Allozyme differentiation of European populations and cultivars of Lolium perenne L., and the relation to ecogeographical factors

Summary Sixty Lolium perenne populations were screened for allozyme diversity at five loci. Objective was to determine whether allozyme diversity could be used as selection criterion for genebank accessions of L. perenne. Subsampling of locations was tested with allozyme analysis to determine whether genetically different populations could be collected at one location. Correlations between allelic frequencies and environmental factors and morphological data were established, to find ecogeographical patterns in the observed variation. Results indicated that with few examples each allelic variant could be observed in each population screened; very few unique alleles were found. Differences between populations were largely due to differences in allelic frequencies. Few correlations were found with environmental factors and morphological data. For some allele frequencies a north-south cline was observed. Generally, allozymic data of the five screened loci did not appear to be useful for the selection of accessions for genebank storage. Significant genotypic differences between populations collected at one location could be established. In general these results agreed with earlier results concerning phenotypic variation in the same populations.     

Genetic diversity of natural populations of Atriplex halimus L. in Morocco: An isoenzyme-based overview

Based on polyacrylamide gel electrophoresis, nine natural populations of Atriplex halimus L., a perennial shrub, collected in different regions of Morocco, were studied for their genetic variation using isoenzyme polymorphism of the highly active enzyme systems: esterases (EST), acid phosphatases (ACP) and glutamate oxaloacetate transaminase (GOT). Different allozyme frequencies from 7 different loci were obtained for all populations of this halophyte species. High levels of genetic diversity were revealed. The mean number of alleles per locus (A = 1.9–2.0), the percentage of polymorphic loci (p = 71.4–85.7) and the mean expected heterozygosity (H_e = 0.339–0.385) showed an important variability in all populations. Gene diversity was essentially explained by the within population component. The between populations differentiation accounted for 8% of the whole diversity (F_ST, averaged over all loci, is 0.08). The relationships among the 9 populations were inferred from the Nei’s genetic distances. Four major groups were formed. The northern population ‘Tanger’, forming a unique group, was highly divergent from the other groups. It appeared that the genetic distance between all groups was related to the geographic distance that separates them. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pure-lining of flax (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.) genebank accessions for efficiently exploiting and assessing seed character diversity

Genetic heterogeneity within genebank accessions of a self-pollinating crop has great implications for their preservation, diversity assessment, utilization and exploitation. This study investigated the intra-accession diversity of 38 flax (Linum usitatissimum L.) genebank accessions preserved by Plant Gene Resources of Canada. Seven quantitative seed characters were studied: 1000 seed weight, seed oil concentration and fatty acid proportion of palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and α-linolenic acid (18:3). Ten additional morphological characters of high heritability were also recorded. In 13 accessions the intra-accession diversity was assessed based on 4 years of observation of 7–12 pure lines selected randomly from the original accessions. From these 13 accessions, seven showed significant variation among the pure lines for all seven seed characters. In five accessions most seed characters showed significant variation and only in one accession the homogeneity of seed characters dominated. Variation was found in landraces, breeding material and cultivars. In 40 cases an attempt was made to identify pure lines within an accession that significantly exceeded the mean value of the original accession in a seed character, for which the original accession had shown high or low values. In 25 cases and for all seven seed characters such pure lines could be detected. In six cases, the most extreme performing pure lines were not from the same accession, that had the highest or lowest accession mean for the considered character. There was no association of morphological variation with variation of the quantitative seed characters. For genebanks, separation of mixed accessions based on obvious phenotypic diversity and in particular based on characters that create a selective pressure is probably the most efficient way to ensure preservation of a wide range of diversity at reasonable cost. For exploitation of diversity in breeding programmes, pure-lining is very useful. J. Philip Raney—Deceased.     

Genetic variability among Turkish pop, flint and dent corn (Zea mays L. spp. Mays) races: Enzyme polymorphism

To determine magnitude and pattern of genetic variation, 32 Turkish corn accessions available from the USDA/ARS North Central T-Regional Plant Introduction Station (NCRPIS) collections (Ames, Iowa), representing pop, flint and dent corn races, different climatic, geographic and topographic areas in Turkey, were identified and, 19 isozyme systems were studied. Thirty-nine alleles were detected by 19 isozyme loci in 32 accessions. The PGD-2, Mmm-1, GOT-3 and IDH-1 loci were found to be monomorphic in all accessions. Mean number of alleles per locus varied between 1.2 in Balıkesir-167949 accession to 1.7 in Ankara-177600, Trabzon-185062, Eskişehir-204822 and Samsun-239573 accessions. The proportion of polymorphic loci ranged from as low as 15.8% in Balıkesir-167949 to as high as 57.9% in Trabzon-185049 accession. Observed heterozygosity was the highest in Adana-183779 and the lowest in Ankara-204800 accession. Genetic identities ranged from 0.823 for 170881-Kocaeli and 167949-Balikesir pair to 0.997 for 182327-Iğdır and 168008-Kırklareli pair. Dendrogram constructed by using Nei's genetic distances (1978) revealed three clustering groups, though one of the clusters included only 170881-Kocaeli accession. It is concluded that170881-Kocaeli accession must have experienced with intensive selection, inbreeding and/or bottleneck effects in the past. Corn germplasm managers and breeders could use the results of the present studies for monitoring genetic resources, accession identification, and sampling genetic diversity, but agronomic characteristics of these corn accessions are also needed for a better utlization of Turkish corn land races evolved over the years. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetics of some in vitro characters in pearl millet

Summary Genetics of four in vitro characters was studied using immature inflorescence-derived callus from four inbred lines. The number of genes controlling total callus quantity varied from 5–11, those for embryogenic(E) callus quantity from 4–7, for callus growth rate from 3–12 and those for regeneration frequency varied from 4–9 in the three segregating F₂ populations. Callus derived from the dwarf plants (d₂ d₂) was phenotypically distinguishable from that of the tall accessions. The association between d₂ locus and the in vitro characters was analysed using three different approaches. It is suggested that d₂ locus might be closely linked to majority of the loci governing E callus production.     

Diversity in the wild potato species Solanum chacoense Bitt.

Summary The morphological and isozyme variation was studied in 22 accessions of Solanum chacoense from Paraguay and Argentina. Clear geographic groups were identified through the use of multivariate analyses. S. chacoense from mountain sites in Argentina could be readily distinguished from plains forms from Paraguay, on the basis of several correlated morphological characters. Three isozyme systems, namely phosphoglucose isomerase (PGI), glutamate-oxaloacetate transaminase (GOT) and peroxidase (PRX) were studied using starch gel electrophoresis. The banding patterns indicated that for each isozyme there were several loci, which were polymorphic. A genetic interpretation of one of the PGI loci was made, and indices of genetic diversity and genetic identity calculated. Principal components analysis, cluster analysis and genetic diversity indicated a close relationship between the geographical groups. These results are discussed in the context of in situ genetic conservation.     

Genetic variation for isozyme genes and proteins in spanish primitive cultivars and wild subspecies of Lens

Summary An analysis of the variability for genes encoding seven isozyme systems and storage proteins in a collection of cultivated and wild accessions of Lens is reported. The collection, which is part of the Spanish INIA Cenebank, contains the ssp. culinaris, orientalis, odemensis, nigricans and ervoides, and presents a high degree of genetic diversity both within and between the accessions. A total 25 loci were examined; of these, 18 were polymorphic (the 7 genes encoding storage proteins, and the following isozyme loci: Acp-1, Acp-2, Cpx-1, Cpx-2, Aat-p, Aat-m, Lap-1, Mdh-2, Mdh-3, Mdh-4 and 6pgd-p) and 7 were monomorphic (Aat-mb, Aat-c, Mdh-1, Mdh-5, 6pgd-2, Pgm-c and Pgm-p). The phylogenetic relationships between subspecies were analyzed using the allelic frequencies. The study suggests that orientalis and odemensis share more biochemical characters than the other subspecies, and that those subspecies keep an intermediate position between Lens culinaris and Lens nigricans.     

The European Prunus mapping project Progress in the almond linkage map

Summary Six European research groups are collaborating to develop genetic markers and linkage maps for use inPrunus breeding programmes. A basic map with 200 RFLPs and 50 more markers including isozymes and RAPDs will be constructed using two highly segregating populations: an interspecific peach × almond F₂ and a cherry F₂. Then, the parents of eleven almond, cherry, peach or plum breeding progenies segregating for target characters will be screened for polymorphisms at the marker loci, and a set of reduced maps, one per progeny, will be constructed with markers spaced 20–30 cM and covering the whole genome. Cosegregation analysis of markers and characters of interest will allow us to find linkages between markers and major genes or quantitative trait loci responsible for the expression of these traits. A map with 72 markers, 7 isozymes and 65 RFLPs, has been developed at the IRTA-Cabrils laboratory using an intraspecific almond progeny, ‘Ferragnes’ × ‘Mono’. Probes for the analysis of RFLPs were obtained from almond genomic and cDNA libraries. The level of polymorphism for RFLPs and the distribution of markers in the chromosomes of almond are discussed.     

Evaluation and variation in response to infection with Puccinia striiformis and Puccinia recondita of local wheat landraces

The purpose of this study was to identify the species of local landraces of wheat (Triticum spp.), held in the Israel Gene Bank, to evaluate them for basic characters and to assess their response to infection by two rust fungi under artificial inoculation conditions. One-hundred-thirty one seed samples were collected from local or Beduin farmers during 1978–1981 throughout the Galilee, Mt. Gilboa. Judean Desert and the south Negev. The samples were collected and stored in the Israel Gene Bank without any characterization or evaluation. Each accession was planted in a 1 m row at Bet Dagan and grown under favorable conditions for plant growth and rust development. Determination of the species, data of plant height, days to heading and reaction of the landraces to artificial inoculation with a composite inoculum of Puccinia recondita and P. striiformis were collected from each row. A small part of the landraces collection consisted of mixed populations of T. durum and T. aestivum plants, where one of the two species was predominant. One-hundred-fourteen and 17 accessions from this collection represented, respectively, Triticum durum and T. aestivum Israel landraces. Large variations were found for all the characters examined. Of the total accessions, 6.5% (8 accessions) and 32% (42 accessions) were resistant, respectively, to yellow- and leaf-rust. It was concluded that the diversified populations of the local landraces of wheat can be used as a source not only for genes affecting basic characters such as plant height and heading date, but also for resistance to leaf rust and yellow rust. This revised version was published online in July 2006 with corrections to the Cover Date.     

Association of isozyme markers with quantitative trait loci in random single seed descent derived lines of lentil (Lens culinaris Medik.)

Summary Polymorphism at isozyme loci was used to locate factors responsible for variation in quantitative traits of lentil. Eight sets of random single seed descent (RSSD) derived lines were developed by advancing individual F₃ plants of interspecific (L. culinaris Medik. × L. orientalis Boiss.) hybrids to the F₆. The RSSD lines in each of the eight sets differed for alleles at 2–8 isozyme loci. In each set, association of isozyme loci with variation in seven quantitative traits (days to flower, days to mature, plant height, biomass, seed yield, harvest index, seed weight) was determined for each pairwise combination of a quantitative trait with a marker locus. Loci affecting variation in all seven quantitative traits were detected by their association with 14 isozyme markers (Aat-c, Aat-m, Aat-p, Adh-1, Fk, Gal-1, Gal-2, Lap-1, Lap-2, Pgd-p, Pgi, Pgm-c, Pgm-p, Skdh). The known position of 10 the 14 isozyme loci on the lentil genetic map was used to mark the genomic regions for possible location of associated quantitative trait loci (QTL). Detected QTL were found to be located in six of the seven linkage groups on lentil genetic map. Regions of the genome represented by linkage groups, 1, 5 and 7 appeared to affect a greater number of traits than other genomic regions represented by linkage groups 2, 3 and 4. Results indicated that the mean expression of quantitative traits at segregating marker locus classes can be used to locate the genetic factors in lentil which influence the behavior of economically important traits.     

Estimation of the Isozymatic Genetic Variability of a Spanish A vena sativa Germplasm Collection

An analysis of Spanish land races of Avena sativa from the Germplasm Bank of the I.N.I.A. was carried out using starch gel electrophoresis. Samples of 15 seedlings per accession were used for estimating the intra- and interpopulational genetic variability. Electrophoretic analyses were performed with crude extracts from 15 day-old seedling leaflets by means of eight isozymatic systems representing a total of ten genetic loci. Two systems were mono-morphic in all the accessions, while the other six showed polymorphism both within and between populations. Intrapopulational variability was particularly high in this collection, only 16 out of the 137 accessions being uniform for all the isozyme systems. Thus the material was shown to represent a high potential of genetic variability for breeding programmes.     

用SSR标记研究不同耐盐特性四倍体紫花苜蓿的遗传多样性

用8个微卫星位点，63个等位基因，研究了8个栽培型紫花苜蓿品种（Medicago sativa L.），320个单株群体内和群体间的遗传变异。首次提出用四倍体方法统计紫花苜蓿微卫星位点的等位基因频率，以等位基因频率为基础，得到遗传杂合度、有效等位基因数、标准遗传距离和NJ（Neighbor Jointing）聚类图。结果表明这8个材料在群体内和群体间均具较高变异。通过0/1法和四倍体方法得到的遗传杂合度之间的显著性检验（P<0.01），表明四倍体方法更适于紫花苜蓿微卫星多态性研究。     

Application of isozyme electrophoresis for purity testing and cultivar identification of F1 hybrids of Brassica oleracea

Summary Six isozyme genes were analyzed in seed samples of 65 commercial F₁ hybrids of four horticultural groups of Brassica oleracea (cabbage, Brussels sprouts, sprouting broccoli and cauliflower). Results obtained from electrophoretic assays led to the following conclusions: 1) the electrophoretic test of F₁ hybrid purity was possible in 59 (91%) of the hybrids analyzed, since their inbred parents were apparently fixed each for a different allele in at least on of the loci studied; 2) forty-eight (74%) of the hybrids were individually distinguished by their isozyme phenotype; 3) high levels of segregation in the inbred parents were inferred from the analysis of a sample of seeds of each hybrid.     

Isozyme polymorphism in Festuca rubraL.

Summary Six Festuca rubra populations from Europe and Scandinavia were studied for variation at three isozyme loci; phosphoglucoisomerase (PGI-2), glutamate oxaloacetate transaminase (GOT-3) and superoxide dismutase (SOD-1). Seven alleles were found at the Pgi-2 locus, four at the Got-3 locus and five at the Sod-1 locus. Most plants were heterozygous and up to five alleles were found in the same plant at the Pgi-2 locus. Each population could be distinguished by the presence or absence of certain alleles or by differences in the frequencies of the alleles present. Values for the Shannon diversity index were calculated which showed that there was considerable heterogeneity both within and between loci. In general, 53% of this diversity could be attributed to within population variation.     

Molecular characterization of genetic diversity in European germplasm of perennial ryegrass

Summary Perennial ryegrass (Lolium perenne L.) is the most important grass species for temperate grassland agriculture. The level and distribution of genetic variation in gene bank ecotype collections is still largely unknown but of great interest for the planning of breeding programs. The objectives of this study were to (i) assess the molecular diversity of Polish ecotypes of perennial ryegrass, and (ii) compare the relationship between this group and German ecotypes and European cultivars investigated previously. A total number of 166 polymorphic marker bands were detected among the 171 individual plants of the 9 Polish ecotypes. In a joint analysis with 9 Polish and 22 German ecotypes, and 22 European cultivars 172 polymorphic RAPD markers could be found. Genetic distance among the Polish ecotypes ranged from 0.31 to 0.51, while for all 53 populations a broader range was detected (0.25–0.67). An analysis of molecular variance (AMOVA) revealed a much larger variation within populations (71%) than among them (29%). The Polish ecotypes contained the highest within population variation (74%). The largest among group difference (15%) was found between the Polish ecotypes versus all other accessions. We conclude that the Polish ecotypes represent a valuable genetic resource for enlarging the genetic variation in the West European germplasm pool of perennial ryegrass.     

Conservation of diversity in bulk populations of barley (hordeum vulgare L.)

Summary A series of experiments was conducted to investigate the effectiveness of mass propagated heterogeneous populations (bulk populations) in preserving genetic and phenotypic diversity. Five genetically broad-based bulk populations of cultivated barley (Hordeum vulgare L.) were employed. All of them were produced originally through mass hybridization that was facilitated by male sterility. Four of these populations had a common origin, but were subsequently propagated in different North American locations. Comparisons between early and advanced generations of these bulk populations revealed loss of variability in all populations for morphological and agronomic characters and very little to none for eight isozyme characters. Populations propagated in different locations differed in levels of residual diversity. The bulk populations were less variable than a random sample of parental accessions. The rapid loss of diversity in bulk populations was considered detrimental to genetic conservation.     

Association of agronomic traits with isozyme loci in perennial ryegrass (Lolium perenne L.)

Summary Inbred lines possessing different genotypes at three isozyme loci, PGI-2, GOT-3 and ACP-1, were produced by two generations of selfing from two contrasting cultivars of perennial ryegrass. Four separate F₂ populations were obtained by selfing or crossing F₁ plants from crosses between cultivar inbred lines. F₂ plants were scored for genotype at the three isozyme loci and for eight agronomic traits. The three isozyme loci showed independent segregation confirming that they belonged to separate linkage groups. A consistent association was observed between water soluble carbohydrate content and genotype at the PGI-2 locus. Some association was also detected between genotype at the ACP-1 locus and heading date. The relevance of these observations to ryegrass breeding programmes is discussed.     

QTLs identification of crude fat content in brown rice and its genetic basis analysis using DH and two backcross populations

Fat content is a concern for the enhancement of rice for eating, cooking, and storage qualities. To clarify its genetic mechanism, a double haploid (DH) population derived from anther hybrid F₁ of Zhenshan 97B (indica) and Wuyujing 2 (japonica) and two backcross F₁ (BCF₁) populations, which came from the DH lines backcrossing to two parents, were used to scan quantitative trait loci (QTLs) and dissect gene effects for the crude fat content (CFC) in brown rice. Fourteen QTLs were resolved, distributing on chromosomes 1, 3, and 5–9. Three loci were detected repeatedly in two populations, DH or BCF₁. Among these loci, a major QTL, qCFC5, flanking markers RM87 and RM334, was located on chromosome 5, which was detected simultaneously among three populations. The main QTLs had a major role in controlling CFC in brown rice and were modified by several mini-effect QTLs and epistatic affection. Wenjun Liu and Jing Zeng are contributed equally to this paper.     

Genetic mapping of quantitative resistance to race 5 of Pseudomonas syringae pv. phaseolicola in common bean

Halo-blight is an important worldwide bacterial disease of common bean (Phaseolus vulgaris L.) caused by Pseudomonas syringae pv. phaseolicola. Nine races of the pathogen and five race-specific resistance genes have been previously described. However, a quantitative response to this pathogen has also been described. The objective of this study was to identify halo-blight resistance loci linked to molecular markers that could be used in resistance breeding. Chromosomal regions related to race 5 halo-blight resistance were localized on a genetic map of RAPD and AFLP molecular markers and constructed by the analysis of a “Jules” × “Canela” F₂ progeny. “Jules” shows quantitative resistance to halo-blight and “Canela” is a very appreciated but susceptible Spanish bean landrace. Two QTL for resistance to halo-blight were mapped in two linkage groups. There were four large groups, with 14–22 molecular markers each, five with 4–8 markers each, and three with 2 or 3 markers each.     

Determination of the frequencies of restorer- and maintainer-alleles involved in CMS1 and CMS2 in German chive varieties

There are two cytoplasmic male sterility (CMS)‐systems in chives (Allium schoenoprasum L.), which can be employed in hybrid breeding. However, the probability for selection of maintainer genotypes from German open pollinated varieties is not known. Therefore, the allelic frequencies of the restorer genes X and T involved in CMS¹ were determined in 12 German commercial chive varieties by test crossing single plants to male sterile, temperature‐insensitive genotypes [(S₁)xxT] for segregation analyses of offspring. Temperature sensitive genotypes [(S₁)xxT_] are able to produce pollen at higher temperatures, and should therefore be excluded from hybrid breeding to avoid self‐pollination of the maternal parent. The mean value of the frequency of the non‐restoring allele x in the populations examined was 0.62. The mean value of the allele t, which is responsible for the temperature insensitivity, was 0.9. As a consequence of these allelic frequencies about one‐third of all plants of the chive varieties examined were designated CMS₁ maintainer genotypes, leading to the production of temperature insensitive male sterile lines. The incidence of CMS₂ maintainers in the German varieties examined was nearly four times lower than CMS₁ maintainers. The mean value of the frequency of the non‐restoring allele _st2 involved in the CMS₂‐system was 0.29.