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Winter Injury to Grapevine Secondary Phloem and Cambium Impairs Budbreak, Cambium Activity, and Yield Formation

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Abstract

Vitis vinifera is a species of temperate origin that reactivates the dormant secondary phloem from the previous year at the resumption of growth in spring. Following harsh winters, grapevines may display a set of symptoms including delayed and heterogeneous budbreak, dieback with shoot renewal from the trunk base or sudden death of the plant. Although it was suggested that these symptoms may be associated with freeze damage to the secondary phloem, there is no experimental evidence that quantifies tissue responses to freezing and their consequences for the plant. This work evaluated how different severities of cold damage to the secondary phloem during the dormant season impacted the anatomical, physiological, and agronomic responses of grapevines during the subsequent growing season. Single-node cane sections were subjected to a range of freezing temperatures that damaged only the phloem, and changes in anatomy and physiology were monitored. In addition, the consequences of natural winter freezes for yield formation of field-grown plants were evaluated. Our results suggest that the more severe a freeze event is, the greater will be the degree of secondary phloem disorganization, leading to delays in budbreak and subsequent phenological stages, and in cambial activity. Winter freezes also led to a loss of plant vigor and a reduction in cluster number, berries per cluster, and fruit sugar content. We conclude that winter freeze events can produce hidden damage in grapevine perennial tissues, which may compromise subsequent growth and productivity depending on the severity of the damage.

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Acknowledgements

This work was supported by Agencia Nacional de Promoción Científica y Tecnológica, Argentina (ANPCyT); Universidad Nacional de Cuyo (UNCuyo) [PRH, 2007]; and Concejo Nacional de Investigaciones Científicas y Técnicas (CONICET) [PhD fellowship, 2016–2018]. We thank the staff of the Plant Physiology Department and Biological Chemistry of Agronomy Faculty of UNCuyo Mendoza (especially Bruno Cavagnaro and Emiliano Malovini); Viticulture Laboratory at Washington State (especially Lynn Mills, John Ferguson, and Alan Kawakami), DAAC (especially Inés Krause, Laura Ventura, Daniel Ferrero, Vanina Gonzalez, and Natalia Astorga); INTA EEA Mendoza Ecophysiology Department (especially Jorge Perez Peña, Eugenia Galat Giorgi, and Jorge Prieto); IANIGLA CCT-Mendoza (especially Federico Gonzalez and Silvina Lassa); and Facundo Bonamaizon for sharing their knowledge, equipment, and technical support. Moreover, we thank all the students that participated in this research. Mercier Plant Nursery supplied the plant material for this research, and Floralis provided access to laboratory infrastructure and equipment.

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344_2019_10051_MOESM1_ESM.tif

Supplementary material 1 (TIFF 4843 kb) Supplementary Figure 1. Photographs of phloem damage due to winter freeze events in field-grown V. vinifera grapevines in the province of Mendoza, Argentina. A) Plant with successive retraining attempts following trunk death. B) plant with dead cordons requiring renewal. C) Young plant and D) mature plant with dead trunk and regrowth of suckers at the base of the trunk; E) trunk with dead (brown) phloem and live (green) xylem. Authors photos.

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Supplementary material 2 (TIFF 2383 kb) Supplementary Figure 2. Sketch of methodology used to determine the effects of simulated freezing events on anatomical and phenological responses of single-node cuttings of V. vinifera cv. Malbec. Tests were conducted before budbreak in samples collected from the field. DAY 1: a) Twenty dormant canes were collected from a commercial vineyard and kept in airtight bags with moist paper in a Styrofoam box for transport to the laboratory, where the canes were processed immediately. b) Basal cane portions (buds 1 through 8) were divided into 3-cm sections and mixed to obtain a composite sample. Five sections were randomly selected from the composite sample and wrapped in aluminum foil. In total, ten packages were made. c) Each package was tagged and assigned to a simulated freezing event from -2 to -18 °C at 2 °C increments. A control treatment was maintained at room temperature. d) Packages were placed inside a freezer controlled by a data-logger-computer. e) Once the temperature had dropped from ambient to 10 °C, a drop rate of -2 °C/h was maintained down to -19 °C. The temperature was recorded in real-time with an integrated DS18B20 sensor. f) Once a package reached its target temperatures it was removed from the freezer and deposited in a box at 7 °C for 24 hours. Then, packages were maintained at 20 °C for another 24 h. g) Samples were examined for brown discoloration under a stereoscope to determine if phloem and xylem tissues were alive or dead. The proportion of live tissue was recorded for each target temperature to determine phloem LT50. DAY 2: Eighty dormant canes were collected as described for DAY 1. h) Based on the LT50 obtained on DAY 1, three temperature treatments were applied, one coincident with LT50 (FTMed) and two other treatments below and above this measurement (sublethal: FTLow; and superlethal: FTHigh). Untreated samples were considered the control (Ctr). i) Single-node cuttings (n = 50) were subjected to the same type of freezing simulation but with different target temperatures. After the simulated freezing events, the cuttings were kept in trays with water at 20 °C and a photoperiod of 16 h to assess phenological development (n = 20) and anatomical changes (n = 30). j) Phenology was evaluated visually every 3 days. k) Anatomy was studied in cuttings sampled every 7 days and fixed prior to sectioning. FT means Freezing treatment.

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Supplementary material 3 (TIFF 1446 kb) Supplementary Figure 3. Classification criteria for different phases of cambium activity used in this study. Phase 0: dormant cambium; Phase 1: beginning of cell division; Phase 2: cell elongation; Phase 3: cell wall thickening and lignification. Micrographs in 40× magnification, bars indicate 200 μm. p = phloem; c = cambium; x = xylem. Authors photos.

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Supplementary material 4 (TIFF 1419 kb) Supplementary Figure 4. Sketch of sampling protocol in a commercial vineyard to evaluate trunk damage and vegetative and reproductive responses of V. vinifera cv. Malbec grapevines in the province of Mendoza, Argentina. a) Grapevines prior to budbreak. b) Tangential cuts in vine trunks to determine phloem damage by visual observation. c) Healthy plants with live xylem and live (green) phloem. d) Damaged plants with dead (brown) phloem and live xylem. e and f) Plants were tagged for subsequent evaluation at harvest. g and h) Yield and vigor measurements were made at harvest.

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Supplementary material 5 (TIFF 9426 kb) Supplementary Figure 5. Photos of typical differences in vigor and yield of V. vinifera cv. Malbec grapevines tagged prior to budbreak and grown in a commercial vineyard in the province of Mendoza, Argentina. A and C) “No cold damage”: Healthy plants with shoots of normal vigor and yield formation. B and D) “Phloem damage”: Plants affected by cold damage to the trunk phloem with stunted shoots and reduced yield. Authors photos.

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Gonzalez Antivilo, F., Paz, R.C., Tognetti, J. et al. Winter Injury to Grapevine Secondary Phloem and Cambium Impairs Budbreak, Cambium Activity, and Yield Formation. J Plant Growth Regul 39, 1095–1106 (2020). https://doi.org/10.1007/s00344-019-10051-w

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