Pascale Maillard

Non-structural carbohydrates in woody plants compared among laboratories

Non-structural carbohydrates (NSC) in plant tissue are frequently quantified to make inferences about plant responses to environmental conditions. Laboratories publishing estimates of NSC of woody plants use many different methods to evaluate NSC. We asked whether NSC estimates in the recent literature could be quantitatively compared among studies. We also asked whether any differences among laboratories were related to the extraction and quantification methods used to determine starch and sugar concentrations. These questions were addressed by sending sub-samples collected from five woody plant tissues, which varied in NSC content and chemical composition, to 29 laboratories. Each laboratory analyzed the samples with their laboratory-specific protocols, based on recent publications, to determine concentrations of soluble sugars, starch and their sum, total NSC. Laboratory estimates differed substantially for all samples. For example, estimates for Eucalyptus globulus leaves (EGL) varied from 23 to 116 (mean = 56) mg g(-1) for soluble sugars, 6-533 (mean = 94) mg g(-1) for starch and 53-649 (mean = 153) mg g(-1) for total NSC. Mixed model analysis of variance showed that much of the variability among laboratories was unrelated to the categories we used for extraction and quantification methods (method category R(2) = 0.05-0.12 for soluble sugars, 0.10-0.33 for starch and 0.01-0.09 for total NSC). For EGL, the difference between the highest and lowest least squares means for categories in the mixed model analysis was 33 mg g(-1) for total NSC, compared with the range of laboratory estimates of 596 mg g(-1). Laboratories were reasonably consistent in their ranks of estimates among tissues for starch (r = 0.41-0.91), but less so for total NSC (r = 0.45-0.84) and soluble sugars (r = 0.11-0.83). Our results show that NSC estimates for woody plant tissues cannot be compared among laboratories. The relative changes in NSC between treatments measured within a laboratory may be comparable within and between laboratories, especially for starch. To obtain comparable NSC estimates, we suggest that users can either adopt the reference method given in this publication, or report estimates for a portion of samples using the reference method, and report estimates for a standard reference material. Researchers interested in NSC estimates should work to identify and adopt standard methods.

Do trees use reserve or newly assimilated carbon for their defense reactions? A 13C labeling approach with young Scots pines inoculated with a bark-beetle-associated fungus (Ophiostoma brunneo ciliatum)

Three-year-old saplings of Pinus sylvestris L. were labeled with 13CO2 prior to inoculating the trunk with Ophiostoma brunneo ciliatum, a blue-staining fungus usually associated to Ips sexdentatus. During incubation, half the trees were submitted to a severe drought that decreased photosynthesis and natural 13C content in non-labeled saplings. A large 13C-excess was obtained in wood and phloem, especially in the fractions of soluble proteins, starch and soluble sugars of labeled saplings. Drought increased 13C-excess, due to reduced photosynthesis and smaller dilution of 13C by the addition of newly assimilated 12C. The induced-reaction zones in inoculated saplings displayed large total C (58 g 100 g−1) because of the accumulation of secondary metabolites. They also showed much larger 13C-excess than any other compartment: the contribution of stored C to the reaction zones was much higher than that of currently assimilated C. Moreover, drought lowered the contribution of the latter, as shown by the increase of 13C in the reaction zones. We conclude that stored C was readily mobilized for the construction of reaction tissues, and that the contribution of currently assimilated C was only minor.RésuméDe jeunes pins sylvestres (Pinus sylvestris L.) âgés de trois ans ont été marqués avec du 13CO2 puis inoculés dans le tronc avec Ophiostoma brunneo ciliatum, un champignon habituellement associé au scolyte Ips sexdentatus. Pendant l’incubation, la moitié des arbres a été soumise à une sécheresse sévère qui a fortement réduit la photosynthèse et l’abondance naturelle en 13C des individus non marqués. Un fort excès en 13C a été détecté dans le bois et le phloème ainsi que dans les protéines solubles, l’amidon et les sucres solubles des individus marqués. La sécheresse a amplifié cet excès, du fait d’une photosynthèse réduite et donc d’une moindre dilution du 13C par du 12C récemment assimilé. Les zones de réaction induite autour des points d’inoculation présentaient de fortes teneurs en C (58 g 100 g−1), du fait de l’accumulation massive de métabolites secondaires. Elles présentaient également un excès de 13C plus marqué que n’importe quel autre tissu : ces zones de réaction étaient donc essentiellement constituées à partir de C provenant des réserves avec une faible contribution de C récemment assimilé. De plus, la sécheresse a augmenté la contribution du C de réserve, comme le montre l’augmentation de l’excès de 13C dans les zones de réaction.

Seasonal changes of C and N non-structural compounds in the stem sapwood of adult sessile oak and beech trees

We assessed the pools of non-structural nitrogen compounds (NSNC) through a year, thereby addressing the question of whether mature sessile oak [Quercus petraea (Matt.) Liebl.] and beech (Fagus sylvatica L.), which differ in wood anatomy and growth patterns, exhibit contrasting seasonal dynamics of NSNC pools as previously shown for non-structural carbohydrate (NSC) pools. Seasonal fluctuations of NSNC (amino acids and soluble proteins) and NSC (starch and soluble sugars) pools were analyzed in the inner and the outer stem sapwood. In oak, NSC showed marked seasonal variation within the stem sapwood (accumulation during winter and decrease during bud burst and early wood growth), whereas in beech seasonal fluctuations in NSC were of minor amplitude. Even if the distribution and intensity of the NSNC pools differed between the two species, NSNC of the stem sapwood did not show seasonal variation. The most significant change in NSNC pools was the seasonal fluctuation of protein composition. In both species, two polypeptides of 13 kDa (PP13) and 26 kDa (PP26) accumulated during the coldest period in parallel with starch to sugar conversion and disappeared with the onset of spring growth. The absence of seasonal changes in total soluble protein concentration suggests that the polypeptides are involved in the internal nitrogen (N) cycling of the stem rather than in N storage and remobilization to the other growing organs of the tree.

Inflorescence of grapevine (Vitis vinifera L.): a high ability to distribute its own assimilates

The distribution of carbon (C) into whole grapevine fruiting cuttings was investigated during flower development to determine the relative contribution of inflorescence and leaf photoassimilates in the total C balance and to investigate their partitioning towards other plant organs. A (13)C labelling procedure was used to label C photoassimilates by leaves and inflorescences in grapevine. Investigations were carried out at various stages of flower/berry development, from separated cluster to fruit set, using grapevine fruiting cuttings with four leaves (Vitis vinifera L. cv. Chardonnay). This is the first study reporting that, during its development, (i) the carbon needs of the inflorescence were met by both leaf and inflorescence photosynthesis, and (ii) the inflorescence amazingly participated significantly to the total C balance of grapevine cuttings by redistributing an important part of its own assimilates to other plant organs. With regard to flowering, 29% of C assimilated by the inflorescence remained in the inflorescence, while partitioning towards the stem reached 42% and, as a lower proportion, 15% in leaves, and 14% in roots.

Nitrogen sources for current-year shoot growth in 50-year-old sessile oak trees: an in situ 15N labeling approach

We used long-term in situ (15)N labeling of the soil to investigate the contribution of the two main nitrogen (N) sources (N uptake versus N reserves) to sun shoot growth from bud burst to full leaf expansion in 50-year-old sessile oaks. Recovery of (15)N by growing compartments (leaves, twigs and buds) and presence of (15)N in phloem sap were checked weekly. During the first 2 weeks following bud burst, remobilized N contributed ~90% of total N in growing leaves and twigs. Nitrogen uptake from the soil started concomitantly with N remobilization but contributed only slightly to bud burst. However, the fraction of total N due to N uptake increased markedly once bud burst had occurred, reaching 27% in fully expanded leaves and 18% in developed twigs. In phloem sap, the (15)N label appeared a few days after the beginning of labeling and increased until the end of bud burst, and then decreased at full leaf expansion in June. Of all the shoot compartments, leaves attracted most of the absorbed N, which accounted for 68% of new N in shoots, whereas twigs and new buds accounted for only 28 and 3%, respectively. New N allocated to leaves increased from unfolding to full expansion as total N concentration in the leaves decreased. Our results underline the crucial role played by stored N in rapid leaf growth and in the sustained growth of oak trees. Any factors that reduce N storage in autumn may therefore impair spring shoot growth.

Mathematical analysis and comparison of growth fluctua- tions of the aerial system of young Terminalia superba Englers et Diels (Combretaceae)

HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Mathematical analysis and comparison of growth fluctuations of the aerial system of young Terminalia superba, Englers et Diels (Combretaceae) P. Maillard, Marie Agnes Jacques, Emile Miginiac, B. Millet, . Inra, . Universite