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1 Jour. Korean For. Soc. Vol. 99, No. 6, pp. 922~928 (2010) JOURNAL OF KOREAN FOREST SOCIETY ye ƒ ù xÿ Ÿw» e w wá Á½ Á½ w Effect of Calcium Chloride(CaCl 2 ) on Chlorophyll Fluorescence Image and Photosynthetic Apparatus in the Leaves of Prunus sargentii Joo Han Sung, Sun Mi Je, Sun-Hee Kim and Young-Kul Kim Department of Forest Conservation, Korea Forest Research Institute, Seoul , Korea : š ye (CaCl 2 ) ù (P. sargentii) ƒ e w w» w, ye ƒ 2z z xÿ Ÿ -Ÿw Ÿw», w. 3 ù ye 0.5%(9 mm), 1.0%(18 mm), 3.0%(54 mm) 2(1 L)z w. ye, ye ƒ w ye w a/b, Ÿw,, y w. Ÿ ye ƒ ƒ. Ÿw, y, Ÿ Ÿ ùkû (p<0.05). wr, xÿ(f M ) xÿ(f 0 ) Fv xÿ mw w ƒ y w ùkù, Ÿ y (Fv ) Ÿyw (NPQ) 80 w w. ye w ù Ÿw, y» š. Abstract: There is a little information on the effect of calcium cloride (CaCl 2 ) which is used as deicing salt in Korea on the physiological responses of the street trees. Prunus sargentii is one of the most widespread tree species of street vegetation in Korea. In this study, the effect of CaCl 2 on photosynthetic apparatus such as chlorophyll fluorescence image and light response curve of P. sargentii in relation to their leaf and root collar growth responses were investigated. To study the effect of CaCl 2 treatment in the early spring, we irrigated twice in rhizosphere of P. sargentii (3-year-old) planted plastic pots with solution of 0.5%, 1.0%, 3.0% CaCl 2 concentration before leaf expansion. Results after treatments, total chlorophyll contents and the chlorophyll a/b, photosynthetic rate, quantum yield, dark respiration decreased with increasing CaCl 2 concentration. On the contrary, light compensation point increased with increasing CaCl 2 concentration. Through the linear regressions of correlation of photosynthetic rate with photosynthetic parameters (quantum yield, dark respiration and light compensation point), we found a significant relationship (p<0.05) between photosynthetic rate and quantum yield and light compensation point except dark respiration. Calcium cloride (CaCl 2 ) induced inhibition of photochemical efficiency (F V ) and non-photochemical quenching (NPQ) were found in treatments of CaCl 2, and these reduction rates between control and CaCl 2 treatments were drastically showed at 80 days. We suggest that physiological activities are limited from treatment of CaCl 2. These reductions of photosynthetic apparatus ability caused eventually the reduction of leaf and diameter at root collar growth. Key words : Prunus sargentii, Calcium chloride, photosynthetic rate, dark respiration, chlorophyll contents, Fluorescence image *Corresponding author jhs033@forest.go.kr 922

2 ye ƒ ù xÿ Ÿw» e w 923 ƒ, w m» w y w» p w ƒ v w. p, s w w w. y yw NaCl, KCl, K 2 SO 2, Na 2 SO 4, MgCl (Jonsson and Magnusdottir, 2007; Goodrich et al., 2009). ù CaCl 2 ( ye ) wš ( m, 2003). š p e w ƒ w, m sl ww ù(greenway and Munns, 1980; Hagemann and Murata, 2003), w w vw s ù vw y w (Wang et al., 2008). w» w w CO 2 š w Ÿw v. 1 yw œ sü w ƒ kƒ k k» w Ÿ w. Ÿ xÿ xk, xÿ w Ÿ» Ÿw» p w w t w. xÿ w vw w w NaCl w. (F V ) ù Ÿyw (NPQ, non-photochemical quenching) xÿ w NaCl w ü w w ü j yƒ w (F V ) w ù Ÿyw (NPQ)ƒ ƒw š šw (Lu et al., 2002; Maricle et al., 2007). ù ù y w sw. üw w ù wš p ƒ, v y wš ƒ v œ ù ƒ w w ( ƒ l, 2009) ƒ xy 4,527 ù, wù, v k, pù ƒ 75% w, ù ƒ 30% w (, 2009). ƒ ù ye w szwš CO 2, š w Ÿw» y xÿ mw š wš y mw «ye ù y wš w. 1. œ ye (CaCl 2 )» y w w» w w ( p z» 57) ƒ e(4 m 4 m) w 3 ù šƒ w, ye 0.5%, 1.0%, 3.0% ƒ 3 w ew. ye CaCl 2 H 2 Oƒ 74% t z ƒ f. ye» œ y 3 29 w. x» l 6 17 ¾ 80 ƒ vw y l w. x» 3, 4, 5, 6 s³ ƒƒ 6.1, 11.4, 18.1, 23.2 o C, 59.7, 52.9, 62.1, 60.9% ùkû (», 2009). ye w œ ye w ƒ ww w z, ù» ml 1 wš z 2 w. ye z, k» w y e j w 300 ml w w, x» w w. 2. Ÿw» d 1) w ƒƒ ye w š l w e ƒ ƒ 4~5 y w w z, r 0.1 g 10 ml DMSO(Dimethyl Sulfoxide) 65 o C» 6 30 w z, ŸŸ (UV/Visible Diode Array, Walden Precision Apparatus Ltd., UK) w q 663 nm 645 nm d w Arnon(1949) (Chlorophyll a=12.7 A A 645, Chloyophyll b=22.9 A A 663, Total Chlorophyll = Chlorophyll a + Chlorophyll b) w. 3. ƒ y d Ÿw (Net photosynthesis rate) { Ÿ w d»(licor-6400, Li-cor Inc., USA) w

3 924 ªƒžª 99«6y (2010) d w w w e d w. Ÿ š red-blue LED w PPFD(photosynthetic photon flux density) 0, 30, 50, 100, 300, 700, 1000, 1500, 2000 µmol photons m 2 s Ÿ ƒ œ û œ w. chamber œ» 500 mol s, chamber 25 o C, CO µmol mol 60-70% w. Ÿw (φ) PPFD 100 µmol m 2 s w Ÿ Ÿw z w w, z y = a + bx x r Ÿ, Ÿ ƒ 0, z y r a y w (½q», 2001; Timm et al., 2002; Muraoka et al., 2003). z Ÿw / (Ashraf et al., 2002) PPFD 1000 µmol m 2 s Ÿ w. 4. xÿ d xÿ œ z, 690 nm ƒ¾ xÿ CCD(Charge-Coupled Device) e xÿ d w Portable HandyCam(FluorCam, Photon System Instruments Ltd., Brno, Czech Republic) w y w. t 7 cm d w w. 15 z FluorCam vp quenching analysis d w v mg lv w (10 s pulses intensity µmol m 2 s ) û Ÿ d F 0, z, actinic saturating light( 2000 µmol photons m 2 s ) d š xÿ F M d w. xÿ F 0 Ÿ II a ƒ z ƒ w» ù xÿ w, xÿ F M F 0 F V (=F M -F 0 ) š w. F V Ÿ II y ùkü t, sz ùkü. xÿ Ÿyw (nonphotochemical quenching, NPQ) (F M ')-1 w (Lawson et al., 2008). 5. sp ü m l 1 cm t w x» x» 80 d w w. (( x )-( x» )/( x )) 100 w. 6. m ü Ÿw» ( w, a bw, a/b, Ÿw,, y, Ÿ, F V, NPQ) w ye GLM(General Linear Model) w w š, Duncan multiple range test mw p<0.05 s³ w. w ye Ÿ w, y, Ÿ Pearson mw wš, ƒ z ww. m SAS 9.1 v w. š 1. w ye z 80 w ye ƒ w (Chl. T), a (Chl. a) b (Chl. b)w w. w ye 0.5% 1.0% ƒƒ w 62%, 85% w (Table 1). 3.0% ù. w ye a s b s w j ùkû» a/b w 0.5% 32%, 1.0% 55% w. w w y yƒ w ù kú, p w t (, 2007; Abreu and Munné-Bosch, 2008; Zhang et al., 2008). 2. Ÿw» ye Ÿ y Ÿ w y r Ÿ ƒ ƒw Ÿw ƒw w ƒ, 40, 0.5%, 1.0%, 3.0% Ÿ w ƒ û % š w š, w Ÿw ƒ x w û ùkû (Figure 1). Ÿ š mw Ÿw» w Table 1. Change of Chlorophyll contents and Chlorophyll a/ b ratio at 80 days after CaCl 2 concentration treatments in the leaves of P. sargentii. unit: mg g F.W Treatment Chl.a Chl.b Chl.T Chl.a : Chl.b Control 1.31±0.16a 0.58±0.08a 1.89±0.24a 2.26±0.04a 0.5% 0.43±0.15b 0.29±0.08b 0.72±0.21b 1.53±0.39b 1.0% 0.14±0.06c 0.14±0.01c 0.29±0.05c 1.02±0.49b 3.0%

4 ye ƒ ù xÿ Ÿw» e w 925 Figure 1. The light response curve of net photosynthetic rate in the leaves of P. sargentii at 40 days(a) and 80 days(b) after treatment of different CaCl 2 concentration. Table 2. Photosynthetic apparatuses in the leaves of P. sargentii at 40 and 80 days after treatment with different CaCl 2 concentration. Treatment Φ µmol ) D resp m 2 s ) P n m 2 s ) L comp (µmolm 2 s ) 40days 80days 40days 80days 40days 80days 40days 80days Control 1.68±0.17a 1.26±0.44a 0.89±0.16a 0.92±0.41a 6.28±0.96a 4.54±1.93a 1.46±0.10bc 1.38±0.28b 0.5% 0.74±0.17b 0.83±0.38ab 0.84±0.19a 1.21±0.26a 2.32±0.67b 1.88±0.72b 1.36±0.33c 2.13±0.14a 1.0% 0.54±0.15bc 0.35±0.07b 0.86±0.06a 0.20±0.06b 1.53±0.56b 1.40±0.47b 1.86±0.37b 2.31±0.13a 3.0% 0.28±0.13c ±0.47a ±0.18c ±0.10a - *Φ: Quantum yield, D resp : Dark respiration, Pn : photosynthetic rate at PPFD 1000 µmolm 2 s, L comp : Light compensation point (Table 2), Ÿ w yk ùkü (Φ, quantum yeild) 40 w 0.5% 56% ùkþ, 1.0% w 68%, 75% ƒw. Ÿw w» w w ye w ùkü ye w syÿ w 1000 µmol m 2 s Ÿw. y (Dresp, dark respiration) j ùkù %. y s v» w v w k, y š ATP œ w w», y k 30~80% y w CO 2 ƒ (Amthor, 2000). p y y w (Poorter et al., 1990). p w y y ù kú (Galmés et al., 2007; Gratani et al., 2008), y Ÿw w (Cannell and Thornley, 2000), % y w û Ÿw Ÿw y k ƒ. Ÿ CO 2 w Ÿ, Ÿ û j û Ÿ l CO 2 w w. Ÿ % ƒ, p 80 w 0.5% 54% ƒ ùkþ. w ù ƒ û ye j q. 3. xÿ xÿ xÿ d w, vwƒ w w q. 15 z xÿ (F ) M xÿ (F 0 ) w Figure 2 xÿ mw ù ye û xÿ t.

926 ªƒžª 99«6y (2010) 40 80 Ÿ II y (F V ) yƒ 80 0.5%. Figure 2. The represent fluorescence image in the leaves of P. sargentii at 40 and 80 days after CaCl 2 treatment.

wr Ÿyw w NPQ(non-photochemical quenching) ƒ ü, p w, (NaCl) w NPQ ƒ NaCl w ùkù, ü NPQ ƒ ƒw (Masojidek et al., 2000; James et al., 2002). ù 40 j yƒ ù 80 ƒ ƒw NPQ ƒ 0.5% 39%, 1.0% 68% w w (Figure 3).

5 926 ªƒžª 99«6y (2010) Ÿ II y (F V ) yƒ %. Figure 2. The represent fluorescence image in the leaves of P. sargentii at 40 and 80 days after CaCl 2 treatment. Above fluorescence image's fluorescence values are F M -F % 30% Ÿ II y (F V ) w (Figure 3). F V Ÿ II y ùkü, z p w t. NaCl w ü w š F V w (Glynn et al., 2003), F V w š w (Maricle et al., 2007). wr Ÿyw w NPQ(non-photochemical quenching) ƒ ü, p w, (NaCl) w NPQ ƒ NaCl w ùkù, ü NPQ ƒ ƒw (Masojidek et al., 2000; James et al., 2002). ù 40 j yƒ ù 80 ƒ ƒw NPQ ƒ 0.5% 39%, 1.0% 68% w w (Figure 3). w ù ƒ û ye l vw. 4. ye x 80 d z w, w Ÿw xÿ š, Ÿw» n w š Figure 3. The maximum F V (F M -F 0 ) and amount of NPQ (none-photochemical quenching) in the leaves of P. sargentii 80 days after CaCl 2 treatment was measured following 15 min of dark adaptation. Three replicates are given for each species/treatment combination. Bars indicate standard deviation (SD). Figure 4. Growth rate of diameter at root collar in the P. sargentii at the last day of experiment compare of initial value. Bars indicate standard deviation (SD). Table 3. Correlations of photosynthetic rate with photosynthetic parameters in the leaves of P. sargentii at 40 and 80 days after treatment with different CaCl 2 concentration. Division Φ µmol ) D resp m 2 s ) L comp (µmolm 2 s ) equation r 2 P-value equation r 2 P-value equation r 2 P-value 40days y=0.2351x < y=0.0018x y=-0.222x days y=0.2356x y=0.0554x y= x Φ: Quantum yield, D resp : Dark respiration, L comp : Light compensation point

6 ye ƒ ù xÿ Ÿw» e w 927 ye w. w 0.5%, 1.0%, 3.0% ƒƒ 45%, 49%, 89%ƒ w ùkû (Figure 4). ye ƒ szwš CO 2 š w Ÿw» vw w w e q. ye w w sl w ³x y w p w y ƒ vwƒ ùkú. ù ye w Ÿw xÿ p ye 40 (Φ) Ÿw ƒ ù kû ù, F V NPQ yƒ % š w, 0.5% l w a/b ƒ ùkû. w, ye (Φ) syÿ Ÿw š Ÿ ƒ mw ù ƒ s zwš CO 2 š w w. xÿ mw F V NPQ 80 ƒ 0.5% l ƒƒ 30% ùkþ. w szwš w CO 2 š wš w Ÿw ƒ ye w vw. ye ƒ ù y w ye ƒ v w. x 1. ƒ l nature.go.kr 2.» ». pp m w. pp ƒ Á. pp ½q»,,,, w, Ÿy ü Ÿw e w. w wz 90(4): ,, ƒ»œ y, w w y z y. w wz 96(4): Abreu, M.E. and Munné-Bosch, S Salicylic acid may be involved in the regulation of drought-induced leaf senescence in perennials: A case study in field-grown Salvia officinalis L. plants. Environmental and Experimental Botany 64(2): Amthor, J.S The McCree-de Wit-Penning de Vries- Thornley Respiration Paradigms: 30 Years Later. Annals of Botany 82: Arnon, D.I Copper enzymes in isolated chloroplasts, polyphenol-oxidase in Betula vulgaris. Plant Physiology 24: Ashraf, M., Arfan, M., Shahbaz, M., Ahmad, M. and Jamil, A Gas exchange characteristics and water relation in some elite skra cultivars under water deficit. Photosynthetica 40(4): Cannell, M.G.R., Thornley, J.H.M Modelling the components of plant respiration: Some guiding principles. Annals of Botany 85: Galmés, J., Ribas-Carbó, M., Medrano, H. and Flexas, J Response of leaf respiration to water stress in Mediterranean species with different growth forms. Journal of Arid Environments 68: Glynn, C.P., Gillia, A.F. and Oxenham, G Foliar salt tolerance of Acer genotypes using chlorophyll fluorescence. Journal of Arboriculture 29(2): Gratani, L., Varone, L. and Catoni, R Relationship between net photosynthesis and leaf respiration in Mediterranean evergreen species. Photosynthetica 46(4): Goodrich, B.A., Koski, R.D. and Jacobi, W.R Condition of soils and vegetation along roads treated with magnesium chloride for dust suppression. Water, Air, Soil Pollution 198: Greenway, H. and Munns, R Mechanisms of salt tolerance in non-halophytes. Annual Review Plant Physiology 31: Hagemann, M. and Murata., N Glucosylglycerol, a compatible solute, sustains cell division under salt stress. Plant Physiology 131: James, A.R Factors affecting CO 2 assimilation, leaf injury and growth in salt-stressed durum wheat. Functional plant biology 29(12): Jonsson, T.H. and Magnusdottir, M.L Salt-related suppression of bud break in Populus trichocarpa: Cost of inclusion, ion-specific or osmotic effects? Icelandic Agricultural Sciences 20: Lawson, T., Lefebvre, S., Baker, N.R., Morison, J.I.L. and Baines, C.A Reductions in mesophyll and guard cell photosynthesis impact on the control of stomatal responses to light and CO 2. Journal of Experimental Botany 59(13): Lu, C.M., Qiu, N.W., Lu, Q.T., Wang, B.S. and Kuang, T.Y Dose salt stress lead to increased susceptibility of photosystem to photoinhibition and changes in photosynthetic pigment composition in halophyte Suaeda salsa grown outdoors? Plant Science 163:

7 928 ªƒžª 99«6y (2010) 21. Maricle, B.R., Lee, R.W., Hellquist, C.E. Kirasts, O. and Edwards, G.E Effects of salinity on chlorophyll fluorescence and CO 2 fixation in C4 estuarine grasses. Photosynthetica 45(3): Masojidek, J., Torzillo, G., Kopecký, J., Koblžek, M., Nidiaci, L., Komenda, J., Lukavska, A. and Sacchi, A Change in chlorophyll fluorescence quenching and pigment composition in the green Chlorococcum sp. Grown under nitrogen deficiency and salinity stress. Journal of Applied Phycology 12: Muraoka, H., Koizumi, H. and Pearcy, R.W Leaf display and photosynthesis of tree seedlings in a cool temperate deciduous broad leaf forest understory. Oecologia 135: Poorter, H., Remkes, C. and Lambers, H Carbon and nitrogen economy of 24 wild species differing in relative growth rate. Plant physiology 94: Timm, H.C., Stegemann, J. and Küppers, M Photosynthetic induction strongly affects the light conpensation point of net photosynthesis and coincidentally the apparent quantum yield. Trees 16: Wang, R., Chen, S., Zhou, X., Shen, X., Deng, L., Zhu, H., Shao, J., Shi, Y., Dai, S., Fritz, E., Hüttermann, A., and Polle, A Ionic homeostasis and reactive oxygen species control in leaves and xylem sap of two poplars subjected to NaCl stress. Tree physiology 28: Zhang, X., Wollenweber, B., Jiang, D., Liu, F. and Zhao, J Water deficits and heat shock effects on photosynthesis of a transgenic Arabidopsis thaliana constitutively expressing ABP9, a bzip transcription factor. Journal of Experimental Botany 59(4): ( ; k)

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