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1.
Diversity in 20 microsatellite loci of wild emmer wheat, Triticum dicoccoides, was examined in 15 populations (135 genotypes) representing a wide range of ecological conditions of soil, temperature, and water availability, in Israel and Turkey. An extensive amount of diversity at microsatellite loci was observed despite the predominantly selfing nature of this plant species. The 20 Gatersleben wheat microsatellites (GWM), representing 13 chromosomes of genomes A and B of wheat, revealed a total of 364 alleles, with an average of 18 alleles per GWM marker (range: 5–26). The proportion of polymorphic loci per population averaged 0.90 (range: 0.45– 1.00); genic diversity, He, averaged 0.50 (range 0.094– 0.736); and Shannon’s information index averaged 0.84 (range 0.166–1.307). The coefficients of genetic distance between populations were high and averaged D=1.862 (range 0.876–3.320), an indication of sharp genetic divergence over short distances. Interpopulation genetic distances showed no association with geographic distance between the population sites of origin, which ruled out a simple isolation by distance model. Genetic dissimilarity values between genotypes were used to produce a dendrogram of the relationships among wild wheat populations by the unweighted pair-group method with arithmetic averages (UPGMA). The results showed that all the wild emmer wheat populations could be distinguished. Microsatellite analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides, originating from diverse ecogeographical sites in Israel and Turkey, with 88% of the 135 genotypes correctly classified into sites of origin by discriminant analysis. Our present microsatellite results are non-random and in agreement with the previously obtained allozyme and RAPD patterns, although the genetic-diversity values obtained with microsatellites are much higher. Significant correlates of microsatellite markers with various climatic and soil factors suggest that, as in allozymes and RAPDs, natural selection causes adaptive microsatellite ecogeographical differentiation, not only in coding, but most importantly in non-coding genomic regions. Hence, the concept of ”junk DNA” needs to be replaced by at least partly regulatory DNA. The obtained results suggest that microsatellite markers are useful for the estimation of genetic diversity in natural populations of T. dicoccoides and for the tagging of agronomically important traits derived from wild emmer wheat. Received: 27 February 2001 / Accepted: 22 March 2001  相似文献   

2.
 Stripe rust (yellow rust), caused by Puccinia striiformis f. sp. tritici, is one of the most important diseases of wheat throughout the world. Wild emmer wheat, Triticum dicoccoides, the progenitor of cultivated wheat, was found to be a valuable source for novel stripe-rust-resistance genes. The objective of the present study was to estimate the extent of genetic diversity among the wild emmer wheat accessions, previously identified as highly resistant to stripe rust, in order to select suitable parents for genetic-mapping studies. Twenty three wheat microsatellite (WMS) markers were used to detect DNA polymorphism among 21 accessions of T. dicoccoides, which included 19 resistant and two susceptible accessions originating mainly from the center of origin and diversity in the Upper Galilee and Hermon Mountain in northern Israel. In addition, two Triticum durum and one Triticum aestivum lines were also included in the analysis. The 23 WMS markers used were located on 23 chromosome arms, representing all 14 chromosomes of genomes A and B of wheat, and revealed a total of 230 alleles. The number of alleles ranged from 5 to 18, with an average of ten alleles per WMS. Genetic dissimilarity values between genotypes, calculated by the WMSderived data, were used to produce a dendrogram of the relationships among accessions using the unweighted pair-group method with arithmetic averages (UPGMA). The results showed that all of the wild emmer wheat accessions could be distinguished. Most of the resulting groups were strongly related to the ecogeographical origin of the accessions, indicating that the genetic diversity of T. dicoccoides is correlated with geographic distribution. The three major groups were the Rosh Pinna group (north of the Sea of Galilee), the Mount Hermon group (north of the Golan Heights) and Mount Kena’an group (Upper Galilee). The genetic similarity (GS) of the 21 T. dicoccoides accessions based on WMS results averaged 0.31. As expected, the T. durum and T. aestivum lines were grouped separately from the T. dicoccoides accessions. The results obtained suggest that a relatively small number of microsatellites can be used for the estimation of genetic diversity in wild material of T. dicoccoides. These results will be useful in the identification of suitable parents for the development of mapping populations for tagging yellow-rust resistance genes derived from T. dicoccoides. Furthermore, future work could test the adaptive evolutionary significance of microsatellites in natural populations of wild emmer wheat. Received: 8 August 1997 / Accepted: 25 August 1997  相似文献   

3.
 Genetic diversity was examined by RAPD-PCR analysis in 118 registered individuals of wild emmer wheat, Triticum dicoccoides, from a microsite at Yehudiyya, northeast of the Sea of Galilee, Israel. The test involved two climatic microniches in the open oak-park forest of Quercus ithaburensis: (1) sunny between trees and (2) shady under the trees’ canopies. Comparisons were based on 97 loci amplified by 20 oligonucleotide primers. Significant genetic differentiations were found at single-, two- and multilocus structures between the neighbouring shady and sunny niches. These DNA polymorphisms appear to be associated with microclimatic stresses. The pronounced niche-effect on the significance of linkage disequilibrium and niche-specific linkage disequilibrium may suggest that natural selection directed the two-locus associations. The structure of the multilocus associations also mainly results from natural selection, and not by chance from population subdivision, or founder effects. These findings are largely parallel to the previous allozymic results at single-locus and multilocus levels. Both the DNA and the allozymic results suggest that microclimatic selection appears to play an important role in DNA differentiation as well as in protein polymophism. Received: 30 October 1998 / Accepted: 2 November 1998  相似文献   

4.
抗条锈病基因Yr69对我国小麦条锈菌(Puccinia striiformis f. sp. tritici)小种具有广谱抗性,在小麦抗条锈病育种中具有重要价值。为提高分子标记辅助选择育种的效率,加快Yr69在小麦抗病育种中的应用,本研究利用条锈菌小种CYR34对包含340个小麦家系的‘Taichung29/CH7086’F9代RIL(Recombinant inbred line)群体进行接种鉴定,并利用BSA-SNP(Bulked segregant analysis-single nucleotide polymorphism)技术对其抗条锈病基因进行了重新定位。抗病鉴定结果显示,RIL群体中抗感病家系的数量呈双峰分布,‘CH7086’的条锈病抗性受一个主效位点控制。BSA-SNP基因分型结果表明,多态性SNP主要集中于小麦2AS染色体末端0~30Mb的染色体区段。在该基因组区段开发了208个SSR分子标记,利用抗感病小群体从中筛选到14个与Yr69连锁的分子标记。利用14个标记对340个RIL家系进行PCR扩增和分子作图,将Yr69定位于2AS111和2AS171之间约7.76...  相似文献   

5.
Near-isogenic lines (NILs) for the leaf rust resistance gene Lr9 were screened for polymorphisms at the molecular level. RAPD (random amplified polymorphic DNA) primers as well as RFLP (restriction fragment length polymorphism) markers were used. Out of 395 RAPD primers tested, three showed polymorphisms between NILs, i.e., an additional band was found in resistant lines. One of these polymorphic bands was cloned and sequenced. Specific primers were synthesized, and after amplification only resistant lines showed an amplified product. Thus, these primers define a sequence-tagged site that is specific for the translocated fragment carrying the Lr9 gene. A cross between a resistant NIL and the spelt (Triticum spelta) variety Oberkulmer was made, and F2 plants were analyzed for genetic linkage. All three polymorphisms detected by the PCR (polymerase chain reaction) and one RFLP marker (cMWG684) showed complete linkage to the Lr9 gene in 156 and 133 plants analyzed, respectively. A second RFLP marker (PSR546) was closely linked (8±2.4 cM) to the Lr9 gene and the other four DNA markers. As this marker maps to the distal part of the long arm of chromosome 6B of wheat, Lr9 and the other DNA markers also map to the distal region of 6BL. All three PCR markers detected the Lr9 gene in independently derived breeding lines and varieties, thus proving their general applicability in wheat breeding programs.  相似文献   

6.
Twenty eight microsatellite markers were used to analyze genetic divergence in tandem dinucleotide repeated DNA regions between two edaphic subpopulations of Triticum dicoccoides growing on the contrasting terra rossa and basalt soilsfrom a microsite at Tabigha, north of the Sea of Galilee, Israel. The terra rossa soil niche was drier and more stressful than the basalt throughout the growing season (November to May). Significant microsatellite divergence in allele distribution, repeat length, genetic diversity, and linkage disequilibria were found between emmer wheat from the two soil types over two short transects of 100 m each. Soil-specific and -unique alleles and linkage disequilibria were observed in the terra rossa and basalt subpopulations. A permutation test showed that the effects of random genetic drift were very low for the significant genetic diversity at microsatellite loci between the two subpopulations, suggesting that an adaptive molecular pattern derived by edaphic selection may act upon variation of the microsatellites. Received: 4 February 2000 / Accepted: 31 March 2000<@head-com-p1a.lf>Communicated by H.F. Linskens  相似文献   

7.
 Genetic diversity in random amplified polymorphic DNAs (RAPDs) was studied in 110 genotypes of the tetraploid wild progenitor of wheat, Triticum dicoccoides, from 11 populations sampled in Israel and Turkey. Our results show high level of diversity of RAPD markers in wild wheat populations in Israel. The ten primers used in this study amplified 59 scorable RAPD loci of which 48 (81.4%) were polymorphic and 11 monomorphic. RAPD analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides originating from diverse ecogeographical sites in Israel and Turkey, with 95.5% of the 100 genotypes correctly classified into sites of origin by discriminant analysis based on RAPD genotyping. However, interpopulation genetic distances showed no association with geographic distance between the population sites of origin, negating a simple isolation by distance model. Spatial autocorrelation of RAPD frequencies suggests that migration is not influential. Our present RAPD results are non-random and in agreement with the previously obtained allozyme patterns, although the genetic diversity values obtained with RAPDs are much higher than the allozyme values. Significant correlates of RAPD markers with various climatic and soil factors suggest that, as in the case of allozymes, natural selection causes adaptive RAPD ecogeographical differentiation. The results obtained suggest that RAPD markers are useful for the estimation of genetic diversity in wild material of T. dicoccoides and the identification of suitable parents for the development of mapping populations for the tagging of agronomically important traits derived from T. dicoccoides. Received: 13 July 1998 / Accepted: 13 August 1998  相似文献   

8.
Two stripe-rust resistance genes, YrH52 and Yr15, derived from the Israeli wild emmer wheat, Triticum dicoccoides, have been located on chromosome 1B. The main objectives of the present study were to increase marker density in the vicinity of YrH52 gene by means of AFLP, RAPD and microsatellite markers, to improve the map of another T. dicoccoides-derived stripe-rust resistance gene Yr15 using microsatellite markers, and to preliminarily discriminate these two genes. Additional 26 marker loci comprising 20 AFLPs, three RAPDs, and three microsatellites were found to be linked to YrH52 gene. An updated genetic map consisting of 45 marker loci, in the region of YrH52 gene, was constructed with a total map length of 107.7 cm. The mean interval length was 0.96 cm in the region Xgwm359b–P55M53b carrying YrH52 gene. YrH52 was bracketed by Xgwm413 (Nor1 and UBC212a) and Xgwm273a (Xgwm273d) with map distance of 1.3 and 2.7 cm from either side, respectively. Eight additional microsatellite markers were found to be linked with Yr15, and the linkage map of Yr15 gene was thus obviously improved. In the YrH52-mapping population, no crossover was detected in the interval UBC212a (Xgwm413)–Yr15Nor1, and YrH52 was located distally outside this interval. It may suggest that YrH52 is different from Yr15 even though both of them are derived from T. dicoccoides and are mapped on chromosome 1BS. The large number of molecular makers revealed in the present study would be helpful for the marker-assisted introgression of the T. dicoccoides-derived YrH52 and Yr15 stripe-rust resistance genes into elite cultivars of wheat, and the high-density map would accelerate the map-based cloning of the two genes. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

9.
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.  相似文献   

10.
11.
 A sequence-tagged-site (STS) marker is reported linked to Lr28, a leaf rust resistance gene in wheat. RAPD (random amplified polymorphic DNA) analysis of near-isogenic lines (NILs) of Lr28 in eight varietal backgrounds was carried out using random primers. Genomic DNA enriched for low-copy sequences was used for RAPD analysis to overcome the lack of reproducibility due to the highly repetitive DNA sequences present in wheat. Of 80 random primers tested on the enriched DNA, one RAPD marker distinguished the NILs and the donor parent from the susceptible recurrent parents. The additional band present in resistant lines was cloned, sequenced, and STS primers specific for Lr28 were designed. The STS marker (Indian patent pending: 380 Del98) was further confirmed by bulk segregation analysis of F3 families. It was consistently present in the NILs, the resistant F3 bulk and the resistant F3 lines, but was absent in recurrent parents, the susceptible F3 bulk and the susceptible F3 lines. Received: 20 February 1998 / Accepted: 4 March 1998  相似文献   

12.
Stripe rust resistance was identified in Triticum vavilovii (T. vavilovii Aus22498)-derived Russian wheat aphid (RWA)-resistant germplasm. Inheritance studies indicated monogenic control of resistance. The resistance gene was tentatively designated as Yrvav and was located on chromosome 1B by monosomic analysis. A close association (1.5±0.9% recombination) of Yrvav with a T. vavilovii-derived gliadin allele (Gli-B1vav) placed it in chromosome arm 1BS. Yrvav was allelic with Yr10. Tests with Yr10 avirulent and virulent pathotypes showed that Yrvav and Yr10 possess identical pathogenic specificity. Yrvav and Yr10 showed close genetic associations with alternate alleles at the Xpsp3000 (microsatellite marker), Gli-B1 and Rg1 loci. Based on these observations Yrvav was named as Yr10vav. The close association between Xpsp3000 and Gli-B1 was also confirmed. The Yr10vav-linked Xpsp3000 allele (285 bp) was not present in 65 Australian cultivars, whereas seven Australian wheats lacking Yr10 carried the same Xpsp3000 allele (260 bp) as Yr10 carrying wheat cultivar Moro. Xpsp3000 and/or Gli-B1 could be used in marker-assisted selection for pyramiding Yr10vav or Yr10 with other stripe rust resistance genes. Yr10vav was inherited independently of the T. vavilovii-derived RWA resistance. Received: 5 December 2000 / Accepted: 3 April 2001  相似文献   

13.
 Geographical variation in vernalization response and narrow-sense earliness was investigated for accessions of wild emmer wheat, Triticum dicoccoides, collected in Israel. Wide variation between and within populations was observed in both characters. The analysis of vernalization response showed that 2 accessions from Tabigha were of a strong spring growth habit, and thus wild emmer wheat was classified into four types, i.e., strongly spring type, moderately spring type, moderately winter type, and strongly winter type, according to their vernalization response. Whereas winter types were frequently found in most populations except that of Tabigha, the distribution of spring types was sporadic and restricted to warmer areas. It was thus suggested that spring type in T. dicoccoides might have evolved from a winter prototype as an adaptation to warmer conditions. Within moderately winter and moderately spring types, quantitative differences in vernalization response, measured as Dof70/Dof20 and Dof20/Dof0, were observed between populations. Inter- and intra-population variation in vernalization response could be explained to some extent by the difference in growing conditions at each habitat. It was clearly indicated that environmental heterogeneity caused ecogenetic differentiation in wild emmer wheat in Israel. Wild emmer wheat also varied considerably for narrow-sense earliness, ranging from 32.9 days to 69.5 days among accessions. However, it was difficult to explain its geographical variation simply by a linear relationship with environmental factors, and a nonlinear relationship and/or unknown microgeographic heterogeneity may be responsible. Received: 18 March 1996/Accepted: 13 December 1996  相似文献   

14.
 White rust, caused by Albugo candida (Pers.) Kuntze, is an economically important disease of Brassica juncea (L.) Czern. and Coss mustard, particularly in India. The most efficient and cost-effective way of protecting mustard plants from white rust disease is through genetic resistance. The objective of this study was to identify RAPD markers for white rust resistance in an F1-derived doubled-haploid (DH) population originating from a cross between white rust-susceptible and white rust-resistant breeding lines of B. juncea from the canola-quality B. juncea breeding project of the Agriculture and Agri-Food Canada-Saskatoon Research Centre. The DH population was used to screen for RAPD markers associated with white rust resistance/susceptibility using bulked segregant analysis. Two markers, WR2 and WR3, linked to white rust resistance, flanked the resistance locus Ac2 1 and were highly effective in identifying the presence or absence of the resistance gene in the DH population. These two markers were shown to be specific to the Russian source of white rust resistance utilized in this project. It is concluded that the availability of these RAPD markers will enhance the breeding for white rust resistance in B. juncea. Received: 17 December 1997 / Accepted: 7 April 1998  相似文献   

15.
 Stripe rust caused by Puccinia striifomis West. is one of the most devastating diseases relating to wheat production. Wild emmer wheat, Triticum dicoccoides, the tetraploid progenitor of cultivated wheat, has proven to be a valuable source of novel stripe-rust resistance genes for wheat breeding. For example, T. dicoccoides accessions from Mt. Hermon, Israel, are uniformly and highly resistant to stripe-rust. The main objective of the present study is to map a stripe-rust resistance gene, derived from the unique Mt. Hermon population of wild emmer, using microsatellite markers. An F2 mapping population was established by crossing stripe-rust resistant T. dicoccoides accession H52 from Mt. Hermon with the Triticum durum cultivar Langdon. The stripe-rust resistance derived from accession H52 was found to be controlled by a single dominant gene which was temporarily designated as YrH52. Out of 120 microsatellite markers tested, 109 (91%) showed polymorphism between the parental lines. Among 79 segregating microsatellite loci generated from 56 microsatellite primer pairs, nine were linked to YrH52 with recombination frequencies of 0.02–0.35, and LOD scores of 3.56–54.22. A genetic map of chromosome 1B, consisting of ten microsatellite loci and the stripe-rust resistance gene YrH52, was constructed with a total map length of 101.5 cM. YrH52 is also closely linked to RFLP marker Nor1 with a map distance of 1.4 cM and a LOD value of 29.62. Apparent negative crossover interference was observed in chromosome 1B, especially in the region spanning the centromere. Negative crossover interference may be a common characteristic of gene-rich regions or gene clusters in specific chromosomes. Received: 30 October 1998 / Accepted: 2 November 1998  相似文献   

16.
Spatiotemporal diversity at 35 allozyme loci was assayed over 6 years in 1,207 individuals of wild emmer wheat (Triticum dicoccoides)from a microgeographic microsite, Ammiad, north Israel. This analysis used new methods and two additional sample sets (1988 and 1993) and previous allozymic data (1984–1987). This microsite includes four major habitats (North-facing slope, Valley, Ridge, and Karst) that show topographic and ecological heterogeneity. Significant temporal and spatial variations in allele frequencies and levels of genetic diversity were detected in the four subpopulations. Significant associations were observed among allele frequencies and gene diversities at different loci, indicating that many allele frequencies change over time in the same or opposite directions. Multiple regression analysis showed that variation in soil-water content and rainfall distribution in the growing season significantly affected 10 allele frequencies, numbers of alleles at 8 loci, and gene diversity at 4 loci. Random genetic drift and hitchhiking models may not explain such locus-specific spatiotemporal divergence and strong allelic correlation or locus correlation as well as the functional importance of allozymes. Natural ecological selection, presumably through water stress, might be an important force adaptively directing spatiotemporal allozyme diversity and divergence in wild emmer wheat at the Ammiad microsite. Received: 3 July 2000 / Accepted: 1August 2000  相似文献   

17.
18.
The Yr17 gene, which is present in many European wheat cultivars, displays yellow rust resistance at the seedling stage. The gene introduced into chromosome 2A from Aegilops ventricosa was previously found to be closely linked (0.5 cM) to leaf and stem rust resistance genes Lr37 and Sr38, respectively. The objective of this study was to identify molecular markers linked to the Yr17 gene. We screened with RAPD primers, for polymorphism, the DNAs of cv. Thatcher and the leaf rust-resistant near-isogenic line (NIL) RL 6081 of cv. Thatcher carrying the Lr37 gene. Using a F2 progeny of the cross between VPM1 (resistant) and Thésée (susceptible), the RAPD marker OP-Y15580 was found to be closely linked to the Yr17 gene. We converted the OP- Y15580 RAPD marker into a sequence characterized amplified region (SCAR). This SCAR marker (SC-Y15) was linked at 0.8 ± 0.7 cM to the Yr17 resistance gene. We tested the SC-Y15 marker over a survey of 37 wheat cultivars in order to verify its consistency in different genetic backgrounds and to explain the resistance of some cultivars against yellow rust. Moreover, we showed that the Xpsr150-2Mv locus marker of Lr gene described by Bonhomme et al. [6] which possesses A. ventricosa introgression on the 2A chromosome was also closely linked to the Yr17 gene. Both the SCAR SC-Y15 and Xpsr150-2Mv markers should be used in breeding programmes in order to detect the cluster of the three genes Yr17, Lr37 and Sr38 in cross progenies. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

19.
The objective of this work was to develop a marker for the adult plant leaf rust resistance gene Lr35. The Lr35 gene was originally introgressed into chromosome 2B from Triticum speltoides, a diploid relative of wheat. A segregating population of 96 F 2 plants derived from a cross between the resistant line ThatcherLr35 and the susceptible variety Frisal was analysed. Out of 80 RFLP probes previously mapped on wheat chromosome 2B, 51 detected a polymorphism between the parents of the cross. Three of them were completely linked with the resistance gene Lr35. The co-segregating probe BCD260 was converted into a PCR-based sequence-tagged-site (STS) marker. A set of 48 different breeding lines derived from several European breeding programs was tested with the STS marker. None of these lines has a donor for Lr35 in its pedigree and all of them reacted negatively with the STS marker. As no leaf rust races virulent on Lr35 have been found in different areas of the world, the STS marker for the Lr35 resistance gene is of great value to support the introgression of this gene in combination with other leaf rust (Lr) genes into breeding material by marker-assisted selection. Received: 14 December 1998 / Accepted: 30 January 1999  相似文献   

20.
The leaf rust resistance gene Lr47 confers resistance to a wide spectrum of leaf rust strains. This gene was recently transferred from chromosome 7S of Triticum speltoides to chromosome 7A of hexaploid wheat Triticum aestivum. To facilitate the transfer of Lr47 to commercial varieties, the completely linked restriction fragment length polymorphism (RFLP) locus Xabc465 was converted into a PCR-based marker. Barley clone ABC465 is orthologous to the type-I wheat sucrose synthase gene and primers were designed for the conserved regions between the two sequences. These conserved primers were used to amplify, clone and sequence different alleles from T. speltoides and T. aestivum. This sequence information was used to identify the T. speltoides sequence, detect allele-specific mutations, and design specific primers. Cosegregation of the PCR product of these primers and the T. speltoides chromosome segment was confirmed in four backcross-populations. To complement this dominant marker, a cleavage amplified polymorphic sequence (CAPS) was developed for the 7A allele of Xabc465. This CAPS marker is useful to select homozygous Lr47 plants from F2 or backcross-F2 segregating populations, and in combination with the T- speltoides specific primers is expected to facilitate the deployment of Lr47 in new bread wheat varieties. Received: 11 October 1999 / Accepted: 30 December 1999  相似文献   

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