Genetically different isolates of Trypanosoma cruzi elicit different infection dynamics in raccoons (Procyon lotor) and Virginia opossums (Didelphis virginiana)

Trypanosoma cruzi is a genetically and biologically diverse species. In the current study we determined T. cruzi infection dynamics in two common North American reservoirs, Virginia opossums (Didelphis virginiana) and raccoons (Procyon lotor). Based on previous molecular and culture data from naturally-exposed animals, we hypothesised that raccoons would have a longer patent period than opossums, and raccoons would be competent reservoirs for both genotypes T. cruzi I (TcI) and TcIIa, while opossums would only serve as hosts for TcI. Individuals (n = 2 or 3) of each species were inoculated with 1 × 10⁶ culture-derived T. cruzi trypomastigotes of TcIIa (North American (NA) – raccoon), TcI (NA – opossum), TcIIb (South American – human), or both TcI and TcIIa. Parasitemias in opossums gradually increased and declined rapidly, whereas parasitemias peaked sooner in raccoons and they maintained relatively high parasitemia for 5 weeks. Raccoons became infected with all three T. cruzi strains, while opossums only became infected with TcI and TcIIb. Although opossums were susceptible to TcIIb, infection dynamics were dramatically different compared with TcI. Opossums inoculated with TcIIb seroconverted, but parasitemia duration was short and only detectable by PCR. In addition, raccoons seroconverted sooner (3–7 days post inoculation) than opossums (10 days post inoculation). These data suggest that infection dynamics of various T. cruzi strains can differ considerably in different wildlife hosts.     

Interactive signal transfer between host and pathogen during successful infection of barley leaves by Blumeria graminis and Bipolaris sorokiniana

Using ion-selective microprobes, interactive signalling between barley and Blumeria graminis or Bipolaris sorokiniana has been investigated. The question was raised whether a biotrophically growing fungus manipulates the electrical driving forces (membrane potential, transmembrane pH), required for H⁺ cotransport of energy-rich compounds. Electrodes were positioned in the substomatal cavity of open stomata or on the leaf surface, and pH was measured continuously up to several days during fungal development. We demonstrate that surface and apoplastic fluids are electrically coupled and respond in a similar manner to stimuli. Apoplastic pH, monitored from the moment of inoculation with conidia, reveals several phases: 2–4 h after inoculation of the barley leaf with either fungus, the host displays rapid transient responses after its first contact with the fungal cell wall; apoplastic pH and pCa increases, cytoplasmic pH and pCa decreases. About 1 day after inoculation, the apoplastic pH increases by up to 2 pH units, which is thought to reflect a resistance response against the intruder. Whereas barley leaf cells possess a membrane potential of −152±5 mV, hyphae of B. graminis yield −251±8 mV, indicative of a substantial driving force advantage for the fungus. Although the resting membrane potential of barley remains constant during the first days after inoculation, leaves infected with B. sorokiniana get confronted with an energy problem, indicated by a retarded repolarization following a “light-off” stimulus. Five days after inoculation, apoplastic pH has increased to 5.97±0.47 (n=11) and does no longer respond to “light-off” when measured within lesions. In contrast, it stays at near normal values outside the lesions and responds to “light-off”.It is concluded that biotrophically growing fungi do not manipulate the cotransport driving forces since (i) any change in apoplastic pH would be experienced by both partners; (ii) the resting membrane potential is not changed. It is suggested that measured pH changes reflect defence responses of the host against the fungus rather than fungal action to increase compatibility.     

Leaf scarring by the weevils Neochetina eichhorniae and N. bruchi enhances infection by the fungus Cercospora piaropi on waterhyacinth, Eichhornia crassipes

Additive or synergistic effects among introduced and native insect and plant pathogen agents are necessary to achieve biological control of waterhyacinth (Eichhornia crassipes), a globally damaging aquatic weed. In field plots, plants were infested with waterhyacinth weevils (Neoechetina bruchi and N. eichhorniae) and leaves were scarred by weevil feeding. Subsequent infection by the fungal pathogen Cercospora piaropi caused necrotic lesions to form on leaves. Necrosis development was 7.5- and 10.5-fold greater in plots augmented with both weevils and C. piaropi and weevils alone, respectively, than in plots receiving only C. piaropi. Twenty-four days after weevil infestation, the percentage of laminar area covered by lesions on third-youngest and oldest live leaves was elevated 2.3–2.5-fold in plots augmented with weevils. Scar density and necrosis coverage on young leaf laminae were positively correlated, even though antipathogenic soluble peroxidases were elevated 3-fold in plots augmented with weevils alone or weevils and C. piaropi. Combined weevil and fungal augmentation decreased shoot densities and leaves per plant. In a no-choice bioassay, weevil feeding on oldest but not young leaves was reduced 44 two weeks after C. piaropi inoculation. Protein content and peroxidase activities were elevated 2–6-fold in oldest leaves three weeks after inoculation. Augmentation with both waterhyacinth weevils and C. piaropi led to the development of an additive biological control impact, mediated by one or more direct interactions between these agents, and not plant quality effects.     

Translocation and multiplication ofSpiroplasma citri in turnip (Brassica rapa)

Following inoculation of designated leaves of turnip plants withSpiroplasma citri byCirculifer tenellus, spiroplasmas were cultured first from roots (four days) and then from youngest leaves (eight days), but almost never from oldest leaves. In experiments using enzyme-linked immunosorbent assay to monitor changes in titer in turnip leaves during the course of plant infection,S. citri was detected seven days after inoculation and reached peak titers of 10¹⁰–10¹¹ colony-forming units/g 12–20 days after inoculation, declining thereafter. Spiroplasmas were detected 5–9 days before symptoms appeared.     

High-level expression of recombinant phospholipase C from Bacillus cereus in Pichia pastoris and its characterization

The phospholipase c (plc) gene from Bacillus cereus was cloned into the pPICZC vector and integrated into the genome of Pichia pastoris. The phospholipase C (PLC) when expressed in P. pastoris was fused to the -factor secretion signal peptide of Saccharomyces cerevisiae and secreted into a culture medium. Recombinant P. pastoris X-33 had a clear PLC band at 28.5 kDa and produced an extracellular PLC with an activity of 678 U mg^–1 protein which was more than a recombinant P. pastoris GS115 (552 U mg^–1 protein) or KM71H (539 U mg^–1 protein). The PLCs were purified using a HiTrap affinity column with a specific activity of 1335 U mg^–1 protein by P. pastoris GS115, 1176 U mg^–1 protein by P. pastoris KM71H and 1522 U mg^–1 protein by P. pastoris X-33. The three recombinant PLCs had high PLC activity in the low pH range of 4-5 and higher thermal stability (e.g. stable at 75 °C) than the wild-type PLC from B. cereus. Some organic solvents, surfactants and metal ions, e.g. methanol, acetone, Co²⁺ and Mn²⁺ etc., also influenced the activity of the recombinant PLCs.     

Agrobacterium-mediated transient expression in citrus leaves: a rapid tool for gene expression and functional gene assay

In this study, we present a method for transient expression of the type III effector AvrGf1 from Xanthomonas citri subsp. citri strain A^w in grapefruit leaves (Citrus paradisi) via Agrobacterium tumefaciens. The coding sequence of avrGf1 was placed under the control of the constitutive CaMV 35S promoter in the binary vectors pGWB2 and pGWB5. Infiltration of grapefruit leaves with A. tumefaciens carrying these constructs triggered a hypersensitive response (HR) in grapefruit 4 days after inoculation. When transiently expressed in grapefruit leaves, two mutants, AvrGf1ΔN116 and AvrGf1ΔC83, failed to induce an HR. Moreover, using bioinformatics tools, a chloroplast transit signal was predicted at the N terminus of AvrGf1. We demonstrated chloroplast localization by using an AvrGf1::GFP fusion protein, where confocal images revealed that GFP fluorescence was accumulating in the stomatal cells that are abundant in chloroplasts. Transient expression in citrus has the potential for aiding in the development of new disease defense strategies in citrus.     

3种外源诱导剂预处理对甜瓜抗病特征及其Fom 2和CHT基因表达的影响

该研究选用水杨酸(SA)、茉莉酸甲酯(MeJA)、Ca~(2+)、无菌水(对照)作为外源预处理诱导剂,以抗、感枯萎病甜瓜品种为材料,分别于诱导预处理2d后接种甜瓜枯萎菌,并于接种5、7、9d时观察发病情况,进行病情调查;在接种后1、3、5、7、9d取甜瓜叶片,分析抗病甜瓜(MR-1)和感病甜瓜(M1-15)叶片中甜瓜抗枯萎病基因(Fom-2)、几丁质酶基因(CHT)的表达变化,以探寻提高防治甜瓜枯萎病菌侵染的技术途径。结果显示:(1)外源MeJA和SA预处理接种后2品种的病情指数显著低于对照,但Ca~(2+)处理后的病情指数与对照无显著差异。(2)经外源诱导预处理接种后,MR-1和M1-15品种叶片的Fom-2和CHT基因均出现差异表达,但Ca~(2+)诱导其上调表达的效果微弱。(3)经SA、MeJA诱导预处理接种后,2品种叶片的Fom-2和CHT基因表达总体均显著高于对照;Fom-2基因的表达抗病甜瓜MR-1分别在接种后5d、7d时达到峰值,而感病甜瓜M1-15则均在接种9d时达到峰值;CHT基因的表达抗病甜瓜MR-1则均在接种后7d时达到峰值,而感病甜瓜M1-15分别在接种后7d、9d时达到峰值。(4)Ca~(2+)处理对抗、感甜瓜叶片的Fom-2和CHT基因的表达均无显著影响。(5)相关分析表明,经SA、MeJA诱导预处理接种后,甜瓜枯萎病病情指数与Fom-2和CHT基因表达量有显著的相关性;而Ca~(2+)处理效果不显著。研究表明:SA、MeJA通过诱导Fom-2、CHT基因上调表达,进而使甜瓜的抗病性提高,而Ca~(2+)处理对两基因表达和甜瓜抗病性均无显著影响。     

The Effect of Arachidonic Acid and Viral Infection on the Phytohemagglutinin Activity during the Development of Tobacco Acquired Resistance

The effects of arachidonic acid (AA) on the development of viral infection and the activity of phytohemagglutinins in Nicotiana tabacum L. plants were studied. Cv. Samsun NN was used, which displayed a genotypically determined hypersensitive response to tobacco mosaic virus (TMV) infection. When tobacco leaf disks were treated with 10^–9 to –10^–7 M AA, viral reproduction was suppressed by 90–100%. The AA concentration of 10^–8 M was optimal for the improvement of plant virus resistance. Tobacco leaves maintained virus resistance for at least two weeks. Both AA treatment and TMV inoculation were accompanied by an enhanced lectin activity, which may indicate the involvement of lectins in the development of plant defense responses. Lectin accumulation was observed in the intact plants developing systemic resistance and in the detached leaves characterized by local resistance.     

The effect of seasonality on oxidative metabolism in Nacella (Patinigera) magellanica

We studied the seasonal variation on aerobic metabolism and the response of oxidative stress parameters in the digestive glands of the subpolar limpet Nacella (P.) magellanica. Sampling was carried out from July (winter) 2002 to July 2003 in Beagle Channel, Tierra del Fuego, Argentina. Whole animal respiration rates increased in early spring as the animals spawned and remained elevated throughout summer and fall (winter: 0.09 ± 0.02 μmol O₂ h^− 1 g^− 1; summer: 0.31 ± 0.06 μmol O₂ h^− 1 g^− 1). Oxidative stress was assessed at the hydrophilic level as the ascorbyl radical content / ascorbate content ratio (A / AH⁻). The A / AH⁻ ratio showed minimum values in winter (3.7 ± 0.2 10^− 5 AU) and increased in summer (18 ± 5 10^− 5 AU). A similar pattern was observed for lipid radical content (122 ± 29 pmol mg^− 1 fresh mass [FW] in winter and 314 ± 45 pmol mg^− 1 FW in summer), iron content (0.99 ± 0.07 and 2.7 ± 0.6 nmol mg^− 1 FW in winter and summer, respectively) and catalase activity (2.9 ± 0.2 and 7 ± 1 U mg^− 1 FW in winter and summer, respectively). Since nitrogen derived radicals are thought to be critically involved in oxidative metabolism in cells, nitric oxide content was measured and a significant difference in the content of the Fe–MGD–NO adduct in digestive glands from winter and summer animals was observed. Together, the data indicate that both oxygen and nitrogen radical generation rates in N. (P.) magellanica are strongly dependent on season.     

Thermoregulation of water foraging wasps (Vespula vulgaris and Polistes dominulus)

A comparison of the thermoregulation of water foraging wasps (Vespula vulgaris, Polistes dominulus) under special consideration of ambient temperature and solar radiation was conducted. The body surface temperature of living and dead wasps was measured by infrared thermography under natural conditions in their environment without disturbing the insects’ behaviour. The body temperature of both of them was positively correlated with T_a and solar radiation. At moderate T_a (22–28 °C) the regression lines revealed mean thorax temperatures (T_th) of 35.5–37.5 °C in Vespula, and of 28.6–33.7 °C in Polistes. At high T_a (30–39 °C) T_th was 37.2–40.6 °C in Vespula and 37.0–40.8 °C in Polistes. The thorax temperature excess (T_th–T_a) increased at moderate T_a by 1.9 °C (Vespula) and 4.4 °C (Polistes) per kW⁻¹ m⁻². At high T_a it increased by 4.0 °C per kW⁻¹ m⁻² in both wasps. A comparison of the living water foraging Vespula and Polistes with dead wasps revealed a great difference in their thermoregulatory behaviour. At moderate T_a (22–28 °C) Vespula exhibited distinct endothermy in contrast to Polistes, which showed only a weak endothermic activity. At high T_a (30–39 °C) Vespula reduced their active heat production, and Polistes were always ectothermic. Both species exhibited an increasing cooling effort with increasing insolation and ambient temperature.     

Survival and photosynthetic activity of different Dinophysis acuminata populations in the northern Baltic Sea

This study deals with a recently found phenomenon in the northern Baltic Sea: the occurrence of the dinoflagellate Dinophysis acuminata in the deep water below the thermocline. This was first observed in July 2001 at the station BY 15 in the Gotland Deep, where a sharp and intensive chlorophyll fluorescence signal was encountered at 77 m depth. The fluorescence peak was due to a dinoflagellate community dominated by Dinophysis acuminata (approximately 18 000 cells l⁻¹). The survival of this community was followed in laboratory incubations in low light (20 μE m⁻² s⁻¹) and low temperature (+5 °C). After 5 weeks incubation, 67–84% of the initial cell abundance was lost, while few D. acuminata cells survived up to 24 weeks in the original sample. During the incubation, the fluorescence signal of the cells became fainter and the chloroplasts smaller and aggregated. On two occasions a D. acuminata cell was found attached to a smaller cell by a thin cytoplasm strand, possibly indicating mixotrophic behavior. During the following summer (2002), the photosynthetic efficiency of D. acuminata collected from thermocline layers of few stations and from the nitracline (75–80 m) at one station was studied in photosynthesis irradiance (P–E) incubations. Photosynthetic activity occurred in all populations, with differences in their photosynthetic carbon uptake rates. Photosynthesis of D. acuminata populations was saturated between 250 and 500 μE m⁻² s⁻¹; maximum cell-specific carbon uptake rates (P_m) ranged from 160–925 pg C cell⁻¹ h⁻¹. The P_m-rates in populations originating below the thermocline and in an artificially darkened population were markedly lower than in populations from upper water layers. The varying maximum photosynthetic rates of these populations may reflect their history, e.g. time spent in different light environments.     

Candidate Agtr2 influenced genes and pathways identified by expression profiling in the developing brain of Agtr2 mice

Intellectual disability (ID) is a common developmental disability observed in 1 to 3% of the human population. A possible role for the Angiotensin II type 2 receptor (AGTR2) in brain function, affecting learning, memory, and behavior, has been suggested in humans and rodents. Mice lacking the Agtr2 gene (Agtr2^−/y) showed significant impairment in their spatial memory and exhibited abnormal dendritic spine morphology. To identify Agtr2 influenced genes and pathways, we performed whole genome microarray analysis on RNA isolated from brains of Agtr2^−/y and control male mice at embryonic day 15 (E15) and postnatal day one (P1). The gene expression profiles of the Agtr2^−/y brain samples were significantly different when compared to profiles of the age-matched control brains. We identified 62 differently expressed genes (p ≤ 0.005) at E15 and in P1 brains of the Agtr2^−/y mice. We verified the differential expression of several of these genes in brain samples using quantitative RT-PCR. Differentially expressed genes encode molecules involved in multiple cellular processes including microtubule functions associated with dendritic spine morphology. This study provides insight into Agtr2 influenced candidate genes and suggests that expression dysregulation of these genes may modulate Agtr2 actions in the brain that influences learning and memory.     

Effects of polyphenol oxidase on lipolysis and proteolysis of red clover silage with and without a silage inoculant (Lactobacillus plantarum L54)

Polyphenol oxidase (PPO) has been shown to reduce proteolysis and lipolysis in red clover through deactivation of proteolytic and lipolytic enzymes and/or through formation of protein–phenol–lipid complexes. This experiment investigated the time course of both lipolysis and proteolysis in two red clover lines with different PPO activities with and without addition of a silage inoculant to help understand the action of PPO in the silo, and its potential effects on protein and glycerol-based lipid conservation, and to determine effects of a more rapid pH reduction with inoculation on PPO activity. Four silages were prepared from high or low PPO precision chopped red clover in 60 test-tube-silos, each containing 120 g fresh weight: (a) high PPO red clover without inoculation (H−), (b) low PPO red clover without inoculation (L−), (c) high PPO red clover with inoculation (H+), and (d) low PPO red clover with inoculation (L+). Each treatment had three replicates for each time point of 1, 2, 4, 8 and 90 days. The inoculant used was Lactobacillus plantarum strain L54 applied at a rate of 10⁶ CFU/g fresh weight. Silage pH was reduced (P < 0.001) by inoculation with no effect of PPO. Inoculation had no effect on either lipolysis or free amino acid release, although more (P < 0.01) soluble protein and less (P < 0.01) ammonia-N was in inoculated silages. H silages had a lower level of both proteolysis (release of free amino acids, P < 0.05) and lipolysis (loss of membrane lipid, P < 0.01) than L red clover silages. Results indicate that PPO reduced proteolysis and lipolysis in the silo and that inoculation had no adverse effects on PPO activity.     

Characteristics of an endophytic pyrene-degrading bacterium of Enterobacter sp. 12J1 from Allium macrostemon Bunge

One pyrene-degrading endophytic bacterium was isolated from plants grown in polycyclic aromatic hydrocarbon-contaminated soils and identified as Enterobacter sp. 12J1 based on the 16S rDNA gene sequence analysis. Heavy metal and antibiotic resistance, degradation of pyrene, solubilization of inorganic phosphate and cell surface hydrophobicity characteristics of the isolate were further characterized. The isolate was also evaluated for promoting plant growth of wheat and maize and pyrene removal from pyrene-amended soil in pot experiments. High-performance liquid chromatograph (HPLC) analysis showed that the degradation rate of pyrene (5 mg l⁻¹) by the endophytic bacterial strain 12J1 was 83.8% under 28 °C for 7 days. The Enterobacter sp. 12J1 could produce indole acetic acid (IAA), siderophore and solubilize inorganic phosphate. The Enterobacter sp. 12J1 also has a cell surface hydrophobicity. In the live bacterial inoculation experiment, an increase in pyrene removal varying from 60% to 107% was observed in the planted soils treated with 100 mg kg⁻¹ of pyrene compared with the unplanted soils. The rate of pyrene removal increased by 43–65% in the live bacterium-inoculated planted soils compared with the dead bacterium-inoculated planted soils. Although there were no significant differences in the total culturable bacterial numbers between live and dead bacterial inoculation, the numbers of pyrene-degrading bacteria were significantly greater in the live bacterium-inoculated planted or unplanted soils. The isolate could colonize the tissue (root and stem) interiors and rhizosphere soils of wheat and maize after root inoculation.     

ICChI, a glycosylated chitinase from the latex of Ipomoea carnea

A multi-functional enzyme ICChI with chitinase/lysozyme/exochitinase activity from the latex of Ipomoea carnea subsp. fistulosa was purified to homogeneity using ammonium sulphate precipitation, hydrophobic interaction and size exclusion chromatography. The enzyme is glycosylated (14–15%), has a molecular mass of 34.94 kDa (MALDI–TOF) and an isoelectric point of pH 5.3. The enzyme is stable in pH range 5.0–9.0, 80 °C and the optimal activity is observed at pH 6.0 and 60 °C. Using p-nitrophenyl-N-acetyl-β-d-glucosaminide, the kinetic parameters K_m, V_max, K_cat and specificity constant of the enzyme were calculated as 0.5 mM, 2.5 × 10⁻⁸ mol min⁻¹ μg enzyme⁻¹, 29.0 s⁻¹ and 58.0 mM⁻¹ s⁻¹ respectively. The extinction coefficient was estimated as 20.56 M⁻¹ cm⁻¹. The protein contains eight tryptophan, 20 tyrosine and six cysteine residues forming three disulfide bridges. The polyclonal antibodies raised and immunodiffusion suggests that the antigenic determinants of ICChI are unique. The first fifteen N-terminal residues G–E–I–A–I–Y–W–G–Q–N–G–G–E–G–S exhibited considerable similarity to other known chitinases. Owing to these unique properties the reported enzyme would find applications in agricultural, pharmaceutical, biomedical and biotechnological fields.     

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

Control of green mold and sour rot of stored lemon by biofumigation with Muscodor albus

Control of postharvest lemon diseases by biofumigation with the volatile-producing fungus Muscodor albus was investigated. In vitro exposure to M. albus volatile compounds for 3 days killed Penicillium digitatum and Geotrichum citri-aurantii, causes of green mold and sour rot of lemons, respectively. Lemons were wound-inoculated with P. digitatum and placed in closed 11-L plastic boxes with rye grain cultures of M. albus at ambient temperature. There was no contact between the fungus and the fruit. Biofumigation for 24–72 h controlled green mold significantly, even when treatment began 24 h after inoculation. Effectiveness was related to the amount of M. albus present. In tests conducted inside a 11.7-m³ degreening room with 5 ppm ethylene at 20 °C, green mold incidence on lemons was reduced on average from 89.8 to 26.2% after exposure to M. albus for 48 h. Ethylene accelerates color development in harvested citrus fruit. M. albus had no effect on color development. Biofumigation in small boxes immediately after inoculation controlled sour rot, but was ineffective if applied 24 h later. G. citri-aurantii may be less sensitive to the volatile compounds than P. digitatum or escapes exposure within the fruit rind. Biofumigation with M. albus could control decay effectively in storage rooms or shipping packages.     

Regulation of expression of nif and hut operons in Klebsiella pneumoniae by glnA linked genes of Escherichia coli

Summary Recombinant DNA plasmids containing inserts from the glnA region of Escherichia coli were used to study the expression of gln, hut, and nif operons in a regulation defective mutant (Gln^–Hut^–Nif^–) of Klebsiella pneumoniae, KP5060. Genes adjacent to the C-terminal end of glnA on the E. coli chromosome were able to derepress hut and nif operons in K. pneumoniae in the absence of glnA product. However, complete derepression of nif operons required inclusion of the segment adjacent to the N-terminal end of the glnA region of the E. coli chromosome along with the C-terminal end segment. In the absence of functional glnA, such a fully derepressed strain expressed nif and hut constitutively indicating a role for the catalytic activity of glutamine synthetase in repression of the genes under nitrogen control.     

Gas exchange of Ginkgo biloba leaves at different CO2 concentration levels

One and a half year-old Ginkgo saplings were grown for 2 years in 7 litre pots with medium fertile soil at ambient air CO₂ concentration and at 700 μmol mol⁻¹ CO₂ in temperature and humidity-controlled cabinets standing in the field. In the middle of the 2nd season of CO₂ enrichment, CO₂ exchange and transpiration in response to CO₂ concentration was measured with a mini-cuvette system. In addition, the same measurements were conducted in the crown of one 60-year-old tree in the field. Number of leaves/tree was enhanced by elevated CO₂ and specific leaf area decreased significantly.CO₂ compensation points were reached at 75–84 μmol mol⁻¹ CO₂. Gas exchange of Ginkgo saplings reacted more intensively upon CO₂ than those of the adult Ginkgo. On an average, stomatal conductance decreased by 30% as CO₂ concentration increased from 30 to 1000 μmol mol⁻¹ CO₂. Water use efficiency of net photosynthesis was positively correlated with CO₂ concentration levels. Saturation of net photosynthesis and lowest level of stomatal conductance was reached by the leaves of Ginkgo saplings at >1000 μmol mol⁻¹ CO₂. Acclimation of leaf net CO₂ assimilation to the elevated CO₂ concentration at growth occurred after 2 years of exposure. Maximum of net CO₂ assimilation was 56% higher at ambient air CO₂ concentration than at 700 μmol mol⁻¹ CO₂.