Isozyme variation in Silene pratensis: a response to different environments

The isozymes of nine enzyme systems were screened and the frequencies of the flavone glycosylating genes were determined in an outdoor experiment with 70 populations of European S. pratensis and an indoor experiment with 30 populations of the same species. Cluster analysis (using Ward's cluster criterion) were performed on all data sets. In the outdoor experiment, cluster analysis of the flavonoid data gave the same pattern that we obtained in an ealier survey of a larger set of populations and showed clearly that there are eight chemical races in European S. pratensis. No comparable geographic distributiion could be found in the isozyme data set, although the dendrogram showed two very clearly separated groups. The two groups represented the two years in which the populations were grown. This result indicates that isozyme variation in European S. pratensis is largely determined by environmental factors. Observations on changes in isozyme patterns during ontogeny and on differences between indoor and outdoor grown plants of the same F2 crosses confirm this. Differences in isozyme patterns can be caused by very small differencs in environment as is shown by the results of the indoor experiment, in which a slight gradient in environmental conditions was present in the greenhouses. The cluster analysis of the isozyme data from the indoor experiment revealed three distinct groups of populations that could be related to location within the greenhouses. As in the outdoor experiment, the dendrogram for the flavonoids gave the same geographic pattern as found with the earlier survey.     

Alien introgression in the grasses Lolium perenne (perennial ryegrass) and Festuca pratensis (meadow fescue): the development of seven monosomic substitution lines and their molecular and cytological characterization

Background and Aims

To address the issues associated with food security, environmental change and bioenergy in the context of crop plants, the production, identification and evaluation of novel plant phenotypes is fundamental. One of the major routes to this end will be wide hybridization and introgression breeding. The transfer of chromosomes and chromosome segments between related species (chromosome engineering or alien introgression) also provides an important resource for determining the genetic control of target traits. However, the realization of the full potential of chromosome engineering has previously been hampered by the inability to identify and characterize interspecific introgressions accurately.

Methods

Seven monosomic substitution lines have been generated comprising Festuca pratensis as the donor species and Lolium perenne as the recipient. Each of the seven lines has a different L. perenne chromosome replaced by the homoeologous F. pratensis chromosome (13 L. perenne + 1 F. pratensis chromosome). Molecular markers and genomic in situ hybridization (GISH) were used to assign the F. pratensis chromosomes introgressed in each of the monosomic substitutions to a specific linkage group. Cytological observations were also carried out on metaphase I of meiosis in each of the substitution lines.

Results

A significant level of synteny was found at the macro-level between L. perenne and F. pratensis. The observations at metaphase I revealed the presence of a low level of interspecific chromosomal translocations between these species.

Discussion

The isolation of the seven monosomic substitution lines provides a resource for dissecting the genetic control of important traits and for gene isolation. Parallels between the L. perenne/F. pratensis system and the Pooideae cereals such as wheat, barley, rye, oats and the model grass Brachypodium distachyon present opportunities for a comparison across the species in terms of genotype and phenotype.     

Intraspecies comparison of Streptomyces pratensis genomes reveals high levels of recombination and gene conservation between strains of disparate geographic origin

Background

Streptomyces are widespread bacteria that contribute to the terrestrial carbon cycle and produce the majority of clinically useful antibiotics. While interspecific genomic diversity has been investigated among Streptomyces, information is lacking on intraspecific genomic diversity. Streptomyces pratensis has high rates of homologous recombination but the impact of such gene exchange on genome evolution and the evolution of natural product gene clusters remains uncharacterized.

Results

We report draft genome sequences of four S. pratensis strains and compare to the complete genome of Streptomyces flavogriseus IAF-45-CD (=ATCC 33331), a strain recently reclassified to S. pratensis. Despite disparate geographic origins, the genomes are highly similar with 85.9% of genes present in the core genome and conservation of all natural product gene clusters. Natural products include a novel combination of carbapenem and beta-lactamase inhibitor gene clusters. While high intraspecies recombination rates abolish the phylogenetic signal across the genome, intraspecies recombination is suppressed in two genomic regions. The first region is centered on an insertion/deletion polymorphism and the second on a hybrid NRPS-PKS gene. Finally, two gene families accounted for over 25% of the divergent genes in the core genome. The first includes homologs of bldB (required for spore development and antibiotic production) while the second includes homologs of an uncharacterized protein with a helix-turn-helix motif (hpb). Genes from these families co-occur with fifteen pairs spread across the genome. These genes have evidence for co-evolution of co-localized pairs, supporting previous assertions that these genes may function akin to a toxin-antitoxin system.

Conclusions

S. pratensis genomes are highly similar with exceptional levels of recombination which erase phylogenetic signal among strains of the species. This species has a large core genome and variable terminal regions that are smaller than those found in interspecies comparisons. There is no geographic differentiation between these strains, but there is evidence for local linkage disequilibrium affecting two genomic regions. We have also shown further observational evidence that the DUF397-HTH (bldB and hpb) are a novel toxin-antitoxin pair.     

Flavonoid glycosides and the chemosystematics of Eucalyptus camaldulensisis

Leaf flavonoid glycosides of Eucalyptus camaldulensis were identified as kaempferol 3-glucoside and 3-glucuronide; quercetin 3-glucoside, 3-glucuronide, 3-rhamnoside, 3-rutinoside and 7-glucoside, apigenin 7-glucuronide and luteolin 7-glucoside and 7-glucuronide. Two chemical races were observed based on the flavonoid glycosides. These races correspond to the northern and southern populations of species growing in Australia. The Middle Eastern species examined were found to belong to the southern Australian chemical race. The major glycosides of E. occidentalis proved to be quercetin and myricetin 3-glucuronide.     

The geographic distribution of flavone-glycosylating genes in Silene pratensis (Rafn.) Godron and Gren. (Caryophyllaceae)

In Silene pratensis three loci (g, gl and fg) control the glycosylation of isovitexin. Three alleles are known for both the g-locus (g, g ^G and g ^X ) and the gl-locus (gl, gl ^A and gl ^R ); for the fg-locus there are only two alleles (fg and Fg). The distribution of these alleles over 285 European populations of S. pratensis has been investigated. It was concluded that there are three different chemical races within S. pratensis in Europe. The first race contains the populations in western and southern Europe and displays high frequencies of g ^G , gl and fg. The frequencies of g ^G and gl ^R are very high in the second chemical race, which can be found in the USSR, Scandinavia and eastern Poland. The third chemical race occurs in central Europe and in this race the frequencies of both g and gl ^R are high, Fg has low to moderate frequencies in the second and third groups. The alleles gl ^A and g ^X are seldom found in S. pratensis, but are present in the closely related S. dioica. They do occur with low frequencies in some populations of S. pratensis, possibly as a result of hybridization with S. dioica.     

Craniometric differences between karyotypic races of the common shrew Sorex araneus (Mammalia) as a result of limited hybridization

The contact points of four karyotypic races (St. Petersburg, Moscow, Seliger and West Dvina) of the common shrew Sorex araneus L. were studied at the Valdai Hills (European Russia) in an area unimpeded by geographic barriers. The populations of the races are separated by narrow hybrid zones that represent the most complex heterozygous hybrid karyotypes. At these points of contact, the morphometric differentiation of karyotype races was examined in 12 cranial measurements in 190 shrews of a known karyotype. A comparison of the mean values in studied samples of immature shrews revealed statistically significant differences and the correlation of some measurements which describe the level of musculus temporalis. It has been proposed that morphometric differences in the karyotypic races were preserved and accumulated because of a 50% reduction of the frequencies of hybrids. The deviation from the Hardy-Weinberg ration in the frequencies of the genotype and haploid sets of chromosomes in the hybrid zones can be attributed to a number of fatalities of hybrid embryos or the nonrandom mating of karyotypic races. The ethological isolation might arise in the evolution of some karyotypic races from the reduced fitness of the hybrids.     

Genetic Structure of Races of Heterodera glycines and Inheritance of Ability to Reproduce on Resistant Soybeans

Four field populations of Heterodera glycines tested for ability to reproduce on three host differentials were each classified into one of the recognized races. A fifth population represented a new race. Genetic analysis indicated that the designated races are actually field populations that differ from each other primarily in the frequencies of three groups of genes (genes for parasitism) that act quantitatively and control the ability of the nematode to reproduce on resistant P.I. 88788, Pickett, and P.I. 90763 soybeans. Populations of race-3 have none of these genes for parasitism, or they have some in low frequency that results in an index of parasitism of less than 10 on any one of the resistant soybeans. Race-1 has a high frequency of one group of genes that enable it to reproduce on P.I. 88788. Race-2 has two groups of genes for parasitism in high frequency; one for P.I. 88788, and one for Pickett. Based on these findings, it was assumed that race-4 has three groups of genes for parasitism; one for P.I. 88788, one for Pickett, and one for P.I. 90763. Additional races may be recognized when new genes are identified, or when new gene combinations are discovered. The ability to reproduce on P. I. 88788 is inherited independently from the ability to reproduce on Pickett. Although the genetic structure of field populations does not provide a solid foundation for race designation, recognizing races under the present system may be useful when it clearly characterizes the behavior of field populations. Race designations, however, should be regarded as provisional since gene frequencies change with time in response to selection forces and, therefore, the race status of a population may change accordingly.     

Differential expression of VRN1 and other MADS-box genes in Festuca pratensis selections with different vernalization requirements

Most perennial and winter annual temperate grasses have a vernalization requirement (VR) for flowering, that is, they require a cold period before they can flower in response to long days. From a F1 mapping population of the outbreeding perennial forage grass Festuca pratensis Huds. (meadow fescue) previously used to map several quantitative trait loci (QTLs) for VR, we produced two F2 populations divergently selected for high or low VR. The two populations were characterised for flowering behaviour and gene expression of VRN1 as well as other MADS-box genes with a putative function in the induction of flowering. Expression of FpVRN1 and the VRN1-like genes FpMADS2 and FpMADS3 was associated with flowering but the response of gene expression to vernalization differed between genes and populations. The expression of the SVP-like genes FpMADS10 and FpMADS16 was not affected by vernalization and did not differ between the two F2 populations.     

Characterization of CYP1A2, CYP2C19, CYP3A4 and CYP3A5 polymorphisms in South Brazilians

Potential causes of variability in drug response include intrinsic factors such as ethnicity and genetic differences in the expression of enzymes that metabolize drugs, such as those from Cytochrome P450 (CYPs) superfamily. Pharmacogenetic studies search for genetic differences between populations since relevant alleles occur with varying frequencies among different ethnic populations. The Brazilian population is one of the most heterogeneous in the world, resulting from multiethnic admixture of Amerindians, Europeans, and Africans across centuries. Since the knowledge of CYP allele frequency distributions is relevant to pharmacogenetic strategies and these data are scarce in the Brazilian population, this study aimed to describe genotype and allele distributions of 15 single nucleotide polymorphisms (SNPs) at CYP 1A2, 2C19, 3A4, and 3A5 genes in African and European descents from South Brazil. A sample of 179 healthy individuals of European and African ancestry was genotyped by the MassARRAY SNP genotyping system. CYP3A5*3, CYP1A2*1F, CYP3A4*1B, and CYP2C19*2 were the most frequent alleles found in our sample. Significant differences in genotype and allelic distribution between African and European descents were observed for CYP3A4 and CYP3A5 genes. CYP3A4*1B was observed in higher frequency in African descents (0.379) than in European descents (0.098), and European descents showed higher frequency of CYP3A5*3 (0.810) than African descents (0.523). Our results indicate that only a few polymorphisms would have impact in pharmacogenetic testing in South Brazilians. Further studies with larger sample sizes are required also among other Brazilian regions.     

Distribution of Races of Heterodera glycines in the Central United States

A total of 62 populations of Heterodera glycines were collected in 10 states along the Mississippi and Missouri rivers, and 206 populations were collected in Arkansas. Among the 62 populations, races 2, 3, 4, 5, 6, 9, and 14 were found south of 37°N latitude, and races 1 and 3 were found north of 37°N latitude. In Arkansas samples, races 2, 4, 5, 6, 9, and 14 comprised 87% of the populations. In both groups of samples, H. glycines populations with genes that enabled the population to parasitize cv. Pickett occurred the most frequently, followed by those with genes for parasitism of cv. Peking, then PI88.788, and the fewest with genes for parasitism of PI90.763. The diversity of races in this study raises questions about the effectiveness of race-specific cultivars for the management of soybean cyst nematodes. The greater diversity of races of H. glycines in the southern United States may be because of a longer history of planting resistant cultivars.     

Effects of salinity and clipping on biomass and competition between a halophyte and a glycophyte

Global climate change will likely result in the reduction of water levels in intermountain wetlands and ponds, and the vegetation communities associated with these wetlands are an important forage source for livestock. Lowered water levels will not only constrict wetland plant communities, it will potentially change aquatic and soil salt concentrations. Such an increase in salinity can reduce plant growth and potentially affect competitive interactions between plants. A greenhouse experiment examined the effects of salinity and competition on the growth of two wet meadow grass species, Poa pratensis (a glycophyte) and Puccinellia nuttalliana (a halophyte). The following hypotheses based on published data were tested: (1) Biomass of both species will decrease with increasing concentration of salt; (2) root:shoot (R:S) ratio of P. pratensis will decrease with increasing salt concentration while R:S ratio of P. pratensis and P. nuttalliana will increase with clipping; (3) competitive importance will decrease for P. pratensis and P. nuttalliana with increasing salt concentration because salt induces a stress response and competitive importance is reduced in stressed environments. A factorial design included 3 plant treatments (P. nuttalliana alone, P. pratentsis alone, P. nuttalliana + P. pratensis) × 4 salinity rates (control; 5, 10, 15 g/L NaCl) × 2 clipping intensities (plants clipped or not clipped) for a total of 24 combinations replicated 6 times over a period of 90 days. We found a reduction in dry biomass as salinity increased, and this effect was greatest for P. pratensis. (1.94 g (SE 0.13) at 0 g/L NaCl to 0.22 g (SE 0.11) at 15 g/L NaCl). The R:S ratio of P. pratensis was reduced by salinity, but not for P. nuttalliana. Competitive importance of both species was reduced by clipping and by salinity, but the effect was greater and more consistent for P. pratensis. We conclude that salt concentration reduces plant growth and the effect of competition.     

Biochemical properties and level of expression of alcohol dehydrogenases in the allotetraploid plant Tragopogon miscellus and its diploid progenitors

This study demonstrates that homoeologous genes in two diploid plant species that specify different amounts of an enzyme maintain the same relative level of expression in an allotetraploid derivative. The three predominant alcohol dehydrogenase (ADH) isozymes (DD, DP, PP) in seeds of the recently evolved allotetraploid plant Tragopogon miscellus (Compositae) are dimers specified by Adh3-D and Adh3-P genes derived from its diploid progenitors T. dubius and T. pratensis. Seeds of T. pratensis contain twice as much ADH activity as those of T. dubius, while T. miscellus is intermediate. The three isozymes were similar in a number of catalytic properties; the densitometric ratio of the isozymes purified from T. miscellus was 1 DD:4DP:4PP for both ADH activity and protein; and dissociation-reassociation of the DP enzyme gave a 1:2:1 ratio of the three isozymes. Therefore, the enzymes were similar in specific activity, but twice as many P as D subunits were present in active enzymes in T. miscellus, precisely the difference in activity between the parents. In T. miscellus, the specific activity of ADH and its activity per mg tissue are intermediate to those of the diploids, because relative expression of the Adh gene in each genome is not influenced by the presence of the other genome.     

Flavonoid glycosides of Lathyrus pratensisleguminosae

Eight flavonoid glycosides have been identified in the leaves of Lathyrus pratensis, two of which are new glycosides.     

Hybridization of Races of Heterodera glycines

Progeny from single females of four known races of Heterodera glycines Ichinohe were used to establish relatively uniform populations. Single females from these populations were mated with males of other races in all possible combinations to study compatibility and inheritance patterns. When race 1 or 3 was crossed with either race 2 or 4, there was a significant reduction in number of females and a greater number of eggless females than in crosses of races 1 × 3 and 2 × 4. More females matured and fewer were eggless when matings were of the same race. Parasitic capabilities of races 2 and 4 were dominant or partially dominant over those of races 1 and 3, based on parasitism of F₁ hybrids. Segregation patterns were generally similar for reciprocal crosses between races. There appeared to be either one or two major genes segregating for parasitism of ''Pickett'' soybean in the different crosses. A hybrid isolate (race 3 × 4) that differed in parasitic capability from the four known races produced as many females on the resistant soybean genotype, PI 90,763, as on the susceptible Lee cultivar. Those data indicate that isolates of H. glycines with a different parasitic capability may develop from gene recombination.     

Chromosome relationships between Lolium and Festuca (Gramineae)

With a view to eclucidating chromosome relationships between Lolium perenne (Lp), L. multiflorum (Lm) and Festuca pratensis (Fp), chromosome pairing in different diploid (2n=14), auto-allotriploid (2n=3x=21), trispecific (2n=3x=21), amphidiploid (2n=4x=28) and auto-allohexaploid (2n=6x=42) hybrids between them was analysed. At all these levels of ploidy there was very good chiasmate pairing between the chromosomes of the three species and, on the whole, there was little evidence of preferential pairing of the chromosomes of a particular species in the triploid, tetraploid and hexaploid hybrids. A critical test for this also came from the synaptic ability of the chromosomes of the single genome with those of the duplicated genome in the auto-allotriploids which formed predominantly trivalents with 2, 3 or even 4 chiasmata. Moreover, the homology between the Lp and Lm chromosomes seems strong enough to pass the discrimination limits of the B-chromosomes which do not suppress homoeologous pairing in the Lp LmLm triploid and LpLm diploid hybrids. — The triploids having two genomes of a Lolium species and one of F. pratensis had some male and female fertility which suggested genetic compatibility of the parental chromosomes resulting, presumably, in compensation at the gametic level. Also, the occurrence of comparable chiasma frequencies in the auto-allotriploids and trispecific hybrids showed that they were not markedly affected whether two doses of one genome and one of the other or all the three different genomes from the three species were present. From the trend of chromosome pairing in all these hybrids it is concluded that there is little structural differentiation between the chromosomes of the three species, no effective isolation barrier to gene-flow between them, and that they are closely related phylogenetically, having possibly evolved from a common progenitor. Taxonomic revision of the two Lolium species is suggested.     

Development of chickpea near-isogenic lines for fusarium wilt

Four pairs of near-isogenic lines (NILs) of chickpea with resistance/susceptibility to Fusarium oxysporum f. sp. ciceris (Foc) have been developed in this study. These lines were produced by searching in advanced recombinant inbred lines (RILs) that are segregating for Foc race 5 based on a phenotypic screening. The sequence tagged microsatellite (STMS) marker TA59, closely linked to wilt resistance genes on linkage group 2 (LG2) of the chickpea map, was used to assist the selection of resistant or susceptible genotypes. The NILs were also characterized for disease reaction to Foc races 1A, 2, 3 and 4. Resistance, susceptibility and slow wilting reactions were found in these NILs. Our results suggest that more than one gene controls the resistance to race 5. Combination of the major gene foc-5 linked to TA59 with other gene/s appears to be required to complete resistance, and the absence of these unknown genes leads to slow wilting reactions. The independent differential responses to races 2 and 3 observed in three NILs could be explained as recombination events. This result suggests that foc-2 and foc-3 are delimiting points at opposite ends of a genomic region that includes the remaining foc genes and the TA59 marker. This set of NILs has great potential for studying the genetics and mechanisms of wilt resistance. In addition, the NIL RIP8-94-11 can be used as differential line for Foc race 3; it showed a clear resistance reaction to race 3 and susceptibility to the other Foc races.     

Olfactory associative behavioral differences in three honey bee Apis mellifera L. races under the arid zone ecosystem of central Saudi Arabia

Apis mellifera jemenitica is the indigenous race of honey bees in the Arabian Peninsula and is tolerant to local drought conditions. Experiments were undertaken to determine the differences in associative learning and memory of honey bee workers living in the arid zone of Saudi Arabia, utilizing the proboscis extension response (PER). These experiments were conducted on the indigenous race (A. m. jemenitica) along with two introduced European races (A. m. carnica and A. m. ligustica). The data revealed that A. m. jemenitica is amenable to PER conditioning and may be used in conditioning experiments within the olfactory behavioral paradigm. The results also demonstrated that the three races learn and retain information with different capacities relative to each other during the experimental time periods. Native Arabian bees (A. m. jemenitica) exhibited significantly lower PER percentage during second and third conditioning trials when compared to exotic races. Apis mellifera jemenitica also exhibited reduced memory retention at 2?h and 24?h when compared to A. m. carnica and A. m. ligustica. Therefore, the native Arabian bees were relatively slow learners with reduced memory retention compared to the other two races that showed similar learning and memory retention. Three or five conditioning trials and monthly weather conditions (October and December) had no significant effects on learning and memory in A. m. jemenitica. These results emphasized a novel line of research to explore the mechanism and differences in associative learning as well as other forms of learning throughout the year among bee races in the harsh arid conditions of Saudi Arabia. This is the first study in Saudi Arabia to demonstrate inter-race differences regarding olfactory associative learning between native Arabian bees and two introduced European honey bee races.     

Photomorphogenically Defined Light and Resistance of Poa pratensis to Drechslera sorokiniana

Photomorphogenic light definitions were derived by mathematical determination of the estimated phytochrome photoequilibrium for each light treatment spectrum. A wide range of photomorphogenic light treatments represented by spectra with estimated phytochrome photoequilibria of 0.45, 0.54, 0.60, 0.67, and 0.71 was utilized to determine the influence of photomorphogenically defined light on resistance of Poa pratensis L. to pathogenesis by Drechslera sorokiniana. Accurate resolution of D. sorokiniana leaf spot development required evaluation of separate leaf ages due to the sequential appearance, development, and senescence of P. pratensis leaves. Disease development (all light treatments) was greatest on leaf 4 (oldest, postmature) followed by leaf 1 (youngest, premature). Low levels of disease occurred on leaves 2 and 3 (mature). Photomorphogenic light defined by estimated phytochrome photoequilibria greater than 0.60 (natural light 0.60) was most disease promotive on leaf 1. Conversely, photomorphogenic light defined by estimated phytochrome photoequilibria of less than 0.60 was most disease promotive on leaf 4. These responses indicate that inherent resistance or susceptibility expressed by P. pratensis to pathogenesis by D. sorokiniana is regulated in part by leaf age (developmental senescent stage) and by photomorphogenically defined light quality. A hypothesis is presented and discussed which integrates and speculates on these observations with respect to the literature.