Alex Beharav

Retrotransposon BARE-1 and Its Role in Genome Evolution in the Genus Hordeum

The replicative retrotransposon life cycle offers the potential for explosive increases in copy number and consequent inflation of genome size. The BARE-1 retrotransposon family of barley is conserved, disperse, and transcriptionally active. To assess the role of BARE-1 in genome evolution, we determined the copy number of its integrase, its reverse transcriptase, and its long terminal repeat (LTR) domains throughout the genus Hordeum. On average, BARE-1 contributes 13.7 × 103 full-length copies, amounting to 2.9% of the genome. The number increases with genome size. Two LTRs are associated with each internal domain in intact retrotransposons, but surprisingly, BARE-1 LTRs were considerably more prevalent than would be expected from the numbers of intact elements. The excess in LTRs increases as both genome size and BARE-1 genomic fraction decrease. Intrachromosomal homologous recombination between LTRs could explain the excess, removing BARE-1 elements and leaving behind solo LTRs, thereby reducing the complement of functional retrotransposons in the genome and providing at least a partial “return ticket from genomic obesity.”

RAPD polymorphism of wild emmer wheat populations, Triticum dicoccoides, in Israel

Abstract Genetic diversity in random amplified polymorphic DNAs (RAPDs) was studied in 110 genotypes of the tetraploid wild progenitor of wheat, Triticum dicoccoides, from 11 populations sampled in Israel and Turkey. Our results show high level of diversity of RAPD markers in wild wheat populations in Israel. The ten primers used in this study amplified 59 scorable RAPD loci of which 48 (81.4%) were polymorphic and 11 monomorphic. RAPD analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides originating from diverse ecogeographical sites in Israel and Turkey, with 95.5% of the 100 genotypes correctly classified into sites of origin by discriminant analysis based on RAPD genotyping. However, interpopulation genetic distances showed no association with geographic distance between the population sites of origin, negating a simple isolation by distance model. Spatial autocorrelation of RAPD frequencies suggests that migration is not influential. Our present RAPD results are non-random and in agreement with the previously obtained allozyme patterns, although the genetic diversity values obtained with RAPDs are much higher than the allozyme values. Significant correlates of RAPD markers with various climatic and soil factors suggest that, as in the case of allozymes, natural selection causes adaptive RAPD ecogeographical differentiation. The results obtained suggest that RAPD markers are useful for the estimation of genetic diversity in wild material of T. dicoccoides and the identification of suitable parents for the development of mapping populations for the tagging of agronomically important traits derived from T. dicoccoides.

Ecological-genomic diversity of microsatellites in wild barley, Hordeum spontaneum, populations in Jordan.

Abstract.We analyzed the ecological-genomic diversity of microsatellites of wild barley, Hordeum spontaneum (C. Koch) Thell., at 18 loci in 306 individuals of 16 populations from Jordan across a southward transect of increasing aridity. The 18 microsatellites revealed a total of 249 alleles, with an average of 13.8 alleles per locus (range 3–29), with nonrandom distribution. The proportion of polymorphic loci per population averaged 0.91 (range 0.83–1.00); gene diversity, He, averaged 0.512 (range 0.38–0.651). We compared the number of alleles of the 18 loci to those found in Israel populations by Turpeinen et al.. Out of the 280 alleles, 138 (49.3%) were unique (i.e. occurred in only one of the countries). The percentage of unique alleles in Jordan and Israel populations was 43.0% and 17.9%, respectively, suggesting that Jordan is an important center of origin and diversity of wild barley. Estimates of mean gene diversity were highest in the populations collected near the Golan Heights, such as Shuni North, Shuni South and Jarash. Sixty nine percent of the microsatellite variation was partitioned within populations and 31% between populations. Associations between ecogeographical values and gene diversity were established for eight microsatellite loci. The cluster produced by simple sequence repeat (SSR) data is mostly coincidence with the result of the dendrogram of the Spalax ehrenbergi superspecies of subterranean mole rats in Jordan based on allozyme gene loci. The major soil type in the wild barley habitat of each ecological group was different. Stepwise multiple regression analysis indicated that the variance of gene diversity was explained by altitude (R2 = 0.362**). These observations suggest that microsatellites are at least partly adaptive and subject to natural selection. Electronic Supplementary Material is available if you access this article at http://dx.doi.org/10.1007/s00122-002-1029-7. On that page (frame on the left side), a link takes you directly to the supplementary material.

Adaptive Melanin Response of the Soil Fungus Aspergillus niger to UV Radiation Stress at “Evolution Canyon”, Mount Carmel, Israel

Background Adaptation is an evolutionary process in which traits in a population are tailored by natural selection to better meet the challenges presented by the local environment. The major discussion relating to natural selection concerns the portraying of the cause and effect relationship between a presumably adaptive trait and selection agents generating it. Therefore, it is necessary to identify trait(s) that evolve in direct response to selection, enhancing the organisms fitness. “Evolution Canyon” (EC) in Israel mirrors a microcosmic evolutionary system across life and is ideal to study natural selection and local adaptation under sharply, microclimatically divergent environments. The south-facing, tropical, sunny and xeric “African” slope (AS) receives 200%–800% higher solar radiation than the north-facing, temperate, shady and mesic “European” slope (ES), 200 meters apart. Thus, solar ultraviolet radiation (UVR) is a major selection agent in EC influencing the organism-environment interaction. Melanin is a trait postulated to have evolved for UV-screening in microorganisms. Here we investigate the cause and effect relationship between differential UVR on the opposing slopes of EC and the conidial melanin concentration of the filamentous soil fungus Aspergillus niger. We test the working hypothesis that the AS strains exhibit higher melanin content than strains from the ES resulting in higher UV resistance. Methodology/Principal Findings We measured conidial melanin concentration of 80 strains from the EC using a spectrophotometer. The results indicated that mean conidial melanin concentration of AS strains were threefold higher than ES strains and the former resisted UVA irradiation better than the latter. Comparisons of melanin in the conidia of A. niger strains from sunny and shady microniches on the predominantly sunny AS and predominantly shady ES indicated that shady conditions on the AS have no influence on the selection on melanin; in contrast, the sunny strains from the ES displayed higher melanin concentrations. Conclusions/Significance We conclude that melanin in A. niger is an adaptive trait against UVR generated by natural selection.

RAPD divergence caused by microsite edaphic selection in wild barley

Random amplified polymorphic DNA polymerase chain reaction (RAPDPCR) was used to assess genetic diversity in four subpopulations (86 individuals) of wild barley, Hordeum spontaneum, sampled from Tabigha microsite near the Sea of Galilee, Israel. The microsite consists of two 100 m transects that are topographically separated by 100 m, each equally subdivided into 50 m of basalt and terra rossa soil types. Despite the same macroclimate characterizing the area around the Sea of Galilee, the microsite offers two edaphically different microhabitats, with basalt being a more ecologically heterogeneous and broader-niche than the relatively drier but more homogeneous and narrow-niche terra rossa. Analysis of 118 putative loci revealed significant (P<0.05) genetic differentiation in polymorphism (P0.05) between the two soils across the transects with P being higher in the more heterogeneous basalt (mean P0.05 = 0.902), than in terra rossa (mean P0.05 = 0.820). Gene diversity (He) was higher in basalt (mean He=0.371), than in terra rossa (mean He=0.259). Furthermore, unique alleles were confined to one soil type, either in one or both transects. Rare alleles were observed more frequently in terra rossa than basalt, and in transect II only. Gametic phase disequilibria showed a larger multilocus association of alleles in basalt than terra rossa, and in transect I than II. Spearman rank correlation (rs) revealed a strong association between specific loci and soil types, and transects. Also, analysis of multilocus organization revealed soil-specific multilocus-genotypes. Therefore, our results suggest an edaphically differentiated genetic structure, which corroborates the niche width-variation hypothesis, and can be explained, in part, by natural selection. This pattern of RAPD diversity is in agreement with allozyme and hordein protein diversities in the same subpopulations studied previously.

Polymorphism at rDNA loci in barley and its relation with climatic variables

Abstract The variation in length of the intergenic spacer (IGS) region of the ribosomal DNA repeat unit was examined in 63 accessions of wild barley, Hordeum spontaneum, and seven accessions of cultivated barley, Hordeum vulgare. The accessions of wild barley were collected from ecologically diverse climatic and edaphic microsites in Israel, and the barley cultivars were those grown in India. Sixteen spacer-length variants (slvs) observed in the present study presumably belonged to two known rDNA loci (Rrn1 and Rrn2). Each accession had one or more variants, which together represented the rDNA phenotype. The rDNA phenotypes of wild barley accessions were widely diverse and differed substantially from those of cultivated barley. The slv phenotypes and the corresponding alleles were shown to be largely correlated with different climatic, edaphic and ecogeographical microsites and niches (the ”Evolution Canyon” at Lower Nahal Oren, Mount Carmel; and Tabigha, Eastern Upper Galilee Mountains), so that a particular rDNA phenotype of an accession could be used to predict the climate and soil to which the accession belonged. This sharp microsite ecogeographic variation in ribosomal DNA appears adaptive in nature, and is presumably driven by climatic and edaphic natural selection.

Microscale ecological stress causes RAPD molecular selection in wild barley, Neve Yaar microsite, Israel

Genetic micro-differentiation was studied in wild barley at the Neve Yaar microsite, Israel, an area of 3182 m2, which includes a mosaic of sun-rock, sun-soil, shade-rock, and shade-soil microniches. Fifty-six genotypes were tested for ecological-genetic microniche correlates. Analysis of 75 RAPD loci revealed a significantly higher polymorphism and gene diversity (P0.05 = 0.920; He = 0.411) on the more stressful sun-rock microniche, compared to the least stressful shade-soil microniche (P0.05 = 0.653; He = 0.188). Fifty-six (74.7%) out of the 75 loci varied significantly in allele frequencies. On the average, of the 75 loci, 75.7% of the total genetic diversity exists within the four microniches, while 24.3% exists between the microniches. The highest genetic distance was between shade-soil and sun-rock (D = 0.220), which confirmed their sharp microecological contrasts. Gametic phase disequilibria (LD) differed significantly between sunny and shady microniches. The levels of correct classification of individual genotypes into their respective microniche, based on the presence/absence of bands of the 2, 3, 4, 5, and 7 best differentiating loci, obtained by stepwise discriminant analysis, were 79, 84, 89, 95, and 100%, respectively. We conclude that ecological contrasts at the Neve Yaar microsite cause molecular diversifying selection (presumably at both coding and noncoding genomic regions of wild barley) overriding gene flow.

AFLP analysis of Lactuca saligna germplasm collections from four European and three Middle Eastern countries

Lactuca saligna L. (least lettuce, willow-leaf lettuce) is widely distributed around the Mediterranean basin and extends to the Caucasus and temperate Europe; its extra-European distribution covers the Middle East. The aim of this study was to estimate the level of molecular variation and the extent of genetic differentiation of L. saligna in accessions collected in natural and semi-natural habitats in Europe and the Middle East. In sum, 67 accessions of L. saligna, originating in four European countries (Czech Republic, France, Italy, Portugal) and three Middle Eastern countries (Israel, Jordan, Turkey), were compared on the basis of AFLP polymorphism. AFLP analysis of the accessions revealed 415 (84.7%) polymorphic fragments out of 490 fragments amplified by means of seven primer pair combinations. The number of bands produced by individual primer pair combinations ranged from 44 to 101. The UPGMA dendrogram generated from Jaccards similarity matrix showed a similar genetic background of accessions from the Middle East (Turkey, Israel, Jordan), which were quite distinct from the group of accessions from the European part of the Mediterranean basin (Italy, France, Portugal). Surprisingly, two accessions, one from the Czech Republic (central Europe) and one from Israel, proved genetically wholly separate from all other accessions of L. saligna and are themselves wholly separate. Accessions originating in various eco-geographical conditions were found to differ significantly in their genetic polymorphism.

Mammalian microevolution in action: adaptive edaphic genomic divergence in blind subterranean mole–rats

Genomic diversity of anonymous regions across the genome, most probably including coding and noncoding amplified fragment length polymorphisms (AFLPs), was examined in 20 individuals of the blind mole–rat, Spalax galili, one of four allospecies of the Spalax ehrenbergi superspecies of blind subterranean mole–rats in Israel. We compared 10 individuals from two nearby populations in Upper Galilee, separated by only a few dozen to hundreds of metres and living in two sharply contrasting ecologies: white chalk and rendzina soil with Sarcopterium spinosum and Majorana syriaca versus black volcanic basalt soil with Carlina hispanica-Psorelea bitominosa and Alhagi graecorum plant formations. The microsite tested ranged in an area of less than 10 000 m2. Out of 729 AFLP loci, 433 (59.4%) were polymorphic, with 211 soil unique alleles. Genetic polymorphism was significantly higher on the ecologically more xeric and stressful chalky rendzina soil than on the neighbouring mesic basalt soil. This is a remarkable pattern for a mammal that can disperse each generation between tens to hundreds of metres. These results cannot be explained by migration (which causes homogenization) or by chance (which will exclude sharp genomic soil divergence). Natural selection is the only evolutionary adaptive force that can cause genetic divergence across the genome matching the sharp microscale ecological contrast.

Root tubers of Lactuca tuberosa as a source of antioxidant phenolic compounds and new furofuran lignans.

From the root tubers of Lactuca tuberosa, a wild edible plant species, nine phenolic compounds were isolated, including two new furofuran lignan glucosides, named lactuberin A and lactuberin B. Their structures were elucidated by spectroscopic methods, especially HRESIMS and 2D NMR techniques. This is the first time that compounds belonging to the epi series of 2,6-diaryl-3,7-dioxabicyclo[3.3.0]octane type furofuran lignans have been found in Lactuca species. The total phenolic content of the root tuber extract was evaluated and its major phenolic constituents, caffeic acid, chlorogenic acid and 3,5 dicaffeoylquinic acid, known to possess antioxidant activity, were quantified. Additionally, the root tuber extract showed DPPH radical scavenging activity implying its potential as functional food.