Bradyrhizobium japonicum mutants defective in root-nodule bacteroid development and nitrogen fixation

The genome of the slow-growing Bradyrhizobium japonicum (strain 110) was mutagenized with transposon Tn5. A total of 1623 kanamycin/streptomycin resistant derivatives were screened in soybean infection tests for nodulation (Nod) and symbiotic nitrogen fixation (Fix). In this report we describe 14 strains possessing a stable, reproducible Nod⁺Fix^- phenotype. These strains were also grown under microaerobic culture conditions to test them for free-living nitrogen fixation activity (Nif). In addition to strains having reduced Fix and Nif activities, there were also strains that had reduced symbiotic Fix activity but were Nif⁺ ex planta.Analysis of the genomic structure revealed that the majority of the strains had a single Tn5 insertion without any further apparent physical alteration. A few strains had additional insertions (by Tn5 or IS50), or a deletion, or had cointegrated part of the vector used for Tn5 mutagenesis. One of the insertions was found in a known nif gene (nifD) whereas all other mutations seem to affect different, hitherto unknown genes or operons.Several mutant strains had an altered nodulation phenotype, inducing numerous, small, widely distributed nodules. Light and electron microscopy revealed that most of these mutants were defective in different stages of bacteroid development and/or bacteroid persistence. The protein patterns of the mutants were inspected by two-dimensional gel electrophoresis after labelling microaerobic cultures with l-(³⁵S)methionine. Of particular interest were mutants lacking a group of proteins the synthesis of which was known to be under oxygen control. Such strains can be regarded as potential regulatory mutants.     

The effect of oxygen on nitrate and nitrite assimilation in leaves of Zea mays L. under dark conditions

The assimilation of nitrate under dark-N₂ and dark-O₂ conditions in Zea mays leaf tissue was investigated using colourimetric and ¹⁵N techniques for the determination of organic and inorganic nitrogen. Studies using ¹⁵N indicated that nitrate was assimilated under dark conditions. However, the rate of nitrate assimilation in the dark was only 28% of the rate under non-saturating light conditions. No nitrite accumulated under dark aerobiosis, even though nitrate reduction occurred under these conditions. The pattern of nitrite accumulation in leaf tissue in response to dark-N₂ conditions consisted of three phases: an initial lag phase, followed by a period of rapid nitrite accumulation and finally a phase during which the rate of nitrite accumulation declined. After a 1-h period of dark-anaerobiosis, both nitrate reduction and nitrite accumulation declined considerably. However, when O₂ was supplied, nitrate reduction was stimulated and the accumulated nitrite was rapidly reduced. Anaerobic conditions stimulated nitrate reduction in leaf tissue after a period of dark-aerobic pretreatment.     

Nitrogen supply as a factor influencing photoinhibition and photosynthetic acclimation after transfer of shade-grown Solanum dulcamara to bright light

We have compared the ability of shadegrown clones of Solamum dulcamara L. from shade and sun habitats to acclimate to bright light, as a function of nitrogen nutrition before and after transfer to bright light. Leaves of S. dulcamara grown in the shade with 0.6 mM NO ₃ ^- have similar photosynthetic properties as leaves of plants grown with 12.0 mM NO ₃ ^- . When transferred to bright light for 1–2 d the leaves of these plants show substantial photoinhibition which is characterized by about 50% decrease in apparent quantum yield and a reduction in the rate of photosynthesis in air at light saturation. Photoinhibition of leaf photosynthesis is associated with reduction in the variable component of low-temperature fluorescence emission, and with loss of in-vitro electron transport, especially of photosystem II-dependent processes.We find no evidence for ecotypic differentiation in the potential for photosynthetic acclimation among shade and sun clones of S. dulcamara, or of differentiation with respect to nitrogen requirements for acclimation. Recovery from photoinhibition and subsequent acclimation of photosynthesis to bright light only occurs in leaves of plants provided with 12.0 mM NO ₃ ^- . In these, apparent quantum yield is fully restored after 14 d, and photosynthetic acclimation is shown by an increase in light-saturated photosynthesis in air, of light-and CO₂-saturated photosynthesis, and of the initial slope of the CO₂-response curve. The latter changes are highly correlated with changes in ribulose-bisphosphate-carboxylase activity in vitro. Plants supplied with 0.6 mM NO ₃ ^- show incomplete recovery of apparent quantum yield after 14 d, but CO₂-dependent leaf photosynthetic parameters return to control levels.Symbols and abbreviations F_o initial level of fluorescence at 77 K - F_m maximum level of fluorescence at 77 K - F_v variable components of fluorescence at 77 K (F_v=F_m-F_o) - PSI, PSII photosystem I and II, respectively - RuBP ribulose-1,5-bisphosphate - RuBPCase ribulose-1,5-bisphosphate carboxylase-oxygenase (EC 4.1.1.39)     

Diffusion du15N2 dans le sol pendant la mesure de fixation biologique de l'azote

Summary The kinetic of¹⁵N₂ diffusion has been measured in a system similar to that for the estimation of N₂ fixation in plant microorganism associations cultivated in soil. The¹⁵N₂ enrichment of the soil atmosphere reached an homogenous value one hour after injection of¹⁵N₂ and is identical to that obtained by calculation, indicating that no adsorption occurs in the soil particles.

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Diffusion du¹⁵N₂ dans le sol pendant la mesure de fixation biologique de l'azote

Résumé La cinétique de diffusion du¹⁵N₂ est mesurée sur un système identique à ceux pouvant être utilisés pour la mesure de fixation de l'azote chez les associations plantes-microorganismes cultivées sur sol. L'enrichissement homogène de l'atmosphère du sol est obtenu une heure environ après l'injection de¹⁵N₂ et correspond à l'enrichissement calculé, ce qui indique qu'aucune adsorption n'a lieu dans les particules du sol.

     

Genetic specificity of nitrogen nutrition in leguminous plants

Summary Mineral nitrogen did not increase grain yield and seed protein levels ofVicia faba L. andLupinus luteus L. in field trials and pot experiments. Fixed N₂ was substituted by mineral nitrogen in these cases because of inhibition of N₂ fixation by mineral nitrogen. Contrary to these results mineral nitrogen increased grain yields and seed protein amounts ofLupinus albus L.,Pisum sativum L., andGlycine max. (L.) Merr. The nitrogen effect was caused at an early stage by saving energy due to inhibition of N₂ fixation (measurement of gas exchange by means of IRGA). In case of the N application after flowering grain, yields and seed protein levels increased because the mineral N was an additional nitrogen source for plants. At this stage the plants had ceased fixing atmospheric nitrogen. The high sink activity of growing fruits induced a lack of assimilates in nodules (determined by means of¹⁴CO₂ application). The N effect was therefore the consequence of the lower assimilate pool for supplying root nodules in these plants in comparison withVicia faba L. andLupinus luteus L. Hence it follows that response to mineral nitrogen can be a criterion for discovering more effective Rhizobium-host combinations.     

Interrelationships between Ethylene Production,Gall Formation,and Root-knot Nematode Development in Tomato Plants Infected with Meloidogyne javanica

Ethylene production was determined in excised tomato (Lycopersicon esculentum) root cultures of Meloidogyne javanica susceptible and resistant cultivars infected with M. javanica. Uninfected cultivars produced very low amounts of ethylene. Relatively high amounts of ethylene were produced by the infected susceptible cultivars. Peak production of 1.6 n moles * g root⁻¹ * h¹⁻ occurred between 9 and 16 days after inoculation (DAI). The period of high ethylene production coincided with that of rapid increase in gall weight. Low amounts of ethylene were also released by the infected resistant cultivar between 9 and 12 DAI, which follows the hypersensitivity reaction. Ethylene production in infected intact plants during the period of rapid gall growth was twice as much as in uninfected plants during the same time. Exposing excised root cultures to 0.5 or l0 ppm ethylene accelerated the rate of increase in gall weight of M. javanica infected roots. In contrast, overall root growth was inhibited by these treatments, compared to infected roots which were not exposed to ethylene.     

Trypanosoma congolense. I. Clinical observations of experimentally infected cattle

The course of disease was studied in 8 cattle infected with Trypanosoma congolense. Although the onset of patency was dependent on the numbers of infecting organisms, the duration of the infection was not. High fevers were present on the day of or the day after initial patency. Succeeding peaks of parasitemia, and a progressive weight loss of over 30% occurred. A decrease in packed cell volume (PCV) beginning the first week after infection was observed. Early in the course of the developing anemia, many polychromatophilic erythrocytes and occasional normoblasts were found in the blood. A leucopenia persisted for the duration of the disease. Total serum protein concentrations fell sharply during the first 5 weeks of infection, then gradually increased to low normal levels. Serum albumin levels followed a similar pattern for the first 5 weeks, and remained at a relatively low level. Although gamma globulin levels also declined during the first 5 weeks, their levels gradually surpassed those of preinfection samples. No marked changes in serum glucose were noted. A mild elevation of serum urea nitrogen values occurred early during infection, but subsided. The animals dying early after infection developed elevated total bilirubin levels.     

Nutrition et élevage du prédateur polyphageMacrolophus caliginosus [Heteroptera,Miridae] sur milieux artificiels

La possibilité de nourrir la punaiseMacrolophus caliginosus Wagner à l'aide de milieux artificiels a été testée. En l'absence de végétal, les témoins nourris avec des ?ufs d'Ephestia kuehniella Zeller ont une survie médiocre et fournisent 33% d'adultes. Avec le meilleur milieu artificiel la survie est comparable à celle des témoins et 21 % d'adultes sont obtenus. Avec ce même milieu, mais en présence de feuille de géranium, la production d'adultes atteint 62%. Les compositions en acides aminés des punaises élevées avec le milieu montrent des écarts inférieurs à 20% par rapport aux témoins. Le développement complet deM. caliginosus nourri à l'aide de milieu artificiel a été obtenu. Le végétal joue un rôle important dans la biologie de ce prédateur.     

虫生真菌蝉拟青霉的研究

观察了蝉拟青霉的无性世代,在24℃下载片培养结果,分生孢子经8hr萌发,24hr普遍形成菌丝,36hr出现产孢结构和产生次代分生孢子。该菌生长合适温度24—26℃,分生孢子萌芽要求相对湿度在90%以上。pH4—12范围均见生长,但以5—6为佳。对10种碳源和9种无机氮源利用检测结果,用葡萄糖作碳源孢子产量高,用果糖作碳源菌丝体产量高。不利用菊糖、L-山梨糖、L-鼠李糖。对KNO_3利用佳,但不能利用NaNO_2,和硫脲。该菌能较强抗紫外辐射。