Evidence for pre-zygotic reproductive barrier between the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae).

The degree of reproductive isolation between the B and Q biotypes of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is currently not clear. Laboratory experiments have shown that the two biotypes are capable of producing viable F1 hybrids but that these females are sterile as their F2 generation failed to develop, indicating, most likely, a post-zygotic reproductive barrier. Here, we confirm, by molecular and ecological tools, that the B and Q biotypes of Israel are genetically isolated and provide two independent lines of evidence that support the existence of a pre-zygotic reproductive barrier between them. Firstly, monitoring of mating behaviors in homogeneous and heterogeneous couples indicated no copulation events in heterogeneous couples compared to approximately 50% in homogeneous B and Q couples. Secondly, we could not detect the presence of sperm in the spermathecae of females from heterogeneous couples, compared to 50% detection in intra-B biotype crosses and 15% detection in intra-Q biotype crosses. The existence of pre-zygotic reproductive barriers in Israeli B and Q colonies may indicate a reinforcement process in which mating discrimination is strengthened between sympatric taxa that were formerly allopatric, to avoid maladaptive hybridization. As the two biotypes continued to perform all courtship stages prior to copulation, we also conducted mixed cultures experiments in order to test the reproductive consequences of inter-biotype courtship attempts. In mixed cultures, a significant reduction in female fecundity was observed for the Q biotype but not for the B biotype, suggesting an asymmetric reproductive interference effect in favour of the B biotype. The long-term outcome of this effect is yet to be determined since additional environmental forces may reduce the probability of demographic displacement of one biotype by the other in overlapping niches.

The emergence of life on Earth.

Combined top-down and bottom-up research strategies and the principle of biological continuity were employed in an attempt to reconstruct a comprehensive origin of life theory, which is an extension of the coevolution theory (Lahav and Nir, Origins of Life Evol. Biosphere (1997) 27, 377-395). The resulting theory of emergence of templated-information and functionality (ETIF) addresses the emergence of living entities from inanimate matter, and that of the central mechanisms of their further evolution. It proposes the emergence of short organic catalysts (peptides and proto-ribozymes) and feedback-loop systems, plus their template-and-sequence-directed (TSD) reactions, encompassing catalyzed replication and translation of populations of molecules organized as chemical-informational feedback loop entities, in a fluctuating (wetting-drying) environment, functioning as simplified extant molecular-biological systems. The feedback loops with their TSD systems are chemically and functionally continuous with extant living organisms and their emergence in an inanimate environment may be defined as the beginning of life. The ETIF theory considers the emergence of bio-homochirality, a primordial genetic code, information and the incorporation of primordial metabolic cycles and compartmentation into the emerging living entities. This theory helps to establish a novel measure of biological information, which focuses on its physical effects rather than on the structure of the message, and makes it possible to estimate the time needed for the transition from the inanimate state to the closure of the first feedback-loop systems. Moreover, it forms the basis for novel laboratory experiments and computer modeling, encompassing catalytic activity of short peptides and proto-RNAs and the emergence of bio-homochirality and feedback-loop systems.

A model for proto-tRNA loading.

A kinetic model for loading of proto-tRNA is presented. The kinetic parameters were first estimated from the results of Francklyn & Schimmel (1989;Nature337, 478--481), who studied the aminoacylation of both tRNA and its minihelix. Then these parameters were reduced several-fold, as is more appropriate for the prebiotic world. The simulations revealed a very slow time course of the loading reaction. We also consider a possibility for the proto-tRNA loading without a catalyst and discuss the feasibility of such processes. Analytical approximations are presented for the kinetics of proto-tRNA loading with and without enzyme. An estimate is given for the time required for the development of template- and sequence-directed systems.

Simulation of non-enzymatic template-directed synthesis of oligonucleotides and peptides.

A simulation model is proposed for the template- and sequence-directed (TSD) condensation of two trideoxyribonucleotide 3'-phosphate molecules into a hexameric template with palindromic sequence studied experimentally by von Kiedrowski (1986;Angew. Int. Ed. Engl.25, 932--935). The model simulates reasonably well the kinetics of synthesis of both the template, and the pyrophosphate product which is not directly involved in the autocatalytic reaction. It offers quantitative approximation of the different rate constants of the processes involved in the reaction. The model simulates and gives predictions for the influence of factors such as the initial concentrations of the trimers and the template, and gives predictions for the effect of temperature on the dynamics of the autocatalytic reaction. The model also simulates well the production rate of a different self-replicating system (coiled coil peptide) used in the experiments of Lee et al. (1997;Nature390, 591--594). Comparing the different rate constants, it seems that chain elongation occurs at higher rates in the peptide system (at 23 degrees C) than in the nucleotide one (at 0 degrees C), but that the relative contribution of template-directed synthesis is significantly larger with the nucleotides.

Infrared Study of the Intercalation of Kaolinite by Caesium Bromide and Caesium Iodide.

CsBr- and CsI-kaolinite intercalation complexes were synthesized by gradually heating caesium halide disks of the DMSO-kaolinite intermediate up to 330 degreesC. Infrared spectroscopy revealed two types of complexes with the caesium salts: almost nonhydrous, obtained during thermal treatment of the DMSO complex, and hydrated, produced by regrinding the disk in air. Comparison of band positions for CsBr-kaolinite and CsI-kaolinite with those for the CsCl complex (observed in a previous study) shows that the strength of the hydrogen bond between the intercalated halide and the inner surface hydroxyl decreases on the order CsCl > CsBr > CsI. The nonreactivity of CsI in mechanochemical intercalation may arise from weak interaction between I- and inner surface hydroxyl groups, resulting from the fact that caesium is a very soft acid and iodide is a very soft base. Consequently, the very strong interaction between the two ions in the crystal is not disrupted during mechanochemical treatment. Copyright 1998 Academic Press and Minister of Natural Resources, Canada.

Emergence of template-and-sequence-directed (TSD) syntheses: I. A bio-geochemical model.

A biogeochemical model for the evolution of template-and-sequence-directed (TSD) syntheses of biological templates (proto-RNAs) and catalysts (peptides) is described. A fluctuating environment characterized by hydrating (cool) and dehydrating (warm) phases with cycles of consecutive organic reactions, as well as a constant supply of the polymeric building blocks is assumed. The scenario starts with the catalyzed formation of a primordial population of small random peptides, based on the relatively-ineffective mineral catalysts. The resulting peptides initiate a catalytic takeover process, during which the catalytic functions are gradually taken over by peptides. The evolution of TSD peptides is based on a combination of Lahav's (1991) co-evolution and Moller and Janssen's (1990) specific recognition sites hypotheses. During the emergence of TSD systems the fraction of TSD peptides and proto-RNA constituents rises from almost insignificance to dominance in a TSD Reactions Takeover. The TSD system is characterized by autocatalysis, positive feedback loops and a primordial genetic code. The model is the basis for a computer program (Part II of present series).

Emergence of template-and-sequence-directed (TSD) syntheses: II. A computer simulation model.

The initiation of the bio-geochemical scenario described in Part I serves in the present work as the basis for computer modeling, where the central process of the simulation algorithm. i.e., peptide-catalyzed oligomeric growth, is based on mass action equations. The computer model starts with a minimal system in which catalyzed growth processes of proto-RNA templates and small peptides take place, starting from their building blocks. The emerging populations of random oligomers also include a very small fraction of proto-tRNAs and a small fraction of catalytic peptides. Using simplifying assumptions regarding catalyzed proto-RNA template-replication, as well as selectivity of certain molecules and processes, the proportion of proto-tRNA in the proto-RNA molecular population increases rapidly; it is followed by TSD peptide synthesis, based on an ad hoc genetic code and specific peptide catalysts allocated for this synthesis. Consequently, a feedback system is initiated in which TSD peptides involved in the relevant catalytic reactions of the TSD syntheses also start to accumulate. The initial sporadic formation of TSD peptides is thus replaced gradually by cycles of positive feedback and autocatalysis characterized by accumulation of catalytic peptides and Proto-tRNAs and TSD-Reaction-Takeover. The model system which can be considered a 'toy model' can synthesize its templates and catalysts under a wide range of reaction parameters and initial concentrations, thus demonstrating a robustness which is essential for molecular evolution processes. The critical stage of the buildup of a molecular mechanism for the initiation of a minimal TSD reaction cycle has thus been described; because of the centrality of TSD reaction cycles in biology, it is assumed to be central also in the origin of life processes.

The RNA-world and co-evolution hypotheses and the origin of life: implications, research strategies and perspectives.

The applicability of the RNA-world and co-evolution hypotheses to the study of the very first stages of the origin of life is discussed. The discussion focuses on the basic differences between the two hypotheses and their implications, with regard to the reconstruction methodology, ribosome emergence, balance between ribozymes and protein enzymes, and their major difficulties. Additional complexities of the two hypotheses, such as membranes and the energy source of the first reactions, are not treated in the present work. A central element in the proposed experimental strategies is the study of the catalytic activities of very small peptides and RNA-like oligomers, according to existing, as well as to yet-to-be-invented scenarios of the two hypotheses under consideration. It is suggested that the novel directed molecular evolution technology, and molecular computational modelling, can be applied to this research. This strategy is assumed to be essential for the suggested goal of future studies of the origin of life, namely, the establishment of a 'Primordial Darwinian entity'.

Prebiotic co-evolution of self-replication and translation or RNA world?

A prebiotic scenario is proposed, based on the recent "domain hypothesis" model (Lahav, 1989, J. molec. Evol. 29, 475-479), suggested for domain propagation of RNA-like molecules in a fluctuating environment. The same system is suggested now not only for the evolution of ribozymes, but also for the evolution of directed peptide synthesis, as follows: Short, self-structured strands (termed prebioectons), each possessing a templatable domain which is chargeable by an amino acid, are the predecessors of tRNA (proto-tRNA). Complementary domains are formed on these prebioectons during an environmental cycle such as wetting-drying, followed by their dissociation from their template domain and ligation, to form the predecessor of mRNA (proto-mRNA). The evolution of directed peptide synthesis is suggested to be based on the ability of the charged prebioectons to attach preferentially to their complementary domains on the proto-mRNA. Two stages of this process are envisioned, namely: (a) Template-directed, random peptide synthesis taking place when non-specifically-charged prebioectons are sequentially attached each to its complementary domain on the proto-mRNA, followed by peptide bond formation. (b) Template-and-sequence-directed peptide synthesis, which can be realized after the "invention" of a catalytic molecule capable of specifically charging a proto-tRNA by an amino acid; this is the crucial evolutionary stage, where a crude genetic code becomes functional. Gradually, catalytic peptides and ribozymes are selected for their functions and evolve, while being encoded in the primitive "memory" of the emerging system. Thus, rather than the RNA monopoly postulated by the RNA World hypothesis, an early co-evolution of primitive enzymes and ribozymes is suggested.(ABSTRACT TRUNCATED AT 250 WORDS)

Exon-intron-like pattern of the first propagating molecule?

A biogeochemical scenario is proposed, according to which prebiotic self-replication of RNA-like molecules evolved gradually from domain propagation to entire strand replication. The hypothetical replicating entities that formed an evolutionary continuity in time, the prebioectons, were characterized by self-structuring, and started their evolution from strands possessing one or more templatable domains. The inherent self-structuring implies a very early coevolution of the replicating molecules with prebiotic peptides, in line with the cassette model of Cedergren and Grosjean (1987). The proposed approach can be partially tested in the laboratory.

The biogeochemical cycle of the adsorbed template. II: Selective adsorption of mononucleotides on adsorbed polynucleotide templates.

Experimental results are presented for the verification of the specific interaction step of the 'adsorbed template' biogeochemical cycle, a simple model for a primitive prebiotic replication system. The experimental system consisted of gypsum as the mineral to which an oligonucleotide template attaches (Poly-C or Poly-U) and 5'-AMP, 5'-GMP, 5'-CMP and 5'-UMP as the interacting biomonomers. When Poly-C or Poly-U were used as adsorbed templates, 5'-GMP and 5'-AMP, respectively were observed to be the most strongly adsorbed species. Moreover, there exists a direct quantitative relationship between the quantity of cytidine or uracil residues in the adsorbed state and the amount of the complementary mononucleotide that is attached to it. NaCl added to the system in order to create conditions of high ionic strength seems to enhance the selectivity of the adsorption of the monmucleotides to these adsorbed templates.

Soluble minerals in chemical evolution. II. Characterization of the adsorption of 5'-AMP and 5'-CMP on a variety of soluble mineral salts.

The adsorption of 5'-AMP and 5'-CMP was studied in saturated solutions of several soluble mineral salts (NaCl, Na2SO4, MgCl2 X 6H2O, MgSO4 X 7H2O, CaCl2 X 2H2O, CaSO4 X 2H2O, SrCl2 X 6H2O, SrSO4, and ZnSO4 X 7H2O) as a function of pH, ionic strength, and surface area of the solid salt. The adsorption shows a pH dependence; this can be correlated with the charge on the nucleotide molecule which is determined by the state of protonation of the N-1 nitrogen of 5'-AMP or N-3 nitrogen of 5'-CMP and the phosphate oxygens. The adsorption which results from the binding between the nucleotide molecule and the salt surface is proposed as being due to electrostatic forces. It was concluded that the adsorption was reversible in nature. The adsorption shows a strong dependence upon ionic strength and decreases with increasing ionic strength. Surface area is shown to be an important factor in evaluating and comparing the magnitude of adsorption of nucleotides onto various mineral salts. The implications of the results of the study are discussed in terms of the importance of soluble mineral salts as adsorption sites in the characterization of the adsorption reactions of an adsorbed template in biogeochemical cycles.

The biogeochemical cycle of the adsorbed template. I: Formation of the template.

Experimental results are presented for the verification of the first adsorption step of the 'adsorbed template' biogeochemical cycle, a simple model for a primitive prebiotic replication system. The adsorption of Poly-C, Poly-U, Poly-A, Poly-G, and 5'-AMP, 5'-GMP, 5'-CMP and 5'-UMP onto gypsum was studied. It was found that under the conditions of the experiment, the polymers have a very high affinity for the mineral surface, while the monomers adsorb much less efficiently.

The synthesis of primitive 'living' forms: definitions, goals, strategies and evolution synthesizers.

The arbitrariness of the definition of life is discussed in relation to both the archaic biological entities that preceded cells during the Molecular Evolution era, and the hypothetical, primitive, 'living' entities that presumably can be synthesized in the laboratory. Several experimental approaches to the synthesis, detection, and characterization of 'living' entities are discussed. The experimental approaches considered for the synthesis are the constructionist strategy, the whole-environment strategy, and the modular strategy, which is a combination of the first two. The whole-environment strategy is discussed in more detail and the establishment of an Evolution Synthesizer, based on this strategy, is proposed and rationalized. The guidelines for the detection and characterization of populations and processes of 'living' entities include chemical and physical analyses, but are based mainly on the reproductive characterization of these entities. It is expected that the higher the evolutionary level of the 'living' entities, the longer and more difficult it will be to synthesize them, but the easier it will be to detect them.

Soluble minerals in chemical evolution. I. Adsorption of 5'-AMP on CaSO4--a model system.

The adsorption of 5'-AMP onto solid CaSO4 2H2O was studied in a saturated suspension as a function of pH and electrolyte concentration. The adsorption is pH-dependent and is directly correlated with the charge on the 5'-AMP molecule which is determined by the state of protonation of the N-1 nitrogen of the purine ring and the phosphate oxygens. It is proposed that the binding occurs between the nucleotide and the salt is electrostatic in nature. The adsorption decreases with increasing ionic strength of the solution which means that in a fluctuating environment of wetting and drying cycles, a biomolecule similar to 5'-AMP could be expected to desorb during the drying phase. The results indicate that CaSO4 2H2O can serve as a concentrating surface for biomolecules. The significance of this is discussed with regard to the possible role of soluble minerals and their surfaces in a geochemical model consistent with the evolution of the Earth and the origin of life.

Characterization of dehydration-induced luminescence of kaolinite.

Dehydration-induced luminescence (DIL), the emission of light from a clay paste upon dehydration, was characterized experimentally for a colloidal kaolinite. The relationship between total photon count of the emitted light and film thickness is linear up to a thickness of 30 micrometers. The photon emission was obtained over a critical range of water contents (25-60%) of the oven-dry clay, and the kinetics of photon emission was presumed to be closely associated with the kinetics of film dehydration. Whether drying proceeded throughout the bulk or via a moving front was undetermined, but in either mode it was preceded by the formation of a thin dry film at the interface with the atmosphere. Grinding of the kaolinite for several minutes by mortar and pestle before paste preparations resulted in an overall increase of photon emission compared to unground kaolinite and in the formation of more than one emission peak, as well as a prolongation of the light emission. This effect on the kinetics of light emittance was observed for about two months after the application of the mechanical stress and suggests a means of detecting the mechanical stress history of a clay. An estimate was made of the spectral characteristics of the emitted light using optical filters and by incorporating tryptophan and salicylic acid into the kaolinite paste where they acted as fluorescent probes. The latter technique shifted the frequency of the light emitted by the kaolinite from the ultraviolet to the visible range where it was less effectively reabsorbed. The first method showed that the wavelengths of 97% of the emitted light was <460 nm and that 75% of the light had wavelengths < 410 nm. The second method showed that the total intensity of DIL increased in the presence of fluorescence molecules, suggesting that the emittance was in the ultraviolet range.

Interaction between ATP, metal ions, glycine, and several minerals.

The adsorption of ATP and ADP on montmorillonite, kaolinite, and A1(OH)3 was studied as a function of pH and, for montmorillonite and kaolinite, as a function of the ionic composition of the system. The three minerals exhibit different adsorption characteristics. Mg2+- and Zn2+-montmorillonite adsorb ATP and ADP more than Na+-montmorillonite, presumably because of complex formation. In kaolinite, the effect of these divalent cations is small. Pure ATP decomposes upon heating, and the rate of the decomposition is accelerated by the presence of glycine. Drying and heating glycine to 70 degrees C under vacuum in the presence of ATP results in abiotic peptide formation with yields up to 0.25%. This peptide formation also occurs when kaolinite or montmorillonite is added to the system. The presence of kaolinite, Mg2+- or Zn2+-kaolinite, or Mg2+-montmorillonite results in a reduction in the rate of the ATP decomposition in the abiotic peptide synthesizing system. These results suggest that one role for clays and metal ions in chemical evolution may have been the stabilization of nucleotides during prebiotic peptide synthesis.

A possible role of fluctuating clay-water systems in the production of ordered prebiotic oligomers.

A model is proposed for the intermediate stages of prebiotic evolution, based on the characteristics of the adsorption and condensation of amino acids and nucleotides on the surface area of clay minerals in a fluctuating environment. Template replication and translation of adsorbed oligonucleotides and catalytic effects by peptide products on further condensation are proposed, due to specific properties of hypohydrous clay surfaces as well as the biomolecules themselves. Experimental evidence supports some of the proposed interactions, and all of them can be tested experimentally.

Prebiotic nucleotide oligomerization in a fluctuating environment: effects of kaolinite and cyanamide.

The day kaolinite was tested for its ability to promote nucleotide oligomerization in model prebiotic systems. Heterogeneous mixtures of clay, water and nucleotide were repeatedly evaporated to dryness at 60 degrees C and redissolved in water in cyclic fashion in the presence or absence of cyanamide and/or ammonium chloride. With or without cycling, kaolinite alone did not promote the oligomerization of nucleotides at detectable levels. Cycling of clay in combination with cyanamide, however, promoted high levels of condensation to a mixture of oligonucleotides and dinucleotide pyrophosphate without requiring ammonium chloride. Although cycling with clay favored synthesis of dinucleotide pyrophosphate, cycling without clay enhanced formation of oligonucleotides. These results support the hypothesis that the presence of clays in fluctuating environments would have influenced the -ourse of prebiotic condensation reactions.