Improved identification of methanogenic bacteria by fluorescence microscopy.

Methanogenic bacteria can be tentatively identified by fluorescence microscopy. This technique was improved by carefully selecting a series of excitation and barrier filters that matched the excitation and emission spectra of some unique coenzymes viz., F420 and F350, in methanogenic bacteria.     

Primary fluorescence (Autofluorescence) of fruiting bodies of the wood-rotting fungusFomes fomentarius

Autofluorescence of fruiting bodies of the wood-rotting fungus Fomes fomentarius has been observed and is described among native macrofungi for the first time. The strongest yellow autofluorescence with blue excitation was displayed by pith sets, a weaker yellow, yellow-green to yellow-red fluorescence was due to generative thin-walled hyphae while the weakest yellow-reddish fluorescence was emitted by thick-walled skeletal hyphae (though their parts may emit a more intensive yellow fluorescence). This yellow, yellow-green to yellow-red autofluorescence was assessed to be more intensive than the emission described so far in bacteria and fungi (except for lysed hyphae of the fungus Trametes versicolor). With green excitation all F. fomentarius cells emitted strong red autofluorescence.     

The long-wavelength chlorophyll states of plant LHCI at room temperature: a comparison with PSI-LHCI

The red antenna states of the external antenna complexes of higher plant photosystem I, known as LHCI, have been analyzed by measurement of their preequilibrium fluorescence upon direct excitation at 280 K. In addition to the previously detected F735 state, a hitherto undetected low-energy state with emission maximum around 713 nm was observed. The 280 K bandwidths (FWHM) are 55 nm for the F735 state and approximately 27 nm for the F713-nm state, much greater than for non-red-shifted antenna chlorophylls. The origin absorption band for the F735-nm state was directly detected by determination of its excitation (action) spectrum and lies at 709-710 nm. The absorption spectrum for F735, calculated using the Stepanov expression, closely overlaps the excitation spectrum, indicating that the very large Stokes shift (25 nm) is due to vibrational relaxation within the excited-state manifold and solvent effects can be excluded. Fluorescence anisotropy measurements, with direct excitation of F735, indicate that the transition dipoles of the two red states are parallel. Similar experiments performed in the long-wavelength absorbing tail of PSI-LHCI indicate the presence of emission state(s) that are red-shifted with respect to F735 of isolated LHCI. It is suggested that these are brought about by interactions between the complexes in PSI-LHCI, which occur in some yet undefined way, and which are broken upon solubilization of the component parts.     

Heat stress induces an inhibition of excitation energy transfer from phycobilisomes to photosystem II but not to photosystem I in a cyanobacterium Spirulina platensis.

The effects of high temperature (30-52.5 degrees C) on excitation energy transfer from phycobilisomes (PBS) to photosystem I (PSI) and photosystem II (PSII) in a cyanobacterium Spirulina platensis grown at 30 degrees C were studied by measuring 77 K chlorophyll (Chl) fluorescence emission spectra. Heat stress had a significant effect on 77 K Chl fluorescence emission spectra excited either at 436 or 580 nm. In order to reveal what parts of the photosynthetic apparatus were responsible for the changes in the related Chl fluorescence emission peaks, we fitted the emission spectra by Gaussian components according to the assignments of emission bands to different components of the photosynthetic apparatus. The 643 and 664 nm emissions originate from C-phycocyanin (CPC) and allophycocyanin (APC), respectively. The 685 and 695 nm emissions originate mainly from the core antenna complexes of PSII, CP43 and CP47, respectively. The 725 and 751 nm band is most effectively produced by PSI. There was no significant change in F725 and F751 during heat stress, suggesting that heat stress had no effects on excitation energy transfer from PBS to PSI. On the other hand, heat stress induced an increase in the ratio of Chl fluorescence yield of PBS to PSII, indicating that heat stress inhibits excitation energy transfer from PBS to PSII. However, this inhibition was not associated with an inhibition of excitation energy transfer from CPC to APC since no significant changes in F643 occurred at high temperatures. A dramatic enhancement of F664 occurring at 52.5 degrees C indicates that excitation energy transfer from APC to the PSII core complexes is suppressed at this temperature, possibly due to the structural changes within the PBS core but not to a detachment of PBS from PSII, resulting in an inhibition of excitation energy transfer from APC to PSII core complexes (CP47 + CP43). A decrease in F685 and F695 in heat-stressed cells with excitation at 436 nm seems to suggest that heat stress did not inhibit excitation energy transfer from the Chl a binding proteins CP47 and CP43 to the PSII reaction center and the decreased Chl fluorescence yields from CP43 and CP47 could be explained by the inhibition of the energy transfer from APC to PSII core complexes (CP47 + CP43).     

Stochastic Models for Phototaxis

This work studies two mathematical models for describing the motion of phototactic bacteria, i.e., bacteria that move toward light. Based on experimental observations, we conjecture that the motion of the colony toward light depends on certain group dynamics. These group dynamics are hypothesized to be coordinated through an individual property of each bacterium, which we refer to as excitation. The excitation of each individual bacterium is assumed to change based on the excitation of the neighboring bacteria. Under these assumptions, we propose a (discrete) cellular automaton model and derive an analogous stochastic model for describing the evolution in time of the location of bacteria, the excitation of individual bacteria, and a surface memory effect. We provide simulation results and discuss in detail the role of the various model parameters in controlling the emerging dynamics.     

Pharmacological characterisation of neuropeptide F (NPF)-induced effects on the motility of Mesocestoides corti (syn. Mesocestoides vogae) larvae

Neuropeptide F is the most abundant neuropeptide in parasitic flatworms and is analogous to vertebrate neuropeptide Y. This paper examines the effects of neuropeptide F on tetrathyridia of the cestode Mesocestoides vogae and provides preliminary data on the signalling mechanisms employed. Neuropeptide F (>/=10 microM) had profound excitatory effects on larval motility in vitro. The effects were insensitive to high concentrations (1 mM) of the anaesthetic procaine hydrochloride suggesting extraneuronal sites of action. Neuropeptide F activity was not significantly blocked by a FMRFamide-related peptide analog (GNFFRdFamide) that was found to inhibit GNFFRFamide-induced excitation indicating the occurrence of distinct neuropeptide F and FMRFamide-related peptide receptors. Larval treatment with guanosine 5'-O-(2-thiodiphosphate) trilithium salt prior to the addition of neuropeptide F completely abolished the excitatory effects indicating the involvement of G-proteins and a G-protein coupled receptor in neuropeptide F activity. Addition of guanosine 5'-O-(2-thiodiphosphate) following neuropeptide F had limited inhibitory effects consistent with the activation of a signalling cascade by the neuropeptide. With respect to Ca(2+) involvement in neuropeptide F-induced excitation of M. vogae larvae, the L-type Ca(2+)-channel blockers verapamil and nifedipine both abolished neuropeptide F activity as did high Mg(+) concentrations and drugs which blocked sarcoplasmic reticulum Ca(2+)-activated Ca(2+)-channels (ryanodine) and sarcoplasmic reticulum Ca(2+) pumps (cyclopiazonic acid). Therefore, both extracellular and intracellular Ca(2+) is important for neuropeptide F excitation in M. vogae. With respect to second messengers, the protein kinase C inhibitor chelerythrine chloride and the adenylate cyclase inhibitor MDL-2330A both abolished neuropeptide F-induced excitation. The involvement of a signalling pathway that involves protein kinase C was further supported by the fact that phorbol-12-myristate-13-acetate, known to directly activate protein kinase C, had direct excitatory effects on larval motility. Although neuropeptide F is structurally analogous to neuropeptide Y, its mode-of-action in flatworms appears quite distinct from the common signalling mechanism seen in vertebrates.     

Nitrite regulates distribution of excitation energy between the two photosystems by causing state transition.

The mechanism of distribution of absorbed excitation energy between the two photosystems in the presence of nitrite has been investigated in spinach (Spinacia oleracea L.) thylakoid membranes. Nitrite inhibited PS II activity (H(2)O --> DCPIP reaction) and enhanced PS I activity (DCPIPH(2) --> MV reaction). Nitrite decreased the F(v)/F(m) ratio measured at room temperature and increased the F(730)/F(685) ratio measured at low temperature (77 K). These results suggested that nitrite caused a decrease in the excitation energy available to PS II and transferred more energy to PS I by the mechanism of state transition. Measurement of fluorescence excitation spectra at 77 K showed that nitrite increased the absorption cross-section of PS I antenna at the expense of chlorophyll b and LHC II. Based on these observations we have suggested a role of nitrite in causing state transition.     

Exciton Annihilation in the Two Photosystems in Chloroplasts at 100°K

The fluorescence yield (F) of spinach chloroplasts at 100°K measured at 735 nm (photosystem I fluorescence—F 735) and at 685 nm (photosystem II fluorescence—F 685) has been determined with different modes of laser excitation. The modes of excitation included a single picosecond pulse, sequences of picosecond pulses (4, 22, and 300 pulses spaced 5 ns apart) and a single nonmode-locked 2-μs pulse (MP mode). The F 735/F 685 intensity ratios decrease from 1.62 to 0.61 when a single picosecond pulse (or low-power continuous helium-neon laser) is replaced by excitation with the 300-ps pulse train (PPT mode) or MP mode. In the PPT mode of excitation, the 735-nm fluorescence band is quenched by a factor of 45 as the intensity is increased from 10¹⁵ to 10¹⁸ photons/cm² per pulse train and the 685-nm fluorescence is quenched by a factor of 10. In the MP mode, the quenching factors are 25 and 7, respectively, in the same intensity range. Fluorescence quantum yield measurements with different picosecond pulse sequences indicate that relatively long-lived quenching species are operative, which survive from one picosecond pulse to another within the pulse train. The excitonic processes possible in the photosynthetic units are discussed in detail. The differences in the quenching factors between the MP and PPT modes of excitation are attributed to singlet-singlet annihilation, possible when picosecond pulses are utilized, but minimized in the MP mode of excitation. The long-lived quenchers are identified as triplets and/or bulk chlorophyll ions formed by singlet-singlet annihilation. The preferential quenching in photosystem I is attributed to triplet excitons. The influence of heating effects, photochemistry, bleaching, and two-photon processes is also considered and is shown to be negligible.     

Excitation energy trapping in anoxygenic photosynthetic bacteria*

Various aspects of excitation energy conversion in anoxygenic photosynthetic bacteria are surveyed. This minireview discusses different models that have been proposed during the past 60 years to describe excitation energy transfer from an antenna molecule to the reaction center. First, a simple one-dimensional model was suggested, but over time the models became more detailed when structural and dynamic information was included. This review focuses mainly on the picture of purple bacteria and green sulfur bacteria developed during the past decades. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparative analysis of the immune response of a rabbit to antigens to live and killed Francisella species bacteria]

Serum antibodies were analyzed in rabbits immunized with live and formalin-killed Francisella (F. tularensis, F. novicida, F. novicida-like, and F. philomiragia). Passive hemagglutination test with erythrocytes sensitized by these bacteria' LPS showed much higher titers of species-specific antibodies in all sera to live microorganisms than sera to killed bacteria. The results of immunoblotting with purified LPS and bacterial lysates indicate that sera to live bacteria contained mainly immunoglobulins to species-specific antigenic epitopes of LPS O-polysaccharide chain and few antibodies to the protein component of the cell. By contrast, killed bacterial cells induced weak production of antibodies to S-LPS and a pronounced antibody response to protein antigens. Besides the quantitative differences, live and killed bacteria differed by the qualitative spectrum of immunodominant proteins. Serum to live F. tularensis 15/10 contained antibodies to at least 3 immunodominant antigens of the cell, while serum to killed bacteria contained antibodies to only two of these. Immunoglobulins to protein antigens, absent in homologous sera to live bacteria, were detected in the sera to killed F. novicida and F. novicida-like bacteria. Both sera to F. philomiragia had antibodies reacting with LPS epitopes and immunodominant complex containing protein. In contrast to other Francisella, F. philomiragia was found to synthesize an uncommon LPS representing two major lipooligosaccharides with different molecular weights and antigenic specificity. Therefore, immune response of the host to live and killed Francisella is different: live cells more effectively induce the production of antibodies to S-LPS epitopes, while killed ones to protein antigens.     

Fluorescence spectral properties of troponin C mutant F22W with one-, two-, and three-photon excitation.

We report the first measurements of protein fluorescence with three-photon excitation, using a mutant of troponin C (TnC) that contains a single tryptophan residue F22W. From the emission intensity dependence on laser power we determine that TnC F22W displays one-, two-, and three-photon excitation at 285, 570, and 855 nm, respectively. The emission spectra and intensity decays are identical for one-, two-, or three-photon excitation. The steady-state and time 0 anisotropies are distinct for each mode of excitation, but the correlation times were the same, suggesting that three-photon excitation of proteins can be accomplished without significant effects of the locally intense illumination. The excitation anisotropy spectrum from 830 to 900 nm displays only negative values, suggesting dominant excitation via the 1Lb state of tryptophan from 830 to 900 nm.     

The possible site of action of 5-hydroxytryptamine, 6-hydroxytryptamine, tryptamine and dopamine on identified neurons in the central nervous system of the snail, Helix aspersa

Intracellular recordings were made from identified neurons in the right parietal ganglion of the snail, Helix aspersa. Cells F 4, 5 and 6 were excited by 5-hydroxytryptamine (5-HT) and inhibited by dopamine while cells in the F 30 area were inhibited by both compounds. Low doses of both tryptamine and 6-HT produced weak excitation of cells F 4, 5 and 6 while higher doses of both compounds inhibit the activity of these cells. In terms of the inhibitory responses, tryptamine and 6-HT are approximately equipotent but between 10 and 100 times less potent than dopamine. d-Tubocurarine reversibly antagonized the excitatory action of 5-HT on cells F 4, 5 and 6 and converted tryptamine and 6-HT excitation to inhibition. In the presence of the antagonist, ergometrine, the dopamine inhibitory response was almost completely abolished while the inhibitory responses to tryptamine and 6-HT were converted to weak excitation. All four agonists inhibited cells in the F 30 area with the following potency ratios: dopamine much greater than tryptamine/6-HT greater than 5-HT. Tubocurarine had no antagonist effects on these responses while ergometrine reduced or blocked all four, often irreversibly. In potassium-free Ringer the inhibitory responses to all four agonists were enhanced. It is concluded that on cells F 4, 5 and 6, low concentrations of tryptamine and 6-HT act on 5-HT receptors while higher concentrations of both agonists act on dopamine receptors. On cells in the F 30 area, 5-HT, 6-HT and tryptamine all act on a dopamine receptor.     

Transducing phages of Pseudomonas aeruginosa related to phage F116L

New phages K104 and B26, which are relative to F116L by a number of biological characters, appeared to show general transducing activity. Phage K104 transduces all tested markers with higher frequency than the phage B26. Linkage of the bacterial markers pair ilv202--met28 durspectively. When recipient bacteria lysogenic for phages K104 and B26 are used, frequencies of transduction by phage F116L are decreased. In the presence of F116L prophage the frequency of transduction by phage B26 is 10-fold increased. Phages B26 and F116L do not grow on bacteria lysogenic for these phages. Phage F116L does not grow on the lawn of bacteria, lysogenic for phage K104, while phage B26 grows on the same lawn with the efficiency of plating about 10(-2).     

不同寄主植物西花蓟马内生细菌多样性分析

为探讨不同寄主植物对西花蓟马Frankliniella occidentalis内生细菌分布的影响,应用Illumina MiSeq第二代测序技术对取食不同寄主植物的西花蓟马内生细菌的16S rDNA V5~V7区域进行测定。结果显示,Alpha多样性分析发现,西花蓟马内生细菌在取食不同寄主植物后多样性表现为西葫芦Cucurbita pepo L. >菊花Dendranthema morifolium(Ramat.) Tzvelev >曼陀罗Datura stramonium L. >玫瑰Rosa rugosa Thunb. >牵牛花Pharibitis purpurea (L.) Voigt >辣椒Capsicum annuum L.;在门水平的菌群组成上,变形菌门在各样品中所占比例最高,是西花蓟马内生菌中的优势菌门,厚壁菌门为次优势菌菌门。在属水平上内生细菌也表现出较大的差异,其中假单胞菌属Pseudomonas和欧文氏菌属Erwinia在西花蓟马取食各寄主植物时均有分布,取食辣椒、玫瑰、曼陀罗时,优势菌属均为欧文氏菌属Erwinia,取食菊花和牵牛花时,优势菌属为立克次体Rickettsia,而取食西葫芦时,则为短波毛单胞菌属Brevundimonas;利用KEGG数据库,对内生细菌进行基因功能预测分析发现,内生细菌群富集了高丰度的代谢功能基因,且主要集中于碳水化合物代谢、氨基酸代谢以及能量代谢。综上所述取食不同寄主植物时西花蓟马内生细菌多样性与群落结构具有一定的差异,寄主植物可对西花蓟马内生细菌群落组成产生重要的影响;且各寄主植物上西花蓟马内生细菌基因功能主要集中在碳水化合物代谢、氨基酸代谢和能量代谢。     

Changes in the chlorophyll fluorescence characteristics of chloroplasts from intact pumpkin cotyledons,caused by organ excision and kinetin treatment

The chlorophyll (Chl) fluorescence emission as well as excitation and polarization characteristics of chloroplasts from intact cotyledons were determined in pumpkin seedlings after removal of one cotyledon (co-cotyledon) or apical bud or primary root, or after kinetin treatment of derooted seedlings. Qualitatively, the fluorescence emission and excitation spectra of chloroplasts were similar. The fluorescence emission spectra showed a maximum at 685 (F685) and a hump at 735 nm (F735), whereas the excitation spectra showed peaks at 439, 471, 485, and 676 nm. The fluorescence intensities at F685 and F735 differed in various groups of seedlings, as indicated by changes in their ratios. Similarly, the ratios of 471/439, 485/439, and 676/439 nm were also different. Variability in the Chl fluorescence intensity values and the fluorescence polarization of chloroplasts prepared from various seedling types may suggest a different degree of binding between the pigment complexes and light-harvesting Chl-protein (LHCP), resulting in different rates of photoexcitation energy loss in the form of fluorescence emission. Kinetin treatment improved the coupling of pigment complexes with reaction centre, as indicated by low polarization values in derooted and kinetin-treated seedlings, which suggests the development of a suntype chloroplast.     

Modeling of energy migration and trapping in purple bacteria. Analysis of extreme formulae

Homogeneous pigment ensembles similar to those of purple bacteria Rhodospirillum rubrum were studied. Two formulae were advanced for the limiting values of excitation lifetime and quantum yield of excitation trapping in these ensembles, provided all reaction centers are in an active state. It was demonstrated by mathematical modeling that these limiting values strictly depend on three parameters of molecular ensembles: the numbers of core-bacteriochlorophyll molecules per reaction center, the values of rate constants for excitation trapping in reaction centers, and excitation wasteful deactivation in all molecules. The excitation lifetime and quantum yield were proved to approach their limiting values as the rate constants of excitation intermolecular migration increase. The closeness of experimental values for two above mentioned functions to their calculated limiting values proves the migration-limited type of the photosynthetic unit investigated and a high efficiency of excitation trapping in its reaction centers.     

Survival and growth of Francisella tularensis in Acanthamoeba castellanii

Francisella tularensis is a highly infectious, facultative intracellular bacterium which causes epidemics of tularemia in both humans and mammals at regular intervals. The natural reservoir of the bacterium is largely unknown, although it has been speculated that protozoa may harbor it. To test this hypothesis, Acanthamoeba castellanii was cocultured with a strain of F. tularensis engineered to produce green fluorescent protein (GFP) in a nutrient-rich medium. GFP fluorescence within A. castellanii was then monitored by flow cytometry and fluorescence microscopy. In addition, extracellular bacteria were distinguished from intracellular bacteria by targeting with monoclonal antibodies. Electron microscopy was used to determine the intracellular location of F. tularensis in A. castellanii, and viable counts were obtained for both extracellular and intracellular bacteria. The results showed that many F. tularensis cells were located intracellularly in A. castellanii cells. The bacteria multiplied within intracellular vacuoles and eventually killed many of the host cells. F. tularensis was found in intact trophozoites, excreted vesicles, and cysts. Furthermore, F. tularensis grew faster in cocultures with A. castellanii than it did when grown alone in the same medium. This increase in growth was accompanied by a decrease in the number of A. castellanii cells. The interaction between F. tularensis and amoebae demonstrated in this study indicates that ubiquitous protozoa might be an important environmental reservoir for F. tularensis.     

Polyphasic classification of 0.2 microm filterable bacteria from the western Mediterranean Sea

The 0.2 microm filtration of sea water samples from the Mediterranean Sea (Bay of Calvi, Corsica), collected from 10 m and 35 m depth led to the isolation of several gram-negative bacterial strains able to grow on full-strength media as well as on diluted media. The analysis of the 16S rRNA gene sequences and estimation of the phylogenetic relationships of these facultative oligotrophic bacteria indicated that they grouped into two phylogenetic branches. The strains RE10F/2, RE10F/5 (10 m depth samples) and RE35/F12 (35 m depth samples) were assigned to the gamma-subclass, while RE35F/1 (35m depth sample) was assigned to the alpha-4-subclass of the Proteobacteria. The strains RE10/F2 and RE10/F5 were most closely related to species and strains of the Pseudoalteromonas group, whereas the strain RE35F/12 placed adjacent to the family Vibrionaceae. The phylogenetic analysis of strain RE35F/1 revealed that this bacterium clusters with marine strains and species of the aerobic anoxygenic phototrophic bacteria Erythrobacter as well as Erythromicrobium and more distantly to Sphingomonas spp. Supplementary to those genotypic classifications the chemotaxonomic signatures including the major respiratory lipoquinone systems, the cellular fatty acid compositions as well as the polyamine contents of the bacteria were investigated. The isolated organisms displayed differences in their physiological and biochemical properties to already described strains belonging to the same genera or families, as revealed by the comparative 16S rRNA analysis. Despite the fact that these bacteria were isolated from a 0.2 microm filtrate, the cultured organisms which were all rod-shaped, displayed width dimensions ranging from 0.4 up to 0.7 microm, indicating that these bacteria were starvation forms at the time of isolation and not ultramicrobacteria as defined by Torella and Morita (1981) or by Schut et al. (1993). Because our isolated strains represent potentially new taxa, this first investigation on 0.2 pm filterable bacteria from the Western Mediterranean Sea supports the hypothesis that this bacterial fraction contributes to the diversity of marine bacteria.     

Changes in the chlorophyll fluorescence characteristics of chloroplasts from intact pumpkin cotyledons, caused by organ excision and kinetin treatment

The chlorophyll (Chl) fluorescence emission as well as excitation and polarization characteristics of chloroplasts from intact cotyledons were determined in pumpkin seedlings after removal of one cotyledon (co-cotyledon) or apical bud or primary root, or after kinetin treatment of derooted seedlings. Qualitatively, the fluorescence emission and excitation spectra of chloroplasts were similar. The fluorescence emission spectra showed a maximum at 685 (F685) and a hump at 735 nm (F735), whereas the excitation spectra showed peaks at 439, 471, 485, and 676 nm. The fluorescence intensities at F685 and F735 differed in various groups of seedlings, as indicated by changes in their ratios. Similarly, the ratios of 471/439, 485/439, and 676/439 nm were also different. Variability in the Chl fluorescence intensity values and the fluorescence polarization of chloroplasts prepared from various seedling types may suggest a different degree of binding between the pigment complexes and light-harvesting Chl-protein (LHCP), resulting in different rates of photoexcitation energy loss in the form of fluorescence emission. Kinetin treatment improved the coupling of pigment complexes with reaction centre, as indicated by low polarization values in derooted and kinetin-treated seedlings, which suggests the development of a suntype chloroplast. This revised version was published online in June 2006 with corrections to the Cover Date.     

Loss and repair of conjugal fertility and infectivity of the resistance factor and sex factor in Escherichia coli

Hirota, Yukinori (University of Osaka, Osaka, Japan), Toshio Fujii, and Yukinobu Nishimura. Loss and repair of conjugal fertility and infectivity of the resistance factor and sex factor in Escherichia coli. J. Bacteriol. 91:1298-1304. 1966.-The drug-resistance factor, R, and the sex factor, F, have homologous traits, including contagious transmission, mediation of sexuality of the host cell, and autonomous replication in their host bacteria. Cooperation between F and R factors was found with a mutant R factor, which is nontransmissible in F(-) bacteria, becoming transmissible when introduced into bacteria carrying F. Conversely, the chromosome of a sterile male strain carrying the mutant sex factor, F(r), becomes transmissible when an R factor is introduced into the cell. The genetic determinants of R factors have been analyzed by isolation of mutant R factors, by sexual conjugation of the host bacteria, and by transduction of R factors with phage P1kc. The fertility determinant of the R factor, m, is inseparable from the determinant for its infectivity, but can be separated from the loci for autonomous replication of the R factor. R and F thus carry genetic determinants governing the same functions.