The earth and its life: Systems perspective

From the planetary point of view, the Earth can be imagined as a system comprised of interwined natural populations propagated through time via recycling. This recycling, or birth/death process, imposes age patterns on natural populations of the solid earth, hydrosphere, atmosphere and living entities. Mathematically, the concept is analogous to that of population dynamics in living systems. The populations of the Earth system form a hierarchical structure. The hierarchy of geological tectonic realms contains populations of 10²⁴–10²⁶ gr in size, with half-lives in the 10⁷–10⁹ yr time range. The approximate parameters for the oceans are 10²⁴ gr and 10²–10⁷ yr, for the atmosphere 10²¹ gr and 10^–2–10⁷ yr, and for living systems 10¹⁴–10¹⁹ gr and 10^–3–10² yr, respectively. In this perspective, and in departure from the GAIA hypothesis, the subordinate populations, such as those of living systems, are constrained to operate within limits imposed by the larger, and slower, hierarchies. They can overstep the imposed limits only on time scales shorter than the response time of the dominant populations. Isotopic record of past sea water shows that on time scales of 10⁷ yr the solid earth, hydrosphere, atmosphere and life act as a unified system controlled by tectonics, that is by the hierarchy of the solid earth.     

Observation of a distinct transition in the mode of interconversion of ring pucker conformers in non-crystalline d-ribose-2'-d from 2H NMR spin-alignment

Internal motions of d-ribose selectively ²H-labeled at the 2 position were measured using solid state ²H NMR experiments. A sample of d-ribose-2 -d was prepared in a hydrated, non-crystalline state to eliminate effects of crystal-packing. Between temperatures of –74 and –60°C the C2–H2 bond was observed to undergo two kinds of motions which were similar to those of C2–H2/H2 found previously in crystalline deoxythymidine (Hiyama et al. (1989) J. Am. Chem. Soc., 111, 8609–8613): (1) Nanosecond motion of small angular displacement with an apparent activation energy of 3.6 ± 0.7 kcal mol^–1, and (2) millisecond to microsecond motion of large amplitude with an apparent activation energy 4 kcal mol^–1. At –74°C, the slow, large-amplitude motion was best characterized as a two-site jump with a correlation time on the millisecond time scale, whereas at –60°C it was diffusive on the microsecond time scale. The slow, large-amplitude motions of the C2–H2 bond are most likely from interconversions between C2-endo and C3-endo by way of the O4-endo conformation, whereas the fast, small-amplitude motions are probably librations of the C2–H2 bond within the C2-endo and C3-endo potential energy minima.     

Oscillations of algal biomass,nutrients and dissolved oxygen as a measure of ecosystem stability

Graphic presentation of weekly rates of change of algal biomass (expressed as chlorophyll a) and nutrient and dissolved oxygen concentrations can be regarded as harmonic oscillation motion. Maximum amplitudes of these oscillations provide a useful tool to assess the degree of stability of aquatic ecosystems in relation to their trophic state. Data sets from seven different lakes ranging from hypereutrophic to oligo-mesotrophic were processed using a computerized method. The high values of oscillation amplitudes of approximately 150 g l^–1 wk^–1 chlorophyll a, 500 g l^–1 wk^–1 ammonia nitrogen, 50 g l^–1 wk^–1 soluble reactive phosphorus and 10 mg l^–1 wk^–1 dissolved oxygen, indicated strong ecosystem instability, while low values of less than 10 g l^–1 wk^–1 of chlorophyll a, 20 g l^–1 wk^–1 ammonia nitrogen, 2 g l^–1 wk^–1 soluble reactive phosphorus, and 3 mg l^–1 wk^–1 dissolved oxygen represented a stable system. Oscillation amplitudes of the chlorophyll a values were found to be the most representative indicator of ecosystem stability.     

Voltage dependence of the basolateral membrane conductance in theAmphiuma collecting tubule

Summary The basolateral potassium conductance of cells of most epithelial cells plays an important role in the transcellular sodium transport inasmuch as the large negative equilibrium potential of potassium across this membrane contributes to the electrical driving force for Na⁺ across the apical membrane. In the present study, we have attempted to establish, theI-V curve of the basolateral membrane of theAmphiuma collecting tubule, a membrane shown to be K⁺ selective. TransepithelialI-V curves were obtained in short, isolated perfused collecting tubule segments. The shunt conductance was determined using amiloride to block the apical membrane Na⁺ conductance. In symmetrical solutions, the shuntI-V curve was linear (conductance: 2.2±0.3 mS·cm^–2). Transcellular current was calculated by subtracting the shunt current from the transepithelial current in the absence of amiloride. Using intracellular microelectrodes, it was then possible to measure the basolateral membrane potential simultaneously with the transcellular current. The basolateral conductance was found to be voltage dependent, being activated by hyperpolarization: conductance values at –30 and –80 mV were 3.6±1.0 and 6.6±1.0 mS·cm^–2, respectively. BasolateralI-V curves were thus clearly different from that predicted by the constant field model. These results indicate that the K⁺-selective basolateral conductance of an amphibian collecting tubule shows inward (anomalous) rectification. Considering the electrogenic nature basolateral Na–K-pump, this may account for coupling between pump-generated potential and basolateral K⁺ conductance.     

Solution conformations of proline rings in proteins studied by NMR spectroscopy

Summary Three different conformations of proline rings in a protein in solution, Up, Down and Twist, have been distinguished, and stereospecific assignments of the pyrrolidine -, - and -hydrogens have been made on the basis of ¹H-¹H vicinal coupling constant patterns and intraresidue NOEs. For all three conformations, interhydrogen distances in the pairs -³, ³-³, ²-², ²-², and ³-³ (2.3 Å) are shorter than those in the pairs -², ²-³, ³-², ²-³, and ³-² (2.7–3.0 Å), resulting in stronger NOESY cross peaks. For the Up conformation, the ³-² and ²-³ spin-spin coupling constants are small (<3 Hz), and weak cross peaks are obtained in a short-mixing-time (10 ms) TOCSY spectrum; all other vicinal coupling constants are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. For the Down form, the -², ²-³, and ³-² vicinal coupling constants are small, leading to weak TOCSY cross peaks; all other couplings again are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. In the case of a Twist conformation, dynamically averaged coupling constants are anticipated. The procedure has been applied to bovine pancreatic trypsin inhibitor and Cucurbita maxima trypsin inhibitor-V, and ring conformations of all prolines in the two proteins have been determined.     

IS EXTRATERRESTRIAL ORGANIC MATTER RELEVANT TO THE ORIGIN OF LIFE ON EARTH?

I review the relative importance of internal and external sources of prebiotic molecules on Earth at the time of life's origin 3.7Gyr ago. The efficiency ofsynthesis in the Earth's atmosphere was critically dependent on its oxidation state. If the early atmosphere was non-reducing and CO2-dominated, external delivery might havebeen the dominant source. Interplanetary dust grains and micrometeorites currently deliver carbonaceous matter to the Earth's surface at a rate of 3 × 10⁵kg/yr(equivalent to a biomass in 2Gyr), but this may havebeen as high as 5 × 10⁷kg/yr (a biomass in only10Myr) during the epoch of late bombardment. Much ofthe incoming material is in the form of chemically inactive kerogens and amorphous carbon; but if the Earth once had a dense (10-bar) atmosphere, small comets rich in avariety of prebiotic molecules may have been sufficiently air-braked to land non-destructively. Lingering uncertainties regarding the impact history of the Earth and the density and composition of its early atmosphere limit our ability to draw firm conclusions.     

The quantitative analysis of chromosome pairing and chiasma formation based on the relative frequencies of M I configurations

Whilst reliable estimates of chiasma frequencies can usually not be obtained, the probability (b) of a chromosome arm to be bound by at least one chiasma can often be determined. In the absence of interference this probability equals (1–e ^–2), where 2 is the average chiasma frequency of the chromosome arm and the average crossover frequency or map length. In the presence of interference is shown to retain its genetic meaning as an additive metric that may describe the chromosome arm or other distinctive chromosome segment in terms of genetic recombination. It is a form of potential map length, comparable to, but numerically different from the regular map length. It is termed provisionally crossing-over potential.A chromosome with armsm andn with crossing-over potentials and will form ring bivalents with a frequency (1–e ^–2).(1–e ^–2); open bivalents with a frequency (1–e ^–2).e ^–2+(1–e ^–2).e ^–2; univalent pairs with a frequencye ^–2.e ^–2. Estimates of these frequencies yield equations from which and may be solved. In rye (Secale cereale) their ratio (q) is approximately two and differs from the mitotic arm length ratio of 1.4, indicating localization of chiasmata in the long arms.Graphs are given to show how, with constantq, the relation between the probabilitiesb _m andb _n of the two arms being bound changes with changing averageb.Data are presented on chiasma frequencies in M I, and compared with the frequencies expected in the absence of interference to give an impression of the degree of interference. Apparent fusion of chiasmata simulates interference.     

Effect of the insecticides methylpyrimifos and chlorpyrifos on soil microflora in an agricultural loam

A study of the effects of two selected organophosphorus insecticides methylpyrimifos and chlorpyrifos on soil microflora in an agricultural loam was made. The insecticides had concentrations of 10 to 300 g g^–1. The presence of methylpyrimifos at concentrations of 100 to 300 g g^–1 or chlorpyrifos at concentrations from 10 to 300 g g^–1 significantly decreased aerobic dinitrogen fixing bacteria and dinitrogen fixation. Nitrifying bacteria decreased at concentrations of 200 and 300 g g^–1 of methylpyrimifos. The presence of 10 to 300 g g^–1 of chlorpyrifos decreased the total number of bacteria. However, fungal populations and denitrifying bacteria were not affected as a consequence of the addition of the organophosphorus insecticides to the agricultural soil, showing that these microorganisms can tolerate high amounts of those insecticides.     

Distribution and abundance of littoral benthic fauna in Canadian prairie saline lakes

The littoral benthos of 18 lakes in Alberta and Saskatchewan ranging in salinity from 3 to 126 (g1^–1 TDS) were investigated twice, in the spring and in the summer of 1986. Multiple Ekman dredge samples were taken at water depths of about 0.5, 1.0 and 2 metres in each transect. Two to three transects were used in each lake according to its estimated limnological diversity for a total of 114 stations. A total of 76 species was present varying from 29–31 species in the three lakes of lowest salinity (means of 3.1–5.55) to only 2 species in lakes exceeding 100. Species richness decreased rapidly in salinities greater than 15.Biomass maximum mean of 10.91 g m^–2 dry weight (maximum 63.0 g m^–2) occurred in culturally eutrophic Humboldt Lake (3.1) but one third as great in other low salinity lakes. However, biomass again increased to about 4.5 gm^–2 in two lakes of 15 As the salinity increased still further biomass declined steadily until a minimum of 0.0212 g m^–2 was recorded in most saline Aroma Lake (mean 119). Summer biomass (11 lakes) was greater than spring biomass (4 lakes) because some groups such as amphipods, corixids and ostracods became more abundant in summer. Wet weight biomass averaged 15.8 of dry weight biomass.Seasonality (spring or summer), sediment texture and organic matter content, water depth, pH, salinity (TDS) and the presence of aquatic plants ( plant cover) were considered in the matrix involving species dry weight biomass at each of 117 stations. TWINSPAN classification of the samples yielded a dendrogram with 18 indicator species. Successive dichotomies divided these indicator species into four main lake groups based on salinity, i.e., Group I: 3–10 (Gammarus, Glyptotendipes I, Chironomus cf. plumosus), Group II: 10–38%. (Hyalella, Enallagma,Bezzia), Group III: 38–63 (Hygrotus salinarius, Cricotopus ornatus), Group IV: >63 (Dolichopodidae, Ephydra hians). Each of these main groups was subdivided into smaller groups of lakes based on factors such as pH, seasonality (spring or summer species dominance), organic matter and plant cover. Depth of samples played no apparent role.     

Optimization of an electroporation procedure for Kluyveromyces lactis transformation

Summary An electroporation method using a Bio-Rad Gene Pulser has been optimized for introducing heterologous DNA into Kluyveromyces lactis yeasts. The plasmid pCR1, derived from a native Kluyveromyces plasmid, was used to transform K. lactis. This plasmid produces a wheat -amylase and contains both the biosynthetic marker URAA and G418 resistance genes. Transformation was optimal at 4500 V/cm, 25 F, and with 0.2 g plasmid DNA. Transformation efficiencies in the range 10⁴–10⁵ transformants/10⁷ cells/g DNA were obtained.     

Plant Resistance Suppressors in the Pathosystem Formed by Potato and the Causal Agent of Late Blight

The properties and effects of two plant resistance suppressors (1,3--1,6--glucan and a pentasaccharide of xyloglucan origin) involved in the pathosystem of potato (Solanum tuberosum) and the causal agent of blight (Phytophthora infestans(Mont) de Bary) were compared. The microbial 1,3--1,6--glucan suppressed the defense response over a narrow concentration range (10^–2M), whereas the plant pentasaccharide had a broad range of effective concentrations (10^–12to 10^–6M). In the pathosystem of potato and the causal agent of late blight, the -glucan caused a local and race-specific suppressor effect on the plant host defense response. In contrast, the pentasaccharide caused both local and systemic suppression of potato resistance and the presence of terminal fucosyl residue in the xyloglucan oligosaccharine played a decisive role in its effect. The recognition of both suppressors by potato cell membrane sites is discussed.     

Certain withanolides from Iochroma gesnerioides antagonize ecdysteroid action in a Drosophila melanogaster cell line

Sixteen withanolides isolated from Iochroma gesnerioides (Kunth) Miers (Solanaceae) have been assessed for their activities as ecdysteroid agonists and antagonists. None of the compounds showed any agonistic activity, but several showed significant antagonistic activity. With a 20-hydroxyecdysone concentration of 5 × 10^–8 M, the ED₅₀ values for 2,3-dihydro-3-methoxywithaferin A, 2,3-dihydro-3-methoxywithacnistine, 2,3-dihydro-3-methoxyiochromolide and 2,3-dihydro-3-hydroxywithacnistine are 3.5 × 10^-5 M, 1 × 10^–5 M, 5 × 10^–6 M and 2.5 × 10^–6 M, respectively.     

Divalent cation effects on calcineurin phosphatase: differential involvement of hydrophobic and metal binding domains in the regulation of the enzyme activity

Summary The effects of divalent metals, metal chelators (EDTA, EGTA) and sodium dodecyl sulfate were investigated on the phosphatase activity of isolated bovine brain calcineurin assayed in the absence (called intrinsic) and presence of calmodulin. Intrinsic phosphatase was increased by Mn²⁺, was unaffected by Mg²⁺, Ca^2–, and Ba⁺, and was markedly inhibited by Ni^2–, Fe²⁺, Zn²⁺ and Cu^2–. When assayed in the presence of calmodulin, many divalent metals (Ni^2–, Zn²⁺, Pb²⁺, Cd²⁺), besides Mn²⁺, increased modestly the phosphatase activity at low concentrations (10–100 M) and inhibited it markedly at high concentrations. Ca^2–-calmodulin stimulated phosphatase activity was antagonized by Ni²⁺, Zn²⁺, Fe²⁺, Cu²⁺, Pb²⁺, at low concentrations (50 M), and by Ba²⁺, Cd²⁺ at slightly higher concentrations (> 100 M); Mn²⁺ and Co^2– (50 M to 1 mM) in fact augmented it. EDTA and EGTA in a concentration and time dependent fashion inhibited the intrinsic phosphatase activity, particularly that of trypsinized calcineurin. SDS in low concentrations (0.005%) augmented the phosphatase activity and inhibited it at high concentrations. Mn²⁺ (± calmodulin) and Ca²⁺ only with calmodulin present increased the phosphatase activity assayed with low concentrations of SDS. The EDTA dependent inhibition of intrinsic phosphatase was almost abolished in assays containing SDS. Prior exposure of calcineurin to Mn²⁺ led to a high activity conformation state of calcineurin that was long-lived or pseudo-irreversible. Such Mn²⁺-activated state of calcineurin exhibited no discerbible change in the affinity towards myelin basic protein or its inhibition by trifluoperazine. At alkaline pH, Mg²⁺ supported the intrinsic phosphatase activity, although to a lesser degree than Mn²⁺. The latter cation, compared to Mg²⁺ and Ni²⁺, was also a more powerful stimulator of the calcineurin phosphatase assayed with histone (III-S) and myosin light chain as substrates.     

Use of15N/14N rations to evaluate the food source of the mysid,Neomysis intermedia Czerniawsky,in a eutrophic lake in Japan

The stable isotope ratios of nitrogen were measured in the mysid,Neomysis intermedia, together with various biogenic materials in a eutrophic lake, Lake Kasumigaura, in Japan throughout a year of 1984/85. The mysid, particulate organic matter (POM, mostly phytoplankton), and zooplankton showed a clear seasonal change in ¹⁵N with high values in spring and fall, but the surface bottom mud did not. A year to year variation as well as seasonal change in ¹⁵N was found in the mysid. The annual averages of ¹⁵N of each material collected in 1984/85 are as follows: surface bottom mud, 6.3 (range: 5.7–6.9); POM, 7.9 (5.8–11.8); large sized mysid, 11.6 (7.7–14.3); zooplankton, 12.5 (10.0–16.4); prawn, 13.2 (9.9–15.4); goby, 15.1 (13.8–16.7). The degree of¹⁵N enrichment by the mysid was determined as 3.2 by the laboratory rearing experiments. The apparent parallel relationship between the POM and the mysid in the temporal patterns of ¹⁵N with about 3 difference suggests the POM (mostly phytoplankton) as a possible food source ofN. intermedia in this lake through the year.     

Morphological development and gibberellin production by different strains of Gibberella fujikuroi in shake flasks and bioreactor

Strain H-984 of G. fujikuroi grown for 38h in a shake flask with medium containing 20g glucose l^–1, 3g yeast extract l^–1, 2.5g NH4NO3 l^–1, 0.5g KH2PO4 l^–1, 0.1g MgSO4 l^–1, 1g CaCO₃ l^–1, and inoculated into a bioreactor with medium containing 60g glucose l^–1; 1g NH_4Cl l^–1; 3g KH2PO4 l^–1 and 1.5g MgSO₄ l^–1 produced 1100mg gibberellic acid l–1.     

Experimental and computational study of the radiation-induced decomposition of formaldehyde. Implications to cometary nuclei

The radiation-induced decomposition of aqueous solutions of formaldehyde was studied at 298 and 77 K in order to obtain an insight into the possible role of ionizing radiation on cometary chemistry. Aqueous solutions of 1.0 mol dm^–3 formaldehyde were exposed to -radiation in the dose range from 0.01 kGy to 1.2 MGy. The radiation chemical yield of decomposition of formaldehyde was determined to be: G(–CH₂(OH)₂)=26.3±1.2 at 298 K and G0.48 at 77 K.Based on previous estimates of the total dose of ionizing radiation that comets have accumulated over 4.6 billion years, we predict a radiation-induced depletion of formaldehyde as a function of depth in comet nuclei: 100% destruction in the outer layers (0–20 m) and 10% destruction in the interior layers.A Portion of this work was presented at the XXVIII COSPAR Plenary Meeting (MF. 7) held at the Hague, The Netherlands on June 25–26, 1990.     

O-Glycosidically linked oligosaccharides from peptidorhamnomannans ofSporothrix schenckii

-Elimination of peptidorhamnomannans purified from yeast-like and mycelial phases ofSporothrix schenckii released neutral and acidic reduced oligosaccharides that were O linked to serine and/or threonine. Man-(1–2)Man-ol, Rha(1–3)Man(1–2)Man-ol, Rha(1–4)GlcA(1–2)Man(1–2)Man-ol, and Rha(1–4)[Rha(1–2)] GlcA(1–2)Man(1–2)Man-ol were characterized based on methylation analysis, proton magnetic resonance and fast atom bombardment mass spectrometry.Abbreviations FAB fast atom bombardment - GLC gas liquid chromatography - GlcA d-glucopyranosyluronic acid - Man d-mannopyranose - Man-ol d-mannitol - MS mass spectrometry - NMR nuclear magnetic resonance - Rha l-rhamnopyranose     

Four new species of Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from reptiles from the islands of Seychelles

Four new Isospora species (Apicomplexa: Eimeriidae) are described from reptiles collected in Seychelles. Oöcysts of I. gardneri n. sp. from Phelsuma astriata astriata, P. sundbergi sundbergi and P. sundbergi longinsulae are ellipsoid, 28.9 (22–31) × 23.5 (18–24) m with a rough 1.5–2 m thick wall. A micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ellipsoid, 14.9 (12.5–17) × 8.8 (8–9.5) m, with a dome-like Stieda body, globular substieda body and a sporocyst residuum consisting of small granules; and the sporozoites have transversal striations. Oöcysts of I. seychellensis n. sp. from 3/7 Mabuya seychellensis are ellipsoid, 19.8 (17.5–21.5) × 15.3 (14.5–16) m. A micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ovoid to broadly ovoid, 11.2 (10–12) × 7.4 (7–8) m, with Stieda and substieda bodies. A sporocyst residuum is present, consisting of small granules; and the sporozoites have distinct transverse striations. Oöcysts of I. tigris n. sp. from 1/1 Calumma tigris are ellipsoid, 22.5 (19–24) × 18 (16–20) m. A micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ovoid or ellipsoid, 13.6 (12–15) × 7 (6–8) m, with large Stieda body and substieda bodies. A sporocyst residuum is present, consisting of numerous small granules; and the sporozoites are vermiform with distinct transverse striations. Oöcysts of I. ladiguensis n. sp. from Phelsuma sundbergi ladiguensis and P. sundbergi longinsulae are spherical to subspherical, 13.2 (12–13.5) × 12 (9–13) m, without micropyle and oöcyst residuum but with one globular polar granule. Sporocysts ovoid, 9 (8–10) × 5.6 (5–6) m, with dome-like Stieda and subglobular substieda body; and the sporozoites are vermiform with distinct transverse striations.     

Biotechnological production of unnatural L-amino acids from D,L-5-monosubstituted hydantions. II. L-α- and L-β-naphthylalanine

Summary Resting cells ofArthrobacter sp. (DSM 3745) with the ability to form L-tryptophan from D,L-5-(3-indolylmethy)hydantoin were used for the bioconversion of D,L-5-- and D,L-5--naphthylmethylhydantoin (D,L-5-- and D,L-5--NMH) to the corresponding L-amino acids. Under the optimal reaction conditions of pH 9.7 and 40°C specific productivities of 0.2 (-naphtylalanine) and 0.6 (-naphtylalanine) mM amino acid x g cell dry mass^–1 x h^–1 were obtained in a 0.1 M Na₂CO₃/NaHCO₃-buffer in a strirred bioreactor.     

Genetical studies on the heterocyst and nitrogen fixation of the blue-green alga Nostoc muscorum

Summary The short-trichome-forming, non-heterocystous and non-nitrogen-fixing (het ^– nif^–) mutant of the nitrogen-fixing blue-green alga Nostoc muscorum was isolated by N-methyl-N-nitro-N-nitrosoguanidine (NTG)-mutagenesis after penicillin enrichment technique and characterized. The mutant did not grow and fix nitrogen in combined-nitrogen-free medium while in nitrate-containing medium it grew well (K=0.112/day, G=64.27 h), although its growth was comparatively poor than the parent alga (K=0.128/day; G=56.14 h). The mutant was stable and both the het ^– and nif ^– characters reverted to wild type (het ⁺ nif⁺) with the reversion frequency of 2.62×10^-7.The het ^– nif^– mutant tolerated 0.5 g/ml of streptomycin sulphate on the agar medium and its streptomycin resistant mutant capable of growing in presence of 10g/ml of streptomycin was isolated spontaneously with a frequency of 1.45×10^-8. These streptomycin resistant isolates (het ^– nif^– str^R) resisted 100 g/ml of streptomycin sulphate on the agar medium and 200 g/ml in liquid medium. Spontaneous virus-resistant mutant of het ^– nif^– str^R was isolated with a mutation frequency of 4.02×10^-4.The data of genetic recombination experiments suggested that there is transfer of both het and nif genes to het ^– nif^– strain with the frequency of 2×10^-6 to 2×10^-5 simultaneously. There was increase in recombination frequency with increasing the incubation period. The virus-resistance marker is also transferred to the sensitive recipient.Abbreviations CFU colony forming units - C–N Chu-10 medium without combined nitrogen - C+N Chu-10 medium with 0.232 g/l calcium nitrate - G generation time - het heterocyst differentiating genes - K specific growth rate constant - MOI multiplicity of infection - nif nitrogen-fixing genes - NTG N-methyl-N-nitro-N-nitrosoguanidine - PFU plaque forming units - str ^R streptomycin resistance - str ^R streptomycin sensitive