Relació entre fotosíntesi i respiració en cultius sotmesos a estrés hídric mitjançant isòtops estables

Report for the scientific sojourn carried out at the Universite Paris Sud – XI, France, from July until october 2007. The relationship between photosynthesis and respiration were studied in crops using carbon and nitrogen stable isotopes under well watered and water-stressed conditions. The analyses of the 13C isotopic composition (δ13C) of total organic matter (TOM) recently fixed of well-watered plants revealed that it was mainly delivered to apical tissues and tap root. The fact that that the apical leaf and stems together with the inflorescences were d13C depleted, suggests that those tissues were newly formed and had a larger sink strength and metabolic activity. Secondly, the analyses of δ13C of respired CO2 immediately after the labelling (T=0) showed that a significant part of the C respired by leaves and nodules proceeded of the recently fixed CO2. In the following harvests (T=7 and T=14) such percentage tended to decrease, especially in apical leaves. Interestingly, the respiration d13C data also highlighted that even if at T=0 part of the respired proceeded from the CO2 fixed during the labelling, this percentage was even larger at T=7. Finally, the d15N also revealed that, similarly to what described for 12C, immediately after the 15N2 labelling (T=0), apical leaf and stems, together with tap root and in this case the nodules, were the tissues with larger sink strength. It is noteworthy the fact that the largest amount of N2 newly fixed was delivered to the tap roots where it was stored until it was required for the aboveground regrowth period.

Comparison of flag leaf and ear photosynthesis with biomass and grain yield of durum wheat under various water conditions and genotypes

Photosynthetic activity of cereals has traditionally been studied using leaves, thus neglecting the role of other organs such as ears. Here, we studied the effects of water status and genotypes on the photosynthetic activity of the flag leaf blade and the ear of durum wheat. The various parameters related to the photosynthetic activity were analysed in relation to the total above-ground plant biomass and grain yield at maturity. Four local varieties plus two cultivars adapted to the semiarid areas of South Morocco were grown in pots in a greenhouse. Five different water treatments were maintained from the beginning of stem elongation to maturity, when shoot biomass and grain yield were recorded. The net photosynthesis (A), stomatal conductance (gs) and transpiration (T) of the ear and the flag leaf were measured at anthesis. In both organs these factors decreased significantly with water deficit, whereas the A/T and A/gs ratios increased. The genotype effect was also significant for all traits studied. Whole-organ photosynthesis was much higher in the ear than in the flag leaf in well-watered conditions. As water stress developed, photosynthesis decreased less in the ear than in the flag leaf. Whole-ear photosynthesis correlated better than flag leaf photosynthesis with biomass and yield. Nevertheless, the relationships of the whole flag leaf with biomass and yield improved as the water stress became more severe, suggesting a progressive shift of yield from sink to source limitation. For all water regimes the ratios A/gs and A/T of the ear also showed a higher (negative) correlation with both biomass and yield than those of the flag leaf. The results indicate that the ear has a greater photosynthetic role than the flag leaf in determining grain yield, not only in drought but also in the absence of stress.

Errores conceptuales en Biología. La fotosíntesis de las plantas verdes

This paper tries to point out the existente of conceptual errors in the learning of photosynthesis. They seem to arise from the same sources as those present in more deeply studied fields, such as mechanics.

On the relationship between C and N fixation in nodulated alfalfa (Medicago sativa)

Legumes such as alfalfa (Medicago sativa L.) are vital N2-fixing crops accounting for a global N2 fixation of ~35 MtNyear-1. Although enzymatic and molecular mechanisms of nodule N2 fixation are now well documented, some uncertainty remains as to whether N2 fixation is strictly coupled with photosynthetic carbon fixation. That is, the metabolic origin and redistribution of carbon skeletons used to incorporate nitrogen are still relatively undefined. Here, we conducted isotopic labelling with both 15N2 and 13C-depleted CO2 on alfalfa plants grown under controlled conditions and took advantage of isotope ratio mass spectrometry to investigate the relationship between carbon and nitrogen turn-over in respired CO2, total organic matter and amino acids. Our results indicate that CO2 evolved by respiration had an isotopic composition similar to that in organic matter regardless of the organ considered, suggesting that the turn-over of respiratory pools strictly followed photosynthetic input. However, carbon turn-over was nearly three times greater than N turn-over in total organic matter, suggesting that new organic material synthesised was less N-rich than pre-existing organic material (due to progressive nitrogen elemental dilution) or that N remobilisation occurred to sustain growth. This pattern was not consistent with the total commitment into free amino acids where the input of new C and N appeared to be stoichiometric. The labelling pattern in Asn was complex, with contrasted C and N commitments in different organs, suggesting that neosynthesis and redistribution of new Asn molecules required metabolic remobilisation. We conclude that the production of new organic material during alfalfa growth depends on both C and N remobilisation in different organs. At the plant level, this remobilisation is complicated by allocation and metabolism in the different organs. Additional keywords: carbon exchange, carbon isotopes, nitrogen fixation, nitrogen 15 isotope

The effects of depleted, current and elevated growth [CO2] in wheat are modulated by water availability

Drought is the main constraint on wheat yield in Mediterranean conditions. The photosynthesis, chlorophyll fluorescence and plant growth parameters of durum wheat (Triticum turgidum, L. var. durum) were compared at three [CO2] (i.e., depleted 260 ppm, current 400ppm and elevated 700 ppm) in plants subjected to twowater regimes (i.e.,well-wateredWW, and mildwater stress by drought orwater deficit WS), during pre-anthesis, post-anthesis and the end of grain filling. We showed that [CO2] effects on plants are modulated by water availability. Plants at depleted [CO2] showed photosynthetic acclimation (i.e., up-regulation) and reduced plant biomass and Harvest Index, but depleted [CO2] combined with WS has a more negative impact on plants with decreases in C assimilation and biomass. Plants at elevated [CO2] had decreased plant growth and photosynthesis in response to a down-regulation mechanism resulting from a decrease in Rubisco and N content, but plants exposed to a combination of elevated [CO2] and WS were the most negatively affected (e.g., on plant biomass).

Rhizodeposition of organic carbon by plants with contrasting traits for resource acquisition: responses to different fertility regimes

Background and aims Rhizodeposition plays an important role in mediating soil nutrient availability in ecosystems. However, owing to methodological difficulties (i.e., narrow zone of soil around roots, rapid assimilation by soil microbes) fertility-induced changes in rhizodeposition remain mostly unknown. Methods We developed a novel long-term continuous 13C labelling method to address the effects of two levels of nitrogen (N) fertilization on rhizodeposited carbon (C) by species with different nutrient acquisition strategies. Results Fertility-induced changes in rhizodeposition were modulated by root responses to N availability rather than by changes in soil microbial biomass. Differences among species were mostly related to plant biomass: species with higher total leaf and root biomass also had higher total rhizodeposited C, whereas species with lower root biomass had higher specific rhizodeposited C (per gram root mass). Experimental controls demonstrated that most of the biases commonly associated with this type of experiment (i.e., long-term steady-state labelling) were avoided using our methodological approach. Conclusions These results suggest that the amount of rhizodeposited C from plants grown under different levels of N were driven mainly by plant biomass and root morphology rather than microbial biomass. They also underline the importance of plant characteristics (i.e., biomass allocation) as opposed to traits associated with plant resource acquisition strategies in predicting total C rhizodeposition.

Effects of priority and emerging pollutants on river biofilms

L'activitat humana representa una de les majors causes d'entrada d'una gran varietat de substàncies en els ecosistemes fluvials. L'objectiu principal d'aquest treball es investigar els efectes que els tòxics orgànics poden exercir en els biofilms fluvials. El riu Llobregat ha estat sotmès a fortes pressions, fet que l'ha portat a uns nivells molt elevats de contaminació. En aquest estudi s'ha observat una influència dels plaguicides presents al riu en la distribució de la comunitat de diatomees, així com efectes en el biofilm a nivell funcional i estructural. Experiments amb canals experimentals han mostrat que l'herbicida diuron i el bactericida triclosan poden ocasionar una cadena d'efectes en els biofilms, incloent efectes directes i també efectes indirectes en les relacions entre els components del biofilm. Experiments amb cultius algals han mostrat que aquests tòxics, aplicats en barreja, poden tenir una major toxicitat de la prevista pels models, resultant en efectes sinèrgics.

Estudi dels carotenoides en espècies marrons de bacteris verds del sofre: diversitat, eco-fisiologia i regulació

El present treball es centra en l'estudi a diferents nivells dels carotenoides de les espècies marrons de Bacteris Verds del Sofre (GSB, de l'anglès Green Sulfur Bacteria). L'objectiu global ha estat el d'esbrinar quina és la funció d'aquests pigments dins l'aparell fotosintètic d'aquests microorganismes i aprofundir en el coneixement de la seva estructura i interaccions amb els altres pigments de l'aparell fotosintètic. En primer lloc es va dissenyar un nou mètode de cromatografia líquida d'alta resolució (HPLC) per analitzar de manera més ràpida i precisa els carotenoides de diferents soques de GSB (Capítol 3). Aquest mètode es basa en una purificació prèvia dels extractes pigmentaris amb columnes d'alúmina per eliminar les bacterioclorofil·les (BCls). Això va permetre analitzar amb una elevada resolució i en tan sols 45 min de carrera cromatogràfica els diferents carotenoides i els seus precursors, així com les configuracions trans i cis dels seus isòmers. El segon mètode utilitzat va consistir en una modificació del mètode de Borrego i Garcia-Gil (1994) i va permetre la separació precisa de tot tipus de pigments, procedents tant de cultius purs com de mostres de caràcter complex. Un exemple concret foren uns paleosediments de la zona lacustre de Banyoles. En aquests sediments (0,7-1,5 milions d'anys d'antiguitat) es van detectar, entre d'altres pigments, carotenoides específics de les espècies marrons de GSB, la qual cosa va permetre confirmar la presència d'aquests bacteris a la zona lacustre de Banyoles ja des del Pleistocè inferior. En aquest primer capítol també es van analitzar els carotenoides de Chlorobium (Chl.) phaeobacteroides CL1401 mitjançant cromatografia líquida acoblada a espectrometria de masses (LC-MS/MS), amb l'objectiu de confirmar la seva identificació i el seu pes molecular. A més, també es va avaluar l'efecte de la temperatura, la llum i diferents agents oxidants i reductors en la composició quantitativa i qualitativa dels carotenoides i les BCls d'aquesta espècie. Això va permetre confirmar el caràcter fotosensible de les BCls i que els isòmers trans/cis dels diferents carotenoides no són artefactes produïts durant la manipulació de les mostres, sinó que són constitutius de l'aparell fotosintètic d'aquests microorganismes. El Capítol 4 inclou els experiments de fisiologia duts a terme amb algunes espècies de GSB, a partir dels quals es va intentar esbrinar la dinàmica de síntesi dels diferents pigments de l'aparell fotosintètic (BCl antena, BCl a i carotenoides) durant el creixement d'aquestes espècies. Aquestes investigacions van permetre monitoritzar també els canvis en el nombre de centres de reacció (CR) durant el procés d'adaptació lumínica. La determinació experimental del nombre de CR es va realitzar a partir de la quantificació de la BCl663, l'acceptor primari en la cadena de transport d'electrons dels GSB. L'estimació del nombre de CR/clorosoma es va realitzar tant a partir de dades estequiomètriques i biomètriques presents a la bibliografia, com a partir de les dades experimentals obtingudes en el present treball. El bon ajust obtingut entre les diferents estimacions va donar solidesa al valor estequiomètric calculat, que fou, com a promig, d'uns 70 CR per clorosoma. En aquest capítol de fisiologia també es van estudiar les variacions en les relacions trans/cis pels principals carotenoides de les espècies marrons de GSB. Aquestes es van determinar a partir de cultius purs de laboratori i de poblacions naturals de GSB. Pel que fa als valors trobats en cultius de laboratori no es van observar diferències destacades entre el valor calculat a alta intensitat de llum i el calculat a baixa intensitat, essent en ambdós casos proper a 2. En els clorosomes aïllats de diferents soques marrons aquest quocient prengué un valor similar tant pels isòmers de l'isorenieratè (Isr) com pels del -isorenieratè (-Isr). En poblacions naturals de Chl. phaeobacteroides aquesta relació va ser també de 2 isòmers trans per cada isòmer cis, mantenint-se constant tant en fondària com al llarg del temps. Finalment, en el Capítol 5 es presenta un marcador molecular que permet la identificació específica d'espècies marrons de GSB. Malgrat que inicialment aquest marcador fou dissenyat a partir d'un gen implicat en la síntesi de carotenoides (crtY, el qual codifica per a una licopè ciclasa) la seqüència final a partir de la qual s'han aconseguit els encebadors selectius està relacionada amb la família de proteïnes de les Policètid-ceto-sintases (PKT). Tot i així, l'eina dissenyada pot ser de gran utilitat per a la discriminació d'espècies marrons de GSB respecte les verdes en poblacions mixtes com les que es troben en ambients naturals i obre la porta a futurs experiments d'ecologia microbiana utilitzant tècniques com la PCR en temps real, que permetria la monitorització selectiva de les poblacions d'espècies marrons de GSB en ecosistemes naturals.