Genotypical features in the expression of allozymes of β-specific hydrolase of carboxylic esters in wild-type <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The expression level of electrophoretically separated S- and F-allozymes of β-specific esterase (EC 3.1.1.2) in genotypes of wild-type Drosophila melanogaster (males and females) that are monozygous or heterozygous with respect to the locus β-Est is determined by means of computerized densitometry; α-naphthylacetate, β-naphthylacetate, and α-naphthylpropionate are used as the substrates. The intensity of the expression of the esterase is judged from the quantity of reaction product created as a result of simultaneous azo coupling between naphthol and diazonium in 4, 24, 44, and 64 min incubation times. Reliable differences in the expressions of the S- and F-allozymes as a function of the structure of the β-Est locus of genotypically distinct individuals are established. In all the variant experiments, a higher level of summary activity of the S- and F-allozymes of the β-esterase of the heterozygotes by comparison with the individual activity of the F-and S-allozymes of the corresponding homozygotes was demonstrated, independently of the sex of the Drosophila individual. A comparative estimate of the temporal dynamics of the expression of in vitro allozymes of the dominant homozygotes (β-Est _S /β-Est _S ), heterozygotes (β-Est _S /β-Est _F ), and recessive homozygotes (β-Est _F /β-Est _F ) is performed. Possible mechanisms for the occurrence of heterosis according to the character of expression of S- and F-allozymes of β-esterase on the basis of the theory of biochemical enrichment of heterozygote genotypes are considered.     

Contribution à l'Etude des Estérases dans Quatre Sous-espéces D'Idotea balthica

In Idotea balthica, α esterase₂ and β esterase₃ have been tested against nine substrates and nine inhibitors. They are dependent on two polyallelic independent loci. Est₂ offers alternative alleles in Mediterranean and Nordic populations. The variation in frequency of alleles at locus Est₃ allows one to characterize the subspecies stagnea compared with tricuspidata and balthica but cannot be used to separate this subspecies of basteri.     

Joint effects of genes underlying a temperature specialization tradeoff in yeast

A central goal of evolutionary genetics is to understand, at the molecular level, how organisms adapt to their environments. For a given trait, the answer often involves the acquisition of variants at unlinked sites across the genome. Genomic methods have achieved landmark successes in pinpointing these adaptive loci. To figure out how a suite of adaptive alleles work together, and to what extent they can reconstitute the phenotype of interest, requires their transfer into an exogenous background. We studied the joint effect of adaptive, gain-of-function thermotolerance alleles at eight unlinked genes from Saccharomyces cerevisiae, when introduced into a thermosensitive sister species, S. paradoxus. Although the loci damped each other’s beneficial impact (that is, they were subject to negative epistasis), most boosted high-temperature growth alone and in combination, and none was deleterious. The complete set of eight genes was sufficient to confer ~15% of the S. cerevisiae thermotolerance phenotype in the S. paradoxus background. The same loci also contributed to a heretofore unknown advantage in cold growth by S. paradoxus. Together, our data establish temperature resistance in yeasts as a model case of a genetically complex evolutionary tradeoff, which can be partly reconstituted from the sequential assembly of unlinked underlying loci.     

Genetic diversity and structure among natural populations of Sindora glabra in Hainan Island,China as revealed by ISSR markers

Sindora glabra, one of the second-class national protective plants in China, has important ecological and commercial values. To understand the genetic diversity and structure of S. glabra, eleven natural populations from four areas of Hainan Island in China were analyzed by inter-simple sequence repeat (ISSR) markers. Thirteen primers were screened out and used to amplify 157 DNA samples. A total of 122 bands were obtained, among which 114 (93.4%) bands were polymorphic. Genetic parameters including average number of effective alleles (1.547), Nei's gene diversity (0.321), Shannon's information index (0.482), and gene differentiation coefficient (0.1944) revealed a high level of genetic diversity maintained in S. glabra populations. The variation within populations accounted for 85.6% of total variation based on analysis of molecular variance. Genetic distance analysis showed that 11 populations could be divided into three groups and populations from the same areas were classified as one group, suggesting that high genetic diversity of S. glabra was attributable to geographic isolation. Together, introduction of germplasm from distant natural distribution areas would be a sound strategy for germplasm resource conservation of S. glabra and selection of elite individuals from populations of far relationship for hybridization is of great importance for breeding improvement programs in future.     

Genetic basis of tristyly in tetraploid Oxalis alpina (Oxalidaceae)

The inheritance of style‐morphs was investigated in tetraploid populations of tristylous Oxalis alpina (Oxalidaceae) to determine if alleles controlling style‐morphs are expressed at duplicated loci. In tetraploid populations, a dominant S allele leads to expression of the short‐styled phenotype at the short/non‐short locus and is epistatic to the M allele at the mid/long locus. The M allele results in expression of the mid‐styled phenotype but only if the S allele is absent. Long‐styled morphs are homozygous recessive at the short and mid loci. Test crosses of many tetraploid short‐styled individuals resulted in segregations of short‐, mid‐ and long‐styled individuals which, because of linkage between the short and mid loci, can only occur with polyploidy and expression of alleles at duplicated loci. Segregation patterns from three crosses suggest the possibility of disomic inheritance via preferential pairing of chromosomes in tetraploid populations of O. alpina. Segregation patterns in the progeny of mid‐styled individuals indicated that only a few individuals had more than one copy of the M allele, despite the potential for accumulation of M alleles via self‐fertilization of partially self‐compatible mid‐styled morphs in some populations. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 308–318.     

Serum esterase genetics in rabbits. II. Genetic analysis of the prealbumin esterase system,including atropinesterase and cocainesterase polymorphism

The zymotypic variation of rabbit prealbumin esterases is controlled by three autosomal loci, each with two alleles: Est-1 ^S and Est-1 ^s, Est-2^F and Est-2 ^f′, Est-3^D and Est-3 ^d. Est-1^S gives rise to the three S zones possessing the cocainesterase activity, Est-2 ^F to the three F zones with atropinesterase activity. Presence of the latter allele is never manifested without the Est-1 ^S allele. Est-3 ^D codes for the D zone. This D esterase reacts with the currently used substrate α-naphthylacetate only in the presence of the F zones. Est-1 and Est-2 loci are closely linked (<0.5% recombination); Est-3 shows no coupling with Est-1 and Est-2. The Est-1 ^S and Est-3 ^D alleles have a complete dominant expression, whereas the Est-2 alleles are codominant. Gene frequencies of the Est-1 and Est-2 loci vary between the examined breeds. A Hardy-Weinberg equilibrium is found in two populations (Cpb:ALU and Cpb:VW). A significant surplus of heterozygotes is demonstrated in a third population (Cpb:CH).     

Efficacy of pyramiding elite alleles for dynamic development of plant height in common wheat

Plant height is an important botanical feature closely related to yield. Two populations consisting of 118 and 262 accessions respectively were used to identify elite alleles for plant height and to validate their allelic effects. Plant height was measured from the early booting to the flowering stages. Simple sequence repeat markers for candidate quantitative trait locus (QTL) regions with large effects identified in a doubled haploid (DH) population (Hanxuan 10 × Lumai 14) were selected for further verification by association analysis. Nine loci significantly (P < 0.001) associated with plant height were detected 13 times in the population with 118 accessions. Three loci (Xgwm11-1B, Xwmc349-4B and Xcfd23-4D) were identified in three, two and two periods of plant height growth, respectively. Markers Xbarc168-2D, Xgwm249-2D, Xwmc349-4B, Xcfd23-4D and Xgwm410-5A located at or near additive QTL regions in the DH population proved to coincide with known Rht loci. The results showed a consistency between linkage analysis and association mapping, and also confirmed the value of fine mapping of QTL through combined linkage and association analyses. For final plant height, the alleles Xgwm11-1B ₂₀₈ , Xwmc349-4B ₁₀₃ and Xcfd23-4D ₂₀₂ exhibited negative effects, i.e. reducing plant height; Xwmc349-4B ₁₀₁ and Xcfd23-4D ₂₀₅ showed significant positive effects. A second larger population (262 accessions) was used to validate the effects of these large-effect alleles and the efficacy of pyramiding in eight environments (year × site × water regime combinations). Strong correlations between final plant height and numbers of large-effect alleles indicated that the alleles contributed additively to plant height. The additive effects showed that pyramiding elite alleles for target traits has significant potential for wheat breeding.     

Genetic analysis of disease resistance to all strains of BaYMV in a Chinese barley landrace, Mokusekko 3

 A Chinese landrace of barley, Mokusekko 3, is unique in being completely resistant against all strains of barley yellow mosaic virus (BaYMV). The present investigation revealed that the resistance of Mokusekko 3 is governed by two recessive genes. As one of the resistance genes was known to be tightly linked with alleles at the Est complex locus, consisting of the Est1, Est2 and Est4 loci for esterase isozymes, each of the resistance genes could be separated by means of marker-assisted selection using an isozyme allelic combination as a marker. One of the resistance genes, ym1, is linked to K (hooded lemma) and gl3 (glossy leaf 3) with recombination values of 25.3% and 9.7% respectively, and these three genes are located in the order K-gl3-ym1 on chromosome 4. Another newly designated resistance gene, ym5, is linked to alleles at the Est complex locus and cu2 (curly growth 2), with recombination values of 1.9% and 19.5% respectively, in the order cu2-Est-ym5 from proximal to distal on the long arm of chromosome 3. The complete resistance of Mokusekko 3 is caused by combining two resistance genes, ym1 and ym5. However, almost all the “resistant” cultivars derived from crosses with Mokusekko 3 are susceptible to the recently detected strain BaYMV-III in Japan, since they contain only one resistance gene, ym5. Marker-assisted selection to combine resistance genes into a cultivar is discussed for the breeding of stabilizing resistance to BaYMV. Received: 23 September 1996 / Accepted: 8 November 1996     

Sporophytic self-incompatibility in Senecio squalidus (Asteraceae): S allele dominance interactions and modifiers of cross-compatibility and selfing rates

Understanding genetic mechanisms of self-incompatibility (SI) and how they evolve is central to understanding the mating behaviour of most outbreeding angiosperms. Sporophytic SI (SSI) is controlled by a single multi-allelic locus, S, which is expressed in the diploid (sporophyte) plant to determine the SI phenotype of its haploid (gametophyte) pollen. This allows complex patterns of independent S allele dominance interactions in male (pollen) and female (pistil) reproductive tissues. Senecio squalidus is a useful model for studying the genetic regulation and evolution of SSI because of its population history as an alien invasive species in the UK. S. squalidus maintains a small number of S alleles (7–11) with a high frequency of dominance interactions. Some S. squalidus individuals also show partial selfing and/or greater levels of cross-compatibility than expected under SSI. We previously speculated that these might be adaptations to invasiveness. Here we describe a detailed characterization of the regulation of SSI in S. squalidus. Controlled crosses were used to determine the S allele dominance hierarchy of six S alleles and effects of modifiers on cross-compatibility and partial selfing. Complex dominance interactions among S alleles were found with at least three levels of dominance and tissue-specific codominance. Evidence for S gene modifiers that increase selfing and/or cross-compatibility was also found. These empirical findings are discussed in the context of theoretical predictions for maintenance of S allele dominance interactions, and the role of modifier loci in the evolution of SI.     

Genome and population dynamics under selection and neutrality: an example of S-allele diversity in wild cherry (Prunus avium L.)

Self-incompatibility, a common attribute of plant development, forms a classical paradigm of balancing selection in natural populations, in particular negative frequency-dependent selection. Under negative frequency-dependent selection population genetics theory predicts that the S-locus, being in command of self-incompatibility, keeps numerous alleles in equal frequencies demonstrating a wide allelic range. Moreover, while natural populations exhibit a higher within population genetic diversity, a reduction of population differentiation and increase of effective migration rate is expected in comparison to neutral loci. Allelic frequencies were investigated in terms of distribution and genetic structure at the gametophytic self-incompatibility locus in five wild cherry (Prunus avium L.) populations. Comparisons were also made between the differentiation at the S-locus and at the SSR loci. Theoretical expectations under balancing selection were congruent to the results observed. The S-locus showed broad multiplicity (16 S-alleles), high genetic diversity, and allelic isoplethy. Genetic structure at the self-incompatibility locus was almost four times lower than at 11 nSSR loci. Analysis of molecular variance revealed that only 5?% of the total genetic variation concerns differentiation among populations. In conclusion, the wealth of S-allelic diversity found in natural wild cherry populations in Greece is useful not only in advancing basic population genetics research of self-incompatibility systems in wild cherry but also in the development of breeding programs.     

Further genetic variation at the esterase loci of Drosophila virilis

Reexamination of the electrophoretic mobilities of esterases encoded by the Est- and the Est- alleles of Drosophila virilis was carried out in detail using both thin-layer agar gel and polyacrylamide slab gel electrophoresis. Many allelic products with fine differences in their electrophoretic mobilities were found and designated by a new system. Some esterases separable by the agar gel method were indistinguishable using the polyacrylamide gel method. But the polyacrylamide gel method uncovered two multiband homozygotes, ^(d).77 and ^(d)1.28. Some allelic frequencies on the basis of the new designation were estimated in two natural populations. As a result, it is proposed that the total scope of allelic variation at the two esterase loci of Drosophila virilis is composed of discrete distribution patterns of gene frequencies, each histogram of which shows a bell-shaped pattern.     

Detection of regional allozyme divergence in the rocky intertidal goby Chaenogobius annularis

Genetic differentiation in the intertidal goby Chaenogobius annularis was studied using allozyme markers. Samples were collected from six localities along the coasts of Japanese islands and the Korean Peninsula. Six out of 13 loci showed allelic variation in at least one population. Although the predominant alleles of 12 loci were the same among all populations, Mdh-1 showed clear differences among the populations located along the coasts of the Pacific Ocean and the Sea of Japan. These two possible geographic groups, the Pacific Ocean and the Sea of Japan groups, were characterized by diagnostic alleles of Mdh-1, namely Mdh-1 ¹⁰⁰ and Mdh-1 ⁷⁰.     

Purple Phototrophic Bacterium Enhances Stevioside Yield by Stevia rebaudiana Bertoni via Foliar Spray and Rhizosphere Irrigation

This study was conducted to compare the effects of foliar spray and rhizosphere irrigation with purple phototrophic bacteria (PPB) on growth and stevioside (ST) yield of Stevia. rebaudiana. The S. rebaudiana plants were treated by foliar spray, rhizosphere irrigation, and spray plus irrigation with PPB for 10 days, respectively. All treatments enhanced growth of S. rebaudiana, and the foliar method was more efficient than irrigation. Spraying combined with irrigation increased the ST yield plant ^-1 by 69.2% as compared to the control. The soil dehydrogenase activity, S. rebaudiana shoot biomass, chlorophyll content in new leaves, and soluble sugar in old leaves were affected significantly by S+I treatment, too. The PPB probably works in the rhizosphere by activating the metabolic activity of soil bacteria, and on leaves by excreting phytohormones or enhancing the activity of phyllosphere microorganisms.     

Synthesis,antimycobacterial and cytotoxic activity of α,β-unsaturated amides and 2,4-disubstituted oxazoline derivatives

The synthesis of six α,β,-unsaturated amides and six 2,4-disubstituted oxazolines derivatives and their evaluation against two Mycobacterium tuberculosis strains (sensitive H37Rv and a resistant clinical isolate) is reported. 2,4-Disubstituted oxazolines (S)-3b,d,e were the most active in the sensitive strain with a MIC of 14.2, 13.6 and 10.8 μM, respectively, and the compounds (S)-3d,f were the most active against resistant strain with a MIC of 6.8 and 7.4 μM. The ex-vivo evaluation of hepatotoxicity on precision-cut rat liver slices was also tested for the α,β-unsaturated amides (S)-2b and (S)-2d,f and for the oxazolines (S)-3b and (S)-3d,f at different concentrations (5, 15 and 30 μg/mL). The results indicate that these compounds possess promising antimycobacterial activity and at the same time are not hepatotoxic. These findings open the possibility for development of new drugs against tuberculosis.     

Dulcosides A and B,new diterpene glycosides from Stevia rebaudiana

New diterpene glycosides, dulcosides A and B were isolated from Stevia rebaudiana Bertoni and their structures were established as 19-O-β-glu     

Quantitative resistance differences between and within natural populations of Solanum chilense against the oomycete pathogen Phytophthora infestans

The wild tomato species Solanum chilense is divided into geographically and genetically distinct populations that show signs of defense gene selection and differential phenotypes when challenged with several phytopathogens, including the oomycete causal agent of late blight Phytophthora infestans. To better understand the phenotypic diversity of this disease resistance in S. chilense and to assess the effect of plant genotype versus pathogen isolate, respectively, we evaluated infection frequency in a systematic approach and with large sample sizes. We studied 85 genetically distinct individuals representing nine geographically separated populations of S. chilense. This showed that differences in quantitative resistance can be observed between but also within populations at the level of individual plants. Our data also did not reveal complete immunity in any of the genotypes. We further evaluated the resistance of a subset of the plants against P. infestans isolates with diverse virulence properties. This confirmed that the relative differences in resistance phenotypes between individuals were mainly determined by the plant genotype under consideration with modest effects of pathogen isolate used in the study. Thus, our report suggests that the observed quantitative resistance against P. infestans in natural populations of a wild tomato species S. chilense is the result of basal defense responses that depend on the host genotype and are pathogen isolate‐unspecific.     

Occurrence of some mono-cis-isomers of asymmetric C40-carotenoids

The 9-cis-isomers of antheraxanthin [(3S,5R,6S)-5,6-epoxy-5,6- dihydro-β, β-carotene-3,3′-diol] and lutein epoxide [(3S,5R,6S, 3′R,6′R)-5,6-epoxy-5,6-dihydro-β, ε-carotene-3,3′-diol] were found to occur without their 9′-cis counterparts in the non-photosynthetic tissues of several higher plants. 9-cis-lutein [(3R,3′R,6′R)- β,ε-carotene-3,3′-diol], on the other hand, was observed together with its 9′-cis counterpart in the samples investigated. The qualitative distribution of carotenoids is also reported.     

Synthesis of anomeric sulfonamides and their behaviour under radical-mediated bromination conditions

O-Peracetylated methyl 3-(d-glycopyranosylthio)propanoates of β-d-gluco, and α- and β-d-galacto configurations were oxidized to the corresponding S,S-dioxides (sulfones) by Oxone^® or MCPBA. Oxidation of the β-d-gluco derivative with H₂O₂/Na₂WO₄ gave the corresponding S-oxide (sulfoxide). DBU-induced elimination of methyl acrylate from the β-d-gluco and β-d-galacto configured S,S-dioxides (sulfones) gave O-peracetylated β-d-glycopyranosyl-1-C-sulfinates which, on treatment with H₂NOSO₃H, furnished the corresponding β-d-glycopyranosyl-1-C-sulfonamides. Radical-mediated bromination of the protected methyl 3-(β-d-glycopyranosylthio)propanoate S,S-dioxides gave mixtures of 1-C- and 5-C-bromoglycosyl compounds. Similar brominations of the O-peracetylated β-d-glycopyranosyl-1-C-sulfonamides resulted in the formation of α-d-glycopyranosyl bromides and 1-C- and 5-C-bromoglycosyl sulfonamides. A rationale for these observations was proposed. Methyl 3-(β-d-glucopyranosylthio)propanoate, its S,S-dioxide, and β-d-glucopyranosyl-1-C-sulfonamide proved inefficient when tested as inhibitors of rabbit muscle glycogen phosphorylase b.     

Evidence for the formation of a steroid S-glutathione conjugate from an epoxysteroid precursor.

Biotransformation of [4-¹⁴C]cholesterol α-epoxide (5α,6α-epoxycholestan-3β-ol) to the S-glutathione conjugate, 3β,5α-dihydroxycholestan-6β-yl-S-glutathione, by S-glutathione transferase of the rat liver soluble supernatant fraction, has been described. After the isolation from the biological incubation mixture by n-butanol extraction, followed by the Amberlite XAD-2 column treatment, the conjugate was chromatographically identified with a synthetic specimen which was prepared from cholesterol α-epoxide and glutathione in an ethanolic solution of sodium hydroxide. Further identification of the biologically formed conjugate with the synthetic one, including structural assignment, was established from the result that they yielded the same desulfrization product, 3β,5α-dihydroxycholestane, by the treatment with Raney nickel in an atmosphere of hydrogen.