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QTL mapping of grain quality traits from the interspecific cross Oryza sativa x O. glaberrima.

International rice export markets are increasing demands for rapid improvements in grain quality characteristics. The African rice Oryza glaberrima is a new potential source of genes that will enhance the eating, cooking, and milling properties of the rice grain. The objective of this research was to identify and characterize quantitative trait loci (QTLs) among 312 doubled haploid lines derived from the BC3F1 of an interspecific cross of O. sativa x O. glaberrima. Genetic material was planted in replicated plots and evaluated for ten grain quality traits in 2001 in Colombia. A linkage map was constructed with 100 polymorphic microsatellite markers using the mapdisto software program to adjust for segregation distortion. Transgressive segregation was observed for all traits. Interval and composite interval analyses identified 27 QTLs for nine characters located on 11/12 chromosomes. The chromosomal positions of QTLs for percentage amylose, alkali-spreading score, and percentage protein were in agreement with data reported by others, whereas QTL markers for percentage head rice, percentage milled rice, percentage protein, and percentage brown rice were different in our mapping population. Five major QTLs were found to be associated with improved percentage rice bran, percentage amylose, and alkali-spreading score. Seven QTLs for improved percentage rice bran, percentage milled rice, alkali-spreading score, percentage protein, and grain length/width ratio were derived from the O. glaberrima accession. Three new QTLs for percentage rice bran are reported here for the first time. Results from this study suggest that the African rice might be a valuable new source for introgression and improvement of several traits that affect quality traits demanded by the different rice export markets.

Chromosome Mapping↗

Biochemical markers in a species endangered by introgression: the red wolf.

The red wolf (Canis rufus), native to much of the southeastern United States, is endangered by man's activities and by hybridization with other species of the genus Canis. The absence of diagnostic morphological markers to distinguish the red wolf from its hybrids has led to the application of the methods of biochemical genetics to this problem. The finding of a unique electrophoretically determined allele with a distribution congruent with the geographical distribution of the remaining red wolf population is reported.

Acid Phosphatase↗

Defining the blood pressure QTL on chromosome 7 in Dahl rats by a 177-kb congenic segment containing Cyp11b1.

Previously we reported that there is a blood pressure quantitative trait locus (QTL) on rat Chromosome (Chr) 7 seen when comparing Dahl salt-sensitive (S) rats and Dahl salt-resistant (R) rats. Evidence was also presented that this QTL was due to genetic variants in the adrenal steroidogenic enzyme 11beta-hydroxylase ( Cyp11b1). A series of congenic strains supported this contention. In the present work we have constructed a final congenic substrain that retains a blood pressure effect and that has an introgressed congenic segment which includes Cyp11b1 and is < 177 kb in size. None of the other genes in the congenic region (eight known genes) have known biological functions for influencing blood pressure. We believe that we have reached the limit of resolution for congenic analysis of a QTL in a rodent animal model, and we conclude that Cyp11b1 causes the observed QTL on rat Chr 7 in Dahl rats.

Animals↗

A loss of genome buffering capacity of Dahl salt-sensitive model to modulate blood pressure as a cause of hypertension.

Essential hypertension is a complex trait influenced by multiple genes known as quantitative trait loci (QTLs) for blood pressure (BP). It is not clear, however, what roles these QTLs play in maintaining normotension. Insights gained toward the maintenance of normotension will shed light on how hypertension can result from a deficiency or malfunctioning of this maintenance. Currently, congenic strains were systematically constructed using Dahl salt-sensitive (DSS) and Lewis (LEW) rats not only to define QTLs (i.e. in DSS background), but also to ascertain effects of the same QTLs in preserving normotension (i.e. in LEW background), a first such study. Results showed that although LEW alleles for two QTLs on Chromosome (Chr) 18 lowered BP on the DSS background, their BP-increasing DSS alleles failed to influence BP in the LEW background. To further prove that the LEW background is resistant and the DSS background is susceptible to the effects of QTLs, BP-increasing alleles of a QTL on Chr 2 were introgressed into the DSS background, and its BP-decreasing alleles into the LEW background. Indeed, there was no BP-decreasing effect on the LEW background while demonstrating a BP-increasing effect on the DSS background. Thus, a genetic regulation of BP QTLs in the LEW genome inhibits BP changes by nullifying the effects of BP-altering QTLs. In comparison, the DSS genome must have lost the buffering capacity for stabilizing BP. The current work presents good evidence that a lack of regulation for functions of BP QTLs is a potential underlying cause of hypertension.

Animals↗

Wheat cytogenetics in the genomics era and its relevance to breeding.

Hexaploid wheat is a species that has been subjected to most extensive cytogenetic studies. This has contributed to understanding the mechanism of the evolution of polyploids involving diploidization through genetic restriction of chromosome pairing to only homologous chromosomes. The availability of a variety of aneuploids and the ph mutants (Ph1 and Ph2) in bread wheat also allowed chromosome manipulations leading to the development of alien addition/substitution lines and the introgression of alien chromosome segments into the wheat genome. More recently in the genomics era, molecular tools have been used extensively not only for the construction of molecular maps, but also for identification/isolation of genes/QTLs (including epistatic QTLs, eQTLs and PQLs) for several agronomic traits. It has also been possible to identify gene-rich regions and recombination hot spots in the wheat genome, which are now being subjected to sequencing at the genome level, through development of BAC libraries. In the EST database also, among all plants wheat ESTs are the highest in number, and are only next to those for human, mouse, Ciona intestinalis (a chordate), rat and zebrafish genomes. These ESTs and sequences of several genomic regions have been subjected to a variety of applications including development of perfect markers and establishment of microcollinearity. The technique of in situ hybridization (including FISH, GISH and McFISH) and the development of deletion stocks also facilitated the preparation of physical maps. Molecular markers are also used for marker-assisted selection in wheat breeding programs in several countries. Construction of a wheat DNA chip, which will also become available soon, may further facilitate wheat genomics research. These enormous resources, knowledge base and the fast development of additional molecular tools and high throughput approaches for genotyping will prove extremely useful in future wheat research and will lead to development of improved wheat cultivars.

Agriculture↗

[Molecular mapping of the S-a locus for F1 pollen sterility in cultivated rice (Oryza sativa L.)].

F1 pollen sterility in cultivated rice (Oryza sativa L.) was found to be caused by at least six loci of F1 pollen sterility genes. At the S-a locus, one of the six loci for F1 pollen sterility, the allelic interaction of S-ai and S-aj causes the male gametes carrying S-aj allele abortive. To map the S-a locus, Taichung 65(T65), a Keng (japonica) variety with S-aj/S-aj, its isogenic F1 sterile line TISL4 with S-ai/S-ai from Chin-tsao, a Hsien (indica) variety, and the F2 population from cross T65 x TISL4 were used as materials. The polymorphism between T65 and TISL4 detected by RFLP and RAPD analysis was less than 1%. This result indicated that short segments from Chin-tsao were introgressed into the isogenic F1 sterile line, since the TISL4 was developed by repeatedly backcrossing for thirteen times. By linkage analysis of S-a and the marker loci, the S-a locus was mapped on chromosome 1. The genetic distances between S-a and RFLP markers CDO548 and RG146 are 6.4 cM and 7.2 cM respectively, and those between S-a and RAPD markers O11-1000 and Y13-500 are 6.8 cM and 11.2 cM respectively. The mapping of the S-a locus is an important step towards marker-aided selection for overcoming the hybrid sterility in rice.

Chromosome Mapping↗

Genetic identification of wild and domestic cats (Felis silvestris) and their hybrids using Bayesian clustering methods.

Crossbreeding with free-ranging domestic cats is supposed to threaten the genetic integrity of wildcat populations in Europe, although the diagnostic markers to identify "pure" or "admixed" wildcats have never been clearly defined. Here we use mitochondrial (mt) DNA sequences and allelic variation at 12 microsatellite loci to genotype 128 wild and domestic cats sampled in Italy which were preclassified into three separate groups: European wildcats (Felis silvestris silvestris), Sardinian wildcats (Felis silvestris libyca), and domestic cats (Felis silvestris catus), according to their coat color patterns, collection localities, and other phenotypical traits, independently of any genetic information. For comparison, we included some captive-reared hybrids of European wild and domestic cats. Genetic variability was significantly partitioned among the three groups (mtDNA estimate of F(ST) = 0.36; microsatellite estimate of R(ST) = 0.30; P < 0.001), suggesting that morphological diversity reflects the existence of distinct gene pools. Multivariate ordination of individual genotypes and clustering of interindividual genetic distances also showed evidence of distinct cat groups, partially congruent with the morphological classification. Cluster analysis, however, did not enable hybrid cats to be identified from genetic information alone, nor were all individuals assigned to their populations. In contrast, a Bayesian admixture analysis simultaneously assigned the European wildcats, the Sardinian wildcats, and the domestic cats to different clusters, independent of any prior information, and pointed out the admixed gene composition of the hybrids, which were assigned to more than one cluster. Only one putative Sardinian wildcat was assigned to the domestic cat cluster, and one presumed European wildcat showed mixed (hybrid) ancestry in the domestic cat gene pool. Mitochondrial DNA sequences indicated that three additional presumed European wildcats might have hybrid ancestry. These four cats were sampled from the same area in the northernmost edge of the European wildcat distribution in the Italian Apennines. Admixture analyses suggest that wild and domestic cats in Italy are distinct, reproductively isolated gene pools and that introgression of domestic alleles into the wild-living population is very limited and geographically localized.

Alleles↗

About the origin of European spelt ( Triticum spelta L.): allelic differentiation of the HMW Glutenin B1-1 and A1-2 subunit genes.

To investigate the origin of European spelt ( Triticum spelta L., genome AABBDD) and its relation to bread wheat ( Triticum aestivum L., AABBDD), we analysed an approximately 1-kb sequence, including a part of the promoter and the coding region, of the high-molecular-weight (HMW) glutenin B1-1 and A1-2 subunit genes in 58 accessions of hexa- and tetraploid wheat from different geographical regions. Six Glu-B1-1 and five Glu-A1-2 alleles were identified based on 21 and 19 informative sites, respectively, which suggests a polyphyletic origin of the A- and B-genomes of hexaploid wheat. In both genes, a group of alleles clustered in a distinct, so-called beta subclade. High frequencies of alleles from the Glu-B1-1 and Glu-A1-2 beta subclades differentiated European spelt from Asian spelt and bread wheat. This indicates different origins of European and Asian spelt, and that European spelt does not derive from the hulled progenitors of bread wheat. The conjoint differentiation of alleles of the A- and B-genome in European spelt suggests the introgression of a tetraploid wheat into free-threshing hexaploid wheat as the origin of European spelt.

Alleles↗

The current status and environmental impacts of glyphosate-resistant crops: a review.

Glyphosate [N-(phosphonomethyl) glycine]-resistant crops (GRCs), canola (Brassica napus L.), cotton (Gossypium hirsutum L.), maize (Zea mays L.), and soybean [Glycine max (L.) Merr.] have been commercialized and grown extensively in the Western Hemisphere and, to a lesser extent, elsewhere. Glyphosate-resistant cotton and soybean have become dominant in those countries where their planting is permitted. Effects of glyphosate on contamination of soil, water, and air are minimal, compared to some of the herbicides that they replace. No risks have been found with food or feed safety or nutritional value in products from currently available GRCs. Glyphosate-resistant crops have promoted the adoption of reduced- or no-tillage agriculture in the USA and Argentina, providing a substantial environmental benefit. Weed species in GRC fields have shifted to those that can more successfully withstand glyphosate and to those that avoid the time of its application. Three weed species have evolved resistance to glyphosate in GRCs. Glyphosate-resistant crops have greater potential to become problems as volunteer crops than do conventional crops. Glyphosate resistance transgenes have been found in fields of canola that are supposed to be non-transgenic. Under some circumstances, the largest risk of GRCs may be transgene flow (introgression) from GRCs to related species that might become problems in natural ecosystems. Glyphosate resistance transgenes themselves are highly unlikely to be a risk in wild plant populations, but when linked to transgenes that may impart fitness benefits outside of agriculture (e.g., insect resistance), natural ecosystems could be affected. The development and use of failsafe introgression barriers in crops with such linked genes is needed.

Argentina↗

[Genetic analysis of the hybridization zone between two subspecies Mus musculus domesticus and Mus musculus musculus in Bulgaria].

The hybrid zone between the two subspecies of mice Mus musculus domesticus and Mus musculus musculus, which has been studied extensively in Denmark, crosses Europe to the Black Sea through the Alps and the Balkans. Two hundred and seventy-nine animals were captured in 22 localities along a transect across the Balkans. The animals were characterized for seven diagnostic nuclear loci by protein electrophoresis and by restriction pattern analysis of their mitochondrial DNA. The nuclear data show a sharp transition between the two subspecies, most of the variations in allele frequencies (from 0.9 to 0.1) occurring within a 36-km section of the transect. The introgression varies from one locus to the other and is more pronounced, in terms of distance, in M. m. musculus territory. Mitochondrial DNA introgression is important but occurs in one direction only, i.e. from M. m. musculus to M. m. domesticus, while a cytoplasmic transfer from M. m. domesticus to M. m. musculus has been reported. A previous study showed that no Y chromosome introgression occurs. The different behaviour of these three types of markers could be due to the interaction between selection against hybrid genomes and meiotic recombination. Objectively, it would appear that the genes that can introgress are neutral or nearly so and have been separated from deleterious genes they were linked to by recombination. This could explain the differential introgression between autosomal loci. The mitochondrial and Y chromosomes undergo no or very little recombination and each is transmitted as a whole. Their degree of introgression is thus indicative of the intensity of selection resulting from the amount of functional differentiation between the two taxa, which seems to be strong for the Y chromosome and weak for mitochondrial DNA. We propose that the asymmetry of nuclear introgression is due to different population structures. As M. m. musculus is relatively less structured, the rapid spreading of introgressed genes would be favoured. Such a scheme, however, can hardly account for the unidirectionality of the mitochondrial flow, which could be due to sex-dependent behaviour.

Alleles↗

Genetic interactions underlying hybrid male sterility in the Drosophila bipectinata species complex.

Understanding genetic mechanisms underlying hybrid male sterility is one of the most challenging problems in evolutionary biology especially speciation. By using the interspecific hybridization method roles of Y chromosome, Major Hybrid Sterility (MHS) genes and cytoplasm in sterility of hybrid males have been investigated in a promising group, the Drosophila bipectinata species complex that consists of four closely related species: D. pseudoananassae, D. bipectinata, D. parabipectinata and D. malerkotliana. The interspecific introgression analyses show that neither cytoplasm nor MHS genes are involved but X-Y interactions may be playing major role in hybrid male sterility between D. pseudoananassae and the other three species. The results of interspecific introgression analyses also show considerable decrease in the number of males in the backcross offspring and all males have atrophied testes. There is a significant positive correlation between sex - ratio distortion and severity of sterility in backcross males. These findings provide evidence that D. pseudoananassae is remotely related with other three species of the D. bipectinata species complex.

Animals↗

Characterization of a hybrid zone between two chromosomal races of the weta Hemideina thoracica following a geologically recent volcanic eruption.

Two chromosomal races (2n=17 and 2n=15; XO) of the weta Hemideina thoracica meet at the centre of a volcanic region in North Island, New Zealand. Five independent polymorphic genetic markers showed broadly coinciding, steep frequency clines from north to south across this zone beside the flooded crater, Lake Taupo. Three unlinked nuclear gene markers provide estimates of zone width that are at least twice the width of the chromosomal and mitochondrial clines, with cline centres displaced at least 2.5 km. The different zone widths and centres suggest that this hybrid zone is a semipermeable barrier reducing the introgression of the chromosomal markers more than genic markers. We estimate that this species of weta must have a dispersal rate of at least 100 m per generation using the time since the last Taupo eruption (1850 years ago), which covered an area of about 20 000 km2 with pyroclastic flow.

Animals↗

An updated 'Essex' by 'Forrest' linkage map and first composite interval map of QTL underlying six soybean traits.

DNA marker maps based on single populations are the basis for gene, loci and genomic analyses. Individual maps can be integrated to produce composite maps with higher marker densities if shared marker orders are consistent. However, estimates of marker order in composite maps must include sets of markers that were not polymorphic in multiple populations. Often some of the pooled markers were not codominant, or were not correctly scored. The soybean composite map was composed of data from five separate populations based on northern US germplasm but does not yet include 'Essex' by 'Forrest' recombinant inbred line (RIL) population (E x F) or any southern US soybean cultivars. The objectives were, to update the E x F map with codominant markers, to compare marker orders among this map, the Forrest physical map and the composite soybean map and to compare QTL identified by composite interval maps to the earlier interval maps. Two hundred and thirty seven markers were used to construct the core of the E x F map. The majority of marker orders were consistent between the maps. However, 19 putative marker inversions were detected on 12 of 20 linkage groups (LG). Eleven marker distance compressions were also found. The number of inverted markers ranged from 1 to 2 per LG. Thus, marker order inversions may be common in southern compared to northern US germplasm. A total of 61 QTL among 37 measures of six traits were detected by composite interval maps, interval maps and single point analysis. Seventeen of the QTL found in composite intervals had previously been detected among the 29 QTL found in simple interval maps. The genomic locations of the known QTL were more closely delimited. A genome sequencing project to compare Southern and Northern US soybean cultivars would catalog and delimit inverted regions and the associated QTL. Gene introgression in cultivar development programs would be accelerated.

Chromosome Mapping↗

Phylogeographical variation of chloroplast DNA in cork oak (Quercus suber).

BACKGROUND AND AIMS: In the last decades, the geographical location of the centre of origin of Quercus suber (cork oak), a strictly western Mediterranean oak species, has been the subject of controversy. METHODS: RFLP variation over the whole chloroplast DNA molecule and PCR-RFLPs over seven specific cpDNA fragments were analysed phylogeographically to reconstruct the evolutionary history of cork oak. KEY RESULTS: Nine chlorotypes of the 'suber' cpDNA lineage were identified throughout the species range. Using closely related Mediterranean oak species as outgroup, the chlorotypes showed a clear phylogeographical pattern of three groups corresponding to potential glacial refuges in Italy, North Africa and Iberia. The most ancestral and recent groups were observed in populations located in the eastern and western parts of the species range, respectively. Several unrelated chlorotypes of the 'ilex' cpDNA lineage were also identified in specific western areas. CONCLUSIONS: The results support a Middle-Eastern or a central Mediterranean origin for cork oak with subsequent westward colonization during the Tertiary Period, and suggest that the 'ilex' chlorotype variation does not reflect entirely cytoplasmic introgression by Q. ilex but originated partly in Q. suber.

DNA, Chloroplast↗

Genetic relationships and population structure of the endangered Steamboat buckwheat, Eriogonum ovalifolium var. williamsiae (Polygonaceae).

Eriogonum ovalifolium var. williamsiae (Steamboat buckwheat) is a narrow endemic subshrub, known from a single locality in Washoe County, Nevada. We examined genetic structure of the only known population by analyzing patterns of allozyme variation. Our results suggest that Steamboat buckwheat has high genetic variability, with levels of variation similar to that typical of a widespread species rather than a narrow endemic. Genotype frequencies suggest that mating is random. We detected no genetic subdivision of the population. Several clones spanning up to 67 cm were found, but we do not know if such clones are common. We used allozyme data to assess the genetic similarity of var. williamsiae to five other varieties of E. ovalifolium. All six varieties are very similar allozymically with var. williamsiae being the most similar to the widespread var. ovalifolium. Although var. williamsiae and var. ovalifolium are morphologically distinct, their genetic similarity warrants further study to determine whether or not they should be treated as separate taxa. Evidence of male sterility in var. williamsiae plus other data leads us to hypothesize that this taxon might be either a hybrid or undergoing cytoplasmic introgression. Information gathered from this study, in concert with ongoing work on the breeding system of Steamboat buckwheat, should be helpful in forming management strategies for this plant.

Journal Article↗

Cytology and fertility of viable hybrids of Nicotiana tabacum L. cv. TB-566 with N. alata Link et Otto.

A vigorous hybrid (N. tabacum cv. TB-566 tetra x N. alata) x N. alata was obtained by backcrossing a partly viable sesquidiploid hybrid N. tabacum cv. TB-566 tetra x N. alata to N. alata. The hybrid was a 35-chromosome near-amphihaploid with a pair of N. alata chromosomes in disomic condition. It was completely self- and cross-sterile and formed from 7 to 8 bivalents in pollen mother cells. By using stem pith culture, polyploidized regenerants were obtained from the 35-chromosome hybrid with somatic chromosome numbers from 65 to 70. These regenerants showed fairly regular meiosis with the number of bivalents in pollen mother cells ranging from 27.3 to 30.4. Irregularities in meiosis included a high number of univalents, aberrant tetrads, and a high frequency of micronuclei. The percentage of acetocarmine-stainable pollen ranged from 22.1 to 78.4. A 66-chromosome regenerant showed fairly regular meiosis and was self-fertile but could not be backcrossed to N. tabacum. This barrier seems to be caused by genic imbalance rather than irregularities of meiotic divisions. Hence transfer programs based on the introgression of entire linkage groups (sexual and somatic hybridization) seem to be of little use in the case of that species.

Chromosomes, Plant↗

Allochronic differentiation among Daphnia species, hybrids and backcrosses: the importance of sexual reproduction for population dynamics and genetic architecture.

Seasonal dynamics of the abundance, sexual reproduction and genetic architecture in a Daphnia hyalina-galeata hybrid complex were studied in the large and deep Lake Constance. We found evidence for the occurrence of first and second order hybridization. Our study revealed strong differences between the parental species not only regarding their seasonal dynamics, genetic architecture and diversity, but also their sexual reproductive behaviour. The overwintering D. hyalina showed low genetic diversity, no genetic differentiation during the season, and reproduced sexually in autumn, whereas D. galeata reached higher levels of genetic diversity, reproduced sexually in early summer, and exhibited changes in genetic structure during the season, but was only present from spring to autumn. However, in both species sexual reproduction was a rare event, and daphnids, including hybrids, reproduced predominantly asexually. This allows long-term persistence of hybrids as well without continuing hybridization events. Within all variables studied, F1 and F2 hybrids showed an intermediate pattern, whereas proposed backcross hybrids were more similar to their respective parentals. These differences in phenotype as well as significant differences in pairwise Fst values between parentals suggest that gene flow seems to be relatively low in the Lake Constance hybrid system. We found evidence for unidirectional introgression by backcrossing from D. galeata to D. hyalina and found a decrease in at least one of the proposed introgressed alleles in the hyalina-backcross while the season progressed. Our findings suggest allochronic differentiation within this hybrid population and different microevolutionary trajectories of the parental species, which will be discussed in the light of the ongoing reoligotrophication process of Lake Constance.

Animals↗