic embryos and mature seeds on selective media. A good agreement between in vitro screening and field resistance on the ear was found for the parental lines, the F₁ and F₄ generation but not for the F₃ generations. This leads to the conclusion that the in vitro screening might be integrated into wheat breeding programs. Populations showing a high susceptibility to the pathogen metabolites in vitro could be discarded. Another promising implementation for wheat breeding would be the screening of advanced breeding material or candidate partners in a crossing program for resistance on the ear. However, the in vitro screening is not precise enough to select single plants in early segregating populations. Received: 18 January 1999 / Accepted: 30 April 1999     

Three stages of meiotic homologous chromosome pairing in wheat: cognition, alignment and synapsis

 Chromosome painting enabled the study of homologous chromosome behaviour prior to and during meiosis. Total genomic DNA from rye, used as a probe for in situ hybridization, identified the rye chromosome arm in a wheat-rye translocation line (T5AS·5RL) at meiotic prophase and the preceding interphase. Accurate staging of the development of the meiocytes was attained by parallel studies of chromatin morphology, nucleolar behaviour and synaptonemal complex formation in electron microscopy thin sections and silver-stained surface spreads. Three stages of pairing were identified for the large cereal genomes that are organized in a Rabl configuration: first, cognition occurs during the long interphase before leptotene, bringing the homologous chromosome domains into close proximity and possibly starting at the centromere; second, homologous chromosome segments align at late leptotene; and third, zygotene synapsis initiates near the telomere, although it was also observed to occur near the centromere. A pairing model is proposed for wheat, with a genome size of 17000 Mbp, that shows prallels to and notable differences from yeast and mammalian models of meiosis. Received: 25 January 1997 / Revision accepted: 14 July 1997     

Identification of inter simple sequence repeat (ISSR) markers associated with seed size in wheat

The feasibility of identifying inter-simple sequence repeat markers associated with seed weight in hexaploid wheat was tested using 113 recombinant inbred lines developed by the single-seed descent method, from a cross between Rye selection111, an Indian genetic stock obtained through the introgression of genes for bold seed size from rye, and Chinese Spring having small seed size. Three markers were associated with low seed size with gene effects of 14.8%, 9.5%, and 6%, while four markers with contributions of 8%, 4.66%, 2.92% and 2.61% were found to be linked to high seed size, together contributing 31% of the phenotypic variance in seed size. Nulli-tetrasomic and di-telosomic analysis revealed the presence of three low seed size QTL-associated markers on three chromosomes, 6BL, 2DL, and 1DS respectively. This study clearly demonstrates that ISSRs are highly useful for finding markers associated with major and minor genes controlling agronomically important traits in wheat. Received: 24 February 2000 / Accepted: 31 March 2000     

Targeted isolation of simple sequence repeat markers through the use of bacterial artificial chromosomes

 Simple sequence repeats (SSRs) are versatile DNA markers that are readily assayed and highly informative. Unfortunately, non-targeted approaches to SSR development often leave large genomic regions without SSR markers. In some cases these same genomic regions are already populated by other types of DNA markers, especially restriction fragment length polymorphisms (RFLPs), random amplified polymorphic DNAs (RAPDs), and amplified fragment length polymorphisms (AFLPs). To identify SSR markers in such regions, bacterial artificial chromosome (BAC) clones can be used as intermediaries. First, one or more BAC clones in a region of interest are identified through the use of an existing DNA marker. BAC clones uncovered in this initial step are then used to create a small insert DNA library that can be screened for the presence of SSR-containing clones. Because BAC inserts are often 100-kb pairs or more in size, most contain one or more SSRs. This strategy was applied to two regions of the soybean genome near genes that condition resistance to the soybean cyst nematode on molecular linkage groups G and A2. This targeted approach to identifying new DNA markers can readily be extended to other types of DNA markers, including single nucleotide polymorphisms. Received: 13 August 1998 / Accepted: 13 October 1998     

QTL analysis of bread-making quality in wheat using a doubled haploid population

A set of 187 doubled haploid lines derived from the cross between cvs. Courtot and Chinese Spring was explored for QTLs for three bread-making quality tests: hardness, protein content and strength of the dough (W of alveograph). The scores of the parental lines were quite different except for protein content, and the population showed a wide range of variation. About 350 molecular and biochemical markers were used to establish the genetic map, and technological criteria were evaluated in 1 to 3 years. QTL detection was performed by the ”marker regression” method. The most significant unlinked markers were used in the model as covariates, and the results were tested by bootstrap resampling. For hardness, we confirmed a previously tagged major QTL on chromosome 5DS, and two additional minor QTLs were found on chromosome 1A and 6D, respectively. For protein content two main QTLs were identified on chromosomes 1B and 6A, respectively. For W, three consistent QTLs were detected: two at the same location as those for hardness, on chromosomes 1A and 5D; the third one on chromosome 3B. Therefore, it appeared that except for the Glu-1A locus, storage protein loci were not clearly involved in the genetic control of the criteria studied in the present work. Despite the reasonable size of the population no QTL with interactive effects could be substantially established as measured. All computations were carried out using home-made programmes in Splus language, and these are available upon request. Received: 16 May 1999 / Accepted: 15 October 1999     

Two simple sequence repeat markers to select for soybean cyst nematode resistance coditioned by the rhg1 locus

The soybean cyst nematode (SCN) (Heterodera glycines Inchinoe) is the most economically significant soybean pest. The principal strategy to reduce or eliminate damage from this pest is the use of resistant cultivars. Identifying resistant segregants in a breeding program is a difficult and expensive process which is complicated by the oligogenic nature of the resistance and genetic variability in the pathogen. Fortunately, resistance at one SCN-resistance locus, rhg1, is generally accepted as a necessity for the development of resistant genotypes using any source of resistance and when challenged by any SCN race. Thus, the development of SCN resistant cultivars would be expedited if an effective and rapid system were available to identify breeding lines carrying a resistance allele at the rhg1 locus. In this study we report two simple sequence repeat (SSR) or microsatellite loci that cosegregate and map 0.4 cM from rhg1. Allelic variation at the first of these loci, BARC-Satt309, distinguished most, if not all, SCN-susceptible genotypes from those carrying resistance at rhg1 derived from the important SCN-resistance sources ’Peking’, PI 437654, and PI 90763. BARC-Satt309 was also effective in distinguishing SCN resistance sources PI 88788 and PI 209332 from many, but not all, susceptible genotypes. BARC-Satt309 cannot be used in marker-assisted selection in populations developed from typical southern US cultivars crossed with the important resistance sources PI 88788 or PI 209332 because these genotypes all carry the identical allele at the BARC-Satt309 locus. A second SSR locus, BARC-Sat_168, was developed from a bacterial artificial chromosome (BAC) clone that was identified using the primers to BARC-Satt309. BARC-Sat_168 distinguished PI 88788 and PI 209332 from southern US cultivars such as ’Lee’, ’Bragg’ and ’Essex’. Both BARC-Satt309 and BARC-Sat_168 were used to assay lines from SCN-susceptible×SCN-resistant crosses and proved to be highly effective in identifying lines carrying rhg1 resistance from those carrying the allele for SCN susceptibility at the rhg1 locus. Received: 5 November 1998 / Accepted: 3 February 1999     

Mapping of simple sequence repeat (SSR) DNA markers in diploid and tetraploid alfalfa

Cultivated alfalfa (Medicago sativa) is an autotetraploid. However, all three existing alfalfa genetic maps resulted from crosses of diploid alfalfa. The current study was undertaken to evaluate the use of Simple Sequence Repeat (SSR) DNA markers for mapping in diploid and tetraploid alfalfa. Ten SSR markers were incorporated into an existing F₂ diploid alfalfa RFLP map and also mapped in an F₂ tetraploid population. The tetraploid population had two to four alleles in each of the loci examined. The segregation of these alleles in the tetraploid mapping population generally was clear and easy to interpret. Because of the complexity of tetrasomic linkage analysis and a lack of computer software to accommodate it, linkage relationships at the tetraploid level were determined using a single-dose allele (SDA) analysis, where the presence or absence of each allele was scored independently of the other alleles at the same locus. The SDA diploid map was also constructed to compare mapping using SDA to the standard co-dominant method. Linkage groups were generally conserved among the tetraploid and the two diploid linkage maps, except for segments where severe segregation distortion was present. Segregation distortion, which was present in both tetraploid and diploid populations, probably resulted from inbreeding depression. The ease of analysis together with the abundance of SSR loci in the alfalfa genome indicated that SSR markers should be a useful tool for mapping tetraploid alfalfa. Received: 10 September 1999 / Accepted: 11 November 1999     

Molecular mapping of the wheat powdery mildew resistance gene Pm24 and marker validation for molecular breeding

Molecular markers were identified in common wheat for the Pm24 locus conferring resistance to different isolates of the powdery mildew pathogen, Erysiphe graminis DM f. sp. tritici (Em. Marchal). Bulked segregant analysis was used to identify amplified fragment length polymorphism (AFLP) markers and microsatellite markers linked to the gene Pm24 in an F₂ progeny from the cross Chinese Spring (susceptible)× Chiyacao (resistant). Two AFLP markers XACA/CTA-407 and XACA/CCG-420, and three microsatellite markers Xgwm106, Xgwm337 and Xgwm458, were mapped in coupling phase to the Pm24 locus. The AFLP marker locus XACA/CTA-407 co-segregated with the Pm24 gene, and XACA/CCG-420 mapped 4.5 cM from this gene. Another AFLP marker locus XAAT/CCA-346 co- segregated in repulsion phase with the Pm24 locus. Pm24 was mapped close to the centromere on the short arm of chromosome 1D, contrary to the previously reported location on chromosome 6D. Pm24 segregated independently of gene Pm22, also located on chromosome 1D. An allele of microsatellite locus Xgwm337 located 2.4±1.2 cM from Pm24 was shown to be diagnostic and therefore potentially useful for pyramiding two or more genes for powdery mildew resistance in a single genotype. Received: 25 August 1999 / Accepted: 16 December 1999     

RAPDs as molecular markers for the detection of Aegilops markgrafii chromatin in addition and euploid introgression lines of hexaploid wheat

 Aegilops markgrafii contains resistance genes to powdery mildew, leaf rust and stripe rust, and also has high crude protein and lysine contents, which can be useful for wheat improvement. These important traits are localized on different chromosomes. Disomic Triticum aestivum-Ae. markgrafii addition lines and euploid introgression lines showing leaf-rust and powdery mildew resistance were screened with RAPDs to detect chromosome-specific markers which can accelerate the breeding process. RAPD markers for all six available disomic addition lines were obtained. The additional chromosomes B, C, D, E, F and G were identified by three, three, three, two, one and seven primers, respectively. All three chromosome-B-specific RAPD markers demonstrated the presence of alien chromatin in the leaf-rust-resistant 42-chromosome introgression lines as well as in the segregating progeny. The three chromosome-C-identifying primers also demonstrated the presence of that chromosome in powdery mildew-resistant euploid introgression lines. The substitution lines (5A)5C and (5D)5C with different genetic backgrounds for both parents, in comparison to the lines mentioned above, showed the chromosome C-specific band with only two of the three primers. The chromosome F-specific primer and a primer evident on all the Ae. markgrafii chromosomes analysed did not generate the expected fragments on the chromosome F_del addition line, indicating that the markers are located on the deleted part of chromosome F. Received: 20 August 1996 / Accepted 17 January 1997     

A leucine to proline mutation in puroindoline b is frequently present in hard wheats from Northern Europe

Endosperm hardness in wheat (Triticum aestivum L.) is determined by one major genetic factor, the Hardness (Ha) gene on the short arm of chromosome 5D. Grain hardness has previously been reported to result from either a failure to express puroindoline a (Pina–D1b) or a glycine to serine mutation at position 46 in puroindoline b (Pinb–D1b). In this study, which involves a large survey of 343 wheat genotypes of mostly Northern European origin, we report two new mutations in puroindoline b associated with hard endosperm. These were characterized as involving a leucine to proline change at position 60 (Pinb–D1c), and a tryptophan to arginine change at position 44 (Pinb–D1d), respectively. While the former seems to be widely distributed in germplasm around the world, the latter was only found in three winter wheats from Sweden and Netherlands. As discussed in the paper, the three known mutations in puroindoline b can be considered ”loss-of-function” mutations (i.e. soft to hard), and structural analysis may serve to predict that their dramatic effect on wheat grain texture is a result of reduced lipid–binding ability. Received: 10 June 1999 / Accepted: 21 September 1999