Nodulation and nitrogen-fixation byHippophaë rhamnoides L. in the field

Summary 1. The extent of nodulation and nitrogen fixation by a population ofHippophaë rhamnoides bushes occurring on a sand dune system on the east coast of England has been studied.2. Nodules were found on all plants (the latter ranged from 3 to 16 years in age) and using the isotope N¹⁵ it was confirmed that these nodules fixed nitrogen. The nodules on the youngest plants were most active in fixation but because of the greater total nodule mass older plants fixed the largest quantities of nitrogen.3. Large increases in total nitrogen (i.e. soil plus plant nitrogen) occurred with increase in age of the plants, such nitrogen increases ranging from 27 kg per hectare per annum in the presence of bushes 0 to 3 years old to 179 kg per hectare per annum in the presence of bushes 13 to 16 years old.4. The N¹⁵-technique indicated that a proportion of the nitrogen accumulated was due to fixation byHippophaë but nitrogen contributions from other sources could not be ruled out.5. The levels of nitrate-nitrogen and ammonium-nitrogen in the soil underHippophaë showed a marked seasonal variation, increasing to a maximum in the winter and to a minimum in the summer. These levels were in general inversely related to the level of average monthly rainfall. Because of the low levels of combined nitrogen in the soil during the summer months it seems unlikely that combined nitrogen should markedly inhibit fixation in this situation.     

The formation of vesicles in the developmental cycle of the nodular endophyte of Hippophaë rhamnoides L.

Summary The mature vesicle of the Hippophaë rhamnoides root nodule endophyte is spherical, approximately 3–4 m in diameter and exhibits a high degree of apparently random septation. Electron micrographs are presented which show that this vesicular form in the endophytic development within the host cell originates as a swelling of the hyphal tip. The young vesicle is non-septate and in general attains a minimum diameter of 3 m before septation becomes evident. The number of septa then increases with vesicle maturity.     

Feedback regulation of nodule formation in Hippophaë rhamnoides

Seedlings of Hippophaë rhamnoides possessing two equally infectible root systems (split roots) were used in conjunction with specific Frankia strains to investigate plant control over nodulation. When a wild-type Frankia strain was inoculated onto both root systems simultaneously or 1, 2, 4, or 8 weeks apart, an inhibitory response occurred which retarded nodulation on the root exposed to the delayed inoculum. Similar suppressive responses were also observed when two different wild-type Frankia inocula were applied onto opposite sides of a split-root system at different times. The depressed response shown by the delayed inoculum was more pronounced as the delay period was increased. The roots exposed to the delayed inoculum displayed a complete lack of nodulation when the delay was 4 or 8 weeks. The nodulation response on the root inoculated first depended on subsequent inoculation of the second root system of the plant, so that maximum nodulation of the first root was observed when the second root was unnodulated. These results provide evidence that sea buckthorn has an active, systemic mechanism for feedback control of nodulation that suppresses further nodule formation and prevents excessive nodulation. The significance of these results to the understanding of nodule ontogeny is discussed.     

The of gamma-radiation for induction of apomixis in sea-buckthorn (Hippophaë rhamnoides L.)

The results of studying of the induced apomixis in pollen of sea-buckthorn irradiated by the 60Co gamma-Radiation are considered. Was established that the most effective dose for pollination of the experimental plants is 50 k Gy. In total, from 46 seedlings 19 cases of apomictic origin were revealed, 7 individuals were found to be haploid (n = = 12) and 19 ones were diploid (2n = 24) of maternal origin. Was supposed that apomictic plants (19 seedlings) have parthenogenetic origin. The reason for such conclusion is that the irradiated anomalous pollen tubes despite not having spermia, are entering embryo sac and stimulate the development of apomictic embryo from non-fertilized female gametes. Apparently, pollen tubes cause the induction of DNA replication in the ovules and the development of parthenogenetic plants. Consequently, the described method can be used for the regulation of parthenogenesis in sea-buckthorn to change natural ratio (1 : 1) of male to female plants in desirable quantity.     

Observations on the fine structure of the root nodule endophyte of Hippophaë rhamnoides L.

Summary Electron microscopy of ultra-thin sections of Hippophaë rhamnoides root nodules has been carried out in order to elucidate the nature of the endophyte. The organism is seen as a branching, septate filament approximately 0.6 microns in diameter bearing on its terminal ends spherical sub-divided vesicles 3–4 microns in diameter. In the mature nodule the vesicles are the most prominent endophyte form and appear to be formed by swelling of the hyphal tips. It is concluded that the endophyte is an actinomycete closely related to but not identical with that of Alnus glutinosa.     

Nematodes (Longidorus sp. andTylenchorhynchus microphasmis loof) in growth and nodulation of sea buckthorn (Hippophaë rhamnoides L.)

Summary Hippophaë rhamnoides seedlings were grown in sterilized and unsterilized soil from a decliningH. rhamnoides scrub, to which different numbers ofLongidorus sp. andTylenchorhynchus microphasmis were added. In sterilized and unsterilized soil, retardation of growth, content of dry matter in the shoots, and incidence of deformed short lateral roots of test plants were positively correlated with counts of both nematode species. Nitrogen content in the shoots, nodulation on the roots of test plants and increase increase in nematodes were negatively correlated with the initial number of both nematode species in sterilized soil. In unsterilized soil, an unknown biotic factor was present that reduces growth ofH. rhamnoides, nodulation and multiplication of the nematodes. This factor seems to interact with the nematodes in reducing growth ofH. rhamnoides.Deceased.     

Ionic balance,proton efflux,nitrate reductase activity and growth ofHippophaë rhamnoides L. ssp.rhamnoides as influenced by combined-N nutrition or N2 fixation

Growth of 2-month-old nonnodulatedHippophaë rhamnoides seedlings supplied with combined N was compared with that of nodulated seedlings grown on zero N. Plant growth was significantly better with combined N than with N₂ fixation and, although not statistically significant for individual harvests, tended to be highest in the presence of NH ₄ ⁺ , a mixture of NH ₄ ⁺ and NO ₃ ^? producing the highest yields. Growth was severely reduced when solely dependent on N₂ fixation and, unlike the combined-N plants, shoot to root ratios had only slightly increased after an initial decrease. An apparently insufficient nodule mass (nodule weight ratio <5 per cent) during the greater part of the experimental period is suggested as the main cause of the growth reduction in N₂-fixing plants. Thein vivo nitrate reductase activity (NRA) of NO ₃ ^? dependent plants was almost entirely located in the roots. However, when grown with a combination of NO ₃ ^? and NH ₄ ⁺ , root NRA was decreased by approximately 85 per cent.H. rhamnoides demonstrated in the mixed supply a strong preference for uptake of N as NH ₄ ⁺ , NO ₃ ^? contributing only for approximately 20 per cent to the total N assimilation. Specific rates of N acquisition and ion uptake were generally highest in NO ₃ ^? +NH ₄ ⁺ plants. The generation of organic anions per unit total plant dry weight was approximately 40 per cent less in the NH ₄ ⁺ plants than in the NO ₃ ^? plants. Measured extrusions of H⁺ or OH^? (HCO ₃ ^? ) were generally in good agreement with calculated values on the basis of plant composition, and the acidity generated with N₂ fixation amounted to 0.45–0.55 meq H⁺. (mmol N_org)^?1. Without acidity control and in the presence of NH ₄ ⁺ , specific rates of ion uptake and carboxylate generation were strongly depressed and growth was reduced by 30–35 per cent. Growth of nonnodulatedH. rhamnoides plants ceased at the lower pH limit of 3.1–3.2 and deterioration set in; in the case of N₂-fixing plants the nutrient solution pH stabilized at a value of 3.8–3.9 without any apparent adverse effects upon plant performance. The chemical composition of experimental and field-growing plants is being compared and some comments are made on the nitrogen supply characteristics of their natural sites.     

Net photosynthetic rate of sour cherry (Prunus cerasus L. ‘Montmorency’) during the growing season with particular reference to fruiting

Diurnal and seasonal net photosynthetic rates (Pn) of sour cherry were determined. Leaf Pn was not significantly affected by shoot excision. Under constant environmental conditions (PFD, 1200 μmol m^-2s^-1; temp. 25; relative humidity, 80–90%) there was no significant diurnal fluctuation in Pn for individual leaves. However, there was a pronounced fluctuation in Pn for whole trees measured under constant temperature but natural variation in sunlight from sunrise to sunset. Maximum Pn occurred before solar noon, remained constant for 1–2 hr, then declined. Photosynthetic rate of recently expanded leaves fluctuated through out the season but, in general, was greatest in the spring as leaves expanded, reached a peak, remained stable for several weeks, then gradually declined. The Pn of leaves on terminal shoots was not significantly different from the Pn of leaves on spurs of the same physiological age. The presence of fruit did not have a consistent effect on the Pn of sour cherry leaves.     

Net photosynthetic rate of sour cherry (Prunus cerasus L. ‘Montmorency’) during the growing season with particular reference to fruiting

Diurnal and seasonal net photosynthetic rates (Pn) of sour cherry were determined. Leaf Pn was not significantly affected by shoot excision. Under constant environmental conditions (PFD, 1200 mol m^-2 s^-1; temp. 25; relative humidity, 80–90%) there was no significant diurnal fluctuation in Pn for individual leaves. However, there was a pronounced fluctuation in Pn for whole trees measured under constant temperature but natural variation in sunlight from sunrise to sunset. Maximum Pn occurred before solar noon, remained constant for 1–2 hr, then declined. Photosynthetic rate of recently expanded leaves fluctuated through out the season but, in general, was greatest in the spring as leaves expanded, reached a peak, remained stable for several weeks, then gradually declined. The Pn of leaves on terminal shoots was not significantly different from the Pn of leaves on spurs of the same physiological age. The presence of fruit did not have a consistent effect on the Pn of sour cherry leaves.Michigan Agricultural Experiment Station Journal Article No. 10739.     

The significance of abrupt transitions in Lineweaver–Burk plots with particular reference to glutamate dehydrogenase. Negative and positive co-operativity in catalytic rate constants

1. Lineweaver-Burk plots for glutamate dehydrogenase, glucose 6-phosphate dehydrogenase and several other enzymes show one or more abrupt transitions between apparently linear sections. These transitions correspond to abrupt increases in the apparent K(m) and V(max.) with increasing concentration of the varied substrate. 2. The generalized reciprocal initial-rate equation for a multi-site enzyme requires several restrictions to be put on it in order to generate such plots. These mathematical conditions are explored. 3. It is shown that the effective omission of a term in the denominator of the reciprocal initial-rate equation represents a minimal requirement for generation of abrupt transitions. This corresponds in physical terms to negative co-operativity followed by positive co-operativity affecting the catalytic rate constant for the reaction. 4. Previous models for glutamate dehydrogenase cannot adequately account for the results. On the other hand, the model based on both negative and positive co-operativity gives a good fit to the experimental points. 5. The conclusions are discussed in relation to current knowledge of the structure and mechanism of glutamate dehydrogenase.     

Lathyrus diversity: available resources with relevance to crop improvement – L. sativus and L. cicera as case studies

Background

The Lathyrus genus includes 160 species, some of which have economic importance as food, fodder and ornamental crops (mainly L. sativus, L. cicera and L. odoratus, respectively) and are cultivated in >1·5 Mha worldwide. However, in spite of their well-recognized robustness and potential as a source of calories and protein for populations in drought-prone and marginal areas, cultivation is in decline and there is a high risk of genetic erosion.

Scope

In this review, current and past taxonomic treatments of the Lathyrus genus are assessed and its current status is examined together with future prospects for germplasm conservation, characterization and utilization. A particular emphasis is placed on the importance of diversity analysis for breeding of L. sativus and L. cicera.

Conclusions

Efforts for improvement of L. sativus and L. cicera should concentrate on the development of publicly available joint core collections, and on high-resolution genotyping. This will be critical for permitting decentralized phenotyping. Such a co-ordinated international effort should result in more efficient and faster breeding approaches, which are particularly needed for these neglected, underutilized Lathyrus species.     

Changes in behaviour associated with pair formation in the polychaete Harmothoë imbricata (L.)

Spacing between individuals in populations of Harmothoë imbricata has been investigated both on the shore and in the laboratory. Males tend to occur closer together than females, and the mean male‐male individual distance measured was less than the mean distance between females; male—female distances for immature worms were intermediate. When worms mature they pair: the male mounts the female and lies across her dorsal surface. There is evidence that after spawning the members of a pair separate. Contact responses between worms have been investigated in the laboratory. Most encounters between immature worms lead to separation, as a result of one or both worms moving rapidly away from the other or fighting; females show more marked avoidance behaviour than males. The majority of male‐male and female‐female encounters between mature worms also lead to separation but in male‐female encounters the male usually mounts the female. A male which has mounted a female becomes highly aggressive and will attack intruding males but not females.     

Hippophae rhamnoides L. (Sanddorn) als neues Züchtungsobjekt

Theoretical and Applied Genetics - Seit im Jahre 1941 die Beeren des Sanddorns (Hippophae rhamnoides L.) als hochwertige Vitaminträger (C+A) erkannt wurden, sind sie alljährlich aus den...     

Antioxidants in Festuca rubra L. seeds affected by the fungal symbiont Epichloë festucae

Vertically transmitted fungal endophytes can be beneficial for host grasses. While the alkaloid-mediated mechanism for herbivore resistance has been widely studied, underlying physiological mechanisms for increased tolerance to abiotic stress remain scarcely explored. In this study we used three maternal lines of perennial grass Festuca rubra to examine the role of antioxidants in endophyte-mediated effects on seed viability over long-term storage. Uncolonized plants (E?) were generated by removing the endophyte from ramets of naturally endophyte-colonized (E+) plants. The E?+ and E? ramets were planted in a common garden in Salamanca, Spain. Seeds produced in 2009, 2010 and 2011 were harvested at maturity, dried and stored at 10 °C until 2011 when we tested seed and endophyte viability, and measured antioxidants. Seed viability and α-tocopherol antioxidant were negatively affected by the endophyte in two maternal lines. In these same lines, the endophyte viability was lowest at the longest storage time. In the maternal line that showed the highest negative effect of endophyte on seed viability, the pattern of glutathione was opposite to that observed for tocopherols since it was higher for E?+ than for E? seeds. In all maternal lines, the glutathione half-cell reduction potential (E_GSSG/2GSH) and % glutathione disulphide (GSSG) increased with storage time but there was no clear pattern associated with endophyte symbiosis. Whether these parameters are good predictors of seed and endophyte longevity in storage and natural conditions should be further explored.     

Modeling the acid–base properties of glutathione in different ionic media, with particular reference to natural waters and biological fluids

The acid-base properties of γ-L-glutamyl-L-cysteinyl-glycine (glutathione, GSH) were determined by potentiometry (ISE-H(+), glass electrode) in pure NaI((aq)) and in NaCl((aq))/MgCl(2(aq)), and NaCl((aq))/CaCl(2(aq)) mixtures, at T = 298.15 K and different ionic strengths (up to I(c) ~ 5.0 mol L(-1)). In addition, the activity coefficients of glutathione were also determined by the distribution method at the same temperature in various ionic media (LiCl((aq)), NaCl((aq)), KCl((aq)), CsCl((aq)), MgCl(2(aq)), CaCl(2(aq)), NaI((aq))). The results obtained were also used to calculate the Specific ion Interaction Theory (SIT) and Pitzer coefficients for the dependence on medium and ionic strength of glutathione species, as well as the formation constants of weak Mg(j)H( i )(GSH)((i+2j-3)) and Ca(j)H(i)(GSH)((i+2j-3)) complexes. Direct calorimetric titrations were also carried out in pure NaCl((aq)) and in NaCl((aq))/CaCl(2(aq)) mixtures at different ionic strengths (0.25 ≤ I (c )/mol L(-1) ≤ 5.0) in order to determine the enthalpy changes for the protonation and complex formation equilibria in these media at T = 298.15 K. Results obtained are useful for the definition of glutathione speciation in any aqueous media containing the main cations of natural waters and biological fluids, such as Na(+), K(+), Mg(2+), and Ca(2+). Finally, this kind of systematic studies, where a series of ionic media (e.g., all alkali metal chlorides) is taken into account in the determination of various thermodynamic parameters, is useful for the definition of some trends in the thermodynamic behavior of glutathione in aqueous solution.     

New insights into vegetation dynamics and settlement history in Hümmling,north-western Germany,with particular reference to the Neolithic

Palynological investigations on two well-dated peat profiles provide insights into Neolithic vegetation and settlement history from Hümmling in north-western Germany. The site selections allow comparisons between local and regional vegetation changes and are used to estimate the extent of Neolithic influence on the vegetation. The interpretation of the fossil spectra relied on radiocarbon dating, evaluation of pollen indicator taxa, non-pollen palynomorphs and multivariate techniques. During the late Mesolithic the vegetation was dominated by mixed oak forests while openings in forest cover were detected, with a decline in elm reflected in the regional pollen record around 4250 cal. b.c. The presence of humans is shown by settlement indicators that are first recorded at ca. 3800 cal. b.c. Vegetation changes were small between 4300 and 3600 cal. b.c. This suggests that regional vegetation was relatively resilient to small-scale disturbances. Possible indications of grazing were recorded in the spectra of the local pollen profile but there is no clear-cut evidence for Neolithic activity. Between 3520 and 2260 cal. b.c. decreases in forest cover were inferred from both profiles and increases in settlement indicators reflect farming activity. These changes coincide with the emergence in the area of the Funnel Beaker Culture and the subsequent Single Grave Culture. Both profiles suggest that settlement probably ceased between ca. 3230 and 3050 cal. b.c. This lull or cessation in activity was probably regional in character. After 2260 cal. b.c. human impact on the vegetation decreases and woodlands regenerate. The longevity of the regeneration phase—ca. 690 years—was probably connected with the low resilient capability of the vegetation on the poor soils.     

Late Quaternary history of Hippophaë rhamnoides L. (Elaeagnaceae) inferred from chalcone synthase intron (Chsi) sequences and chloroplast DNA variation

Fossil pollen records indicate that Hippophaë rhamnoides (Sea Buckthorn) was widespread on late‐ and early postglacial raw soils throughout much of central and northern Europe, but that Early Holocene reforestation restricted populations to northern coastal habitats, or along mountain streams in the Alps, Pyrenees, and Carpathians. We used sequence variation at the nuclear chalcone synthase intron (Chsi), in conjunction with chloroplast DNA–restriction fragment length polymorphism data, to investigate the intraspecific phylogeny, phylogeographic structure, and expansion demographic history of this dioecious and wind‐pollinated shrub at its range‐wide scale in Europe and Asia Minor. Four major Chsi phylogroups of unresolved relationships were identified with estimated divergences ～172 000 years ago. Large‐scale phylogeographic structures of nuclear and cytoplasmic markers were congruent in identifying (i) southeastern Europe as the most likely source of colonization into central Europe and Scandinavia, and (ii) the area just north of the Alps as a contact zone between populations from the Alps and the east/central European‐Scandinavian lineage. Coalescence‐based analyses (i.e. nested clade analysis and mismatch distributions) of Chsi variation were able to detect at least four major episodes of population growth, all within about the last 40 000 years. In particular, these analyses identified a nearly synchronized timing of population expansions in various parts of the species’ range in central‐eastern Europe/Asia Minor, most likely correlating with the Younger Dryas Stadial (～13 000–11 600 years ago). It remains to be established whether the phylogeographic history of H. rhamnoides, and particularly its rapid response to the rapid environmental changes of the Younger Dryas cold snap, is unique to the species, or whether it is shared with other cold‐tolerant shrub (or grassland) species known from late‐glacial raw soils in Europe.     

Gynoecium development inRubiaceae-Vanguerieae,with particular reference to the “stylar head”-complex and secondary pollen presentation

Gynoecium development in taxa of the tribeVanguerieae (Rubiaceae, subfam.Antirheoideae) was studied, using primarily a 2- and a 5-carpellate genus (Keetia andVangueria) as examples. All investigated taxa, characterized by ovaries with a solitary, ± apically inserted, pendulous anatropous ovule per locule/carpel, showed a very similar gynoecium development. Comparisons with other uniovulateRubiaceae (taxa of subfam.Rubioideae) revealed that notwithstanding the place of insertion and orientation of the solitary ovules (apically vs. basally inserted, pendulous vs. erect, anatropous ovules) the gynoecium development follows the same pattern; differing ovule insertion and orientation can easily be explained by the principle of variable proportions. — Since secondary pollen presentation is characteristic for the tribe, particular attention was paid to the development of the conspicuous “stylar head”-complex, defined here as structural unit comprised of pollen presenting organ, “receptaculum pollinis”, plus stigmatic (i.e., receptive) surfaces. It was found that their morphological differentiation, following the same pattern in all investigated taxa, starts at very early stages of floral development. They already had their final shape at a stage when the ovule development in the ovary had just barely started. The fully developed, actual “receptaculum pollinis” is made up of enormously enlarged, ± cylindrical epidermis cells which have peculiar ring-like thickenings in vicinity of and parallel to the outer tangential walls (a “mechanical barrier” preventing pollen tubes from entering?) and a much smaller-celled subepidermal tissue. With regard to shape, size, and exposure of the stigmatic surfaces, the investigated taxa exhibited certain morphological differences. These were found to be correlated with differences in androecium structure (and, ultimately, pollen presentation).