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The tomato Mi-1 gene confers resistance to both root-knot nematodes and potato aphids.

Mi-1, a Lycopersicon peruvianum gene conferring resistance to the agricultural pests, root-knot nematodes, and introgressed into tomato, has been cloned using a selective restriction fragment amplification based strategy. Complementation analysis of a susceptible tomato line with a 100 kb cosmid array yielded a single cosmid clone capable of conferring resistance both to the root-knot nematode Meloidogyne incognita and to an unrelated pathogen, the potato aphid Macrosiphum euphorbiae. This resistance was stable. The Mi-1 gene encodes a protein sharing structural features with the nucleotide-binding site leucine-rich repeat-containing type of plant resistance genes.

Amino Acid Sequence↗

Susceptibility to streptozotocin-induced diabetes is mapped to mouse chromosome 11.

To study the contribution of beta-cell vulnerability to susceptibility to diabetes, we studied beta-cell vulnerability to a single high dose of streptozotocin (STZ) in an animal model of type 2 diabetes, the NSY mouse, a sister strain of the STZ-sensitive NOD mouse, in comparison with the STZ-resistant C3H mouse. NSY mice were found to be extremely sensitive to STZ. Introgression of a single Chr 11, where STZ-sensitivity was mapped in the NOD mouse, from NSY mice converted STZ-resistant C3H mice to STZ-sensitive. Two nucleotide substitutions were identified in the nucleoredoxin gene, a positional and functional candidate gene for STZ-induced diabetes on Chr 11. These data, together with the co-localization of type 1 (Idd4) and type 2 (Nidd1n) susceptibility genes on Chr 11, suggest that the intrinsic vulnerability of pancreatic beta cells is determined by a gene or genes on Chr 11, which may also contribute to susceptibility to spontaneous diabetes.

Animals↗

Rapid verification of wheat-Hordeum introgressions by direct staining of SCAR, STS, and SSR amplicons.

A range of single tagged site (STS), simple sequence repeat (SSR), and sequence-characterized amplified region (SCAR) markers were screened for their utility in detecting Hordeum vulgare and H. chilense chromosomes in a wheat background. PCR conditions were optimized for specific amplification of the targeted sequences and to avoid cross-species amplification. Two H. vulgare derived STSs, six H. vulgare derived SSRs, and nine H. chilense derived SCARs were usable for the detection of five H. vulgare and three H. chilense chromosomes by direct ethidium bromide staining of the PCR products in test tubes, avoiding the more costly and time-consuming DNA electrophoresis step. The practical application of the method is illustrated by the identification of a monotelosomic substitution of H. vulgare chromosome 6HS in tritordeum and a monosomic addition of H. chilense chromosome 6Hch in durum wheat.

Genetic Markers↗

Evidence for gene flow between wild and cultivated Medicago sativa (Leguminosae)based on allozyme markers andquantitative traits.

Genetic differentiation between co-occurring crops and their wild relatives will be greatly modified by crop-to-weed gene flow and variation between human and natural selective pressures. The maintenance of original morphological features in most natural populations of Medicago sativa in Spain questions the relative extent of these antagonistic forces. In this paper, we measured and compared the pattern of population differentiation within and among the wild and cultivated gene pool with respect to both allozymes and quantitative traits. Patterns of diversity defined three kinds of natural populations. First, some populations were intermediate with respect to both allozymes and quantitative traits. This suggests that crop-to-weed gene flow may have created hybrid populations in some locations. Second, some populations were different from all the cultivated landraces with respect to both allozymes and quantitative traits. This probably results from variable gene flow in space and in time, due to demographic stochasticity in either natural or cultivated populations. Third, differentiation from cultivated landraces was only achieved for the quantitative traits but not for allozymes in two populations. This suggests that natural selection in some locations may oppose gene flow to establish cultivated traits into the natural introgressed populations.

Journal Article↗

Identifying loci for behavioral traits using genome-tagged mice.

Identification of behavioral loci through complex trait mapping remains a widely employed approach but suffers from poor gene localization and low replicability. Genome-tagged mice (GTMs) are overlapping sets of congenic strains spanning the whole genome and offer the possibilities of superior mapping power and reproducibility. In this study, three GTM strains each consisting of an average approximately 27 cM DBA/2J genomic intervals introgressed onto a C57BL/6J background were employed for localization of behavioral traits. These GTMs were chosen because the corresponding chromosomal regions had been previously identified as containing loci for learning and memory. Analysis of the GTMs allowed confirmation of the learning and memory loci, and one on chromosome 3 was in addition fine mapped to an 8.8-cM region of overlap between two of the GTMs. Moreover, loci for prepulse inhibition of the startle response, acoustic startle response, and spontaneous locomotor activity were also mapped. These results suggest that the GTMs should be a valuable resource for mapping and confirmation of loci contributing to complex behavioral traits in the mouse.

Animals↗

Unexpected sensitivity of nonobese diabetic mice with a disrupted poly(ADP-Ribose) polymerase-1 gene to streptozotocin-induced and spontaneous diabetes.

Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme that consumes NAD in response to DNA strand breaks. Its excessive activation seems particularly deleterious to pancreatic beta-cells, as exemplified by the complete resistance of PARP-1-deficient mice to the toxic diabetes induced by streptozotocin. Because of the possible implication of this enzyme in type 1 diabetes, many human trials using nicotinamide, an inhibitor of PARP-1, have been conducted either in patients recently diagnosed or in subjects highly predisposed to this disease. To analyze the role of this enzyme in murine type 1 diabetes, we introgressed a disrupted PARP-1 allele onto the autoimmune diabetes-prone nonobese diabetic (NOD) mouse strain. We showed that these mice were protected neither from spontaneous nor from cyclophosphamide-accelerated diabetes. Surprisingly they were also highly sensitive to the diabetes induced by a single high dose of streptozotocin, standing in sharp contrast with C57BL/6 mice that bear the same inactivated PARP-1 allele. Our results suggest that NOD mice are characterized not only by their immune dysfunction but also by a peculiarity of their islets leading to a PARP-1-independent mechanism of streptozotocin-induced beta-cell death.

Animals↗

Dormancy and proliferation in Saccharum officinarumxS. spontaneum hybrids which differ in the number of the introgressed S. spontaneum chromosomes.

Proliferating cells remain transiently blocked at different cycle compartments until specific stressors are removed or until the cells become adapted to their presence. This paper investigates the efficiency of cycle blocks in three sugarcane hybrids with the full noble cane (Saccharum officinarum) genome (2n=8x=80) but differing in the number of introgressed S. spontaneum (2n=8x=64) chromosomes. The My5514, B42231 and C236-51 cultivars possess 20, 30 and 40 additional S. spontaneum chromosomes, respectively. Flow cytometry showed that over 90% of cells were accumulated with a 2C DNA content in their dormant primordia. The presence of S. spontaneum chromosomes decreased the low stringency of the 4C block. The greater the number of these chromosomes, the lower was the number of quiescent cells with a 4C DNA content (P<0.05). Shortly after stimulation of the primordia (85% relative humidity and 30 degrees C), i.e. in the 2 mm long roots, a negative correlation was found between the number of introgressed S. spontaneum chromosomes and the frequency of cells undergoing replication and mitosis. On the other hand, when roots were already proliferating under steady-state conditions (15 mm long roots) the more S. spontaneum chromosomes the cells possessed, the longer the relative time it took for all chromosomes to replicate and segregate, and the longer the relative time they spent in G(2), with the 4C DNA content. The presence of S. spontaneum chromosomes seems to be recognized by these proliferating cells as a stressor which preferentially activates checkpoint pathways operating at the second half of the cycle, but not at its onset.

Cell Cycle↗

Patterns of DNA sequence variation in chromosomally recognized taxa of Anopheles gambiae: evidence from rDNA and single-copy loci.

Patterns of DNA sequence variation in the ribosomal DNA (rDNA) second internal transcribed spacer (ITS2) and five unlinked single-copy nuclear loci were examined for evidence of reproductive isolation among four chromosomally recognized taxa of Anopheles gambiae from West Africa: Savanna, Bamako, Mopti and Forest, as well as sibling species An. arabiensis and An. merus. Included among the single-copy loci were three sequence-tagged random amplified polymorphic DNA (RAPD) loci, two of which (R15 and R37) had been reported as discriminating between Mopti and other chromosomal forms. Each of the five single-copy sequences were highly polymorphic in most samples. However, the R15 and R37 loci had no diagnostic value, and therefore are not recommended as tools in recognition of field-collected An. gambiae chromosomal forms. Although pairwise comparisons between species generally revealed significant levels of differentiation at all five loci, variation was not partitioned by chromosomal form within An. gambiae at any single-copy locus examined. The few exceptions to these trends appear related to a location either inside or nearby chromosomal inversions. At the tryptophan oxygenase locus inside inversion 2Rb, variation was structured only by inversion orientation and not by taxonomic designation even between An. gambiae and An. arabiensis, providing the first molecular evidence that the 2Rb inversion was transferred between species by introgressive hybridization. By contrast, the rDNA showed fixed differences between species and a difference diagnostic for Mopti, consistent with effective, if not complete, reproductive isolation. The apparent disagreement between the data from this locus and multiple single-copy loci within An. gambiae may be explained by the much lower effective population size of rDNA, owing to concerted evolution, which confers increased sensitivity at much shorter divergence times. Taken together with the accompanying reports by della Torre et al. (2001), Favia et al. (2001) and Gentile et al. (2001), our data suggest that neutral molecular markers may not have the sensitivity required to detect isolation between these recently established taxa.

Animals↗

Sequence variation in the guillemot (Alcidae: Cepphus) mitochondrial control region and its nuclear homolog.

We describe sequence variation in the mitochondrial control region and its nuclear homolog in three species and seven subspecies of guillemots (Cepphus spp.). Nuclear homologs of the 5' end of the control region were found in all individuals. Nuclear sequences were approximately 50% divergent from their mitochondrial counterparts and formed a distinct phylogenetic clade; the mitochondrial-nuclear introgression event must have predated the radiation of Cepphus. As in other vertebrates, the guillemot control region has a relatively conserved central block flanked by hypervariable 5' and 3' ends. Mean pairwise interspecific divergence values among control regions were lower than those in other birds. All individuals were heteroplasmic for the number of simple tandem nucleotide repeats (A(n)C) at the 3' end of the control region. Phylogenetic analyses suggest that black guillemots are basal to pigeon and spectacled guillemots, but evolutionary relationships among subspecies remain unresolved, possibly due to incomplete lineage sorting. Describing molecular variation in nuclear homologs of mitochondrial genes is of general interest in phylogenetics because, if undetected, the homologs may confound interpretations of mitochondrial phylogenies.

Animals↗

Sex, mutation and fitness: asymmetric costs and routes to recovery through compensatory evolution.

A wild-type population of Drosophila melanogaster was used to assess the impact of a known deleterious mutation, nub(1), when it had (1) evolved for up to 180 generations with the mutation or (2) recently had the same mutant allele introgressed into it. Relative to this benchmark, we observed much stronger initial fitness depression in males (-74%) than in females (-38%) and also relatively greater fitness recovery by evolved males (+55%) than females (+17%). Experimental assays revealed amelioration in both juvenile and adult fitness and suggested that the greater relative recovery of male fitness was from gains through sexual selection. These evolutionary changes in male fertility depended on pairing with their coevolved mates for both mate choice and post-copulatory components of sexual selection. Without replication at the population level, these results are used to motivate a general hypothesis rather than definitively test it: Differences in reproductive optima may generally skew mutational effects towards the more strongly sexually-selected sex due to genic capture and condition dependence.

Analysis of Variance↗

Analysis of the chlorophyll catabolism pathway in leaves of an introgression senescence mutant of Lolium temulentum.

Pigments, proteins and enzyme activity related to chlorophyll catabolism were analysed in senescing leaves of wild-type (WT) Lolium temulentum and compared with those of an introgression line carrying a mutant gene from stay-green (SG) Festuca pratensis. During senescence of WT leaves chlorophylls a and b were continuously catabolised to colourless products and no other derivatives were observed, whereas in SG leaves there was an accumulation of dephytylated and oxidised catabolites including chlorophyllide a, phaeophorbide a and 13(2) OH-chlorophyllide a. Dephytylated products were absent from SG leaf tissue senescing under a light-dark cycle. Retention of pigments in SG was accompanied by significant stabilisation of light harvesting chlorophyll-proteins compared with WT, but soluble proteins such as Rubisco were degraded during senescence at a similar rate in the two genotypes. The activity of phaeophorbide a oxygenase measured in SG tissue at 3d was less than 12% of that in WT tissue at the same time-point during senescence and of the same order as that in young pre-senescent WT leaves, indicating that the metabolic lesion in SG concerns a deficiency at the ring-opening step of the catabolic pathway. In senescent L. temulentum tissue two terminal chlorophyll catabolites were identified with chromatographic characteristics that suggest they may represent hitherto undescribed catabolite structures. These data are discussed in relation to current understanding of the genetic and metabolic control of chlorophyll catabolism in leaf senescence.

Cellular Senescence↗

Modification of wheat rDNA loci by rye B chromosomes: a chromatin organization model.

The rDNA loci, and their associated NORs, on chromosomes 1B and 6B of the hexaploid wheat cv. Lindström have been used as a chromatin marker to investigate the functional basis of the phenotype effects of introgressed supernumerary B chromosomes (Bs) of rye. The rye Bs themselves lack genes, other than those which determine their mitotic drive mechanism, and the way in which they can modulate characters determined by the A chromosome background has always been a puzzle. An isogenic line of Lindström plants carrying different numbers of Bs was used as the experimental system to see how different doses of Bs (from 0 to 6) affected the activity of the wheat NORs and the organization of their rDNA loci at interphase. Silver staining on metaphase chromosomes was used to evaluate the previous activity of the NORs, and to reveal variations in their size; and the pTa71 FISH probe from wheat was used to visualize structural modifications to the interphase rDNA loci. A single B had no measurable effect, but, as the B-number increased, there were significant changes in the physical dimensions of the metaphase NORs, reflecting reduced levels of their activity earlier in the cell cycle, and also in the condensation patterns of the interphase rDNA loci. In addition, the higher B-numbers caused a size heteromorphism between the homologous NORs. A model is discussed which interprets the phenotypic effects of Bs generically, in nucleotypic terms, based of their being 'genetically inert' but 'chromosomally active'.

Chromatin↗

Fruit carbohydrate metabolism in an introgression line of tomato with increased fruit soluble solids.

A tomato line (IL9-2-5) of the cultivated species, Lycopersicon esculentum, carrying a 9 cM introgression from the wild species, Lycopersicon pennelli, produces fruit with high soluble solids content (Brix), an important determinant of fruit quality for processing. Two quantitative trait loci (QTLs) relating to fruit soluble solids content have been identified within the introgressed segment. One of these QTLs (PW-9-2-5) is silent under the growth conditions used in this study, while a second (Brix-9-2-5) has been shown to encode a fruit apoplastic invertase (Lin5) with altered kinetic properties. In this study, we have undertaken a detailed biochemical analysis of the introgression line to attempt to gain an understanding of the metabolic changes associated with increased fruit soluble solids. Increased Brix in ripe fruit was shown to be the result of increased sucrose and glucose, with a more minor contribution from aspartate and alanine. The introgression leads to a pronounced increase in apoplastic invertase activity in the columella tissue that extends throughout fruit development. Furthermore, columella tissue from IL9-2-5 fruit has a greater capacity to take up exogenously supplied sucrose, an observation that is consistent with the kinetic properties of the introgressed Lin5 allele. Apart from the increase in mature fruit sugar and increases in some amino acids, metabolite profiling revealed few other metabolic perturbations in fruit from IL9-2-5. The only other major change was a dramatic increase in starch accumulation at earlier stages of fruit metabolism. This occurred without any increase in the activity of the enzymes of sucrose metabolism or starch synthesis and may therefore be driven by increased availability of sucrose. We conclude that the major factor that leads to increased fruit sugar in IL9-2-5 is an increase in the capacity to take up sucrose unloaded from the phloem.

Amino Acids↗

Production and characterization of a complete set of individual chromosome additions from Oryza officinalis to Oryza sativa using RFLP and GISH analyses.

Monosomic alien addition lines (MAALs) are valuable materials for comparative analyses of two distinct genomes, for elucidating introgression mechanisms, and for dissecting genes controlling complex traits. In the study reported here, MAALs of rice containing the complete genome of Oryza sativa and individual chromosomes of Oryza officinalis were produced. Interspecific hybridizations were made between O. sativa L. ssp. Japonica (CV, Hejiang 19, 2 n = 24, AA) and O. officinalis (Acc. HY018, 2 n = 24, CC). Two backcrosses were made to the cultivated rice to obtain BC2F1 plants. Through RFLP and GISH analyses, 25 MAALs (2 n = 25, AA + 1C) were identified and divided into 12 syntenic groups, designated MAALs 1-12. MAALs 1, 2, 3, 5, 7 and 10 were each represented by one plant, MAALs 8, 11 and 12 by two plants, MAALs 6 and 9 by four plants, and MAAL 4 by five plants. An ideogram of the C-genome of O. officinalis was constructed, based on GISH analysis of the interspecific hybrid and the MAALs. Comparative RFLP maps showed strong syntenic associations between the A-genomes and C-genomes. Chromosomal arrangements such as translocations and duplications were detected in different alien chromosomes of the MAALs. The complete set of O. officinalis MAALs generated here provides a novel manipulation platform for exploiting and utilizing the O. officinalis genome and carrying out genetic studies.

Blotting, Southern↗

A general method for identifying major hybrid male sterility genes in Drosophila.

The genes responsible for hybrid male sterility in species crosses are usually identified by introgressing chromosome segments, monitored by visible markers, between closely related species by continuous backcrosses. This commonly used method, however, suffers from two problems. First, it relies on the availability of markers to monitor the introgressed regions and so the portion of the genome examined is limited to the marked regions. Secondly, the introgressed regions are usually large and it is impossible to tell if the effects of the introgressed regions are the result of single (or few) major genes or many minor genes (polygenes). Here we introduce a simple and general method for identifying putative major hybrid male sterility genes which is free of these problems. In this method, the actual hybrid male sterility genes (rather than markers), or tightly linked gene complexes with large effects, are selectively introgressed from one species into the background of another species by repeated backcrosses. This is performed by selectively backcrossing heterozygous (for hybrid male sterility gene or genes) females producing fertile and sterile sons in roughly equal proportions to males of either parental species. As no marker gene is required for this procedure, this method can be used with any species pairs that produce unisexual sterility. With the application of this method, a small X chromosome region of Drosophila mauritiana which produces complete hybrid male sterility (aspermic testes) in the background of D. simulans was identified. Recombination analysis reveals that this region contains a second major hybrid male sterility gene linked to the forked locus located at either 62.7 +/- 0.66 map units or at the centromere region of the X chromosome of D. mauritiana.

Animals↗

The interspecific origin of B chromosomes: experimental evidence.

A centric fragment was generated during the introgression of a chromosome region from Nasonia giraulti into N. vitripennis. This neo B chromosome carries the N. giraulti or 123+ gene for wild-type eye color. Using this phenotypic effect, the transmission of this chromosome was analyzed. The supernumerary chromosome showed less than Mendelian segregation rate in meiosis and some mitotic instability manifested as mosaic phenotype for eye color. However, transmission rate and mitotic stability increased over successive generations. The transmission rate through male gametogenesis was nearly 100%. These results support the interspecific hybridization model for B chromosome origin and reveal that problems in chromosome stability can persist for several generations after "foreign chromosomes" are introduced into a different species. We suggest that hybrid zones should be investigated as possible sites for neo-B chromosome generation.

Animals↗

Periodic triggering of an inducible gene for control of a wild population.

A possible method of control for the management of wild populations consists of continual introgression of an inducible transgene by releasing transgenic individuals, with periodic exposure of the population to a trigger. Exposure to the trigger causes death or sterility in carriers of the transgene, but is otherwise benign. We investigate the effectiveness of various strategies for control. We show that suppression of the population density below any pre-specified level is possible using this technique. At the same time we show that too frequent or too efficient exposure to the trigger can select for non-transgenic genotypes at an intensity such that the population density will be largely unaffected by the trigger. Choices for management parameters can ensure that the latter scenario is avoided. We show that releasing individuals carrying the transgene at more than one locus facilitates density control.

Animals↗

The complex causality of geographical parthenogenesis.

Asexual organisms usually have larger and more northern distributions than their sexual relatives. This phenomenon, called geographical parthenogenesis, has been controversially attributed to predispositions in certain taxa; advantages of polyploidy and/or hybrid origin; better colonizing abilities because of uniparental reproduction; introgression of apomixis into sexuals; niche differentiation of clones; or biotic interactions. This review on apomictic plants demonstrates that each of these factors alone has not been able to explain the observed distributions. Establishment of the complex regulatory system of apomixis requires taxonomic and geographical predispositions; hybridization and/or polyploidization do create diversity, but they do not necessarily result in large distributions; colonizing abilities depend on clonal diversity and are outweighed by sexuals by self-compatibility and higher potentials for speciation; niche differentiation, ploidy levels and selfing keep sympatric sexuals and apomicts separated; and the impact of biotic interactions on distributions is uncertain. In conclusion, the distributional success of apomicts has a complex causality and depends on certain circumstances and combinations of factors. The rare establishment of apomixis may help to explain the predominance of sexuality on the large scale.

Genotype↗