Effect of nitrogen load on growth and photosynthesis of seedlings of the hybrid larch F1 (Larix gmelinii var. japonica × L. kaempferi) grown on serpentine soil

We studied the growth and photosynthesis of the hybrid larch F₁ (Larix gmelinii var. japonica × L. kaempferi) grown on serpentine soil and the effects of soil N load, to determine the performance of this species as reforestation material in serpentine regions. We prepared 16 experimental plots (2 m × 4 m each), eight on serpentine and eight on brown forest soil, and planted one-year-old cutting seedlings of the hybrid larch F₁ in each plot, in May 2007. Ammonium sulfate was supplied to half of the plots of each soil type in 2008 and 2009, at a load of 47 kg N ha⁻¹ year⁻¹. Although the growth and photosynthetic capacity of hybrid larch F₁ seedlings in the serpentine soil were limited, the rate of growth in serpentine soil was greater than that of Sakhalin spruce (Picea glehnii) that is dominant species in serpentine regions. There was significant interaction between soil type and N load for the growth and photosynthetic parameters. The N load adversely affected growth and photosynthetic parameters in the serpentine soil, while improved them in brown forest soil. Although the growth rate of hybrid larch F₁ without N loading showed high potential as an afforestation species in serpentine region, increasing deposition of N might be a threat to the growth and photosynthesis of the hybrid larch F₁ in serpentine soil.     

Can CO2 enrichment modify the effect of water and high light stress on biomass allocation and relative growth rate of cork oak seedlings?

To test whether the impact of an enriched-CO₂ environment on the growth and biomass allocation of first-season Quercus suber L. seedlings can modify the drought response under shade or sun conditions, seedlings were grown in pots at two CO₂ concentrations × two watering regimes × two irradiances. Compared to CO₂, light and water treatment had greater effects on all morphological traits measured (height, stem diameter, number of leaves, leaf area, biomass fractions). Cork oak showed particularly large increases in biomass in response to elevated CO₂ under low-watered (W−) and high-illuminated conditions (L+). Allocation shifted from shoot to root under increasing irradiance (L+), but was not affected by CO₂. Changes in allocation related to water limitation were only modest, and changed over time. Relative growth rate (RGR) and net assimilation rate (NAR) were significantly greatest in the L+/W+ treatment for both CO₂ concentrations. Changes in RGR were mainly due to NAR. Growth responses to increased light, water or CO₂ were strongest with light, medium with water availability and smallest for CO₂, in terms of RGR. The rise in NAR for light and water treatments was counterbalanced by a decrease in SLA (specific leaf area) and LMF (leaf mass fraction). Results suggest that elevated CO₂ caused cork oak seedlings to improve their performance in dry and high light environments to a greater extent than in well-irrigated and low light ones, thus ameliorating the effects of soil water stress and high light loads on growth.     

Influence of climatic factors upon tree rings of Larix decidua and L. decidua × L. kaempferi from Pulawy,Poland

Summary Dendroclimatological techniques are used to assess the impact of climatic factors on tree-ring width of Larix decidua and L. decidua × L. kaempferi (= L. x eurolepis) growing in two experimental plots established in 1914 in south-west Poland. One plot included F₁ progeny grown from seeds of an artificial crossing between European and Japanese larch. The other plot included progeny from maternal trees of European larch. Total ring width, earlywood width and latewood widths were dated, standardized and related to monthly climatic data using response function and stepwise multiple regression analyses. Wide rings in larch are associated with high precipitation in May–July, cool conditions in July–September of the preceding year, and cool dry conditions in August. Ring widths in L. x eurolepis are more dependent upon precipitation than ring widths in L. decidua. Latewood widths in L. x eurolepis are more dependent on high temperatures in June and July than latewood in L. decidua as well as total width and earlywood measurements. Variations in latewood were relatively independent of variations in earlywood and total wood. The variability of ring widths in these larches was greater than the variability reported for larches in many alpine sites and for other conifer species in some regions of North America.     

Root growth inhibition and ultrastructural changes in radish root tips after treatment with aqueous extracts of Fallopia japonica and F. ×bohemica rhizomes

Allelopathic compounds released by invasive alien plants can suppress the growth of plants in their vicinity. The aim of this study was to investigate changes in tissue and cell structure in roots of radish seedlings treated with 10% aqueous extracts of rhizomes from the invasive knotweeds Fallopia japonica and F. ×bohemica. After 7 days of growth without and with aqueous extracts from these rhizomes, the anatomical and ultrastructural changes in the radish seedling roots were analyzed with light and transmission electron microscopy, and hydrogen peroxide was localized with diaminobenzidine, to define oxidative stress. The roots of radish seedlings treated with the knotweed extracts were shorter and thicker, due to the shorter and wider shapes of their cortex cells, which were organized in more columns than the control roots. There were signs of cell damage and oxidative stress in the root cap cells, and to a lesser extent in the meristematic zone. As well as the irregularly shaped nuclei and plasma membrane detached from the cell wall, the most prominent ultrastructural effects in the root cap cells of these aqueous rhizome extracts were the ring-shaped form of the mitochondria and large endoplasmic reticulum bodies. Excessive vacuolization was seen for the cells of the root apical meristem.

     

An improved method for somatic plantlet production in hybrid larch (Larix × leptoeuropaea): Part 2. Control of germination and plantlet development

Germination and plantlet development in somatic embryos of Larix x leptoeuropaea were affected by the duration of the maturation treatment and the concentrations of sucrose and abscisic acid in the maturation media. Extension of the maturation period from 3 weeks to 4 weeks resulted in a significant decrease in germination and plantlet development frequencies. There was no significant effect of abscisic acid concentration on either the number of somatic embryos germinated or the number of plantlets obtained, but it affected the rapidity of the epicotyl development. Sucrose at 0.2 M, applied during maturation, was significantly more beneficial in attaining high germination rates than at 0.1 M. High germination rates (92 and 93%) and plantlet development rates (74 and 80%) were achieved when somatic embryos were matured for a 3-week period on media with either 40 or 60 M abscisic acid, respectively, and 0.2 M sucrose prior to transfer to the growth regulator-free germination medium. Two acclimatization methods were applied: the first required 10 to 12 weeks and ensured 97% plantlet survival under greenhouse conditions; the second required 2–3 weeks and ensured 86% plantlet survival. This represents the first detailed study of the effects of maturation regimes on the recovery of somatic embryo-derived plants of Larix.Abbreviations ABA abscisic acid - IBA indolebutyric acid - 2,4-d 2,4-dichlorophenoxyacetic acid - EM embryonal mass     

Red-light-induced changes in the distribution of growth inhibitors A-2α and A-2β in dwarf pea seedlings

The distribution of the endogenous growth inhibitors, A-2 and A-2, was determined in light-grown dwarf pea seedlings (Pisum sativum L. cv. Progress No. 9). Red-light-induced changes in this distribution in dark-grown pea seedlings was monitored. The concentration of both inhibitors in light-grown pea seedlings was maximal in the terminal bud and decreased to the lower parts of the seedlings; their concentration in internodes was 7–10 times that in leaves of the same nodes. After the onset of continuous red-light-irradiation, the concentration of the inhibitors in dark-grown pea seedlings increased rapidly in the terminal bud, epicotyl hook and third internode, and the longitudinal gradients of the inhibitors showed a close correlation with the red-light-induced growth inhibition of the stem. The present results suggest that the endogenous growth inhibitors, A-2 and A-2 may be responsible for the red-light-induced growth inhibition of the dwarf pea seedlings.     

The effect of air containing O2 −,O2 +, CO2 − and CO2 + on the growth of seedlings ofHordeum Vulgaris

Equipment was devised which permitted the addition of specific gaseous ions to the atmosphere of plastic chambers in which seedlings of HORDEUM VULGARIS were grown in sand culture supplied with chemically defined nutrient solutions. Moderate densities of O₂ ^– or O₂ ⁺ ions (1.8×10⁴/cm³)in air containing an added 8% of O₂ accelerated the growth rate. A like number of CO₂ ^– or CO₂ ⁺ ions in air containing 8% of CO₂ inhibited growth, impeded the production of chlorophyll and devitalized the young seedlings. Evidence is presented that O₂ ^– and O₂ ⁺ stimulate the production of cytochromes and other Fe-containing enzymes through their action on the plant regulatory system responsible for the control of Fe metabolism. The toxic effect of CO₂ ^– and CO₂ ⁺ cannot be explained as yet.

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Zusammenfassung Eine Apparatur wurde entwickelt, die die Zufuhr von ionisiertem Gas der AtmosphÄre in Kammern gestattet. Darin wurden Keimlinge von HORDEUM VULGARIS in Sand mit chemisch definierten NÄhrlösungen gezüchtet. Konzentrationen von 1,8×10⁴/cm³ O₂ ^– und O₂ ⁺ in Luft mit zusÄtzlich 8% O₂ beschleunigten die Wachstumsrate. Die gleiche Menge CO₂ ^– und CO₂ ⁺ in Luft mit zusÄtzlich 8% CO₂ hemmte die Wachstumsrate, die Bildung von Chlorophyll und entkrÄftigte die Keimlinge. Es wird gezeigt,dass O₂ ^– und O₂ ⁺ die Bildung von Cytochrom und anderen eisenhaltigen Enzymen anregen durcn ihre Wirkung auf das den Fe-Stoffwechsel regulierende System der Pflanze. Die toxische Wirkung von CO₂ ^– und CO₂ ⁺ lÄsst sich noch nicht erklÄren.

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Resume On a construit un appareil permettant d'introduire dans 1'atmosphères des ions de gaz déterminés. On a alors effectué de telles adjonctions à l'air contenu dans des cellules de plastique dans lesquelles on cultivait HORDEUM VULGARIS sur du sable et dans une solution nutritive chimiquement définie. Des densités modérées d'ions O₂ ^– ou O₂ ⁺ (1,8×10⁴/cm³) dans de l'air additionné de 8% d'O₂ accélèrent la croissance. La meme concentration de CO₂ ^– et CO₂ ⁺ additionnée de 8% de CO₂ a ralenti la croissance et la formation de chlorophylle et a diminué la vitalite des plantes nouvellement germées. On démontre que O₂ ^– et O₂ ⁺ active la formation de cytochrome et d'autres enzymes ferreuses par suite de l'action de ces ions sur le système régularisant le métabolisme du fer dans la plante. L'effet toxique du CO₂ ^– et CO₂ ⁺ reste encore inexpliqué.

     

Ethylene and cytokinin interaction in the morphogenesis of Pelargonium × hortorum L.H. Bailey in vitro

Pelargonium × hortorum ‘Grand Prix’ which is susceptible to leaf yellowing and ‘Bergpalais’ which is not susceptible to leaf yellowing were chosen for the experiments. Ethylene production and action as well as the associated morphological response of Pelargonium shoots grown in the presence of a precursor of ethylene biosynthesis 1-aminocyclopropane-1-carboxylic acid (ACC), ethylene inhibitors: α-aminooxyacetic acid (AOA) and silver nitrate (AgNO₃) and different cytokinins: (meta-topolin) (mT) or 6-benzylaminopurine (BAP) were studied. It was found that ‘Grand Prix’ was more sensitive to ethylene than ‘Bergpalais’ and it showed the leaf yellowing in response to 0.1 mg l^?1 ACC. Moreover, it was noted that ACC added separately or together with cytokinin influenced Pelargonium morphogenesis. Depending on the concentration of ACC (0.1–2.0 mg l^?1), it either stimulated or inhibited shoot and root formation as well as the growth of shoots and leaf blades. ACC-induced leaf yellowing in ‘Grand Prix’ was effectively inhibited by mT. In contrast, BAP did not enhance shoot quality. Simultaneously, the presence of mT in the medium resulted in up to a twofold increase in the ethylene production by ‘Grand Prix’ shoots throughout the culture period compared with the shoots growing on the BAP-medium. The inhibitor of ethylene action (AgNO₃) added with cytokinin prevented the yellowing of Pelargonium shoots, but simultaneously influenced the formation of mature shoots with limited long-term multiplication potential. The shoots of P. × hortorum ‘Grand Prix’ treated with AgNO₃ and mT emitted two- and sevenfold more ethylene after 11th and 21st day of culture compared with those treated with AgNO₃ and BAP. It is suggested that mT inhibits the early senescence of Pelargonium in vitro by decreasing its sensitivity to ethylene.     

Effects of the antibiotic growth promoters flavomycin and florfenicol on the autochthonous intestinal microbiota of hybrid tilapia (Oreochromis niloticus ♀ × O. aureus ♂)

The 16S rDNA PCR-DGGE and rpoB quantitative PCR (RQ-PCR) techniques were used to evaluate the effects of dietary flavomycin and florfenicol on the autochthonous intestinal microbiota of hybrid tilapia. The fish were fed four diets: control, dietary flavomycin, florfenicol and their combination. After 8 weeks of feeding, 6 fish from each cage were randomly chosen for the analysis. The total number of intestinal bacteria was determined by RQ-PCR. The results showed that dietary antibiotics significantly influenced the intestinal microbiota and dramatically reduced the intensity of total intestinal bacterial counts. The intensity of some phylotypes (EU563257, EU563262 and EU563255) were reduced to non-detectable levels by both dietary antibiotics, while supplementation of florfenicol to the diet also reduced the intensity of the phylotypes EU563242 and EU563262, uncultured Mycobacterium sp.-like, uncultured Cyanobacterium-like and uncultured Cyanobacterium (EU563246). Dietary flavomycin only reduced the OTU intensity of one phylotype, identified as a member of the phylum Fusobacteria. The antibiotic combination only reduced the phylotypes EU563242 and EU563262. Based on our results, we conclude that the reduced effect of florfenicol on intestinal microbiota was stronger than that of flavomycin, and when flavomycin and florfenicol were added in combination, the effect of florfenicol overshadowed that of flavomycin.     

Effects of drought stress on the evergreen Quercus ilex L., the deciduous Q. robur L. and their hybrid Q. × turneri Willd.

Five-year-old trees of deciduous Quercus robur L., evergreen Q. ilex L., and their semideciduous hybrid, Q. × turneri Willd. (var. pseudoturneri), growing in pots, were subjected to drought stress by withholding water for 18–22 days, until leaf water potentials decreased below ?2 MPa. Gas-exchange rates, oxygen evolution, and modulated chlorophyll (Chl) fluorescence measurements revealed that by strong stomata closure and declining photosynthetic capacity down to approximately 50%, all three taxa responded with strongly reduced photosynthesis rates. In Q. robur, photochemical quenching of the drought-stressed plants was much lower than in nonstressed controls. Dissection of the occurring events in the photosynthetic electron transport chain by fast Chl fluorescence induction analysis with the JIP-test were discussed.     

The production of β-carotene inMortierella ramanniana var.ramanniana M29: the effect of changes in the environment upon growth and pigmentation

The production of -carotene in a strain ofMortierella ramanniana var.ramanniana (M29) has been shown to have many features characteristic of secondary biosynthetic processes. It is formed at the end of the primary growth phase in amounts greater than those reported inPhycomyces blakesleeanus. It has no apparent function in primary growth. The production has an optimum temperature lower than that of the primary growth phase. The amount of pigment formed is greatest with those carbon sources which do not support maximum growth within a given incubation period. Changes in the initial carbon, nitrogen, thiamine, phosphate, magnesium, trace elements, hydrogen ion concentration or oxygen tension of the medium did not abolish or delay the production of the pigment. It is suggested that the production of -carotene in this strain is an example of secondary metabolite production and that a mutation could be responsible for the production in this one strain.The technical assistance of Mr. D. J. Trott is gratefully acknowledged.     

Apoptotic cell death induces temperature-sensitive lethality in hybrid seedlings and calli derived from the cross of Nicotiana suaveolens×N. tabacum

Hybrid lethality expressed in the interspecific hybrid of Nicotiana suaveolens Lehm. ×N. tabacum L. cv. Hicks-2 is one of the mechanisms for reproductive isolation and it is temperature-sensitive. Apoptotic changes were detected in the cells of hybrid seedlings and calli expressing lethality at 28 °C but not under high-temperature conditions (36 °C), when the lethality is suppressed. Condensation of chromatin, fragmentation of nuclei and cytoplasmic reduction are the cytological changes associated with apoptosis leading to hybrid lethality. Fragmentation of nuclei was correlated with the lethal symptoms in both hybrid seedlings and calli, as confirmed by fluorimetry of the nuclear DNA using laser scanning cytometry. Agarose gel analysis of DNA extracted from hybrid seedlings and calli showing lethal symptoms revealed a specific ladder pattern suggesting nucleosomal fragmentation which is one of the biochemical changes of apoptosis. In-situ detection using terminal deoxyribonucleotidyl transferase-mediated dUTP-fluorescein nick end labeling (TUNEL) showed that this process occurred in distinct stages on each organ of hybrid seedlings and centripetally in hybrid calli. From these results, we confirmed that cell death inducing hybrid lethality was indeed apoptosis. Received: 23 December 1999 / Accepted: 5 April 2000     

In vitro anther culture of sweet orange (Citrus sinensis L. Osbeck) genotypes and of a C. clementina × C. sinensis ‘Hamlin’ hybrid

Citrus, and particularly sweet oranges, are very recalcitrant to anther culture. In this paper it was evaluated for the first time the response of 27 genotypes of Citrus sinensis and of one hybrid C. clementina × C. sinensis, to in vitro anther culture. Ten genotypes of sweet oranges showed embryogenic callus induction, mostly blood sweet oranges genotypes, such as Tarocco, Moro and Sanguinelli. In vitro microspore developmental switches from the gamethophytic to the sporophytic pathway were shown by DAPI staining in microspores of these responsive genotypes, after 10 months in culture. However, microsatellite marker analyses showed that these calli were heterozygous. The flow-cytometric analysis of these embryogenic calli showed the presence of two peaks, corresponding to haploid (n) and diploid (2n) genotypes. Differently, anther cultures of the hybrid C. clementina × C. sinensis produced tri-haploid (3n) embryogenic calli and the embryos obtained were homozygous when analyzed by molecular markers (sample sequence repeats), confirming the more responsive characteristic of clementine to microspore embryogenesis through anther culture.     

The after-effects of temperature and irradiance during early growth of winter oilseed rape (Brassica napus L. var. oleifera, cv. Górczański) seedlings on the progress of their cold acclimation

Experiments performed under controlled conditions showed that level of PPFD (photosynthetic photon flux density) during early seedlings growth (preceding cold acclimation at +2 °C) was not the key factor for the development of frost resistance. It did not modify the beneficial effects of prehardening (Rapacz 1997, in this issue) at moderately low (+12 °C) day temperature. Now I have shown that the increase of PPFD may replace to some extent prehardening in the development of frost resistance. It was particularly seen in non-prehardened plants, which had been grown under warm-day (+20 °C) conditions. Prehardening performed under controlled conditions, as well as seedlings growth under natural autumn conditions in the field, allowed to maintain a high net-photosynthesis rate at chilling temperatures. A net-photosynthesis rate during cold acclimation at +2 °C corresponded well with higher frost resistance. As a result, seedlings non subjected to prehardening and grown before cold acclimation under low PPFD acclimated better, if the cold treatment was applied only at nights (+20/2 °C day/night). Only under such conditions the photosynthetic rate was sufficiently high to allow plants to reach a higher level of frost resistance. All other plants acclimated better when they were exposed to the hardening temperature continuously during days and nights (+2/2 °C day/night).     

Study of the three-genome hybridLycopersicon esculentum Mill. —L. chilense Dun. —L. peruvianum var ‘humifusum’ Mill. and its use as a source for resistance

L. peruvianum var humifusum is reproductively the most isolated of the species of the genusLycopersicon. It can be crossed with the cultivated tomato usingL. chilense as an intermediary. After a series of backcrosses of the three-genome hybrid F₁ (L. esculentum ×L. chilense) ×L. peruvianum var humifusum withL. esculentum, accompanied by selection for resistance to some economically important diseases, several lines were established. One of these lines, Cm 180, which showed resistance toClavibacter michiganensis subsp.michiganensis, was subjected to genetic analysis. This resistance was found to be controlled by a single dominant gene (Cm) that was not allelic to the gene originating fromL. hirsutum f.glabratum. ThisCm gene was genetically mapped on chromosome 4. The germ plasm ofL. peruvianum var humifusum in combination withL. chilense was transferred intoL. esculentum. Different breeding lines possessing resistance to various diseases and pests could be developed from this material.     

Daylength and spatial expression of a gibberellin 20-oxidase isolated from hybrid aspen (Populus tremula L. × P. tremuloides Michx.)

Physiologically active gibberellins (GAs) are key regulators of shoot growth in trees. To investigate this mechanism of GA-controlled growth in hybrid aspen, we cloned cDNAs encoding gibberellin 20-oxidase (GA 20-oxidase), a key, highly regulated enzyme in the biosynthesis of GAs. Clones were isolated from leaf and cambium cDNA libraries using probes generated by polymerase chain reaction, based on conserved domains of GA 20-oxidases. Upon expression in Escherichia coli, the GST-fusion protein was shown to oxidise GA12 as well as oxidising the 13-hydroxylated substrate GA53, successively to GA9 and GA20, respectively. The gene PttGA20ox1 was expressed in meristematic cells and growing tissues such as expanding internodes, leaves and roots. The expression was negatively regulated by both GA4 and overexpression of phytochrome A. RNA analysis also showed that the expression was down-regulated in late-expanding leaf tissue in response to short days (SDs). Actively growing tissues such as early elongating internodes, petioles and leaf blades had the highest levels of C19-GAs. Upon transfer to SDs an accumulation of GA19 was observed in early elongating internodes and leaf blades. The levels of C19-GAs were also to some extent changed upon transfer to SDs. The levels of GA20 were down-regulated in internodes, and those of GA1 were significantly reduced in early expanding leaf blades. In roots the metabolites GA19 and GA8 decreased upon shifts to SDs, while GA20 accumulated slightly. The down-regulation of GA 20-oxidase activity in response to SDs was further indicated by studies of [14C]GA12 metabolism in shoots, demonstrating that the substrate for GA 20-oxidase, [14C]GA53, accumulates in SDs.     

Inhibition of Aspergillus flavus growth and detoxification of aflatoxin B1 by the medicinal plant zimmu (Allium sativum L. × Allium cepa L.)

Aflatoxins are carcinogenic, teratogenic and immunosuppressive secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus. Aflatoxin contamination of peanut is one of the most important constraints to peanut production worldwide. In order to develop an eco-friendly method of prevention of A. flavus infection and aflatoxin contamination in peanut, aqueous extracts obtained from leaves of 30 medicinal plants belonging to different families were evaluated for their ability to inhibit the growth of A. flavus in vitro. Among them the leaf extract of zimmu (Allium sativum L. × Allium cepa L.) was the only one that showed antifungal activity against A. flavus and recorded 73% inhibition of A. flavus growth. The antifungal activity of the zimmu extract was significantly decreased upon dialysis with a dialysis membrane having molecular cut off 12 kDa or autoclaving at 121°C for 20 min or boiling at 100°C for 10 min and recorded inhibition of 52, 16 and 21%, respectively. When A. flavus was grown in medium containing zimmu extract the production of aflatoxin B₁ (AFB₁) was completely inhibited even at a concentration of 0.5%. When AFB₁ was incubated with zimmu extract a complete degradation of AFB₁ was observed 5 days after incubation. When the roots of zimmu were incubated in water containing 70 ng of AFB₁/ml, a reduction (by 58.5%) in AFB₁ concentration was observed 5 days after incubation. A significant reduction in the population of A. flavus in the soil, kernel infection by A. flavus and aflatoxin contamination in kernels was observed when peanut was intercropped with zimmu. The population of the fungal antagonist, Trichoderma viride in the zimmu-intercropped field increased approximately twofold.