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Scientia Horticulturae 110 (2006) 254259 www.elsevier.

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Sap ow in papaya plants: Laboratory calibrations and relationships with gas exchanges under eld conditions
Fabrcio de Oliveira Reis a,*, Eliemar Campostrini a, Elias Fernandes de Sousa b, Marcelo Gabetto e Silva b
Plant Physiology Sector, LMGV, Agricultural Science and Technology Center, State University of North Fluminense, Avenue Alberto Lamego 2000, 28013-602 Campos dos Goytacazes, RJ, Brazil b Laboratory of Agricultural Engineering, Agricultural Science and Technology Center, State University of North Fluminense, Avenue Alberto Lamego 2000, 28013-602 Campos dos Goytacazes, RJ, Brazil Received 14 April 2005; received in revised form 6 July 2006; accepted 10 July 2006
a

Abstract In papaya plants a study to quantify the water ow through the trunk is important for to promote a good water management in commercial orchard. The objective was to study the relationship between water ow through the trunk and temperature measurements determined by probes inserted in the papaya plant stem in laboratory. In addition, was possible to study the relationship between sap ow and instantaneous gas exchange in eld conditions. We constructed an instrument that maintained a stable water ux through a 0.30 m stem section with a constant pressure, simulating the xylem sap ow through the stem. Water ux was adjusted by varying pressure of water in the stem section. The mathematical model used to t the relation between K (Granier heat coefcient) values and sap ow density was the exponential model: u = 0.5511 K1.9104. Field studies was conducted in a commercial orchard located in North of the State of Rio de Janeiro, with 12 plants in October 2002, and eight plants in January 2003. We veried that instantaneous transpiration, measured by a portable system of gas exchange (porometry), presented a good (R2 = 0.75) positive relationship with xylem sap ow. Estimates of papaya sap ow can be obtained by scaling portable photosynthesis system measurements with exposed leaves, however the relationship is non-linear in higher instantaneous transpiration rates. The causes of the non-linear relationship in higher transpiration are discussed. In addition, was possible to obtain a good (R2 = 0.76) relationship between net photosynthesis rate and xylem sap ow in papaya eld-grown. # 2006 Elsevier B.V. All rights reserved.
Keywords: Papaya; Sap ow; Gas exchange

1. Introduction The amount of water required for growth and development of higher plants is larger than those of all other constituents such as mineral ions or carbon dioxide (Steudle, 2002). Stomata are the primary conduit of water vapor to leaf to the atmosphere, and play an important role in water loss control in leaves. Stomata control is coordinated by light intensity and quality, CO2 external concentration, leaf water condition, metabolites produced into foliar mesophyll and into roots, and air humidity (Farquhar and Sharkey, 1982). The leaf-to-air vapor pressure decit inuences stomatal closure, and high values of VPDleaf-to-air cause stomatal closure (Yong et al., 1997). Papaya, which presents stomata only in the leaf abaxial surface (El-Sharkawy et al., 1985), had a decrease of 47 and
* Corresponding author. E-mail address: fareoli@uenf.br (F. de Oliveira Reis). 0304-4238/$ see front matter # 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.scienta.2006.07.010

48% in the stomatal conductance and photosynthesis net rate, when VPDleaf-to-air changed from 1 to 1.5 kPa to 3.54.5 kPa (El-Sharkawy et al., 1985). According to the authors, in 19 species studied, including Carica papaya L., stomata closure caused by VPDleaf-to-air was associated with the peristomatal evaporation (water loss directly from guard cells to environment). In papaya, water limitations reduce stomatal conductance, net rate of photosynthesis and quantum yield, which signicantly affect the crop yield (Marler et al., 1994; Marler and Mickelbart, 1998). These authors demonstrated that although the PS II, was tolerant in relation to the water stress in irrigated plants (csolo = 20 kPa), the maximum photosynthetic rate (measured in 1800 mmol m2 s1 of photosynthetic photon ux densityPPFD) was reduced 40% in plants on a soil with 60 kPa water potential and the quantum yield (A/PPFD) was reduced 50%. Several techniques to estimate the transpiration rate have been developed to improve the irrigation control and elucidate

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plantwater relations. These techniques include xylem sap ow determination and transpiration rate measurements by porometry (Zhang et al., 1997). There are many techniques used to determine the xylem sap ow. All use heat as a tracer for sap movement, and each has advantages and limitations in terms of ease of use, cost and size of plant stem on which they can be used (Smith and Allen, 1996). The heat dissipation method was brought into use by Granier (1985) and it relates the xylem sap velocity to the temperature differences between heated and unheated probes inserted radially into the steam. Heat dissipation increases and the temperature difference between the two probes declines asymptotically with increasing sap velocity (Clearwater et al., 1999). Granier adjusted the equation to: u = 119 106k1.0 231, where u is the sap ow density and k is the Granier heat coefcient, calculated by the equation: k = (DTmax DT)/DT, where DT is temperature differences between the two probes. Heat dissipation probes have been used successfully in a number of studies of tree water use (Breda et al., 1985; Loustau et al., 1996; Andrade et al., 1998). However, Smith and Allen (1996) advised that the technique should be calibrated for each species. Another advantage to this technique is the possibility of automating the irrigation system based on the plant transpiration (Van Bavel et al., 1996). An adjustment of this technique is the use of heat pulse method and not the use of constant heat (Fernandez et al., 2001). In olive, the heat pulse method of sap ow measurement controlled an automated irrigation system, which formed this basis to calculate water movement in the root zone and predict plant water status (Nadezhdina, 1999; Fernandez et al., 2001). There are no reports of constant heat techniques in papaya. Therefore, it is necessary to evaluate the technology in this important tropical fruit crop. The present study evaluated the relationship between water ow through the trunk and the temperature measurements determined by probes inserted in the papaya plant stem in laboratory and eld studies and related the xylem sap ow

values measured under eld conditions with instantaneous gas exchange measurements. 2. Material and methods 2.1. Laboratory calibrations The calibration experiment was performed at the State University of North Fluminense, Agricultural Engineering Laboratory, Campos dos Goytacazes, RJ. A device was construct that maintained a stable water ow through a stem (Fernandez et al., 2001) (Fig. 1). Water ow, simulating the xylem sap ow through the papaya tree stem, was established with constant pressure in a papaya steam 30 cm in length and 5.5 cm in diameter. The water ow difference was measured by pressure changes in water injection into the stem. Two thermocouples probes 2 mm in diameter and 30 mm in length were inserted radially into the node of the papaya trunk section. The node was selected cause this part of the trunk has the highest percentage of vascular bundles and minimal void space. The probes were vertically separated by 50 mm. The higher probe had an electric resistance heater. Three volts was applied to the resistance heater every 10 s, and the temperature difference (DT) between the probes were recorded every 10 min. The trunk and probes were covered with foam insulation to minimize trunk temperature variation. DT was measured for 6 values of water ow and 5 replications of papaya stem sections. The xylem ow density values (u m3 s1 m2 active xylem) were obtained from the relationship between water ux and the active xylem area. The active xylem area was estimated using the red-dye safranin staining from several cut stems of different diameters. The colored area was measured and considered the xylem area in cm2. From these data, it was possible to make the relationship between K (Eq. (1)) and sap ow density (u). K DT m 1 DT (1)

Fig. 1. Schematic representation of the experimental system for analyses of the dynamic sap ow laboratory calibration in papaya trunk (Fernandez et al., 2001).

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where K is the Granier heat coefcient; DTm the maximum temperature difference between sensors in active xylem, 8C (zero ow); DT is the temperature difference between sensors in active xylem, 8C; the sap ow system of Dynamax Co., Houston, TX, USA was used in laboratory and a eld studies. 3. Field experiment 3.1. Plant material and eld conditions The study was carried out in a commercial Formosa papaya orchard, located in the city of Sao Francisco do Itabapoana/RJ (218270 S, 418150 W, 12 m of altitude). The study used 9 and 12-month papaya plants (Carica papaya L.) in October 2002 and January 2003, respectively. The orchard was center pivot irrigated. Tree spacing was 3.8 m between rows and 2.0 m between plants, totalizing 1316 plants ha1. Trees were fertilized. The maximum value of photosynthetic photon ux density in October was 1877 mmol m2 s1 and in January was 2393 mmol m2 s1. 3.2. Xylem sap ow investigation in papaya eld-grown The study was carried out in two periods: October 2002 and January 2003 using eight plants for each period. In each sample period, the xylem sap ow was measured for 4 days; two plants each day. The probes were inserted into the stem to a height of 0.30 m from soil surface similar to the laboratory study. Papaya leaf area was estimated by log AF = 0.315 + 1.85 log NCF, R2 = 0.898; where AF is the leaf area, and NCF is the central vein length (Campostrini and Yamanishi, 2001). The xylem sap ow was expressed by leaf area unit. A 9.83 cm2 area of active xylem was used to estimate the xylem sap ow. In the calibration moment, 9.83 cm2 was the active xylem area of the trunk used. When our equation is applied (0.5511 K1.9104), the ow is expressed in l h1 cm2 (per cm2 of active xylem area). Then, to express it in leaf area, multiply the value by the active xylem area of the measured trunk (the ow is expressed in l h1 plant1), and divide the product by the plant leaf area. Now, the sap ow is expressed in l h1 m2 (leaf area unit). An equation was made to relate the circumference of the trunk with the active xylem area (Y = 3.317X 47.887; R2 = 0.9356); where Y is active xylem area (cm2) and X is circumference of the trunk (cm) where the probes were inserted. Transpiration (E, mmol m2 s1) and net photosynthetic rate (A, mmol m2 s1) were measured at the same time and at the same plant the xylem sap ow was measured (every hour, between 8 a.m. and 5 p.m.), using a portable system of gas exchange; model LI-6200 (LI-COR, Lincoln, NE, USA). Two

Fig. 2. Relationships between Granier heat coefcient and xylem sap ow (u; L h1 cm2 active xylem) estimated by laboratory calibration equation and Granier equation.

illuminated mature leaves (9th and 10th leaf counted from the top) were used. The main water potential of soil (csoil), air temperature, relative humidity, air and vapor pressure decit (VPD), of the October 2002 and January 2003 sampling periods were measured (Table 1). 4. Results and discussion 4.1. Laboratory calibration Sap ow density (u; L h1 cm2 active xylem) was exponentially related to u: 0.5511 K1.9104 (Granier, 1985). The derived laboratory calibration model estimated larger u values than predicted by Granier (1985) for equal K values (Fig. 2). Differences in xylem vessel distribution and diameter, and the higher water content of papaya stems may explain the differences in the results between our results and Granier (1985). As the xylem vessels are more concentrated at the trunk periphery (Fig. 3), this distribution could have provided the passage of a volume of water through the trunk corresponding to the temperature variation of the probes. Although papaya is herbaceous, its stature is not that of typical herbaceous plant. Papaya plants may reach heights of 9 m, and are thus described as giant herbs (Malo and Campbell, 1986), and do not respond like a woody plant (Fig. 4). Therefore, the relationship between K and xylem sap ow need to be established for papaya.

Table 1 Mean air temperature, water potential of soil (csoil), relative humidity and VPD of the air in October 2002 and January 2003 Month October 2002 January 2003 Air temperature (8C) 29.4 2.7 29.9 2.7 Water potential of soil (csoil, kPa) 28.5 8.5 12.1 8.7 Relative humidity (%) 48.3 9.7 68.5 11.15 VPD of the air (kPa) 2.2 0.7 1.4 0.7

Daily medium values obtained from 8 a.m. to 5 p.m., n = 40.

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Fig. 3. Standardized xylem vessel distribution in papaya trunk. Picture obtained from a 6-month papaya plant. Notice that the vessel distribution concentrates near the cambial zone of the trunk. Fig. 5. Relationship between sap ow density (u) (L h1 m2 leaf area) and instantaneous transpiration (E) (mmol m2 s1) measured by LI-6200, in papaya plants cultivated under eld conditions. Sao Francisco do Itabapoana, RJ. October 2002 and January 2003.

4.2. Relationships between xylem sap ow, photosynthesis and instantaneous transpiration The maximum values of xylem sap ow where 0.55 L h1 m2 leaf area (Fig. 5). This value represented 15.6 L m2 d1. A 5-month-old papaya plant has a leaf area of approximately 5 m2 per plant, and therefore, transpires 78 L plant1 d1. In citrus, Steinberg et al. (1990) recorded values of 0.35 L h1 m2 leaf area or approximately 150 L of transpired water plant1 d1. In Prunus serrulata (Lindl., cv. Longipes), xylem sap ow values were 0.041 L h1 m2 (Dugas et al., 1993); young mangosteen (Garcinia mangostana L.), 0.043 L h1 m2 (Wiebel and Chako, 1992), mango (Mangifera indica L.), 0.84 L h1 m2 (Lu and Chacko, 1998) and grapevines 0.09 a 0.05 L h1 m2 (Schmid and Bettner, 1999). The instantaneous transpiration rate (E) was correlated to xylem sap ow (Fig. 5) with the highest values of xylem sap ow between 10 a.m. and 2 p.m.

Fig. 4. Relationship between ow (u; L h1 cm2 active xylem) values and Granier heat coefcient (K) from laboratory study.

The non-linear relationship between transpiration and xylem sap ow may be explained by a lag phase in which transpiration increases without the same change sap ow values. Zhang et al. (1997) observed a higher availability of stored water in Larix and Picea. In Pecan (Carya illinoensis Wichita), the small early morning lag may represent water immediately available in leaves, petioles and small branches (Steinberg et al., 1990). The stored water present in these structures could contribute to transpiration values without xylem water tension development. In addition, the decrease of the hydraulic conductivity (resistance to the movement of water through the xylem vessel and high resistance to water movement in the liquid phase between the epidermis and the mesophyll, El-Sharkawy (1990)) from probe to leave can explain this non-linearity. According to Ansley et al. (1994), the accuracy of scaling E values to u declined with increasing E values suggesting that E is greater than that measured by stem ow or gravimetric techniques during periods of high E. Green et al. (1989) found good correlation between heat pulse and Penman-Monteith adjusted porometer data in kiwifruit vines (Actinidia deliciosa), although at peak transpiration, porometer values exceeded heat pulse values. Gucci et al. (1990) found that porometer measurements of apricot (Prunus armeniaca) leaves overestimated transpiration when compared to gravimetric measurements. Schulze et al. (1985) found that canopy transpiration in Larix and Picea trees scaked from porometer leaf measurements and total needle biomass was very close to that determined by a stem ow technique, although porometer transpiration (of needles) started about 3 h earlier each day than stem ow. Scaling E values to u must recognize the connement of the leaf into a chamber with differences in microclimate (Mcdermitt, 1990). We think that porometers do provide an excellent complement to other measurements because conductance differences owing to the environment can be evaluated over time or between species. The large temporal and spatial variability may require a large number of measurements.

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rate and Rubisco carboxilase sites are limited. For optimum values of water, light and nutrient, from a particular stomatal conductance value, photosynthetic rate remains dependent on the greater CO2 uptake and from the Ribulose-1,5-biphosphate (RuBP) regeneration rate. This fact shows the non-linear response of the photosynthetic rate from sap ow values bigger than 0.28 L h1 m2. At this phase of stability, the net photosynthetic rate remains dependent on RuBP regeneration rate, the increasing of the number of actives sites to CO2 xation and the increasing of the CO2 diffusion to the foliar mesophyll. 5. Conclusion We have showed that mathematical model used to t the relation between K (Granier heat coefcient) values and sap ow density (u, L h1 cm2 active xylem) was the exponential model: u = 0.5511 K1.9104 and instantaneous transpiration (E, mmol m2 s1), measured by a portable system of gas exchange, presented a good positive relationship with xylem sap ow. Estimates of papaya sap ow can be obtained by scaling portable photosynthesis system measurements with exposed leaves, however the relationship is non-linear in higher instantaneous transpiration rates. In addition, it was possible to obtain a good relationship between photosynthesis and sap ow measurements in papaya eld-grown. This fact shows that handling strategies on the orchard, that provide high transpirations rates, can improve the carbon photosynthetic assimilation in papaya plants. Acknowledgements We acknowledge the support of Caliman Agrcola S/A (Brazilian papaya company), Financiadora de Estudos e ` Projetos (FINEP), Fundacao Carlos Chagas de Apoio a Pesquisa no Estado do Rio de Janeiro (FAPERJ) and Dr. Michael Glenn, USDA, ARSAppalachian Fruit Research Station, USA for critical reading of the article. References
Andrade, J.L., Meinzer, F.C., Goldstein, G., Holbrook, N.M., Cavelier, J., Jackson, P., Silvera, K., 1998. Regulation of water ux through trunks, branches and leaves in trees of a lowland tropical forest. Oecologia 115, 463471. Ansley, R.J., Dugas, W.A., Heuer, M.L., Trevino, B.A., 1994. Stem ow and porometer measurements of transpiration from honey mesquite (Prosopis glandulosa). J. Exp. Bot. 275 (45), 847856. Breda, N., Granier, A., Aussenac, G., 1985. Effect of thinning on soil and tree water relations, transpiration and growth in a oak forest (Quercus petraea (Matt.) Liebl.). Tree Physiol. 15, 295306. Campostrini, E., Yamanishi, O.K., 2001. Estimation of papaya leaf area using the central vein length. Scientia Agricola 58, 3942. Clearwater, M.J., Meinzer, F.C., Andrade, J.L., Goldstein, G., Holbrook, N.M., 1999. Potential errors in measurements of nonuniform sap ow using heat dissipation probes. Tree Physiol. 19, 681687. Day, W., Legg, B.J., French, B.K., Johnson, A.E., Lawlor, D.W., Jeffers, W., de, C., 1978. A drought experiment using mobile shelters: the effect of drought on barley yield, water use and nutrient uptake. J. Agric. Sci. 91, 599623. Dugas, W.A., Wallace, J.S., Allen, S.J., Roberts, J.M., 1993. Heat balance, porometer, and deuterium estimates of transpiration from potted trees. Agric. Forest Meteorol. 64, 4762.

Fig. 6. Relationship between net photosynthetic rate (A) (mmol m2 s1) and sap ow density (u) (L h1 m2 leaf area), in papaya plants cultivated under eld conditions. Sao Francisco do Itabapoana, RJ. October 2002 and January 2003.

However, we found a good correlation between E and u in papaya eld-grown. Fig. 6 shows relationships between net photosynthetic rate and xylem sap ow. The CO2 diffusion into the leaf moves in an opposite rote in relation to water molecule movement, in other words, it follows the same direction, but the course of molecule displacement is opposed. Based on Farquhar and Sharkey (1982), photosynthetic rate may be calculated by equation A = (Cext Ci)E/1.6(ei ea), where Cext is the CO2 concentration outside the leaf, Ci the CO2 concentration inside the foliar mesophyll, E the transpiration, ei the vapor pressure inside the leaf and ea is the vapor pressure outside the leaf. Observing Figs. 3 and 4, results justify that high transpiration rates allow high xylem sap ow values, and that high transpiration rates may allow greater CO2 uptake through diffusion in the foliar mesophyll. Day et al. (1978) and Innes and Blackwell (1981) showed a good relationship between yield and water use in barley and wheat. In addition, in grapevines, Schmid and Bettner (1999) showed that high rates of sap ow were always correlated with high rates photosynthesis. However, we have observed a transpiration at a zero water ow (5 mmol m2 s1, Fig. 5). We think that the sap ow is total water movement (ignoring any phloem transport) and that water could be lagging the loss of water. In the morning (sunrise), is possible a measurement transpiration (with LI-6200), the water could be coming from the tissue (including shoot) and thus, little water is moving from the soil part the sap ow probe (lag phase). In analogy with the mathematical equation proposed by Farquhar and Sharkey (1982), to ow values until 0.28 L h1 m2, it was observed linearity between two studied variables (Fig. 6). This part of the curve is associated with Rubisco carboxilase action, in other words, at this stage; there is a greater quantity of enzyme active sites available to CO2 xation (Farquhar and Sharkey, 1982). However, even though xylem sap ow values a high (when gS and E are increased) CO2 diffusion

F. de Oliveira Reis et al. / Scientia Horticulturae 110 (2006) 254259 El-Sharkawy, M.A., 1990. Effect of humidity and wind on leaf conductance of eld grown cassava. Revista Brasileira Fisiologia Vegetal 2 (2), 1722. El-Sharkawy, M.A., Cock, M.J.H., Hernandez, A.D.P., 1985. Stomatal response to air humidity and its relation to stomatal density in a wide range of warm climate species. Photosynth. Res. 7, 137149. Farquhar, G.D., Sharkey, T.D., 1982. Stomatal conductance and photosynthesis. Annu. Rev. Plant Physiol. 33, 317345. Fernandez, J.E., Palomo, M.J., Diaz-Espejo, A., Clothier, B.E., Green, S.R., Giron, I.F., Moreno, F., 2001. Heat-pulse measurements of sap ow in olives for automating irrigation: tests, root ow and diagnostics of water stress. Agricola Water Manage. 51, 99123. ` Granier, A., 1985. Une nouvelle methode pour la mesure du ux de seve brute ` dans le tronc des arbres. Annal. des Sci. Forestieres 42 (2), 193200. Green, S.R., Mcnaughton, K.G., Clothier, B.R., 1989. Observation of night-time water use in kiwifruit vines and apple trees. Agric. Forest Meteorol. 48, 251261. Gucci, R., Call, S.A.R., Xyloyannis, C., 1990. Comparation of gasometric and gravimetric methods of measuring leaf transpiration in potted apricot trees. Acta Horticul. 278, 343349. Innes, P., Blackwell, R.D., 1981. The effect of drought on the water use and yield of two spring wheat genotypes. J. Agric. Sci. 96, 603610. Loustau, D.P., Berbigier, P., Roumaginac, P., Arruda-Pacheco, C., David, J.S., Ferreira, M.I., Pereira, J.S., Tavares, R., 1996. Transpiration of a 64-year-old maritime pine stand in Portugal. I. Seasonal course of Water ux thought maritime pine. Oecologia 107, 3342. Lu, P., Chacko, E., 1998. Evaluation of Graniers sap ux sensor in young mango trees. Agronomie 18, 461471. Malo, S.E., Campbell, C.W., 1986. The papaya. Univ. of Florida Cooperative Extension Service Fruits Crops Fact Sheet FC-11. Gainesville, Fla. Marler, T.E., George, A.P., Nissen, R.J., Anderssen, P.C., 1994. Miscellaneous tropical fruits. In: Schaffer, B., Anderssen, P.C. (Eds.), Handbook of Environmental Physiology of Fruits Crops. VII. Sub-tropical and tropical crops. CRC Press, Florida, pp. 199224.

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Marler, T.E., Mickelbart, M.V., 1998. Drought, leaf gas exchange, and chlorophyll uorescence of eld grown papaya. J. Am. Soc. Horticul. Sci. 123 (4), 714718. Mcdermitt, D.K., 1990. Sources of error in the estimation of stomatal conductance and transpiration from porometer data. Hortscience 25, 1538 1548. Nadezhdina, N., 1999. Sap ow index as an indicator of plant water status. Tree Physiol. 19, 885891. Schulze, E.-D., Cermak, J., Matyssek, R., Penka, M., Zimmerman, R., Vascek, F., Gries, W., Kucera, J., 1985. Canopy transpiration and water uxes in the xylem of the trunk of Larix and Picae TreesA comparison of xylem ow, porometer and cuvette measurements. Oecologia 66, 475483. Schmid, J., Bettner, W., 1999. Experience with granier-type sensors for sap ow measurements in grapevines. Acta Horticul. 493, 177186. Smith, D.M., Allen, S.J., 1996. Measurement of sap ow in plant stems. J. Exp. Botany 305 (47), 18331844. Steinberg, S.L., Mcfarland, M.J., Worthington, J.W., 1990. Comparison of trunk and branch sap ow with canopy transpiration in pecan. J. Exp. Botany 227 (41), 653659. Steudle, E., 2002. Transport of water in plants. Environ. Control Biol. 40 (1), 2937. Van Bavel, C.H.M., Van Bavel, M.G., Lascano, R.J., 1996. Automatic irrigation based on monitoring plant transpiration. In: Proceedings of the International Conference. American Society of Agronomy Engineers, pp. 10881092. Wiebel, J., Chako, E., 1992. Sap Flow Weight Loss and Transpiration in Mangosteen. Csiro Division of Hort., Sap Flow Workshop, Loxton Research Center, Loxton, S.A., 7 p. Yong, W.H., Wong, S.C., Farquha, G.D., 1997. Stomatal responses to changes in vapour pressure difference between leaf and air, plant. Cell Environ. 20, 2131216. Zhang, H., Simmonds, L.P., Morison, J.I.L., Payne, D., 1997. Estimation of transpiration by single trees: comparison of sap ow measurements with a combination equation. Agric. Forest Meteorol. 87, 155169.