Fragaria ananassa. Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, Tsukide, Kumamoto,

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J.SHITA 18 2 115-122. 2006. 115 1 2 1 1 3 1 862-8502 3 1 100 2 884-0003 11609 3 Fragaria ananassa Naotaka MATSUZOE 1, Syuji KAWANOBU 2, Sachiko MATSUMOTO 1, Hirokazu KIMURA 1 and KAZUFUMI ZUSHI 3 1 Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Kumamoto, 862-8502 2 Faculty of Horticulture, Minami Kyushu University, 11609 Takanabe,Miyazaki, 884-0003 3 JSPS Research Fellow, Prefectural University of Kumamoto 2005 8 17 2005 12 16 Coressponding author Naotaka Matsuzoe (matsuzoe@pu kumamoto.ac.jp) 19

116 ( Duch) (Morishita and Honda, 1991; Moing., 2001) (Sone., 1999; Wang and Zheng, 2001; Williner., 2003) (Lee and Kader, 2000) (Stintzing and Carle, 2004) (Amakura., 2000; H kkinen., 1998) (Morishita and Honda, 1992; Ogiwara., 1998) (Sone., 1999; Williner., 2003; Yoshida and Tamura, 2005) (Moing., 2001) (Noguchi, 2002) (Sone., 1999) (Wang and Zheng, 2001) ( ) (Williner., 2003) (Kawanobu., 2000) 1 2004 9 15 cm 1 10 1 (FLI 301N EYELA ( )) 12 200 mol s -1 m -2 (8:00 20:00) 23 (20:00 8:00) 10 15 20 3 9 B ( ( )) 1/4 2 3 2 2-45 2 0.1 g 80 15 m 80 10 (12,074 g 15 min) 50 m 3 0.20 m HPLC Shim pack SCR 101C ( 300 mm 7.9 mm ) SCR(C) 80 0.8 m min -1 (RID 10A ) Shim pack SCR 102H( 300 mm 7.9 mm ) SCR(H) 5 mm 40 0.8 m min -1 0.8 m min -1 20 mm Bis Tris 100 M EDTA 2Na 5 mm (CCD 6A ) (OPA) 20

117 OPA Shim pack FC ODS ( 75 mm 4.6 mm ) GVP ODS ( ) 50 1.0 ml min -1 A 10 mm ( ) (ph 6.9) B A/B = 95/5 2 ; A/B = 92.5/7.5 4 ; A/B = 90/10 8 ; A/B = 88/12 9 ; A/B = 79/21 15 ; A/B = 77/23 17.5 ; A/B = 50/50 18.5 ; A/B = 20/80 (RF 10A ) 350 nm 455 nm 3 3 g 2% 100 m (12,074 g, 4 10 min) 0.20 m HPLC Shim pack SCR 201H ( 300 mm 7.9 mm ) 2 mm 100 mm 100 mm 40 1.0 m min -1 0.5 m min -1 UV (SPD 10AD ) 300 nm 4 0.1 g 50% 20 m 24 (25 ) (12,074 g 13 min) (510 nm) (UVmini 1240 ) 1g (OD510) 0.20 m HPLC Shim pack VP ODS ( 250 mm 4.6 mm ) 35 0.8 m min -1 510 nm ( ) A 1.5% (v/v) B : : : = 1.5 20 25 53.5 (v/v) B 40.5 min 24 85% % 5 0.3 g 30 m 80 1 3 m 0.2 m HPLC Shim pack VP ODS ( 250 4.6 mm ) Paganga. (1999) 1 ( ) Table 1 23/10 23/20 2 Fig 1 ( ) 23/20 (Fig 1A) ( ) (Fig 1B) 23/20 (Fig 1C) 3 Table 2 23/10 23/20 Table 1 Effect of night temperature on days post anthesis until harvest and fruit weight of two strawberrry cultivars. Temperature Days post (day/night, ) anthesis Fruit weight (g) Nyoho 23/10 35 13.0 23/15 30 11.5 23/20 28 9.1 Toyonoka 23/10 39 10.0 23/15 35 9.2 23/20 30 7.5 Significance Cultivar (C) ** z ** Temperature (T) ** ** C T ** ns z ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. 21

118 Fig 1 Effect of night temperature on sugar (A), organic acid (B) and sugar acid ratio (C) of two strawberry cultivars. Vertical bars represent SE (n=6). ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. 4 Fig 2 (ASA) ( DHA) ASA (Fig 2A) DHA (Fig 2B) ASA DHA 5 Fig 3 Table 3 Yamaguchi (., 1999) 3 (Cy3G) 3 (Pg3G) 3 (Pg3MG) Pg3MG Pg3G 6 Fig 4 23/10 23/20 (Table 1) Table 2 Effect of night temperature on amino acid contents (mg g -1 FW) of two strawberrry cultivars. Cultivar Temperature Asparagine (day/night, C) Aspartate Glutamine Glutamate Alanine aminobutyrate Other Total Nyoho 23/10 1.09 0.077 0.504 0.076 0.095 0.025 0.126 1.99 23/15 1.05 0.087 0.393 0.075 0.058 0.026 0.090 1.78 23/20 1.08 0.079 0.499 0.065 0.074 0.033 0.106 1.94 Toyonoka 23/10 0.84 0.109 0.134 0.094 0.067 0.066 0.113 1.43 23/15 0.44 0.090 0.123 0.090 0.030 0.046 0.072 0.89 23/20 0.53 0.097 0.139 0.069 0.036 0.052 0.081 1.00 Significance Cultivar (C) ** z ** ** ** ** ** ** ** Temperature (T) ** ns ns ** ** ns ** ns C T ns ns ns ns ns ** ns ns z ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. 22

119 Fig 2 Effect of night temperature on ascorbic acid (A) and dehydroascorbic acid (B) contents of two strawberry cultivars. Vertical bars represent SE (n = 6). ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. Fig 3 Effect of night temperature on anthocyanin content of two strawberry cultivars. Vertical bars represent SE (n=6). ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. (Miura., 1994) Fig 4 Effect of night temperature on ellagic acid content of two strawberry cultivars. Vertical bars represent SE (n = 6). ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. (Kumakura and Shishido, 1994) 23/20 ( / ) (Fig 1A) ( ) (Ogiwara., 1999) ( ) (Fig 1B) Morishita and Honda (1991) 23/20 ( / ) (Fig 1C) 23

120 Table 3 Effect of night temperature on variation (%) in anthocyanin compositions of two strawberrry cultivars. Temperature (day/night, ) Cyanidin Pelargonidin Pelargonidin 3 glucoside 3 glucoside 3 malonylglucoside Unknown Nyoho 23/10 4.0 64.4 27.1 4.5 23/15 3.1 64.5 28.1 4.3 23/20 1.9 61.9 32.5 3.7 Toyonoka 23/10 2.9 89.7 n.d. z 7.4 23/15 2.7 89.2 n.d. 8.1 23/20 2.2 89.5 n.d. 8.3 Significance Cultivar (C) ns y ** ** Temperature (T) ns ns ns ns C T ns ns ns z Not detected y ns and ** indicate non significant and significant at < 0.05 by ANOVA, respectively. (Fuke and Konosu, 1991; Fuke and Ueda, 1996) (Kajimoto., 2004) (Table 2) Moing. (2001) (Lea and Ireland, 1999) (Conklin, 2001) (Smirnoff and Pallanca, 1996; Conklin, 2001; Toledo., 2003) ASA (Smirnoff and Pallanca, 1996; Smirnoff, 2000; Conklin, 2001) Sone., (1999) ASA Sone., (1999) ASA ASA (Fig 3) Wang and Zheng (2001) (Table 3) (Fig 4) ( ) (Williner., 2003) 24

121 ( : ) 23/10 ( / ) 23/20 23/20 23/20 23/10 23/20 23/10 23/20 Amakura, Y., Okada, M., Tsuji, S., Tonogai, Y. 2000. Determination of ellagic acid in fresh and processed fruits by HPLC. (in Japanese with English abstract) Shokuhin Eiseigaku Zasshi 41: 206-211. Conklin, P. L. 2001. Recent advances in the role and biosynthesis of ascorbic acid in plants. Plant Cell Environ. 24: 383-394. Fuke, S., Konosu, S. 1991. Taste active components in some foods: A review of Japanese research. Physiol. Behav. 49: 863-868. Fuke, S., Ueda, Y. 1996. Interactions between umami and other flavor characteristics. Trends Food Sci. Technol. 7: 407-411. Kajimoto, O., Hirata, H., Nakagawa, S., Kajimoto, Y., Hayakawa, K., Kimura, M. 2004. Hypotensive effect of fermented milk containing aminobutyric acid (GABA) in subjects with high normal blood pressure. (in Japanese with English abstract) Nippon Shokuhin Kagaku Kogaku Kaishi 51: 79-86 Kawanobu, S., Zuhi, K., Matsuzoe, N., Yamaguchi, M. 2000. Effects of dark treatment and temperature on the fruit color of strawberries J. Japan. Soc. Hort. Sci. 69 (supply.2): 192. Kumakura, H., Shishido, Y. 1994. The effect of daytime, nighttime, and mean diurnal temperatures on the growth of Morioka 16 strawberry fruit and plants. (in Japanese with English abstract) J. Japan. Soc. Hort. Sci. 62: 827-832. H kkinen, S. H., K renlampli, S. O., Heinonen, I. M., Mykk nen, H. M., Torronen, A. R. 1998. HPLC method for screening of flavonoids and phenolic acids in berries. J. Sci. Food Agric. 77; 543-551. Lea, P. J., Ireland R. J. 1999. Nitgogen metabolism in higher plants. In: Plant amino acids, ed. by Singh, B. K., Marcel Dekker, Inc., New York, 1-47. Lee, S. K., Kader, A. A. 2000. Preharvest and postharvest factors influencing vitamin C content of horticultural crops. Postharvest Biol. Technol. 20: 207-220. Miura, H., Yoshida, M., Yamashita, A. 1994. Effect of temperature on the size of strawberry fruit. J. Japan. Soc. Hort. Sci. 62: 796-774. Moing, A., Renaud, C., Gaudill re, M., Raymond, P., Roudeillac P., Denoyes-Rothan, B. 2001. Biochemical changes during fruit development of four strawberry cultivars. J. Amer. Soc. Hort. Sci. 126: 394-403. Morishita, M., Honda, F. 1991. Variations of soluble solids content and acidity of strawberry fruits among cultivars, cropping types and harvest dates. (in Japanese with English abstract) Bull. Nat. Res. Ins. Vegetable, Ornamental Plants and Tea, Japan A4: 41-55. Noguchi, Y. 2002. Studies on the stabilization of strawberry forcing culture and breeding of adaptable cultivars in the warmer region of Japan. (in Japanese with English abstract) Bull. Nat. Ins. Vegetable Tea Sci. 1: 37-95. Ogiwara, I., Miyamoto, R., Habutsu, S., Suzuki, M., Hakoda, N., Shimura, I. 1998. Variation in sugar content in fruit of four strawberry cultivars grown in the field and under forced culture, harvest years, and maturation stages. (in Japanese with English abstract) J. Japan. Soc. Hort. Sci. 67: 400-405. Ogiwara, I., Shiraishi, M., Hakoda, N., Shimura, I. 1999. Causes of low sugar levels in forced strawberry fruits. (in Japanese with English abstract) J. Japan. Soc. Hort. Sci. 68: 130-137. Paganga, G., Miller, N., Rice Evans, C. A. 1999. The polyphenolic content of fruit and vegetables and their antioxidant activities. What does a serving constitute? Free Rad. Res. 30: 153-162. Smirnoff, N. 2000. Ascorbate biosynthesis and function in photoprotection. Phil. Trans. R. Soc. Biol. Sci. 355: 1455-1464. Smirnoff, N., Pallanca, J. E. 1996. Ascorbate metabolism in relation to oxidative stress. Biochem. Soc. Trans. 24: 472-478. Sone, K., Mochizuki, T., Noguchi, Y. 1999. Variations in ascorbic acid content among strawberry cultivars and their harvest times. (in Japanese with English abstract) J. Japan. Soc. Hort. Sci. 68: 1007-1014. 25

122 Stintzing, F. C., Carle, R. 2004. Functional properties of anthocyanins and betalains in plants, food, and human nutrition. Trends Food Sci. Technol. 15: 19-38. Toledo, M.E.A., Ueda, Y., Imahori, Y., Ayaki, M. 2003. L ascorbic acid metabolism in spinach ( L.) during postharvest storage in light and dark. Postharvest Biol. Technol. 28: 47-57. Wang S. Y., Zheng, W. 2001. Effect of plant growth temperature on antioxidant capacity in strawberry.j. Agric. Food Chem. 49: 4977-4982 Williner, M. R., Pirovani, M. E., G emes, D. R. 2003. Ellagic acid content in strawberries of different cultivars and ripening stages. J. Sci. Food Agric. 83: 842-845. Yamaguchi, M., Matsuzoe, N., Kawanobu, S., Sone, K., Mochizuki, T. 1999. Identification and distribution of anthocyanins in strawberry Cultivars. J. Japan. Soc. Hort. Sci. 68 (suppl. 2): 134 Yoshida, Y., Tamura, H. 2005. Variation in concentration and composition of anthocyanins among strawberry cultivars. J. Japan. Soc. Hort. Sci. 74: 36-41. 26