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Plant function and evolutionary biology
RESEARCH ARTICLE

Stomatal behaviour of irrigated Vitis vinifera cv. Syrah following partial root removal

V. Zufferey A C and D. R. Smart B
+ Author Affiliations
- Author Affiliations

A Station de recherche Agroscope Changins-Wädenswil ACW, CP 1012, CH-1260 Nyon (Switzerland).

B Department of Viticulture and Enology, University of California, One Shields Avenue, Davis, CA 95616, USA.

C Corresponding author. Email: vivian.zufferey@acw.admin.ch

Functional Plant Biology 39(12) 1019-1027 https://doi.org/10.1071/FP12091
Submitted: 20 March 2012  Accepted: 7 September 2012   Published: 8 October 2012

Abstract

We examined stomatal behaviour of a grapevine cultivar (Vitis vinifera L. cv. Syrah) following partial root removal under field conditions during progressively developing water deficits. Partial root removal led to an increase in hydraulic resistances along the soil-to-leaf pathway and leaf wilting symptoms appeared in the root-pruned plants immediately following root removal. Leaves recovered from wilting shortly thereafter, but hydraulic resistances were sustained. In comparison with the non-root pruned vines, leaves of root-pruned vines showed an immediate decrease in both pre-dawn (ψPD) and midday (ψleaf) leaf water potential. The decline in ψPD was unexpected in as much as soil moisture was not altered and it has been shown that axial water transport readily occurs in woody perennials. Only ~30% of the functional root system was removed, thus leaving the system mainly intact for water redistribution. Stem water potential (ψStem) and leaf gas exchanges of CO2 (A) and H2O (E) also declined immediately following root pruning. The lowering of ψPD, ψleaf, ψStem, A and E was sustained during the entire growing season and was not dependent on irrigation during that time. This, and a close relationship between stomatal conductance (gs) and leaf-specific hydraulic conductance (Kplant), indicated that the stomatal response was linked to plant hydraulics. Stomatal closure was observed only in the root-restricted plants and at times of very high evaporative demand (VPD). In accordance with the Ball-Berry stomatal control model proposed by Ball et al. (1987), the stomatal sensitivity factor was also lower in the root-restricted plants than in intact plants as soil water availability decreased. Although ψPD, ψStem and ψLeaf changed modestly and gradually following root removal, gs changed dramatically and abruptly following removal. These results suggest the involvement of stomatal restricting signals being propagated following removal of roots.

Additional keywords: gas exchanges, grapevine, leaf specific hydraulic conductance, leaf water potential, root pruning, stomatal conductance.


References

Addington RN, Mitchell RJ, Oren R, Donovan LA (2004) Stomatal sensitivity to vapor pressure deficit and its relationship to hydraulic conductance in Pinus palustris. Tree Physiology 24, 561–569.
Stomatal sensitivity to vapor pressure deficit and its relationship to hydraulic conductance in Pinus palustris.Crossref | GoogleScholarGoogle Scholar |

Alsina MM, Smart DR, Bauerle T, de Herralde F, Biel C, Stockert CM, Negron C, Save R (2011) Seasonal changes of whole root system conductance by a drought tolerant grape root system. Journal of Experimental Botany 62, 99–109.
Seasonal changes of whole root system conductance by a drought tolerant grape root system.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhsFamurbK&md5=3dc4bbf5901dfc9cc6491bd9e45a54ddCAS |

Améglio T, Archer P, Cohen M, Valancogne C, Daudet F, Dayau S, Cruiziat P (1999) Significance and limits in the use of predawn leaf water potential for tree irrigation. Plant and Soil 207, 155–167.
Significance and limits in the use of predawn leaf water potential for tree irrigation.Crossref | GoogleScholarGoogle Scholar |

Ball JT, Woodrow IE, Berry JA (1987) A model predicting stomatal conductance and its contribution to the control of photosynthesis under different environmental conditions. In ‘Progress in photosynthesis research’. (Ed. J Biggins) pp. 221–234. (Martinus Nijhoff: Dordrecht, The Netherlands)

Bauerle TL, Richards JH, Smart DR, Eissenstat DM (2008) Importance of internal hydraulic redistribution for prolonging the lifespan of roots in dry soil. Plant, Cell & Environment 31, 177–186.

Breda N, Granier A, Barataud F, Moyne C (1995) Soil water dynamics in an oak stand. I. Soil moisture, water potentials and water uptake by roots. Plant and Soil 172, 17–27.

Brodribb TJ, Holbrook NM (2003) Stomatal closure during leaf dehydratation, correlation with other leaf physiological traits. Plant Physiology 132, 2166–2173.
Stomatal closure during leaf dehydratation, correlation with other leaf physiological traits.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXmsVantbc%3D&md5=8e24cf35910547926f7280757c6f8eaaCAS |

Burgess SSO, Bleby TM (2006) Redistribution of soil water to lateral roots mediated by stem tissues. Journal of Experimental Botany 57, 3283–3291.
Redistribution of soil water to lateral roots mediated by stem tissues.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xps1ygt7w%3D&md5=f05cbe7950e6b8c7aa5b5f815c6549f9CAS |

Chaves MM, Tenhunen JD, Harley P, Lange OL (1987) Gas exchange studies in two Portuguese grapevine cultivars. Physiologia Plantarum 70, 639–647.
Gas exchange studies in two Portuguese grapevine cultivars.Crossref | GoogleScholarGoogle Scholar |

Chaves MM, Zarrouk O, Fransisco R, Costa JM, Santos T, Regalado AP, Rodrigues ML, Lopes CM (2010) Grapevine under deficit irrigation: hints from physiological and molecular data. Annals of Botany 105, 661–676.
Grapevine under deficit irrigation: hints from physiological and molecular data.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3c3ovFGnsg%3D%3D&md5=846d9053fad60ff872755d1c783864cbCAS |

Choat B, Drayton WM, Brodersen CR, Matthews MA, Shackel KA, Wada H, McElrone AJ (2010) Measurement of vulnerability to water stress-induced cavitation in grapevine: a comparison of four techniques applied to long-vesseled species. Plant, Cell & Environment 33, 1502–1512.

Cochard H, Coll L, Le Roux X, Ameglio T (2002) Unraveling the effects of plant hydraulics on stomatal closure during water stress in walnut. Plant Physiology 128, 282–290.
Unraveling the effects of plant hydraulics on stomatal closure during water stress in walnut.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XmvVSqtw%3D%3D&md5=dd9b4cb3bb22d0497d3de57655ce7db8CAS |

Comstock JP (2000) Variation in hydraulic architecture and gas exchange in two desert sub-shrubs, Hymenoclea salsola (T.&G.) and Ambrosia dumosa. Oecologia 125, 1–10.
Variation in hydraulic architecture and gas exchange in two desert sub-shrubs, Hymenoclea salsola (T.&G.) and Ambrosia dumosa.Crossref | GoogleScholarGoogle Scholar |

Correia MJ, Pereira JS, Chaves MM, Rodrigues ML, Pacheco CA (1995) ABA xylem concentrations determine maximum daily leaf conductance of field-grown Vitis vinifera L. plants. Plant, Cell & Environment 18, 511–521.
ABA xylem concentrations determine maximum daily leaf conductance of field-grown Vitis vinifera L. plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXmsVKitr4%3D&md5=2797345d2a39d4bd8444c39e9ec4bbe2CAS |

Davies WJ, Zhang J (1991) Root signals and the regulation of growth and development of plants in drying soil. Annual Review of Plant Physiology and Plant Molecular Biology 42, 55–76.
Root signals and the regulation of growth and development of plants in drying soil.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3MXltFSmsr8%3D&md5=bb3206e6c0e851e911450830afa04db6CAS |

Davies WJ, Tardieu F, Trejo CL (1994) How do chemical signals work in plants that grow in drying soil? Plant Physiology 104, 309–314.

Donovan LA, Richards JH, Linton MJ (2003) Magnitude and mechanisms of predawn disequilibrium between predawn and plant soil water potentials in desert shrubs. Ecology 84, 463–470.
Magnitude and mechanisms of predawn disequilibrium between predawn and plant soil water potentials in desert shrubs.Crossref | GoogleScholarGoogle Scholar |

Ewers BE, Oren R, Sperry JS (2000) Influence of nutrient versus water supply on hydraulic architecture and water balance in Pinus taeda. Plant, Cell & Environment 23, 1055–1066.
Influence of nutrient versus water supply on hydraulic architecture and water balance in Pinus taeda.Crossref | GoogleScholarGoogle Scholar |

Franks PJ (2004) Stomatal control and hydraulic conductance, with special reference to tall trees. Tree Physiology 24, 865–878.
Stomatal control and hydraulic conductance, with special reference to tall trees.Crossref | GoogleScholarGoogle Scholar |

Fuchs EE, Livingston NJ (1996) Hydraulic control of stomatal conductance in Douglas fir (Pseudotsuga mensiesii (Mirb) Franco) and alder (Alnus rubra (Bong)) seedlings. Plant, Cell & Environment 19, 1091–1098.
Hydraulic control of stomatal conductance in Douglas fir (Pseudotsuga mensiesii (Mirb) Franco) and alder (Alnus rubra (Bong)) seedlings.Crossref | GoogleScholarGoogle Scholar |

Harley PC, Baldocchi DD (1995) Scaling carbon dioxide and water vapour exchange from leaf to canopy in a deciduous forest. I. Leaf model parametrization. Plant, Cell & Environment 18, 1146–1156.
Scaling carbon dioxide and water vapour exchange from leaf to canopy in a deciduous forest. I. Leaf model parametrization.Crossref | GoogleScholarGoogle Scholar |

Holbrook NM, Shashidhar VR, James RA, Munns R (2002) Stomatal control in tomato with ABA-deficient roots: response of grafted plants to soil drying. Journal of Experimental Botany 53, 1503–1514.
Stomatal control in tomato with ABA-deficient roots: response of grafted plants to soil drying.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XktlWjtbk%3D&md5=17c333b6ff9673740f07e073673d40daCAS |

Hubbard RM, Bond BJ, Ryan MG (1999) Evidence that hydraulic conductance limits photosynthesis in old Pinus ponderosa trees. Tree Physiology 19, 165–172.
Evidence that hydraulic conductance limits photosynthesis in old Pinus ponderosa trees.Crossref | GoogleScholarGoogle Scholar |

Hubbard RM, Ryan MG, Stiller V, Sperry JS (2001) Stomatal conductance and photosynthesis vary lineary with plant hydraulic conductance in Ponderosa pine. Plant, Cell & Environment 24, 113–121.
Stomatal conductance and photosynthesis vary lineary with plant hydraulic conductance in Ponderosa pine.Crossref | GoogleScholarGoogle Scholar |

Jarvis AJ, Davies WJ (1998) Modeling stomatal responses to soil and atmospheric drought. Journal of Experimental Botany 49, 399–406.
Modeling stomatal responses to soil and atmospheric drought.Crossref | GoogleScholarGoogle Scholar |

Jones HG (1998) Stomatal control of photosynthesis and transpiration. Journal of Experimental Botany 49, 387–398.

Kramer PJ, Boyer JS (1995) ‘Water relations of plants and soils.’ (Academic Press: San Diego, CA)

Leuning R (1990) Modeling stomatal behaviour and photosynthesis of Eucalyptus grandis. Australian Journal of Plant Physiology 17, 159–175.
Modeling stomatal behaviour and photosynthesis of Eucalyptus grandis.Crossref | GoogleScholarGoogle Scholar |

Loveys BR (1991) How useful is a knowledge of ABA physiology for crop improvement? In ‘Abscisic acid physiology and biochemistry’. (Eds WJ Davies, HG Jones) pp. 245–260. (Bios Scientific Publishers: Oxford)

Lovisolo C, Schubert A (1998) Effects of water stress on vessel size and xylem hydraulic conductivity in Vitis vinifera L. Journal of Experimental Botany 49, 693–700.

Lovisolo C, Hartung W, Schubert A (2002) Whole-plant hydraulic conductance and root-to-shoot flow of abscisic acid are independently affected by stress in grapevines. Functional Plant Biology 29, 1349–1356.
Whole-plant hydraulic conductance and root-to-shoot flow of abscisic acid are independently affected by stress in grapevines.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xps1ygtLs%3D&md5=2db6ddddb74b30183990b8ac61749d79CAS |

Meinzer FC (2002) Co-ordination of vapor and liquid phase water transport proprieties in plants. Plant, Cell & Environment 25, 265–274.
Co-ordination of vapor and liquid phase water transport proprieties in plants.Crossref | GoogleScholarGoogle Scholar |

Meinzer FC, Grantz DA (1990) Stomatal and hydraulic conductance in growing sugarcane: stomatal adjustment to water transport capacity. Plant, Cell & Environment 13, 383–388.
Stomatal and hydraulic conductance in growing sugarcane: stomatal adjustment to water transport capacity.Crossref | GoogleScholarGoogle Scholar |

Monteith JL (1995) A reinterpretation of stomatal responses to humidity. Plant, Cell & Environment 18, 357–364.
A reinterpretation of stomatal responses to humidity.Crossref | GoogleScholarGoogle Scholar |

Morison JI (1987) Intercellular CO2 concentration and stomatal response to CO2. In ‘Stomatal function’. (Eds E Zeiger, GD Farquhar, IR Cowan) pp. 229–251. (Stanford University Press: Stanford)

Mott KA (1988) Do stomata respond to CO2 concentrations other than intercellular? Plant Physiology 86, 200–203.
Do stomata respond to CO2 concentrations other than intercellular?Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1cXhsVOgsbg%3D&md5=f8020131fb6dd6ce20f3d699f703dd9aCAS |

Mott KA, Buckley TN (1998) Stomatal heterogeneity. Journal of Experimental Botany 49, 407–417.

Mott KA, Franks PJ (2001) The role of epidermal turgor in stomatal interactions following a local perturbation in humidity. Plant, Cell & Environment 24, 657–662.
The role of epidermal turgor in stomatal interactions following a local perturbation in humidity.Crossref | GoogleScholarGoogle Scholar |

Mott KA, Parkhurst DF (1991) Stomata response to humidity in air and helox. Plant, Cell & Environment 14, 509–515.
Stomata response to humidity in air and helox.Crossref | GoogleScholarGoogle Scholar |

Nardini A, Tyree MT, Salleo S (2001) Xylem cavitation in the leaf of Prunus laurocerasus and its impact on leaf hydraulics. Plant Physiology 125, 1700–1709.
Xylem cavitation in the leaf of Prunus laurocerasus and its impact on leaf hydraulics.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXjtFKqt7k%3D&md5=c498e069dfe4883d96cec9b939e294f3CAS |

Oren R, Sperry JS, Ewers BE, Pataki DE, Phillips N, Megonigal JP (2001) Sensitivity of mean canopy stomatal conductance to vapor pressure deficit in a flooded Taxadium distichum L. forest: hydraulic and non-hydraulic effects. Oecologia 126, 21–29.
Sensitivity of mean canopy stomatal conductance to vapor pressure deficit in a flooded Taxadium distichum L. forest: hydraulic and non-hydraulic effects.Crossref | GoogleScholarGoogle Scholar |

Saliendra NZ, Sperry JS, Comstock JP (1995) Influence of leaf water status on stomatal response to humidity, hydraulic conductance, and soil drought in Betula occidentalis. Planta 196, 357–366.
Influence of leaf water status on stomatal response to humidity, hydraulic conductance, and soil drought in Betula occidentalis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXls1Wqs7s%3D&md5=d58ba73191881f6027b2fca8c4bfd9fdCAS |

Salleo S, Nardini A, Pitt F, LoGullo M (2000) Xylem cavitation and hydraulic control of stomatal conductance in Laurel (Laurus nobilis L.). Plant, Cell & Environment 23, 71–79.
Xylem cavitation and hydraulic control of stomatal conductance in Laurel (Laurus nobilis L.).Crossref | GoogleScholarGoogle Scholar |

Scholander PF, Hammel HT, Bradstreet ED, Hemmingsen EA (1965) Sap pressure in vascular plants. Science 148, 339–346.
Sap pressure in vascular plants.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cvlsVKquw%3D%3D&md5=7a95ab02c16d02693138b3c00d255211CAS |

Schultz HR (2003) Differences in hydraulic architecture account for near-isohydric and anisohydric behaviour of two field-grown Vitis vinifera L. cultivars during drought. Plant, Cell & Environment 26, 1393–1405.
Differences in hydraulic architecture account for near-isohydric and anisohydric behaviour of two field-grown Vitis vinifera L. cultivars during drought.Crossref | GoogleScholarGoogle Scholar |

Schultz HR, Matthews MA (1997) High vapour pressure deficit exacerbates xylem cavitation and photoinhibition in shade-grown Piper auritum H.B.&K. during prolonged sunflecks. I. Dynamics of plant water relations. Oecologia 110, 312–319.
High vapour pressure deficit exacerbates xylem cavitation and photoinhibition in shade-grown Piper auritum H.B.&K. during prolonged sunflecks. I. Dynamics of plant water relations.Crossref | GoogleScholarGoogle Scholar |

Shackel KA, Brinckmann E (1985) In situ measurement of epidermal cell turgor, leaf water potential, and gas exchange in Tradescantia virginia L. Plant Physiology 78, 66–70.
In situ measurement of epidermal cell turgor, leaf water potential, and gas exchange in Tradescantia virginia L.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cnhs1ekug%3D%3D&md5=80e837cb5ca43ed8f9e6039021b53f3aCAS |

Smart RE, Combe B (1983) Water relations of grapevines. In ‘Additional woody crop plants. Water deficit and plant growth. Vol. VII’. (Ed. TT Kozlowski) pp. 138–196. (Academic Press: New York)

Smart DR, Carlisle E, Alonso B (2005) Transverse hydraulic redistribution by a grapevine. Plant, Cell & Environment 28, 157–166.
Transverse hydraulic redistribution by a grapevine.Crossref | GoogleScholarGoogle Scholar |

Smart DR, Breazeale A, Zufferey V (2006) Physiological changes in plant hydraulics induced by partial root removal of irrigated grapevine (Vitis vinifera cv. Syrah). American Journal of Enology and Viticulture 57, 201–209.

Sperry JS, Pockman WT (1993) Limitation of transpiration by hydraulic conductance and xylem cavitation in Betula occidentalis. Plant, Cell & Environment 16, 279–287.
Limitation of transpiration by hydraulic conductance and xylem cavitation in Betula occidentalis.Crossref | GoogleScholarGoogle Scholar |

Sperry JS, Alder NN, Easlack SE (1993) The effect of reduced hydraulic conductance on stomatal conductance and xylem cavitation. Journal of Experimental Botany 44, 1075–1082.
The effect of reduced hydraulic conductance on stomatal conductance and xylem cavitation.Crossref | GoogleScholarGoogle Scholar |

Sperry JS, Hacke UG, Oren R, Comstock JP (2002) Water deficit and hydraulic limits to leaf water supply. Plant, Cell & Environment 25, 251–263.
Water deficit and hydraulic limits to leaf water supply.Crossref | GoogleScholarGoogle Scholar |

Stoll M, Loveys B, Dry P (2000) Hormonal changes induced by partial rootzone drying of irrigated grapevine. Journal of Experimental Botany 51, 1627–1634.
Hormonal changes induced by partial rootzone drying of irrigated grapevine.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXnt12ju7o%3D&md5=1a1b7a9c7891db088e8f5cb02f1dcf52CAS |

Tardieu F, Davies WJ (1993) Integration of hydraulic and chemical signaling in the control of stomatal conductance and water status of droughted plants. Plant, Cell & Environment 16, 341–349.
Integration of hydraulic and chemical signaling in the control of stomatal conductance and water status of droughted plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXlsFKnurw%3D&md5=e69d5493023a17b07101393f6558fdefCAS |

Tardieu F, Simonneau T (1998) Variability among species of stomatal control under fluctuating soil water status and evaporative demand: modeling isohydric and anisohydric behaviours. Journal of Experimental Botany 49, 419–432.

Tardieu F, Lafarge T, Simonneau T (1996) Stomatal control by fed or endogenous xylem ABA in sunflower: interpretation of observed correlations between leaf water potential and stomatal conductance in anisohydric species. Plant, Cell & Environment 19, 75–84.
Stomatal control by fed or endogenous xylem ABA in sunflower: interpretation of observed correlations between leaf water potential and stomatal conductance in anisohydric species.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xhs1ejtbs%3D&md5=1d7dbb0a690b0e8dd3a58fc8575aec18CAS |

Teskey RO, Hincley TM, Grier CC (1983) Effect of interruption of flow path on stomatal conductance of Abies amabilis. Journal of Experimental Botany 34, 1251–1259.
Effect of interruption of flow path on stomatal conductance of Abies amabilis.Crossref | GoogleScholarGoogle Scholar |

Turner NC (1988) Measurement of plant water status by the pressure chamber technique. Irrigation Science 9, 289–308.
Measurement of plant water status by the pressure chamber technique.Crossref | GoogleScholarGoogle Scholar |

Tyree MT, Sperry JS (1988) Do woody plants operate near the point of catastrophic xylem dysfunction caused by dynamic water stress? Answer from a model. Plant Physiology 88, 574–580.
Do woody plants operate near the point of catastrophic xylem dysfunction caused by dynamic water stress? Answer from a model.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cnhvVGjtA%3D%3D&md5=6246ce991fc94f8cf1dd29ac21d1646bCAS |

Tyree MT, Valez V, Dalling JW (1998) Growth dynamics of root and shoot hydraulic conductance in seedling of five neotropical tree species: scaling to show possible adaptation to differing light regimes. Oecologia 114, 293–298.
Growth dynamics of root and shoot hydraulic conductance in seedling of five neotropical tree species: scaling to show possible adaptation to differing light regimes.Crossref | GoogleScholarGoogle Scholar |

von Caemmerer S, Farquhar GD (1981) Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves. Planta 153, 376–387.
Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL38XjtFyjug%3D%3D&md5=93fd9711a50ec487e9478385092d1e9eCAS |

Wong SC, Cowan IR, Farquhar GD (1985) Leaf conductance in relation to rate of CO2 assimilation. I. Influence of nitrogen nutrition, phosphorous nutrition, photon flux density and ambient partial pressure of CO2 during ontogeny. Plant Physiology 78, 821–825.
Leaf conductance in relation to rate of CO2 assimilation. I. Influence of nitrogen nutrition, phosphorous nutrition, photon flux density and ambient partial pressure of CO2 during ontogeny.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cnhs1Gjsw%3D%3D&md5=68aacbf71ce4ea2daaa3aba8211bfacbCAS |

Zufferey V, Cochard H, Ameglio T, Spring J-L, Viret O (2011) Diurnal cycles of embolism formation and repair in petioles of grapevine (Vitis vinifera cv. Chasselas). Journal of Experimental Botany 62, 3885–3894.
Diurnal cycles of embolism formation and repair in petioles of grapevine (Vitis vinifera cv. Chasselas).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXptFGjs74%3D&md5=92ddb390e22b09ad8159327f44274342CAS |