bluuebunny/crossref_metadata_2025 huggingface dataset:
Towards developing intervarietal substitution lines inBrassica napususing marker-assisted selection
Authors: Howell P. M. , Lydiate D. J. , Marshall D. F.
Published: May 2008

Sets of substitution lines have advantages over segregating populations for the rigorous analysis of loci influencing quantitative traits. A general strategy for the rapid production of substitution lines was developed. It involved the systematic application of marker-assisted selection over 2–4 generations of backcrossing. The effectiveness of this strategy was demonstrated by the production of intervarietal substitution lines in Brassica napus. A genetic map containing 158 loci, distributed across all 19 B. napus linkage groups and assayed in 200 B1individuals, was generated. Six complementary B1individuals enriched for recurrent genotype and collectively carrying almost all the donor genome were selected. A total of 288 B2plants derived from the selected B1individuals were analysed and complementary individuals carrying five or fewer donor segments were identified. Similar selection, carried out on 250 B3plants from two distinct B1lineages, identified 74 B3individuals carrying one or two donor segments. Together, 12 of these isolated segments represented 33% of the mapped genome. Lines homozygous for single substituted segments were derived from selfed progeny of selected B3plants. A full set of substitution lines will be used to elucidate the genetic control of quantitative production traits in oilseed rape over several environments. Key words : QTL mapping, quantitative genetics, backcross, genetic linkage map, plant breeding, restriction fragment length polymorphism.
Molecular mapping of quantitative trait loci injaponicarice
Authors: Redoña Edilberto D. , Mackill David J.
Published: May 2008

Rice (Oryza sativa L.) molecular maps have previously been constructed using interspecific crosses or crosses between the two major subspecies: indica and japonica. For japonica breeding programs, however, it would be more suitable to use intrasubspecific crosses. A linkage map of 129 random amplified polymorphic DNA (RAPD) and 18 restriction fragment length polymorphism (RFLP) markers was developed using 118 F2plants derived from a cross between two japonica cultivars with high and low seedling vigor, Italica Livorno (IL) and Labelle (LBL), respectively. The map spanned 980.5 cM (Kosambi function) with markers on all 12 rice chromosomes and an average distance of 7.6 cM between markers. Codominant (RFLP) and coupling phase linkages (among RAPDs) accounted for 79% of total map length and 71% of all intervals. This map contained a greater percentage of markers on chromosome 10, the least marked of the 12 rice chromosomes, than other rice molecular maps, but had relatively fewer markers on chromosomes 1 and 2. We used this map to detect quantitative trait loci (QTL) for four seedling vigor related traits scored on 113 F3families in a growth chamber slantboard test at 18 °C. Two coleoptile, five root, and five mesocotyl length QTLs, each accounting for 9–50% of the phenotypic variation, were identified by interval analysis. Single-point analysis confirmed interval mapping results and detected additional markers significantly influencing each trait. About two-thirds of alleles positive for the putative QTLs were from the high-vigor parent, IL. One RAPD marker (OPAD13720) was associated with a IL allele that accounted for 18.5% of the phenotypic variation for shoot length, the most important determinant of seedling vigor in water-seeded rice. Results indicate that RAPDs are useful for map development and QTL mapping in rice populations with narrow genetic base, such as those derived from crosses among japonica cultivars. Other potential uses of the map are discussed. Key words : QTL mapping, RAPD, RFLP, seedling vigor, japonica, Oryza sativa.
Molecular and biochemical characterization of two nucleoside diphosphate kinase cDNA clones fromFlaveria bidentis
Authors: Ananvoranich Sirinart , Grandmaison Jacques , Gulick Patrick J.
Published: May 2008

Two nucleoside diphosphate kinase cDNA clones have been isolated from Flaveria bidentis by immunoscreening of an expression library with a polyclonal antibody raised against Flaveria chloraefolia flavonol 3-sulfotransferase (F3-ST). The clones represent members of a small multigene family in this species. The nucleotide sequences of the two cDNA clones show a high degree of sequence similarity to other reported nucleoside diphosphate kinases (NDPKs), including the putative human tumor suppressor gene NM23 and the Drosophila regulatory gene. When these cDNA clones were expressed in Escherichia coli, their gene products exhibited NDPK enzymatic activity. The immunocross reaction of the clones with the antibody raised against the F3-ST suggests a common immuno-epitope and a similarity of a nucleotide binding site for the two proteins. Key words : nucleoside diphosphate kinase, cDNA, multigene family.
Genetic analyses of a putativeMutator-induced deletion involving theblue fluorescent1(bf1) andbrown midrib4(bm4) loci in maize
Authors: Robertson Donald S.
Published: May 2008

A putative Mutator-induced alteration (isolate) involving the blue fluorescent1 (Bf1) and the brown midrib4 (bm4) loci is described. This isolate (Bf1-Mu-044-4) was one of the blue fluorescent1 mutants in a large experiment designed to produce Mutator-induced mutations involving this locus. Evidence is presented supporting the conclusion that this alteration deletes all or part of both the bf1 and bm4 loci. The isolate is infrequently transmitted through the pollen, but is more readily transmitted through the egg, although at less than the expected frequency of 50% for a simple mutation. Heterozygotes of this isolate with the bm4-R allele have brown midribs. These brown midrib plants are shorter than homozygous bm4-R plants and approximately 50% of these Bf1-Mu-044-4/bm4-R heterozygotes do not reach maturity. One putative Mutator-induced bm4 mutation, which occurred in the course of the analysis of the Bf1-Mu-044 isolate, is described. Key words : Mutator, deletion, bf1-bm4 loci.
Analysis off cereal chromosomes by atomic force microscopy
Authors: McMaster Terence J. , Miles Mervyn J. , Winfield Mark O. , Karp Angela
Published: May 2008

Atomic force microscopy has been applied to the study of plant chromosomes from cereal grasses Triticum aestivum (bread wheat), Triticum tauschii, and Hordeum vulgare (barley). Using standard mitotic metaphase squashes, high resolution images have been obtained of untreated chromosomes and also of chromosomes after C-banding, N-banding, and in situ hybridization. The true 3-dimensional nature of the images permits detailed analysis of the surface structure and, on untreated uncoated chromosomes, surface features on a length scale consistent with nucleosome structures have been observed. C+and N+regions are manifest as areas of high relief on a slightly collapsed chromosome structure. In situ hybridization leads to a more severe degradation of the native structure, although it is still possible to correlate the optical signal with the topography of the hybridized chromosome. Key words : atomic force microscope, AFM, chromosomes, C-banding, in situ hybridization.
5S rRNA genes in tribe Phaseoleae: array size, number, and dynamics
Authors: Danna Kathleen J. , Workman Rachel , Coryell Virginia , Keim Paul
Published: May 2008

The organization of 5S rRNA genes in plants belonging to tribe Phaseoleae was investigated by clamped homogeneous electric field gel electrophoresis and Southern blot hybridization. Representatives of subtribe Glycininae included the diploid species Neonotonia wightii and Teramnus labialis, as well as three soybean accessions: an elite Glycine max (L.) Merr. cultivar (BSR101), an unadapted G. max introduction (PI 437.654), and a wild Glycine soja (PI 468.916). A cultivar of Phaseolus vulgaris (kidney bean), a member of subtribe Phaseolinae, was also examined. We determined the number of 5S rDNA arrays and estimated the size and copy number of the repeat unit for each array. The three soybean accessions all have a single 5S locus, with a repeat unit size of ~345 bp and a copy number ranging from about 600 in 'BSR101' to about 4600 in the unadapted soybean introduction. The size of the 5S gene cluster in 'BSR101' is the same in roots, shoots, and trifoliate leaves. Given that the genus Glycine probably has an allotetraploid origin, our data strongly suggest that one of the two progenitor 5S loci has been lost during diploidization of soybean. Neonotonia wightii, the diploid species most closely related to soybean, also has a single locus but has a repeat unit of 520 bp and a copy number of about 1300. The more distantly related species T. labialis and P. vulgaris exhibited a more complex arrangement of 5S rRNA genes, having at least three arrays, each comprising a few hundred copies of a distinct repeat unit. Although each array in P. vulgaris exhibits a high degree of homogeneity with regard to the sequence of the repeat unit, heterogeneity in array size (copy number) was evident when individual plants were compared. A cis-dependent molecular drive process, such as unequal crossing-over, could account for both the homogenization of repeat units within individual arrays and the observed variation in copy number among individuals. Key words : pulsed-field gel electrophoresis, rRNA genes, soybean, tandem arrays.
A novel set of uncoordinated mutants inCaenorhabditis elegansuncovered by cold-sensitive mutations
Authors: Hecht Ralph M. , Norman Mary A. , Vu Tammy , Jones William
Published: May 2008

A set of uncoordinated (Unc) cold-sensitive (cs) mutants was isolated at a stringent condition of 11 °C. About half of the 13 independently isolated cs-Unc mutants were alleles of three X-linked Unc mutants that exhibited the "kinker" phenotype. The remaining four isolates identified new mutants that exhibited "kinker," "coiler," or severe paralytic phenotypes. The temperature-sensitive period (TSP) for each gene was determined. As a homozygous or heterozygous dominant, unc-125 exhibited a TSP throughout all stages of development. Its severe paralysis was immediately observed upon a shift down to 11 °C and reversed upon a shift up to 23 °C. The reversible thermolability of the unc-125 gene product indicated that it may function in a multicomponent process involved in neuro-excitation. Key words : Caenorhabditis elegans, cold-sensitive uncoordinated mutants, cs-Unc.
Recombination and chiasmata: few but intriguing discrepancies
Authors: Sybenga J.
Published: May 2008

The paradigm that meiotic recombination and chiasmata have the same basis has been challenged, primarily for plants. High resolution genetic mapping frequently results in maps with lengths far exceeding those based on chiasma counts. In addition, recombination between specific homoeologous chromosomes derived from interspecific hybrids is sometimes much higher than can be explained by meiotic chiasma frequencies. However, almost the entire discrepancy disappears when proper care is taken of map inflation resulting from the shortcomings of the mapping algorithm and classification errors, the use of dissimilar material, and the difficulty of accurately counting chiasmata. Still, some exchanges, especially of short interstitial segments, cannot readily be explained by normal meiotic behaviour. Aberrant meiotic processes involving segment replacement or insertion can probably be excluded. Some cases of unusual recombination are somatic, possibly premeiotic exchange. For other cases, local relaxation of chiasma interference caused by small interruptions of homology disturbing synaptonemal complex formation is proposed as the cause. It would be accompanied by a preference for compensating exchanges (negative chromatid interference) resulting from asymmetry of the pairing chromatid pairs, so that one side of each pair preferentially participates in pairing. Over longer distances, the pairing face may switch, causing the normal random chromatid participation in double exchanges and the relatively low frequency of short interstitial exchanges. Key words : recombination frequency, map length, chiasmata, discrepancy, chromatid interference.
Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences
Authors: Sreenivasaprasad S. , Meehan B. M. , Mills P. R. , Brown A. E.
Published: May 2008

The potential use of the ribosomal DNA internal transcribed spacer (ITS) sequences in understanding the phylogeny and systematics of Colletotrichum species has been evaluated. Sequence data from a limited number of isolates revealed that in Colletotrichum species the ITS 1 region (50.3% variable sites) shows a greater degree of intra- and inter-specific divergence than ITS 2 (12.4% variable sites). Nucleotide sequences of the ITS 1 region from 93 isolates representing 18 Colletotrichum species were determined. Data for 71 of these isolates where molecular and morphological identities concurred were used for phylogenetic analysis. The size of the ITS 1 region varied from 159 to 185 base pairs. Maximum intraspecific divergence was recorded with C. acutatum (5.8%), and C. capsici showed the greatest level of interspecific divergence (8.9–23.3%). Parsimony and distance analyses gave similar tree topologies. The bootstrapped consensus parsimony tree divided the 18 Colletotrichum species into six phylogenetic groups, designated 1–6. These groups, however, are not congruent with species clusterings based on spore shape. For example, the straight cylindrical spored species were represented both in groups 1 and 6; group 6 also included the falcate fusiform spored species C. capsici. The molecular evidence suggests refinement of the species concepts of some of the taxa examined. In group 6, divergence between C. gloeosporioides and C. fuscum (0.6–3.0%) or C. kahawae (0.6–3.0%) or C. fragariae (0.6–4.2%) overlap the divergence (3.6%) within C. gloeosporioides. It is suggested that C. fuscum as well as C. kahawae and C. fragariae fall within the group species C. gloeosporioides. ITS 1 data enabled clear distinction (7.1%) of Colletotrichum isolates from maize and sorghum into C. graminicola and C. sublineolum, respectively (group 2). Species such as C. acutatum, C. coccodes, C. dematium, and C. trichellum can be clearly distinguished based on ITS 1 sequence divergence, but C. destructivum cannot be confidently separated (98% homology) from C. linicola. Colletotrichum dematium f. truncatum is distinct (12.9%) from C. dematium and should probably be called C. truncatum. Key words : Colletotrichum species, rDNA, ITS 1, nucleotide sequence, phylogeny, systematics.
Directional dominance and a developmental model for the expression of the Tda testis-determining autosomal trait of the mouse
Authors: Eales Brenda A. , Nahas Mirna , Biddle Fred G.
Published: May 2008

The POSCH-2 Y chromosome from the poschiavinus variety of Mus musculus domesticus causes incomplete testis development in the recessive autosomal background of the C57BL/6J laboratory mouse strain. Testis development is normal with the POSCH-2 Y in its native strain background as well as in some strains of the laboratory mouse such as DBA/2J. The phenotype or expression of XY gonadal hermaphroditism in a C57BL/6J strain, which was constructed to be consomic for the POSCH-2 Y, is a threshold trait in which liability is normally distributed and thresholds in the development of the testis define the probability of observing XY embryos with different combinations of ovaries, ovotestes, and testes. The difference in this testis-determining autosomal or Tda trait between the C57BL/6J and DBA/2J strain pair has been demonstrated to be multigenic. We conducted a survey among different strains of the laboratory mouse by test mating females with C57BL/6J.Y-POS males that are consomic for the POSCH-2 Y We identified five groups of strains with significantly different response of XY gonadal hermaphroditism in their XY-POS F1 test embryos. In test embryos, four groups of strains produced gonadal hermaphroditism with different distributions of the types of gonad that appear to have the same variance or shape of a normally distributed liability, but the means of the distributions are at different locations on a scale of gonadal development. The fifth group of strains produced only testes in the test embryos. Several additional matings produced results suggesting that a model of dominance, in the direction of more complete testis development, could interpret the strain differences. The differences in response to the POSCH-2 Y chromosome among the five groups of strains may represent the phenotypes of the genetic recombinants in the Tda trait that were suggested previously by a segregation analysis between C57BL/6J and DBA/2J. The strains may also provide the tools to further dissect the allelic differences and locus determinants of the Tda trait. Key words : mouse, Y chromosome, autosomal testis-determining genes, developmental model of XY gonadal hermaphroditism, primary sex determination.
Molecular evidence forTriticum speltoidesas a B-genome progenitor of wheat (Triticum aestivum)
Authors: Daud Hassan Mat , Gustafson J. P.
Published: May 2008

In polyploid wheat, the origin of the B-genome donor has remained relatively unknown in spite of a number of investigations attempting to identify the parental species. A project was designed to isolate and clone a genome-specific DNA sequence from Triticum speltoides L. to determine if that species could be the B-genome donor. A cloning scheme involving the prescreening of 1-kb fragments followed by colony, dot blot, and Southern blot hybridization screenings was used to isolate a speltoides-specific sequence (pSp89.XI). The methods used allowed for rapid isolation of a genome-specific sequence when screened against total DNA from closely related species. Subsequent analyses showed that the sequence was barely detected in any of the other genomes of the annual Sitopsis section. The results of dot blot and Southern blot analyses established that (i) the sequence pSP89.XI, specific to T. speltoides relative to the other species of the Sitopsis section, was present in the genomes of tetraploid and hexaploid wheat, (ii) the relative abundance of pSp89.XI seemed to decrease from the diploid to the polyploid wheats, and (iii) the existence of a related, but modified B genome in polyploid wheat compared with that in modern T. speltoides was probable. Key words : genome-specific, DNA.
A reciprocal translocation and possible insertion(s) tightly associated with host-specific virulence inCochliobolus heterostrophus
Authors: Chang Hsin-Ru , Bronson Charlotte R.
Published: May 2008

A reciprocal translocation and one or more apparent insertions are shown to be tightly associated with Tox1, a locus controlling T-toxin production and host-selective virulence in race T of the maize pathogen Cochliobolus heterostrophus. Chromosome arrangements were examined by separating chromosomal DNAs of a variety of Tox+ and Tox− strains by pulsed-field gel electrophoresis and hybridizing with probes known to detect RFLPs genetically linked to Tox1. The existence of the translocation was demonstrated by chromosome hybridization patterns; the existence of the putative insertion(s) was deduced from chromosome migration rates. Both differences in chromosome arrangement were detected between 8 Tox+ and 8 Tox− near-isogenic laboratory strains, suggesting that the differences are tightly linked to Tox1. The reciprocal translocation was also detected between all 7 Tox+ and 8 Tox− field isolates examined, suggesting that the translocation is common in natural populations. The field isolates may also differ by the insertion(s); however, numerous additional chromosome size polymorphisms in the field isolates prevented a firm conclusion. The tight association of the translocation and insertion(s) with T-toxin production suggests that chromosome rearrangements may have been involved in the evolution of race T and Tox1. These genomic differences may be causally related to the previously reported reduced fitness of race T relative to race O on N-cytoplasm maize. Key words : chromosome rearrangement, southern corn leaf blight, pulsed-field electrophoresis, T-toxin, evolution.
Useful DNA polymorphisms are identified bysnapback, a midrepetitive element inTribolium castaneum
Authors: Stuart Jeffrey J. , Hall Pamela S. , Maxwell Mark E. , Mocelin Giovani , De Gortari Mauricio J. , Muir William M. , Brown Susan J.
Published: May 2008

The red flour bettle, Tribolium castaneum, is both a pest of stored grain products and an important experimental organism. To improve its facility as a genetic model, we are developing DNA fingerprinting methods for this insect. A Tribolium DNA fragment, snapback-1 (SB1), identified among sequences that reassociate before a C0t of 0.03 mol∙s/L, was found to produce a banding pattern in restriction endonuclease digested genomic DNA that is characteristic of a midrepetitive element. DNA fingerprints of individual beetles demonstrated that unvarying inherited DNA polymorphism is revealed, and that polymorphism is inherited in a dominant Mendelian fashion. Linkage between bands was minimal. The sequence of SB1 was determined, and hybridization experiments indicated that SB1 is a fragment of a larger midrepetitive element. Fingerprinting individuals with known inbreeding coefficients indicated that SB1 loci have relatively high mutation rates. The possibility that SB1 is a fragment of a transposable element is discussed. Key words : Tribolium, beetles, DNA fingerprints, repetitive DNA, quantitative genetics.
Regional and racial specificities in sorghum germplasm assessed with DNA markers
Authors: Oliveira Antonio C. de , Richter Todd , Bennetzen Jeffrey L.
Published: May 2008

Three different molecular marker technologies were used to determine the relatedness of 84 different lines of sorghum. Both racial characterization and geographical origin were found to be correlated with relatedness. In some cases, the region of origin was the more significant factor, where samples of different races from the same locality were more closely related than were samples of the same race from different localities. Wild sorghums were shown to have few novel alleles, suggesting that they would be poor sources of germplasm diversity. The results also indicated that Chinese sorghums are a narrow and distinctive group that is most closely related to race bicolor. Key words : Sorghum bicolor, germplasm diversity, random amplified polymorphic DNA, restriction fragment length polymorphism, simple sequence repeats.
The Afrotropical Drosophila montium subgroup: Balbiani ring 1, polytene chromosomes, and heat shock response of Drosophila vulcana
Authors: Pardali E. , Feggou E. , Drosopoulou E. , Konstantopoulou I. , Scouras Z. G. , Mavragani-Tsipidou P.
Published: May 2008

A detailed photographic map of the salivary gland polytene chromosomes of Drosophila vulcana, an Afrotropical species of the montium subgroup of the melanogaster group, is presented, along with chromosomal rearrangements, such as reverse tandem duplications and inversions, the well-formed Balbiani ring 1, and the most prominent puffs during normal larval and white prepupal development and after ecdysone treatment. In addition, the heat inducible protein and puffing pattern and the loci of the major heat shock genes, namely, hsp70, hsp83, the "small" hsps, and a putative hsp68, of this species were studied. In the light of the data revealed by the above studies, phylogenetic relationships among the montium subgroup species are attempted. Key words : Drosophila, Balbiani ring, polytene chromosomes, heat shock, puffs, genes, proteins, hsp70 single locus.
A gene for resistance to a necrosis-inducing isolate of Pyrenophora tritici-repentis located on 5BL of Triticum aestivum cv. Chinese Spring
Authors: Stock W. S. , Brûlé-Babel A. L. , Penner G. A.
Published: May 2008

Several sources of high-level resistance to tan spot caused by Pyrenophora tritici-repentis have been identified in hexaploid wheat (Triticum aestivum L.). This study was conducted to determine the number and chromosome location of a gene(s) in the cultivar Chinese Spring (CS) that confers resistance to a tan necrosis inducing isolate (nec+chl−) of P. tritici-repentis, 86-124, and insensitivity to Ptr necrosis toxin. Reciprocal crosses were made between CS (resistant–insensitive) and 'Kenya Farmer' (KF) (susceptible–sensitive). Analysis of the CS/KF F1and F2 populations and F2-derived F3 families identified a single nuclear recessive gene governing resistance to isolate 86-124 and Ptr necrosis toxin. Evaluation of the CS(KF) substitution series, F2 monosomic analysis, and screening of a series of 19 CS compensating nullitetrasomic and two ditelosomic lines (2AS and 5BL) indicated that the resistance gene was located on chromosome arm 5BL. No linkage exists between Lr18 and the tan necrosis resistance gene on chromosome arm 5BL. It is proposed that the gene for resistance to the tan necrosis inducing isolate 86-124 (nec+chl−) of P. tritici-repentis and Ptr necrosis toxin be named tsn1. Key words : wheat, Triticum aestivum L., tan spot resistance, Pyrenophora tritici-repentis (Died.) Drechs., chromosome location, Ptr necrosis toxin.
RFLP linkage map of asparagus
Authors: Lewis M. E. , Sink K. C.
Published: May 2008

A population resulting from a double pseudotestcross of two outbred-derived asparagus (Asparagus officinalis L.) clones was evaluated by RFLP (restriction fragment length polymorphism) analysis to produce individual maps of the male and female parents. An asparagus PstI genomic library was created and used as the source of probes. Scoring of bands was done by examining SDRFs (single dose restriction fragments) that are present in one parent and absent in the other and segregate 1:1 in the progeny. The data were analyzed as a backcross population; inversion or recoding allowed for the detection of repulsion phase linkage. The male parent map consisted of 33 loci in 10 groups, while the female parent map had 48 loci arranged in 14 groups. Segregation distortion was minimal (5%), and 17% of the markers were found to be unlinked. Loci of the configuration a/b × a/b and a/c × b/c were used to bridge seven homologous linkage groups between the two parents. The sex locus was not found to be associated with any linkage group. Key words : RFLP, bridge loci, repulsion phase linkage, double pseudotestcross.
A high density RFLP linkage map of sugar beet
Authors: Halldén C. , Hjerdin A. , Rading I. M. , Fridlundh B. , Johannisdottir G. , Tuvesson S. , Åkesson C. , Säll T. , Nilsson N.-O.
Published: May 2008

A high density sugar beet RFLP map with an average distance of 1.5 cM between markers has been constructed. The map covers 621 cM and includes 413 markers distributed over the nine linkage groups of sugar beet. The map is based on two F2populations representing two different pairs of parents. The two sets of data were integrated into a single map using 90 markers that were common to both data sets. The quality of the map was assessed in several ways. The common markers were used to investigate how often the loci had been mapped in the same order in the two F2populations. For closely situated markers (<1.5 cM) the order specified in the map is uncertain, but for markers separated by more than 2 cM the locus order is highly reliable. The error rate of the overall process was estimated at 0.3% by independently repeating the analysis of 41 markers. The map is comparatively short, with a map length corresponding to approximately 1.4 crossovers per bivalent. Another feature of the map is a high degree of clustering of markers along the linkage groups. With the possible exception of linkage group 2, each linkage group shows one major cluster, which in most cases is situated in the centre of the linkage group. Our interpretation is that sugar beet, in comparison with most other species, has an extreme localization of recombination. Key words : sugar beet, linkage, RFLP, clustering.
Use of RAPD analyses to estimate population genetic parameters in the alfalfa leaf-cutting bee,Megachile rotundata
Authors: Lu Rui , Rank Gerald H.
Published: May 2008

RAPD analyses were performed on five geographically isolated populations of Megachile rotundata. We used haploid males of the alfalfa leaf-cutting bee, M. rotundata, to overcome the limitation of the dominance of RAPD markers in the determination of population genetic parameters. Sixteen primers gave rise to 130 polymorphic and 31 monomorphic bands. The unbiased estimators calculated in this study include within- and between-population heterozygosity, nucleotide divergence, and genetic distance. The genetic diversity (H = 0.32–0.35) was found to be about 10 times that of previous estimates (H = 0.033) based on allozyme data. Contrary to the data obtained at the protein level, our results suggest that Hymenoptera do not have a lower level of genetic variability at the DNA level compared with other insect species. Regardless of the different assumptions underlying the calculation of heterozygosity, divergence, and genetic distance, all five populations showed a parallel interrelationship for the three parameters. We conclude that RAPD markers are a convenient tool to estimate population genetic variation in haploid M. rotundata and that with an adequate sample size the technique is applicable to the evaluation of divergence in diploid populations. Key words : Megachile rotundata, RAPD, heterozygosity, genetic distance, nucleotide divergence.
A NOR-associated repetitive element present in the genome of two Salmo species (salmo salar and Salmo trutta)
Authors: Abuín M. , Martínez P. , Sánchez L. , Clabby C. , Flavin F. , Wilkins N. P. , Houghton J. A. , Powell R. , Goswami U.
Published: May 2008

A repetitive element was isolated from the genome of Atlantic salmon. Nucleotide sequence analysis revealed the existence of variant monomers that range in length from approximately 200 to 230 bp. Repeat monomers contain regions of cryptic simplicity, internal repetition, and long direct repeats with deletions and insertions between individual units. The repetitive element was shown to have a tandem unit arrangement and was estimated to occupy between two and three percent of the Atlantic salmon genome. Southern blot analysis revealed the repetitive element to be unique to Atlantic salmon and brown trout species. In situ hybridization analysis showed this element to be localized at the main nucleolar organizer region bearing chromosomes of Atlantic salmon (Salmo salar), AS cell line (derived from S. salar), and brown trout (Salmo trutta). Key words : Atlantic salmon, brown trout, AS cell line, repeat DNA, NOR.
Cell synchronization and isolation of metaphase chromosomes from maize (Zea mays L.) root tips for flow cytometric analysis and sorting
Authors: Lee Jai-Heon , Arumuganathan K. , Kaeppler S. M. , Papa C. M. , Kaeppler H. F.
Published: May 2008

Accumulation of cells containing metaphase chromosomes is an important step in cytological analyses and chromosome sorting procedures. The goal of this research was to optimize treatment parameters to synchronize the cell cycle of maize root tip meristem cells. Levels of hydroxyurea, a DNA synthesis inhibitor, were assessed for their utility in accumulating cells at the G1 phase of the cell cycle. Trifluralin, amiprophos-methyl, and colchicine were used to accumulate cells containing metaphase chromosomes upon release from hydroxyurea inhibition. Optimal mitotic indices were achieved by treating seedlings with 5 mM hydroxyurea for 18 h, incubating for 1 h without chemical treatment to release the hydroxyurea block, and then treating emerging roots with 1 μM trifluralin for 4 h. The mitotic index of synchronized maize root tips was over 70%. Uniformity of synchronization depended upon selection of seeds with emerging radicles that were similar in length at the time of treatment. Suspensions of intact chromosomes were prepared by a simple slicing procedure. The chromosome preparations were found to be suitable for flow cytometric characterization and sorting. Chromosome peaks of the observed flow karyotype resembled the predicted flow karyotype calculated on the basis of maize chromosome size. Key words : flow karyotype, hydroxyurea, plant chromosome sorting, trifluralin.
Genetic variation in dosage effects in maize aneuploids
Authors: Lee E. A. , Coe E. H. , Darrah L. L.
Published: May 2008

In maize (Zea mays L.), the consequences of aneuploidy have been well documented, however, genetic variation in the responses to aneuploidy has not been examined. Using simple B–A translocation stocks to generate a dosage series involving segments from 14 chromosome arms, we tested for the presence of genetic variation for dosage responses in maize by examining reciprocal and maternal genotype effects on the dosage responses. Reciprocal effects examined whether there were differences between two distinctly different inbred backgrounds, Mo17Ht and B73Ht, in how they responded to loss or gain of a B73Ht segment in the Mo17Ht × B73Ht (TB) F1cross versus a Mo17Ht segment in the B73Ht × Mo17Ht (TB) F1cross. Maternal genotype effects questioned whether different inbred backgrounds, Sc41R, T8, and either Mo17Ht or B73Ht (depending on the male), when used as females responded differently to the loss or gain of a chromosome arm segment from the same male (either B73Ht TB or Mo17Ht TB) in an F1cross. Numerous examples of reciprocal and maternal genetic effects were identified in this study. Most of the genetic effects were due to differences in magnitude of response rather than direction; however, tassel-branch number involving the 5S chromosome segment in the B73Ht male background and the 7L chromosome segment in the Mo17Ht male background showed a trend toward the maternal genotype effects being due to differences in the direction of the response. Key words : quantitative traits, corn, B–A translocations, dosage analysis.
Pulsed-field gel electrophoresis analysis of the phaseolin locus region inPhaseolus vulgaris
Authors: Llaca Víctor , Gepts Paul
Published: May 2008

Phaseolin is the major seed storage protein of common bean (Phaseolus vulgaris L.). It is encoded by a small multigene family of 6–9 genes that are clustered in a single complex locus (Phs). We have constructed a long-range restriction map of the phaseolin genomic region, including the Phs locus and two flanking marker loci, D1861 and Bng060. Using a combination of high molecular weight DNA isolation, one- and two-dimensional pulsed-field gel electrophoresis of single and double restriction digests followed by Southern hybridization, and PCR analysis of individual fragments, we found that: (i) the maximum size of the Phs locus is 190 kb, (ii) the Phs locus may have increased in size during the evolution of P. vulgaris, (iii) the genomic region marked by D1861–Phs–Bng060 spans 5 cM, which corresponds to a maximum of 1.9 Mb, and (iv) the Phs locus could be oriented with respect to the two adjacent markers. Further progress in determining the gene arrangement in the Phs locus will require cloning and analysis of large DNA fragments containing phaseolin genes via BAC libraries. Key words : multigene family, physical distance, genome mapping, seed protein.
Genetic analysis of oxygen defense mechanisms inDrosophila melanogasterand identification of a novel behavioural mutant with aShakerphenotype
Authors: Humphreys James M. , Duyf Brenda , Joiner Mei-Ling A. , Phillips John P. , Hilliker Arthur J.
Published: May 2008

Mutants of Drosophila melanogaster that lack Cu/Zn superoxide dismutase or urate are hypersensitive to reactive oxygen species (ROS) generated in vivo by the redox-cycling agent paraquat. We have subsequently employed paraquat as a selective agent to identify adult viable mutants potentially defective in other, perhaps unknown, components of ROS metabolism. Paraquat screening of ethyl methanesulfonate-induced second- and third-chromosome mutations yielded 24 paraquat hypersensitive mutants. Two mutants were identified as being new alleles of the previously identified doublesex (dsx) and pink (p) genes. The remainder of the mutations identified previously undescribed genes, including one second chromosome paraquat hypersensitive mutant that was found to exhibit shaking legs, abdomen pulsations, and body shuddering under ether anaesthesia. This recessive mutation was mapped to the polytene chromosome region of 48A5–48B2 and defines a new gene we named quiver (qvr). This mutation is similar in phenotype to the Shaker (Sh), ether-a-gogo (eag), and Hyperkinetic (Hk) mutations, all of which affect potassium channel function in D. melanogaster. Key words : Drosophila, paraquat, EMS-mutagenesis, Shaker, oxidative-stress.
A novel species-specific tandem repeat DNA family from Sinapis arvensis: detection of telomere-like sequences
Authors: Kapila Ritu , Das Sandip , Lakshmikumaran Malathi , Srivastava P. S.
Published: May 2008

DNA sequences representing a tandemly repeated DNA family of the Sinapis arvensis genome were cloned and characterized. The 700-bp tandem repeat family is represented by two clones, pSA35 and pSA52, which are 697 and 709 bp in length, respectively. Dot matrix analysis of the sequences indicates the presence of repeated elements within each monomeric unit. Sequence analysis of the repetitive region of clones pSA35 and pSA52 shows that there are several copies of a 7-bp repeat element organized in tandem. The consensus sequence of this repeat element is 5′-TTTAGGG-3′. These elements are highly mutated and the difference in length between the two clones is due to different copy numbers of these elements. The repetitive region of clone pSA35 has 26 copies of the element TTTAGGG, whereas clone pSA52 has 28 copies. The repetitive region in both clones is flanked on either side by inverted repeats that may be footprints of a transposition event. Sequence comparison indicates that the element TTTAGGG is identical to telomeric repeats present in Arabidopsis, maize, tomato, and other plants. However, Bal31digestion kinetics indicates non-telomeric localization of the 700-bp tandem repeats. The clones represent a novel repeat family as (i) they contain telomere-like motifs as subrepeats within each unit; and (ii) they do not hybridize to related crucifers and are species-specific in nature. Key words : Brassica species, Sinapis arvensis, tandem repeats, telomeres.
Analysis of cultivated potato (Solanum tuberosum) using intermicrosatellite amplification
Authors: Provan Jim , Powell Wayne , Waugh Robbie
Published: May 2008

Twelve potato (Solanum tuberosum) cultivars were analysed using intermicrosatellite amplification with two 5′-anchored repeat primers. Amplification products were resolved on polyacrylamide gels. In total, 24 polymorphic bands were scored and used to construct a dendrogram showing the relationships between the cultivars. The three late 19th – early 20th century cultivars studied were grouped together, as were the three accessions directly descended from S. tuberosum cv. Maris Piper. These results suggest that intermicrosatellite amplification is a potentially useful tool for both phenetic and genotyping studies. Key words : potato, microsatellites, intermicrosatellite amplification, genotyping.
Localized chiasmata and meiotic nodules in the tetraploid onionAllium porrum
Authors: Stack Stephen M. , Roelofs Dick
Published: May 2008

Allium porrum L. (cultivated leek) (2n = 4x = 32) is a fertile tetraploid that forms bivalents with pericentric chiasmata at metaphase I. To investigate the basis of this unusual behavior for a tetraploid, we describe the karyotype, axial cores, synaptonemal complexes (SCs), and meiotic nodules of A. porrum. The karyotype appears to be autotetraploid. This conclusion is also supported by presynaptic alignment of axial cores in groups of four and partner trades between pairs of SCs. Numerous early nodules are distributed all along axial cores and SCs during zygonema, but they are lost by late zygonema – early pachynema. Late (recombination) nodules (RNs) are present on SCs near kinetochores throughout the remainder of pachynema. This pattern of RNs corresponds to the pattern of pericentric chiasmata. Pachytene quadrivalents usually are resolved into bivalents because partner trades between SC lateral elements rarely occur between RNs on the same segment of SC. Thus, the patterns of crossing-over and partner trades promote balanced disjunction and high fertility in autotetraploid A. porrum. Rare quadrivalents observed at metaphase I must be due to infrequent partner trades between RNs. Polycomplexes, unusual in their number and size, were observed during zygonema. Key words : synaptonemal complex, recombination nodules, localized chiasmata, polycomplex, Allium porrum.
Development of a PCR-based allele-specific assay from an RFLP probe linked to resistance to cereal cyst nematode in wheat
Authors: Williams Kevin J. , Fisher John M. , Langridge Peter
Published: May 2008

The RFLP locus Xglk605 identified by the probe Tag605 maps to a proximal position on the long arm of wheat chromosome 2B about 7 cM away from a gene conditioning resistance to cereal cyst nematode in the wheat line AUS10894. The clone Tag605 was partially sequenced and the PCR primer set AWP1 was designed. The 292-bp product, which showed no polymorphism between varieties, was cloned and sequenced. A single base difference was found in the sequence of the AWP1 products amplified and cloned from the wheat lines AUS10894 and 'Spear'. PCR primers were designed with 3′ termini that corresponded to the two alleles. A dual-PCR system was developed in which the primer sets AWP2 and AWP3 produced allele-specific amplification. The concentration of the oligonucleotide primers and the sequence of the primer–template mismatches were critical to the success of discriminatory allele amplification. Key words : Triticum aestivum, STS, cereal cyst nematode, RFLP.
Structure and organization of thePelement related sequences inDrosophila madeirensis
Authors: Paricio N. , Martínez-Sebastián M. J. , Frutos R. de , Miller W. J. , Hagemann S. , Pinsker W.
Published: May 2008

The P element homologous sequences of the two closely related species Drosophila guanche and Drosophila subobscura represent a very special case of transposable-element derivatives. Although they have lost the regions known to be essential for P transposition by random mutations, all of them have selectively conserved the coding capacity for "P-repressor-like" proteins during the past few millions years. In both species, they are tandemly amplified in a single euchromatic gene cluster at equivalent chromosomal positions. In contrast, Drosophila madeirensis, an endemic species that is very closely related to both D. subobscura and D. guanche, harbours an additional P homologous site. Several mechanisms can be invoked to explain the generation of the new site in this species. In this work we present several molecular and cytological data in order to elucidate the possible evolutionary origin of the P derivatives of D. madeirensis. Key words : Drosophila, P elements, molecular evolution, gene cluster.
Identification of RAPD, SCAR, and RFLP markers tightly linked to nematode resistance genes introgressed fromArachis cardenasiiintoArachis hypogaea
Authors: Garcia G. M. , Stalker H. T. , Shroeder E. , Kochert G.
Published: May 2008

Two dominant genes conditioning resistance to the root-knot nematode Meloidogyne arenaria were identified in a segregating F2population derived from the cross of 4x (Arachis hypogaea × Arachis cardenasii)-GA 6 and PI 261942. Mae is proposed as the designation for the dominant gene restricting egg number and Mag is proposed as the designation for the dominant gene restricting galling. The high levels of resistance in GA 6 were introgressed from A. cardenasii and, therefore, a search to identify A. cardenasii specific RAPD markers that are tightly linked to these resistance genes was conducted utilizing bulked segregant analysis. One RAPD marker (Z3/265) was linked at 10 ± 2.5 (SE) and 14 ± 2.9 cM from Mag and Mae, respectively. The marker was mapped to linkage group 1 at 5 cM from Xuga.cr239 in the backcross map in an area where introgression from A. cardenasii had previously been reported. This fragment was cloned and used to generate a pair of primers that specifically amplified this locus (sequence characterized amplified region, SCAR) and as a RFLP probe. Their close linkage with the resistance genes will be useful in marker-based selection while transferring nematode resistance from introgression lines into elite breeding lines and cultivars. The Z3/265 marker associated with the genes Mae or Mag was not found in other highly resistant Arachis species (Arachis batizocoi or Arachis stenosperma), in progenies of interspecific crosses with A. cardenasii that were moderately resistant, or in the resistant A. hypogaea lines PI 259634 and PI 259572. These represent the first molecular markers linked with a resistant gene in peanut and the first report of two physiological responses to nematode attack associated with two genetic factors. Key words : peanut, Arachis hypogaea, Arachis cardenasii, Meloidogyne arenaria, RFLP, RAPD, SCAR, nematode resistance, bulk segregant analysis, introgression.
Genome flux in tomato cell clones cultured in vitro in different physiological equilibria. II. A RAPD analysis of variability
Authors: Bogani Patrizia , Simoni Alessandra , Lio' Pietro , Scialpi Angela , Buiatti Marcello
Published: May 2008

An analysis of the effect of changing physiological conditions on genome evolution in tomato cell populations has been carried out on long-term in vitro cultured clones grown on different auxin–cytokinin equilibria or selected for low–high competence for active defense against Fusarium oxysporum f.sp. lycopersici. RAPD analysis, confirmed through pattern rehybridization, was used as a random tool to measure the genetic variability. Through the use of a modified ANOVA, variation was shown to depend on both the initial genotype and the physiological conditions. Pattern correlation analysis through a mutual information algorithm suggested the fixation of RAPD patterns specific to physiological equilibria. The results are discussed in view of the possible relevance for evolution at hierarchical levels higher than cell populations. Key words : tomato clones, somaclonal variation, RAPD, coadaptation.
The pattern of histone H4 acetylation on the X chromosome during spermatogenesis of the desert locustSchistocerca gregaria
Authors: Wolf Klaus Werner , Turner Bryan M.
Published: May 2008

We have used antibodies directed against histone H4 acetylated at lysine residue 5, 8, 12, or 16 and indirect immunofluorescence microscopy to probe chromosomes from spermatogonia and spermatocytes of the desert locust, Schistocerca gregaria. The autosomes showed bright overall fluorescence, indicative of high levels of H4 acetylation. In contrast, the X chromosome, which is facultatively heterochromatic during spermatogenesis of the locust, remained completely unstained in spermatogonia and secondary spermatocytes and showed only a small terminal fluorescent band in primary spermatocytes. This band probably corresponds to centromere-associated constitutive heterochromatin. Thus, underacetylation is a cytogenetic marker for facultative heterochromatin, but not necessarily constitutive heterochromatin, during spermatogenesis of the locust. Scanning electron microscopy of chromosomes from prophase spermatogonia and prophase I spermatocytes revealed that underacetylation of histone H4 in the X chromosome was not accompanied by a chromatin organization visibly different from that of the autosomes. Transmission electron microscopy of mitotic spermatogonia showed that the X chromosome is separated from the autosomes in a small nuclear compartment of its own in prophase and telophase and associated with membranes in metaphase. In prophase I spermatocytes, autosomes and the sex univalent were in the same compartment. This compartmentalization may be responsible for the underacetylation and (or) transcriptional silencing of the X chromosome in spermatogonial mitosis. Key words : histone acetylation, chromosomes, meiosis, heterochromatin, desert locust.