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51.
Background and Aims
Modellers often define growth as the development of plant structures from endogenous resources, thus making a distinction between structural (WS) and total (W) dry biomass, the latter being the sum of WS and the weight of storage compounds. In this study, short-term C and N reserves were characterized experimentally (forms, organ distribution, time changes) in relation to light and nutrition signals, and organ structural growth in response to reserve levels was evaluated.Methods
Tomato plants (Solanum lycopersicum) were grown hydroponically in a growth room with a 12-h photoperiod and an adequate supply of NO3− (3 mol m−3). Three experiments were carried out 18 d after sowing: [NO3−] was either maintained at 3 mol m−3, changed to 0·02 mol m−3 or to 0 mol m−3. Plants were sampled periodically throughout the light/dark cycles over 24–48 h. Organ WS was calculated from W together with the amount of different compounds that act as C and N resources, i.e. non-structural carbohydrates and carboxylates, nitrate and free amino acids.Key Results
With adequate nutrition, carbohydrates accumulated in leaves during light periods, when photosynthesis exceeded growth needs, but decreased at night when these sugars are the main source of C for growth. At the end of the night, carbohydrates were still high enough to fuel full-rate growth, as WS increased at a near constant rate throughout the light/dark cycle. When nitrate levels were restricted, C reserves increased, but [NO3−] decreased progressively in stems, which contain most of the plant N reserves, and rapidly in leaves and roots. This resulted in a rapid restriction of structural growth.Conclusions
Periodic darkness did not restrict growth because sufficient carbohydrate reserves accumulated during the light period. Structural growth, however, was very responsive to NO3− nutrition, because N reserves were mostly located in stems, which have limited nitrate reduction capacity.Key words: Solanum lycopersicum, tomato, nitrogen, carbon, structural growth, reserves, nitrate, amino acids, carbohydrate, carboxylate 相似文献52.
Giovanni Maga Barbara van Loon Emmanuele Crespan Giuseppe Villani Ulrich H��bscher 《The Journal of biological chemistry》2009,284(21):14267-14275
Abasic (AP) sites are very frequent and dangerous DNA lesions. Their
ability to block the advancement of a replication fork has been always viewed
as a consequence of their inhibitory effect on the DNA synthetic activity of
replicative DNA polymerases (DNA pols). Here we show that AP sites can also
affect the strand displacement activity of the lagging strand DNA pol δ,
thus preventing proper Okazaki fragment maturation. This block can be overcome
through a polymerase switch, involving the combined physical and functional
interaction of DNA pol β and Flap endonuclease 1. Our data identify a
previously unnoticed deleterious effect of the AP site lesion on normal cell
metabolism and suggest the existence of a novel repair pathway that might be
important in preventing replication fork stalling.Loss of purine and pyrimidine bases is a significant source of DNA damage
in prokaryotic and eukaryotic organisms. Abasic (apurinic and apyrimidinic)
lesions occur spontaneously in DNA; in eukaryotes it has been estimated that
about 104 depurination and 102 depyrimidation events
occur per genome per day. An equally important source of abasic DNA lesions
results from the action of DNA glycosylases, such as uracil glycosylase, which
excises uracil arising primarily from spontaneous deamination of cytosines
(1). Although most AP sites are
removed by the base excision repair
(BER)5 pathway, a
small fraction of lesions persists, and DNA with AP lesions presents a strong
block to DNA synthesis by replicative DNA polymerases (DNA pols)
(2,
3). Several studies have been
performed to address the effects of AP sites on the template DNA strand on the
synthetic activity of a variety of DNA pols. The major replicative enzyme of
eukaryotic cells, DNA pol δ, was shown to be able to bypass an AP
lesion, but only in the presence of the auxiliary factor proliferating cell
nuclear antigen (PCNA) and at a very reduced catalytic efficiency if compared
with an undamaged DNA template
(4). On the other hand, the
family X DNA pols β and λ were shown to bypass an AP site but in a
very mutagenic way (5). Recent
genetic evidence in Saccharomyces cerevisiae cells showed that DNA
pol δ is the enzyme replicating the lagging strand
(6). According to the current
model for Okazaki fragment synthesis
(7–9),
the action of DNA pol δ is not only critical for the extension of the
newly synthesized Okazaki fragment but also for the displacement of an RNA/DNA
segment of about 30 nucleotides on the pre-existing downstream Okazaki
fragment to create an intermediate Flap structure that is the target for the
subsequent action of the Dna2 endonuclease and the Flap endonuclease 1
(Fen-1). This process has the advantage of removing the entire RNA/DNA hybrid
fragment synthesized by the DNA pol α/primase, potentially containing
nucleotide misincorporations caused by the lack of a proofreading exonuclease
activity of DNA pol α/primase. This results in a more accurate copy
synthesized by DNA pol δ. The intrinsic strand displacement activity of
DNA pol δ, in conjunction with Fen-1, PCNA, and replication protein A
(RP-A), has been also proposed to be essential for the S phase-specific long
patch BER pathway (10,
11). Although it is clear that
an AP site on the template strand is a strong block for DNA pol
δ-dependent synthesis on single-stranded DNA, the functional
consequences of such a lesion on the ability of DNA pol δ to carry on
strand displacement synthesis have never been investigated so far. Given the
high frequency of spontaneous hydrolysis and/or cytidine deamination events,
any detrimental effect of an AP site on the strand displacement activity of
DNA pol δ might have important consequences both for lagging strand DNA
synthesis and for long patch BER. In this work, we addressed this issue by
constructing a series of synthetic gapped DNA templates with a single AP site
at different positions with respect to the downstream primer to be displaced
by DNA pol δ (see Fig.
1A). We show that an AP site immediately upstream of a
single- to double-strand DNA junction constitutes a strong block to the strand
displacement activity of DNA pol δ, even in the presence of RP-A and
PCNA. Such a block could be resolved only through a “polymerase
switch” involving the concerted physical and functional interaction of
DNA pol β and Fen-1. The closely related DNA pol λ could only
partially substitute for DNA pol β. Based on our data, we propose that
stalling of a replication fork by an AP site not only is a consequence of its
ability to inhibit nucleotide incorporation by the replicative DNA pols but
can also stem from its effects on strand displacement during Okazaki fragment
maturation. In summary, our data suggest the existence of a novel repair
pathway that might be important in preventing replication fork stalling and
identify a previously unnoticed deleterious effect of the AP site lesion on
normal cell metabolism.Open in a separate windowFIGURE 1.An abasic site immediately upstream of a double-stranded DNA region
inhibits the strand displacement activity of DNA polymerase δ. The
reactions were performed as described under “Experimental
Procedures.” A, schematic representation of the various DNA
templates used. The size of the resulting gaps is indicated in nt. The
position of the AP site on the 100-mer template strand is indicated relative
to the 3′ end. Base pairs in the vicinity of the lesion are indicated by
dashes. The size of the gaps (35–38 nt) is consistent with the
size of ssDNA covered by a single RP-A molecule, which has to be released
during Okazaki fragment synthesis when the DNA pol is approaching the
5′-end of the downstream fragment. When the AP site is covered by the
downstream terminator oligonucleotide (Gap-3 and Gap-1 templates) the
nucleotide placed on the opposite strand is C to mimic the situation generated
by spontaneous loss of a guanine or excision of an oxidized guanine, whereas
when the AP site is covered by the primer (nicked AP template), the nucleotide
placed on the opposite strand is A to mimic the most frequent incorporation
event occurring opposite an AP site. B, human PCNA was titrated in
the presence of 15 nm (lanes 2–4 and
10–12) or 30 nm (lanes 6–8 and
14–16) recombinant human four subunit DNA pol δ, on a
linear control (lanes 1–8) or a 38-nt gap control (lanes
9–16) template. Lanes 1, 5, 9, and 13, control
reactions in the absence of PCNA. C, human PCNA was titrated in the
presence of 60 nm DNA pol δ, on a linear AP (lanes
2–4) or 38-nt gap AP (lanes 6–9) template. Lanes
1 and 5, control reactions in the absence of PCNA. 相似文献
53.
A New Basal Sauropod Dinosaur from the Middle Jurassic of Niger and the Early Evolution of Sauropoda
Kristian Remes Francisco Ortega Ignacio Fierro Ulrich Joger Ralf Kosma José Manuel Marín Ferrer for the Project PALDES for the Niger Project SNHM Oumarou Amadou Ide Abdoulaye Maga 《PloS one》2009,4(9)
Background
The early evolution of sauropod dinosaurs is poorly understood because of a highly incomplete fossil record. New discoveries of Early and Middle Jurassic sauropods have a great potential to lead to a better understanding of early sauropod evolution and to reevaluate the patterns of sauropod diversification.Principal Findings
A new sauropod from the Middle Jurassic of Niger, Spinophorosaurus nigerensis n. gen. et sp., is the most complete basal sauropod currently known. The taxon shares many anatomical characters with Middle Jurassic East Asian sauropods, while it is strongly dissimilar to Lower and Middle Jurassic South American and Indian forms. A possible explanation for this pattern is a separation of Laurasian and South Gondwanan Middle Jurassic sauropod faunas by geographic barriers. Integration of phylogenetic analyses and paleogeographic data reveals congruence between early sauropod evolution and hypotheses about Jurassic paleoclimate and phytogeography.Conclusions
Spinophorosaurus demonstrates that many putatively derived characters of Middle Jurassic East Asian sauropods are plesiomorphic for eusauropods, while South Gondwanan eusauropods may represent a specialized line. The anatomy of Spinophorosaurus indicates that key innovations in Jurassic sauropod evolution might have taken place in North Africa, an area close to the equator with summer-wet climate at that time. Jurassic climatic zones and phytogeography possibly controlled early sauropod diversification. 相似文献54.
A. A. Cabrera‐Chávez‐Costa F. Galván‐Magaña O. Escobar‐Sánchez 《Zeitschrift fur angewandte Ichthyologie》2010,26(4):499-503
The objective of this study was to establish the trophic niche of the silky shark and to determine the ecological role of this predator in the ecosystem close to Baja California. The trophic spectrum was analyzed from samples taken during summer and autumn (2000–2002) from the fishing camps of Punta Lobos and Punta Belcher on the western coast of Baja California Sur. A total of 263 stomach contents were analyzed (143 with food; 120 empty). The index of relative importance (IRI) showed that at Punta Lobos, silky sharks fed mainly on red crabs Pleuroncodes planipes (%IRI = 83%), whereas at Punta Belcher the main food item was the jumbo squid Dosidicus gigas (%IRI = 41%), followed by chub mackerel Scomber japonicus (%IRI = 33%). According to the Levin Index (Bi), the trophic niche breadth in silky sharks is low (Bi = <0.6), which means that silky sharks are specialist predators because they mainly consume three prey types: red crab, chub mackerel, and jumbo squid. The Shannon‐Wiener Index indicated that all trophic categories at Punta Belcher (0.85–1.22) had lower diversity than at Punta Lobos (0.50–1.6), because the silky shark feeds more on tropical prey found close to Punta Lobos. The Morisita‐Horn Index (Cλ) showed an overlap in the diet between the two areas analyzed and between sexes (Cλ = >0.6). The juveniles and adult females did not show any overlap. In the caloric analysis of the main prey, the jumbo squid (D. gigas) contributed the most calories to the silky shark diet (76%). 相似文献
55.
Consumption of pasteurized human lysozyme transgenic goats’ milk alters serum metabolite profile in young pigs 总被引:1,自引:0,他引:1
Dottie R. Brundige Elizabeth A. Maga Kirk C. Klasing James D. Murray 《Transgenic research》2010,19(4):563-574
Nutrition, bacterial composition of the gastrointestinal tract, and general health status can all influence the metabolic
profile of an organism. We previously demonstrated that feeding pasteurized transgenic goats’ milk expressing human lysozyme
(hLZ) can positively impact intestinal morphology and modulate intestinal microbiota composition in young pigs. The objective
of this study was to further examine the effect of consuming hLZ-containing milk on young pigs by profiling serum metabolites.
Pigs were placed into two groups and fed a diet of solid food and either control (non-transgenic) goats’ milk or milk from
hLZ-transgenic goats for 6 weeks. Serum samples were collected at the end of the feeding period and global metabolite profiling
was performed. For a total of 225 metabolites (160 known, 65 unknown) semi-quantitative data was obtained. Levels of 18 known
and 4 unknown metabolites differed significantly between the two groups with the direction of change in 13 of the 18 known
metabolites being almost entirely congruent with improved health status, particularly in terms of the gastrointestinal tract
health and immune response, with the effects of the other five being neutral or unknown. These results further support our
hypothesis that consumption of hLZ-containing milk is beneficial to health. 相似文献
56.
57.
López-Lacomba JL García-Cantalejo JM Sanz Casado JV Abarrategi A Correas Magaña V Ramos V 《Biomacromolecules》2006,7(3):792-798
Considering the design and development of biomaterials used in tissue engineering, not only is it important that they are biocompatible but also that they induce the desired cellular response for tissue regeneration. Chitosan, a biocompatible and bioresorbable polymer of N-acetylglucosamine and glucosamine is used in our work combined with recombinant human BMP-2 (rhBMP-2), a potentially useful activation factor for bone repair. In this way, we try to combine the biological and filmogenic properties of this biopolymer with the osseoinductive ability of the rhBMP-2. Results showed that the chitosan films employed, without and with rhBMP-2 activation, are able to support cellular growth and proliferation on them and that only the rhBMP-2 activated ones are able to differentiate from a myoblastic mouse cell line (C2C12) toward osteoblastic phenotype. Osseoinduction properties of rhBMP-2 activated films persist for a long storage time. The in vivo experiments performed confirm the expectative created by the in vitro results obtained and are an indication that rhBMP-2 activated chitosan films could be a very attractive biomaterial for the enhancement of osseointegration of surgical prostheses and implants and for the purpose of tissue engineering bone regeneration. 相似文献
58.
Ramadan K Maga G Shevelev IV Villani G Blanco L Hübscher U 《Journal of molecular biology》2003,328(1):63-72
DNA polymerase lambda is a novel enzyme of the family X of DNA polymerases. The recent demonstration of an intrinsic 5'-deoxyribose-5'-phosphate lyase activity, a template/primer dependent polymerase activity, a distributive manner of DNA synthesis and sequence similarity to DNA polymerase beta suggested a novel beta-like enzyme. All these properties support a role of DNA polymerase lambda in base excision repair. On the other hand, the biochemical properties of the polymerisation activity of DNA polymerase lambda are still largely unknown. Here we give evidence that human DNA polymerase lambda has an intrinsic terminal deoxyribonucleotidyl transferase activity that preferentially adds pyrimidines onto 3'OH ends of DNA oligonucleotides. Furthermore, human DNA polymerase lambda efficiently elongates an RNA primer hybridized to a DNA template. These two novel properties of human DNA polymerase lambda might suggest additional roles for this enzyme in DNA replication and repair processes. 相似文献
59.
60.
Cell cycle-dependent dynamic association of cyclin/Cdk complexes with human DNA replication proteins
Frouin I Montecucco A Biamonti G Hübscher U Spadari S Maga G 《The EMBO journal》2002,21(10):2485-2495
We have previously described the isolation of a replication competent (RC) complex from calf thymus, containing DNA polymerase alpha, DNA polymerase delta and replication factor C. Here, we describe the isolation of the RC complex from nuclear extracts of synchronized HeLa cells, which contains DNA replication proteins associated with cell-cycle regulation factors like cyclin A, cyclin B1, Cdk2 and Cdk1. In addition, it contains a kinase activity and DNA polymerase activities able to switch from a distributive to a processive mode of DNA synthesis, which is dependent on proliferating cell nuclear antigen. In vivo cross-linking of proteins to DNA in synchronized HeLa cells demonstrates the association of this complex to chromatin. We show a dynamic association of cyclins/Cdks with the RC complex during the cell cycle. Indeed, cyclin A and Cdk2 associated with the complex in S phase, and cyclin B1 and Cdk1 were present exclusively in G(2)/M phase, suggesting that the activity, as well the localization, of the RC complex might be regulated by specific cyclin/Cdk complexes. 相似文献