Genetic mapping of a mutation conferring sensitivity to bacteriophage Mu in Salmonella typhimurium LT2.

Two strains of Salmonella typhimurium LT2, SA1475 and MA411, were fortuitously found to be sensitive to bacteriophage Mu. The Mu-sensitivity allele of SA1475 was called musA1 and shown to be linked to the histidine operon both in conjugation and transduction experiments. The Mus allele of MA411 was unlinked to the his region and was tentatively designated musB2. Strains carrying large deletions of the his operon were also tested for Mu sensitivity; those of which the his-rib region is deleted were also sensitive to Mu. Transduction data led to the order zee-2 hisOGDCBAHFIE gnd musA. An Hfr injecting the his operon early (HfrK9) an carrying hisG9424::Tn10 delta 4 delta 11 and musA1 was isolated; this Hfr made it possible to introduce the Mus character into most derivatives of S. typhimurium LT2. Since strain SA1475 is resistant to bacteriophage P1, it could be used to select a new P1-Mu hybrid which has the host range of Mu and the transduction properties of P1.     

A restriction endonuclease map of Streptomyces phage VWB

Summary A physical map of the actinophage VWB has been constructed using the restriction endonucleases BglII, ClaI, EcoRI, EcoRV, HindIII, KpnI and SphI. Phage VWB, genome size 47.3 kb, propagates on Streptomyces venezuelae, and it can also lysogenise this species. The three BglII-generated fragments of VWB DNA were cloned in pBR322, and subsequently mapped. In this manner the restriction map of the VWB phage genome was constructed.Abbreviations dam DNA adenine methylase activity - kb kilobase pairs - :: novel joint     

Frequencies of simultaneous transformation with different T-DNAs and their relevance to the Agrobacterium/plant cell interaction

Summary We investigated whether the efficiency of transformation of plant cells by Agrobacterium tumefaciens during cocultivation is limited by the properties of the plant cells or by the infecting bacteria.Therefore, tobacco protoplasts were infected by cocultivation with two different agrobacteria strains carrying Ti plasmids with distinguishable T-DNAs. These T-DNAs cotransform plant cells at a frequency equal to the product of their independent transformation frequencies, which indicates that all plant cells are equally competent. On the other hand, when these T-DNAs are located on the same Ti plasmid vector within one bacterial strain, the cotransformation frequency is significantly higher than the product of the single transformation frequencies. We interpret these results to indicate that transformation is limited more by the establishment of effective bacteria/plant cell interaction than by (i) the process of DNA integration and (ii) by the number of plant cells capable of being transformed by Agrobacterium. We found that most plant cells are transformed by only one or a few agrobacteria. Analysis of the number of T-DNA copies in these clonally transformed lines indicates amplification of the original, infecting T-region copy.     

Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals.

Alcaligenes eutrophus strain CH34, which was isolated as a bacterium resistant to cobalt, zinc, and cadmium ions, shares with A. eutrophus strain H16 the ability to grow lithoautotrophically on molecular hydrogen, to form a cytoplasmic NAD-reducing and a membrane-bound hydrogenase, and most metabolic attributes; however, it does not grow on fructose. Strain CH34 contains two plasmids, pMOL28 (163 kilobases) specifying nickel, mercury, and cobalt resistance and pMOL30 (238 kilobases) specifying zinc, cadmium, mercury, and cobalt resistance. The plasmids are self-transmissible in homologous matings, but at low frequencies. The transfer frequency was strongly increased with IncP1 plasmids RP4 and pUZ8 as helper plasmids. The phenotypes of the wild type, cured strains, and transconjugants are characterized by the following MICs (Micromolar) in strains with the indicated phenotypes: Nic+, 2.5; Nic-, 0.6; Cob+A, 5.0; Cob+B, 20.0; Cob-, less than 0.07; Zin+, 12.0; Zin-, 0.6; Cad+, 2.5; and Cad-, 0.6. Plasmid-free cells of strain CH34 are still able to grow lithoautotrophically and to form both hydrogenases, indicating that the hydrogenase genes are located on the chromosome, in contrast to the Hox structural genes of strain H16, which are located on the megaplasmid pHG1 (450 kilobases).     

Chromosome transfer and R-prime plasmid formation mediated by plasmid pULB113 (RP4::mini-Mu) in Alcaligenes eutrophus CH34 and Pseudomonas fluorescens 6.2.

Plasmid pULB113 (RP4::mini-Mu), which contains the mini-Mu transposon, promoted both homologous and heterologous gene transfer from Pseudomonas fluorescens 6.2 and Alcaligenes eutrophus CH34. Homologous gene transfer in P. fluorescens 6.2 and A. eutrophus CH34 occurred at a frequency of 10(-4) to 10(-5), and recombinants inherited unselected recessive markers, suggesting a process of chromosome mobilization. Loci involved in autotrophic growth were among those transferred in A. eutrophus. In heterospecific matings, markers were transferred from P. fluorescens to A. eutrophus, Salmonella typhimurium LT2, and Escherichia coli, from A. eutrophus to P. fluorescens, and from Erwinia carotovora subsp. chrysanthemi to A. eutrophus. Heterospecific matings resulted in the formation of R-prime plasmids at frequencies of 10(-7) to 10(-4) per transferred plasmid. When S. typhimurium was the recipient, we observed R-prime plasmids with both restriction-proficient and restriction-deficient strains, although restriction markedly affected the frequency of transfer of pULB113. R-prime plasmids were quite stable, but lost the transposed marker more easily in a rec+ background than in a recA background, suggesting excision of transposed material by reciprocal recombination between flanking copies of mini-Mu. R-prime plasmids could be transferred easily into different recipients and were used in complementation studies. PstI restriction digests of four R-prime plasmids carrying P. fluorescens 6.2 DNA showed a number of additional bands, suggesting that several genes were transposed together with the selected marker on the plasmid.     

Primary production of the periphyton in the littoral of the Danube

Summary The gross primary production of periphyton, grown on artificial substrata in the littoral of the Danube, was measured by the light and dark bottle method from April 1970 to March 1971. The periphyton used for the measurements was sampled from various depths, in order to cover the production of the whole littoral. In connection with this, the littoral area was divided into four zones limited by the heights 600 to 200 cm, according to the local water level gauge. The highest localized zone A (500–600 cm) was inundated only at maximum water levels for rather short period of time, the deepest zone D (200–300 cm) was permanently flooded.The zone A showed an average periphyton primary production of 22.5, the zone B of 54.8, the zone C of 28.9 and the zone D of 9.1 mg O₂/dm²/day. When recalculated to periods when the individual zones were inundated, the annual production was as follows: in zone A: 0.94, zone B: 9.20, zone C: 7.29 and zone D: 3.06g O₂/dm². The highest primary production was always found in the zone just below the water level. Exceptions occurred only when this zone was inundated for a short time as a result of a temporary rise of the water level and the periphyton was insufficiently developed.In order to compare the values of primary production of periphyton obtained from shallower rivers, where the whole bottom is well illuminated, or from rivers that do not exhibit such frequent and extensive level oscillations as the Danube, average value calculated from results obtained from the zone closest to the water level at the time of measurements, were always used. These results represent a special zone following the water level changes which is called surface zone. Primary periphytic production in the surface zone was 43.8 mg O₂/dm²/day. For the annual period it corresponds to the value of 14.74 g O₂/dm². Efficiently of gross photosynthesis in this zone was on the average 1.72%.The height of the water level and the water temperature were higly correlated with gross periphytic production. Close relationships between chlorophyll a concent, biomass and gross primary production of periphyton were found.

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Zusammenfassung Die Bruttoprimärproduktion des auf künstlichem Substrat im litoralen Bereich der Donau wachsenden Periphytons, wurde vom April 1970 bis zum März 1971, mit Hilfe der Hell-Dunkelflaschenmethode, gemessen. Das für die Messung herangezogene Periphyton wurde aus verschiedenen Tiefenzonen entnommen, um die Produktion der gesamten Uferzone festzuhalten. Im Zusammenhang damit wurde der litorale Bereich der Donau in 4 Zonen gegliedert, welche ihre Grenzen bei einem Pegelstand von 600–200 cm des Ortspegels hatten. Die höchste Zone A (500–600 cm) wurde nur bei den höchsten Wasserstanden und nur für kurze Zeit vom Wasser überflutet, die tiefste Zone D (200–300 cm) war dauernd unter Wasser.In der Zone A war die durchschnittliche Produktion des Periphytons 22,5 mg O₂/dm²/Tag, in Zone B 54,8 mg, in Zone C 28,9 mg und in Zone D 9.1 mg. Nach Berechnung für die Zeit, in der die einzelnen Zonen vom Wasser bedeckt waren war folgende Produktion in den einzelnen Zonen festzustellen: in Zone A 0,94 g O₂/dm², in Zone B 9,20 g, in Zone C 7,29 g und in Zone D 3,06 g. Die höchste Periphytonproduktion wurde jeweils in der Zone festgestellt, welche dem Wasserspiegel am nächsten war. Eine Ausnahme bildeten nur jene Fälle, in denen these Zone nach vorübergehender Erhöhung des Wasserspiegels erst kurze Zeit vom Wasser bedeckt war und das Periphyton ungenügend entwickelt war.Zum Vergleich mit den Werten der Primärproduktion des Aufwuchses aus seichteren Gewässern mit gut belichteter Sohle, oder aus Flüssen welche nicht so häufige und weitreichende Spiegelschwankungen aufweisen wie die Donau, wird in der vorgelegten Arbeit der Durchschnittswert aus den Ergebnissen der Zone verwendet, welche zur Zeit der Messungen dem Wasserspiegel am nächsten war. Diese Ergebnisse repräsentieren eine besondere Zone, welche den Schwankungen des Wasserspiegels folgt und von den Autoren durch den Begriff Oberflachenzone bezeichnet wird. Die Primärproduktion des Periphytons in dieser Oberflächenzone war 43,8 mg O₂/dm²/Tag. Dem entspricht im Jahresverlauf ein Wert von 14,74 g O₂/dm². Die Ausnützung der Sonnenenergie war in dieser Zone im Durchschnitt 1,72%.Von den Umweltfaktoren zeigt die Höhe des Wasserspiegels und die Wassertemperatur den engsten Zusammenhang mit der Höhe der Bruttoprimärproduktion des Periphytons. In kürzeren Zeitabschnitten, in denen es zu keinen raschen ausgeprägten Schwankungen des Wasserspiegels des Flusses kommt, wurde eine enge Abhängigkeit zwischen den Werten der Primärproduktion und dem a-Chlorophylgehalt und auch der Biomasse festgestellt.