Bruno Holzapfel

Soil temperature moderates grapevine carbohydrate reserves after bud break and conditions fruit set responses to photoassimilatory stress

In cultivated grapevines (Vitis vinifera L.), suboptimal photoassimilatory conditions during flowering can lead to inflorescence necrosis and shedding of flowers and young ovaries and, consequently, poor fruit set. However, before this study it was not known whether carbohydrate reserves augment fruit set when concurrent photoassimilation is limited. Carbohydrate reserves are most abundant in grapevine roots and soil temperature moderates their mobilisation. Accordingly, we grew potted Chardonnay grapevines in soil at 15°C (cool) or 26°C (warm) from bud break to the onset of flowering to manipulate root carbohydrate reserve status. Then to induce photoassimilatory responses we subjected the plants to low (94µmolmol-1) CO2 or ambient (336µmolmol-1) CO2 atmospheres during fruit setting. Analyses of photoassimilation and biomass and carbohydrate reserve distribution confirmed that fruit set was limited by concurrent photoassimilation. Furthermore, the availability of current photoassimilates for inflorescence development and fruit set was conditioned by the simultaneous demands for shoot and root growth, as well as the restoration of root carbohydrate reserves. Results indicate that great seasonal variability in grapevine fruit set is a likely response of cultivated grapevines to photoassimilatory stresses, such as shading, defoliation and air temperature and to variations in carbohydrate reserve status before flowering.

Rapid monitoring of grapevine reserves using ATR–FT-IR and chemometrics

Predictions of grapevine yield and the management of sugar accumulation and secondary metabolite production during berry ripening may be improved by monitoring nitrogen and starch reserves in the perennial parts of the vine. The standard method for determining nitrogen concentration in plant tissue is by combustion analysis, while enzymatic hydrolysis followed by glucose quantification is commonly used for starch. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FT-IR) combined with chemometric modelling offers a rapid means for the determination of a range of analytes in powdered or ground samples. ATR-FT-IR offers significant advantages over combustion or enzymatic analysis of samples due to the simplicity of instrument operation, reproducibility and speed of data collection. In the present investigation, 1880 root and wood samples were collected from Shiraz, Semillon and Riesling vineyards in Australia and Germany. Nitrogen and starch concentrations were determined using standard analytical methods, and ATR-FT-IR spectra collected for each sample using a Bruker Alpha instrument. Samples were randomly assigned to either calibration or test data sets representing two thirds and one third of the samples respectively. Signal preprocessing included extended multiplicative scatter correction for water and carbon dioxide vapour, standard normal variate scaling with second derivative and variable selection prior to regression. Excellent predictive models for percent dry weight (DW) of nitrogen (range: 0.10-2.65% DW, median: 0.45% DW) and starch (range: 0.25-42.82% DW, median: 7.77% DW) using partial least squares (PLS) or support vector machine (SVM) analysis for linear and nonlinear regression respectively, were constructed and cross validated with low root mean square errors of prediction (RMSEP). Calibrations employing SVM-regression provided the optimum predictive models for nitrogen (R(2)=0.98 and RMSEP=0.07% DW) compared to PLS regression (R(2)=0.97 and RMSEP=0.08% DW). The best predictive models for starch was obtained using PLS regression (R(2)=0.95 and RSMEP=1.43% DW) compared to SVR (R(2)=0.95; RMSEP=1.56% DW). The RMSEP for both nitrogen and starch is below the reported seasonal flux for these analytes in Vitis vinifera. Nitrogen and starch concentrations in grapevine tissues can thus be accurately determined using ATR-FT-IR, providing a rapid method for monitoring vine reserve status under commercial grape production.

The amino acid distribution in rachis xylem sap and phloem exudate of Vitis vinifera ‘Cabernet Sauvignon’ bunches

Amino acids are essential to grape berry and seed development and they are transferred to the reproductive structures through the phloem and xylem from various locations within the plant. The diurnal and seasonal dynamics of xylem and phloem amino acid composition in the leaf petiole and bunch rachis of field-grown Cabernet Sauvignon are described to better understand the critical periods for amino acid import into the berry. Xylem sap was extracted by the centrifugation of excised leaf petioles and rachises, while phloem exudate was collected by immersing these structures in an ethylenediaminetetraacetic acid (EDTA) buffer. Glutamine and glutamic acid were the predominant amino acids in the xylem sap of both grapevine rachises and petioles, while arginine and glycine were the principal amino acids of the phloem exudate. The amino acid concentrations within the xylem sap and phloem exudate derived from these structures were greatest during anthesis and fruit set, and a second peak occurred within the rachis phloem at the onset of ripening. The concentrations of the amino acids within the phloem and xylem sap of the rachis were highest just prior to or after midnight while the flow of sugar through the rachis phloem was greatest during the early afternoon. Sugar exudation rates from the rachis was greater than that of the petiole phloem between anthesis and berry maturity. In summary, amino acid and sugar delivery through the vasculature to grape berries fluctuates over the course of the day as well as through the season and is not necessarily related to levels near the source.

Shifts in biomass and nitrogen allocation of tree seedlings in response to root-zone temperature

Root-zone warming of trees can result in an increase in biomass production but the mechanisms for this increase may differ between evergreen and deciduous species. The leaf gas exchange, carbohydrate and nitrogen (N) partitioning of two Australian evergreens, Acacia saligna and Eucalyptus cladocalyx, were compared to the deciduous Populus deltoides and Acer negundo after exposure to cool or warm soil during spring. The warm treatment stimulated aboveground biomass production in all four species; however, the form of this increase was species dependent. Compared with the evergreens, soluble sugars were mobilised from the above- and belowground components to a greater extent in the deciduous species, especially during root-zone warming. Photosynthesis, stomatal conductance and transpiration were increased in the warm soil treatment for the two evergreens and P. deltoides only. In P. deltoides and A. saligna the new fine roots contained greater starch concentrations when grown in warm soil but only in A. negundo was new root growth greater. Compared with the other three species, the leguminous A. saligna contained the highest N and most of this was concentrated in the phyllodes of warmed plants with no apparent mobilisation from the existing biomass. In the other evergreen, E. cladocalyx, the existing leaves and stems were a N source for new growth, while in the two deciduous species N was derived from the woody components and structural roots. These data show that the carbohydrate movement and N partitioning patterns in response to soil warming differ between perennial and deciduous plants and are likely responsible for the different forms of biomass accumulation in each of these species.

Flowers regulate the growth and vascular development of the inflorescence rachis in Vitis vinifera L.

The rachis, the structural framework of the grapevine (Vitis vinifera L.) inflorescence (and subsequent bunch), consists of a main axis and one or more orders of lateral branches with the flower-bearing pedicels at their fine tips. The rachis is crucial both for support, and transport from the shoot. Earlier suggestions that the flowers per se affect normal rachis development are investigated further in this study. Different percentages (0, 25, 50, 75 or 100) of flowers were removed manually one week before anthesis on field-grown vines. Treatment effects on subsequent rachis development (curvature, vitality, anatomy, starch deposit) were assessed. Sections, both fixed and embedded, and fresh hand-cut were observed by fluorescence and bright-field optics after appropriate staining. Emphasis was on measurement of changes in cross-sectional area of secondary xylem and phloem, and on maturation of fibres and periderm. Specific defects in rachis development were dependent on the percent and location of flower removal one week prior to anthesis. The rachises curved inwards where most of the flowers were removed. When fully de-flowered, they became progressively necrotic from the laterals back to the primary axes and from the distal to the proximal end of those axes, with a concurrent disorganisation of their anatomy. A few remaining groups of flowers prevented desiccation and abscission of the rachis axes proximal to the group, but not distally. Flower removal (50%) reduced rachis elongation, while 75% removal reduced xylem and phloem area and delayed phloem fibre and periderm development. 75% flower removal did not affect starch present in the rachis during berry development. Developing flowers affect the growth and vitality of the rachis and the development of its vascular and support structures. The extent of these effects depends on the cultivar and the number and position of flowers remaining after some are removed one week before anthesis.

Vitis vinifera root and leaf metabolic composition during fruit maturation: implications of defoliation

Grapevine (Vitis vinifera) roots and leaves represent major carbohydrate and nitrogen (N) sources, either as recent assimilates, or mobilized from labile or storage pools. This study examined the response of root and leaf primary metabolism following defoliation treatments applied to fruiting vines during ripening. The objective was to link alterations in root and leaf metabolism to carbohydrate and N source functioning under conditions of increased fruit sink demand. Potted grapevine leaf area was adjusted near the start of véraison to 25 primary leaves per vine compared to 100 leaves for the control. An additional group of vines were completely defoliated. Fruit sugar and N content development was assessed, and root and leaf starch and N concentrations determined. An untargeted GC/MS approach was undertaken to evaluate root and leaf primary metabolite concentrations. Partial and full defoliation increased root carbohydrate source contribution towards berry sugar accumulation, evident through starch remobilization. Furthermore, root myo-inositol metabolism played a distinct role during carbohydrate remobilization. Full defoliation induced shikimate pathway derived aromatic amino acid accumulation in roots, while arginine accumulated after full and partial defoliation. Likewise, various leaf amino acids accumulated after partial defoliation. These results suggest elevated root and leaf amino N source activity when leaf N availability is restricted during fruit ripening. Overall, this study provides novel information regarding the impact of leaf source restriction, on metabolic compositions of major carbohydrate and N sources during berry maturation. These results enhance the understanding of source organ carbon and N metabolism during fruit maturation.

Vascular development of the grapevine ( Vitis vinifera L.) inflorescence rachis in response to flower number, plant growth regulators and defoliation

The grapevine inflorescence is a determinate panicle and as buds emerge, shoot, flower and rachis development occur simultaneously. The growth and architecture of the rachis is determined by genetic and environmental factors but here we examined the role of flower and leaf number as well as hormones on its elongation and vascular development. The consequences of rachis morphology and vascular area on berry size and composition were also assessed. One week prior to anthesis, Merlot and Cabernet Sauvignon field vines were exposed to manual flower removal, exogenous plant growth regulators or pre-bloom leaf removal. Manual removal of half the flowers along the vertical axis of the inflorescence resulted in a shorter rachis in both cultivars. Conversely, inflorescences treated with gibberellic acid (GA3) and the synthetic cytokinin, 6-benzylaminopurine (BAP) resulted in a longer rachis while pre-bloom removal of all leaves on the inflorescence-bearing shoot did not alter rachis length relative to untreated inflorescences. Across the treatments, the cross-sectional areas of the conducting xylem and phloem in the rachis were positively correlated to rachis girth, flower number at anthesis, bunch berry number, bunch berry fresh mass and bunch sugar content at harvest. Conversely, average berry size and sugar content were not linked to rachis vascular area. These data indicate that the morphological and vascular development of the rachis was more responsive to flower number and plant growth regulators than to leaf removal.

Viticulture and Wine Science

The grape and wine industry is commercially attractive because it includes a significant value adding chain. Its structure, size and competitiveness however make it a complex industry. The skills required to produce wine usually distinguish between those associated with the production of the fruit and those associated with processing it. This chapter first looks at the history of wine as well as the global wine industry. The basic botany of grapevines is then reviewed and the more important aspects of viticulture and vineyard management are explored. The production of wine is presented before exploring the microbiological and chemical nature of this agro transformation.