105 1. Introduction In recent years, malignant neoplasm (cancer), cere- brovascular disease and heart disease have become major causes of death among Japanese people, with approxi- mately 75% of deaths in Japan caused by these diseases (Japanese Ministry of Health, Labour and Welfare, 2009 a). Such diseases are generally called lifestyle diseases be- cause they are thought to be strongly related to a lack of exercise and high intake of fat and salt. Therefore, empha- sis has shifted from early diagnosis (secondary prevention) to lifestyle improvement (primary prevention) (Japanese Ministry of Health, Labour and Welfare, 2009 b). In this context, research on foods that have pharmacological ef- fects or physiological functions, such as disease preven- tion and health maintenance, has become more important (Namba, 1999; Tokui et al., 2003). Regarding the phar- macological effects of food, there is a form of medicinal cooking called “Yakuzen” which is based on the philoso- phy of Oriental medicine and is intended to maintain good health and improve physical condition. The preparation of Yakuzen dishes draws from the theory of “Yaku-shoku Dou-gen”, which means that the same principle underlies the daily diet and medical treatment, and on the yin-yang theory, the five-phase theory in Oriental medicine. As a result, Yakuzen has attracted considerable attention for the prevention of lifestyle diseases. To prepare Yakuzen dishes, in-season materials with ap- propriate properties and tastes are selected according to the health status and constitution of each person (Namba, 1995; Lan et al., 2002; Tokui et al., 2003). These properties and tastes are called “Sei-Mi” in Yakuzen theory. Sei-Mi con- sists of four properties (making the human body hot, warm, cool, or cold) and five tastes (salty, bitter, sweet, pungent, and sour), and each is considered to have its own function in the human body (Namba, 1999; Tokui et al., 2003). If the concept of Sei-Mi can be applied to vegetables, a cultivar that has a strong flavor and a high content of functional con- stituents related to the properties and taste is considered to have strong Sei-Mi and is suitable for Yakuzen dishes. However, the inherent flavor of vegetable cultivars has been weakened by breeding because priority has been given to ease of consumption for consumers or ease of production for growers. In the case of eggplant (Solanum melongena L.), popular cultivars in Japan today are F 1 (first filial gen- eration) cultivars derived from a parental line with oval fruit, a deep purple pericarp, and high yield; these cultivars have improved fruit quality with less unpleasant or harsh taste. Evaluation of local eggplant cultivars in terms of the suitability as materials for “Yakuzen” dishes S. Tsukagoshi* (1), K. Kuroda**, M. Hohjo*, F. Ikegami*, N. Kunisaki***, T. Hanamura***, K. Yamada***, T. Hagiwara*** * Center for Environment, Health and Field Sciences, Chiba University, 6-2-1 Kashiwanoha, Kashi- wa, Chiba 277-0882, Japan. ** Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo, Chiba 271-8510, Japan. *** Research and Development Department, Nichirei Foods Inc., 9 Shinminato, Mihama-ku, Chiba 261- 0002, Japan. Key words: functionality, local cultivars, property and taste, Solanum melongena. Abstract: “Yakuzen” is a form of medicinal cooking based on the theories of Oriental medicine. To prepare Yakuzen dishes, in-season materials with appropriate properties and tastes, “Sei-Mi,” are selected according to the health status and consti- tution of each person. In this study, the suitability of eggplant (Solanum melongena L.) cultivars for Yakuzen was evaluated by sensory tests and by analysis of the functional constituents considered to be closely related to the taste and functionality of Yakuzen dishes. Twenty-two eggplant cultivars including 21 Japanese and 1 Italian cultivar, and a Thai species (Solanum xanthocarpum Schrad & Wendl.) were evaluated. Principal component analysis (PCA) was used for comprehensive evalu- ation among the cultivars. From the PCA, many of the cultivars with round or oval fruit were characterized as juicy and sweet and considered easy to eat; most of the long-fruit cultivars were characterized as having higher specific amino acid contents. The small and round fruit cultivar Dewako and the Thai species (Makhuea pro) were considered to contain many functional ingredients, such as ash, polyphenols, and specific amino acids, and to have higher suitability for Yakuzen dishes. Adv. Hort. Sci., 2014 28(2): 105-110 (1) Corresponding author: tsukag@faculty.chiba-u.jp Received for publication 31 March 2014 Accepted for publication 07 July 2014 106 However, eggplant was introduced into Japan more than 1,260 years ago, and it is mentioned in Nara-period docu- ments (The Shõsõin documents) edited in 750 A.D. (Yoshi- da, 2010). Therefore, many cultivars have been developed over the long history of cultivation, and even today there are many cultivars with local origins. The chemical composition, flavor, and texture of the edible parts of plants vary among cultivars. Local cultivars grown for many years may have retained more flavor, and some of them may be more suit- able for Yakuzen dishes than the currently popular cultivars. Regarding the functionality of foods, many studies have focused on certain ingredients and have discussed the relationship between the amounts of those ingredients and the functionality of the food. However, it is important to consider the functionality of foods comprehensively in Yakuzen theory. In previous studies, we used principal component analysis (PCA) to comprehensively evaluate the functionality of local cultivars, and we demonstrated that two local cultivars of Japanese radish (Raphanus sati- vus L.), two of carrot (Daucus carota L.), and one of bitter melon (Momordica charantia L.) were more suitable for Yakuzen dishes than the widely used F 1 cultivar (Saito et al., 2010; Tsukagoshi, et al., 2011 a, b). In this study, two F 1 cultivars and 21 local cultivars of eggplant were evaluated for their suitability as materials for Yakuzen dishes according to their taste and content of functional constituents. 2. Materials and Methods Plant materials and growing condition Twenty-one Japanese eggplant cultivars with different fruit shapes were selected for this study (Table 1). Most of them were purebred cultivars, but two F 1 cultivars were included. In addition, an Italian cultivar and a Thai species (Solanum xanthocarpum Schrad & Wendl.) were used. All cultivars were grown at the Center for Environment, Health and Field Sciences, Chiba University. Seeds of all cultivars were sown in 9-cm plastic pots filled with up- land soil on 17 April 2012, and the seedlings were raised in a glasshouse. On 5 June seedlings were transplanted to the open field at a spacing of 50 cm between plants and 100 cm between rows. Fruits were harvested when they reached the regular size for each cultivar. Harvest began on 31 July and ended on 14 September. All other management was carried out according to the conventional methods in Japan (Chino, 2001). Sensory test Fruits were harvested on 7 and 9 August and the sen- sory test was conducted the following day. The fruits were washed, cut to a size of 1.5×1.5×3.0 cm, and steamed at 100°C for 5 min. The characteristics of steamed samples were evaluated by six panelists. The panelists passed rec- ognition tests for five tastes (sweetness, umami, saltiness, Table 1 - Eggplant cultivars used for the experiment Fruit shape (country) Cultivar Abbreviation Fruit length, weight and color Remark Small, Round (Japan) Dewako De 3-8 cm, 10-15 g, deep purple Minden Min 3-8 cm, 10-15 g, deep purple Round (Japan) Aizu maru AM 8-10 cm, 200-300g, deep purple Kamo nasu KN 12-15 cm, 200-350 g, deep purple Tonosama To approx. 15 cm, 300-450 g, deep purple Yamatoyo maru YM 10-12 cm, 250-350g, deep purple Money pouch (Japan) Nagaoka kinchaku NK 8-10 cm, 300-350g, deep purple (swelling toward the bottom) Saitama ao daimaru SA approx. 15 cm, 300-450 g, green Oval (Japan) Heta murasaki HM approx. 5 cm, 30 g, deep purple Senryo 2 gou Se2 10-12 cm, 80-90 g, deep purple F 1 , Control cultivar in this experiment Se2 (Control) SK 10-11 cm, 150-180 g, deep purple Wase shinkuro WS 10-12cm, 80-120 g, deep purple Yamashina Ya 10-12cm, 80-120 g, deep purple Long (Japan) Chikuyou Chi 20-25 cm, 120-150 g, deep purple F 1 Hakata naga Hak 40-45 cm, 200-300 g, deep purple Hhogo naga Hho 17-18 cm, 85-90 g, deep purple Himo nasu HN 25-30 cm, 150-200 g, pale purple Kitta chunaga KC 10-12cm, 80-120 g, deep purple Kurume oh naga KO 30-35 cm, 250-300 g, deep purple Shikon sendai naga SS 8-10 cm, 20-30 g, bluish purple Shin nagasaki naga SN 35-40 cm, approx. 250 g, deep purple Big, Oval (Italy) Zebra Ze 20-25 cm, 300-400 g, purple and white stripes Small, Round (Thailand) Makhuea pro MP 4-5 cm, approx.10 g, green Solanum xanthocarpum 107 bitterness, and sourness) and the discrimination tests for four solutes (sucrose, sodium chloride, tartaric acid, and sodium glutamate). In addition, they had more than one year experience in evaluating vegetables and were clas- sified as expert assessors (Japanese Society for Sensory Evaluation, 2009). The characteristics listed in Table 2 were evaluated on a scale of -5 (weaker) to 5 (stronger) compared to Se2, the control cultivar in this study. Taste sensor analysis and amino acid content Approximately 50 g of fresh fruit was homogenized in 100 mL of water in an ice bath, then filtered through cot- ton cloth. Although filtrates were prepared separately from three or four fruit samples, the filtrates were mixed to obtain the quantity necessary for measurements. Bitterness, astrin- gency, acridity, and pungency of the filtrates were measured using a taste sensor system (SA402B, Intelligent Sensor Technology, Kanagawa, Japan). Each value was expressed relative to the control cultivar (Se2), which was set at zero. A portion of each filtrate was filtered again through a 0.45- μm filter (DISMIC-25CS, Advantec, Tokyo, Japan) and the amino acid content was measured using an amino acid ana- lyzer (JLC-500/V, JEOL, Tokyo, Japan). Soluble solids content Fresh fruit was cut into small pieces and pressed in gauze to extract the juice. The soluble solids content of the juice was measured using a refractometer (PAL-1, ATA- GO, Tokyo, Japan) and expressed as percent Brix. Ash and polyphenol contents The harvested fruit was stored at -30°C until use. Fresh-frozen fruit samples were freeze-dried and ground into a fine powder. Ash content was determined using the dry ashing method. Briefly, 0.3 g of the powder was put in a crucible and ashed at 550°C for 24-48 hr. After cooling, the weight of the residue was measured. Polyphenol con- tent was determined using iron tartrate spectrophotometry. First, 0.2 g of the powder was mixed with 10 mL of dis- tilled water, and shaken for 10 min at 80°C. After cooling, the sample was centrifuged at 3,000 rpm for 15 min. Then, 3.2 mL of the supernatant was mixed with 1.6 mL of iron tartrate reagent (0.1% (w/v) ferrous sulfate and 0.5% (w/v) potassium sodium tartrate) and 3.2 mL of phosphate buffer (0.1 M, pH7.5). The absorbance at 540 nm was then mea- sured using a spectrophotometer (U-2000, Hitachi, Tokyo, Japan). The polyphenol content was calculated from a Table 2 - Evaluation of the taste of eggplant cultivars by sensory test (z) Fruit shape Cultivar Aroma Softness Juiciness Sweetness Bitterness Astringency Good Grassy Pericarp Flesh & Acridity Small, Round De (y) 0 (x) 0 -1 0 0 0 1 1 Min 1 1 0 -1 0 -1 2 1 Round AM 0 1 0 -1 0 0 0 1 KN 1 1 0 -1 0 0 0 0 To 0 0 -1 0 0 0 1 0 YM 0 0 0 1 1 1 -1 0 Money pouch NK 0 2 -3 -2 -1 -1 0 0 SA 0 2 -4 0 -1 0 0 1 Oval HM 0 0 0 1 0 0 0 0 Se2 (Control) 0 0 0 0 0 0 0 0 SK 0 2 0 1 2 1 0 0 WS 0 0 -1 0 -1 -1 0 0 Ya 1 0 -1 0 0 0 0 0 Long Chi 0 0 0 1 0 -1 0 0 Hak 0 0 1 1 -1 0 -1 0 Hho 0 0 0 0 0 0 0 0 HN 0 1 -1 1 -1 -1 0 0 KC 0 0 -2 1 0 -1 0 1 KO 0 0 -2 1 -1 -1 0 0 SS 0 1 -1 0 0 -1 0 1 SN 0 2 0 1 -2 0 -1 0 Big, Oval Ze 2 2 0 -2 0 -1 0 2 Small, Round MP 0 3 0 -1 0 -1 0 1 (z) Eggplant fruit was cut to the size of 1.5 x 1.5 x 3.0 cm, then steamed at 100 degree C for 5 min before the test. (y) Amino acid which is considered to be important for the functionality of eggplant in Yakuzen theory. (x) Tastes were evaluated on a scale of -5 (weaker) to 5 (stronger) as compared to Senryo 2 gou (Se2). 108 standard curve of ethyl gallate. Principal component analysis and characterization of cul- tivars Data were analyzed by principal component analysis (SPSS for Windows version 13), and the characteristics of cultivars were comprehensively evaluated to determine the suitability of the cultivars as materials for Yakuzen dishes. 3. Results and Discussion Cultivars NK and SA (with money-pouch fruit shape) tended to have harder fruits, and long-fruit cultivars tended to be less sweet (Table 2). The aroma of the Italian cultivar, Ze was characterized as both “good” and “grassy”. How- ever, most cultivars were very similar to Se2 (the control cultivar in this study) in the sensory test. Taste sensor analysis showed that the local cultivars tended to have a less unpleasant taste than Se2 (Table 3). This result did not correspond to the results of the sensory test, and the difference may be due to the heating of sam- ples before the sensory test but not before the taste sensor analysis. Nevertheless, we can conclude that the local cul- tivars were not unpalatable compared with the commonly used cultivar. Min, YM and some other cultivars tended to have higher soluble solids contents, but there was no significant difference between cultivars (Table 4). “Mi” (the taste) of eggplant is “Kan” (sweet). In Yakuzen theory, Mi means not only the taste on the tongue but also specific functions in the human body (Tokui et al., 2003). In this study, we could not discern differences of Kan characteristics from the results of sensory test and soluble solids contents among cultivars; therefore, the suitability of cultivars was evaluated on the basis of other characteristics. Ash content was higher in cultivars De and Min (both of which have small, round fruit) and lower in cultivar WS, and tended to be lower in cultivars with oval fruit. “Sei” (the property) of eggplant is to cool the human body. Potassium accounts for most of the ash of eggplant fruit (USDA, 2013), and the function of this mineral is to release heat inducing diuresis. This function is closely related to the property of eggplant, and higher ash con- tent may be related to greater suitability of cultivars for Yakuzen dishes. Polyphenol content was also higher in De and Min. Eggplant contains polyphenols such as chlorogenic acid and nasunin, which are considered to have antioxidant ac- tivity, and to suppress lipid peroxidation, aging, various lifestyle diseases, and cancer (Kimura et al., 1999; Noda et al., 2000; Kitsuda et al., 2005; Singh et al. 2009). Das et al. (2011) reported that grilled eggplant had a higher poly- phenol content, though the cardioprotective ability was not different. The high polyphenol content in De and Min may increase the pharmacological value of these cultivars. As mentioned above, Mi (the taste) also encompasses specific functions in the human body, and Mi of eggplant is Kan (sweet). Kan is considered to have functions such as supplying nutrition and energy, promoting relaxation, etc. Some amino acids are considered to have Kan func- tions. For example, glutamine is an energy source for di- gestion and plays an important role in the maintenance and improvement of immunity and the repair of organs (Ajinomoto Co. Inc., 2003 a, b), and this may correspond to a Kan function. Alanine has a sweet taste and supplies sugars to the body, and it is also considered to have a Kan function. The amino acids strongly related to the Kan of eggplant include alanine, citrulline, glycine, glutamine, proline and serine. Therefore, these amino acid contents were summed to give specific amino acid content (Table 4): it was highest in Se2 and SN and tended to be higher in the long-fruit cultivars and lower in the round-fruit cul- tivars. Total amino acid content also tended to be higher in the long-fruit cultivars, especially in SN. However, no oth- er trends in amino acid content were observed. The higher content of specific and total amino acids in SN would indi- cate greater suitability of this cultivar for Yakuzen dishes. Table 3 - Evaluation of the taste of eggplant cultivars by taste sensor (z) Fruit shape Cultivar Bitter- ness Astrin- gency Acridity Pungency Small, Round De (y) -0.27 (x) 0.06 -1.74 -0.69 Min -0.38 0.04 -2.27 -0.96 Round AM -0.23 0.20 -1.14 -0.48 KN -0.11 0.21 -2.11 -0.94 To -0.11 0.13 -2.18 -1.18 YM -0.16 -0.02 -1.33 -1.30 Money pouch NK -0.26 0.27 -1.78 -0.64 SA -0.07 0.30 -2.30 -1.07 Oval HM 0.22 0.13 -1.83 -0.99 Se2 (Control) 0.00 0.00 0.00 0.00 SK -0.28 0.05 -1.51 -0.76 WS -0.06 0.23 -2.26 -1.17 Ya -0.09 0.08 -2.57 -1.30 Long Chi -0.33 0.12 -1.39 -0.61 Hak -0.28 -0.09 -2.55 -1.40 Hho -0.15 0.05 -1.61 -0.86 HN -0.01 0.15 -2.87 -1.45 KC -0.12 0.11 -2.24 -1.28 KO -0.43 0.07 -0.52 -0.15 SS -0.26 0.04 -2.03 -1.00 SN -0.47 -0.05 -2.27 -1.09 Big. Oval Ze -0.32 -0.02 -2.73 -1.23 Small. Round MP -0.34 0.19 -2.15 -0.75 (z) Taste sensor was prepared to express the value of Se2 was zero. (y) Amino acid which is considered to be important for the func- tionality of eggplant in Yakuzen theory. (x) Positive and negative value means the taste was stronger and weaker than Se2, respectively. 109 From the PCA, 23 principal components (PCs) were de- rived and 14 PCs were considered to be meaningful (con- tribution rate>1). These 14 meaningful PCs accounted for 96.6% of the total rate (data not shown). Although the first two PCs accounted for only 41.3% of the total rate, a two- dimensional scatter diagram of factor loading was con- structed. Specific and total amino acid contents were in the positive direction along the x-axis, and juiciness, sweetness, percent Brix and some unpleasant taste were in the negative direction. Ash, polyphenol content, and bitterness were in the positive direction along the y-axis, and softness was in the negative direction (data not shown). A two-dimensional scatter diagram constructed from the PC1 and PC2 scores of each cultivar enabled classification of the cultivars (Fig. 1). Many of the common F 1 cultivars of eggplant used for com- mercial production in recent years have been developed to improve ease of consumption and cultivation. Se2 is one of the common cultivars in Japan. It was characterized as juicy and sweet by the PCA and is considered easy to eat. The oth- er cultivars with round or oval fruit were also characterized as juicy and sweet. On the other hand, most of the long-fruit cultivars were distinct from the round-fruit and oval-fruit Table 4 - Soluble solid, ash, polyphenol and amino acid content of eggplant cultivars Fruit shape Cultivar Soluble solids (% Brix) Ash (g 100 g-1 FW) Polyphenol (z) (mg 100 g-1 FW) Specific amino acid (y) (mg 100 g-1 FW) Total amino acid (mg 100 g-1 FW) Small. Round De (x) 5.1 a (w) 0.60 a (w) 673.4 a (w) 120.6 284.9 Min 5.2 a 0.60 a 434.0 a 71.1 186.0 Round AM 4.8 a 0.45 ab 169.0 b 37.2 151.2 KN 5.4v 0.53 (v) 150.0v 48.8 203.1 To 4.8 a 0.39 b 253.7 ab 52.5 196.6 YM 5.3 a 0.50 ab 255.0 ab 36.3 144.3 Money pouch NK 5.0 a 0.45 ab 172.9 b 63.1 197.8 SA 4.6 a 0.50 ab 247.3 ab 39.5 167.3 Oval HM 4.7 a 0.50 ab 300.6 ab 64.5 203.1 Se2 (Control) 4.6 a 0.44 ab 129.1 b 145.6 302.0 SK 4.3 a 0.39 b 158.4 b 48.0 170.6 WS 5.1 a 0.37 b 181.9 b 52.8 199.1 Ya 4.3 a 0.40 b 248.5 ab 58.1 185.4 Long Chi 4.5 a 0.42 ab 188.1 b 34.1 161.2 Hak 4.3 a 0.38 b 127.9 b 116.3 286.5 Hho 4.8 a 0.52 ab 166.2 b 76.8 253.3 HN 4.5 a 0.52 ab 144.1 b 83.6 288.6 KC 4.4 a 0.48 ab 236.6 ab 50.8 211.0 KO 5.1 a 0.45 ab 135.1 b 93.6 314.2 SS 4.1 a 0.38 b 250.6 ab 106.1 281.8 SN 4.5 a 0.41 ab 119.2 b 139.2 342.0 Big. Oval Ze 3.8 a 0.46 ab 124.0 b 95.1 308.9 Small. Round MP 5.1 a 0.56 ab 245.9 ab 63.9 196.7 (z) Polyphenol content was expressed as ethyl gallate equivalent. (y) Amino acid strongly related to Kan of eggplant. (x) Abbreviations as in Table 1. (w) Different letter within the row indicates significant difference by Tukey’s multiple range test at 5% level (n=5). (v) Number of harvested fruit was not enough for statistical analysis. Fig. 1 Two dimensional scatter diagram of the principal component score of eggplant cultivars ■: small, round ◆: round ●: monry pouch ▲: oval : long : Italy : Thailand. Abbreviations as in Table 1. 110 cultivars. They were characterized as having higher specific amino acid contents and little unpleasant taste or sweetness. The small and round fruit cultivar De and the Thai species (MP) were considered to contain many functional ingredi- ents, such as ash, polyphenols, and specific amino acids, and to have greater suitability for Yakuzen dishes. 4. Conclusions Among the local Japanese cultivars used in this study, cv. 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