Food Chemistry 124 (2011) 13691375

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Changes in chemical composition during soybean seed development

Xiaoyu Saldivar a, Ya-Jane Wang a,*, Pengying Chen b, Anfu Hou b
a b

Department of Food Science, University of Arkansas, Fayetteville, AR 72704, USA Department of Crop, Soil, and Environmental Sciences, University of Arkansas, Fayetteville, AR 72701, USA

a r t i c l e

i n f o

a b s t r a c t
This study examined the compositional change of ve specialty soybean genotypes, which are low in oligosaccharides, high in oil, high in protein, large seeded or small seeded, along with two commercial cultivars, Jack and Ozark, during seed development and maturation. Seeds were sampled at 7-day intervals from initial seed formation to full maturity for approximately 8 weeks, and analysed for oil, protein, soluble saccharides, and starch. Although there were signicant differences among the seven soybean genotypes in their chemical compositions, some compositional changes followed similar trends. Protein content decreased during the rst 35 weeks after owering and then gradually increased. Oil was accumulated rapidly during the early stages. The percentage of starch ranged from 6 to 15% in developing seeds, but declined sharply to 0.21% at maturity. Sucrose decreased during seed development and maturation, while non-digestible oligosaccharides (rafnose, stachyose, and verbascose) remained at low levels during early stage until 3 weeks before harvest and increased towards maturity. These ndings provide valuable information for developing and selecting specialty soybean varieties for specic applications. 2010 Elsevier Ltd. All rights reserved.

Article history: Received 30 January 2010 Received in revised form 8 June 2010 Accepted 26 July 2010

Keywords: Soybean seed development Protein Oil Sugar Chemical composition

1. Introduction Soybean [Glycine max (L.) Merrill] is widely used in the food industry because of its high protein and oil contents. Soybean can be classied into oil bean and food bean according to its end uses. Oil soybean, i.e. commodity bean, is the primary source of vegetable oil and soy protein products, such as defatted soy our and soy protein concentrate; food bean, i.e. specialty bean, is either consumed directly or processed into various soy products. Soybean cultivars with large seed size and high sucrose content are desirable in the production of vegetable soybean, which is the food bean harvested at immature stage, also called edamame. On the other hand, cultivars with small seed size and low calcium content are desirable for natto, a traditional fermented soy food in Japan with a rmer texture. For soymilk and tofu production, soybean cultivars with light hilum colour, large seed size, high water absorption, high protein, high sucrose, and low oligosaccharides are desirable. In order to add value to the soybean crop, soybean breeders are developing specialty soybeans with desired quality attributes for specic applications. Therefore, it is important to understand the biosynthesis and changes of seed constituents during seed development and maturation of soybeans from different genetic backgrounds and targeted for different end uses.

* Corresponding author. Tel.: +1 479 575 3871; fax: +1 479 575 6936. E-mail address: yjwang@uark.edu (Y.-J. Wang). 0308-8146/$ - see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodchem.2010.07.091

Changes in seed composition during development and maturation have been investigated previously with commodity soybean. Tada and Kawamura (1963) reported that starch content was 4 5% in immature soybean seeds, but decreased to almost zero at maturity. Reducing sugars decreased, while non-reducing sugars increased during soybean seed development. Rubel, Rinne, and Canvin (1972) reported that the level of oil increased rapidly to 20% at 40 days after owering (DAF), and remained constant thereafter until maturity. Hill and Briedenbach (1974) found that protein accumulated rapidly between 12 and 28 DAF, but declined at the onset of seed desiccation. Yazdi-Samadi, Rinne, and Seif (1977) reported that most oil and protein were accumulated from 20 to 40 DAF; sucrose showed a steady decrease; rafnose and stachyose increased from 40 to 50 DAF; starch reached a peak value at 3040 DAF and then declined sharply at maturity. Lowell and Kuo (1989) studied the relationship between soluble sugar content and enzyme activity during seed development. The formation of rafnose and stachyose in soybean was accompanied by an increase in galactinol synthase activity and galactinol content, and a substantial decrease in myo-inositol, which is the substrate of galactinol synthase. The relationship between oil, protein and sugar contents in mature soybean has also been examined. Hymowitz, Collins, Panczner, and Walker (1972) studied 60 soybean genotypes and reported that sucrose and rafnose contents were positively correlated with oil content, but negatively correlated with protein content. More recently, Hartwig, Kuo, and Kenty (1997) studied 20

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high-oil and 20 high-protein soybeans and reported a negative correlation, although not signicant, between protein content and levels of rafnose and stachyose. Most of the previous seed composition research work focused on oil bean genotypes, and very little research has been done on specialty food bean genotypes. The present work studied the temporal changes in chemical composition during seed development and maturation of both food bean and oil bean genotypes. This work would provide useful information for future development and selection of new food bean varieties with desired traits. 2. Materials and methods 2.1. Plant materials Five soybean genotypes were selected based on their unique seed traits and specialty utilities (Table 1). Two commercial soybean cultivars, Jack and Ozark, were also included as controls, in which Ozark served as a conventional check. Among the seven genotypes, Jack, 03-CB14, and V97-3000 were grown in 2006 for a preliminary experiment, and all seven genotypes were grown in 2007. They were grown in Fayetteville, Arkansas, with 4-row plots arranged in a randomised complete block design with two replications. When each soybean genotype started owering, each ower from the middle two rows of the plot was tagged on the owering date. Tagged pods were sampled randomly at 7-day intervals from initial seed formation to full maturity. Immediately after sampling, soybean pods were shelled by hand, and the length of each seed was measured. Seeds of approximately 40 g were then cut in half and dried in a forced draft oven at 40 C for 24 h. The dried seeds were nely ground in a coffee grinder (Black & Decker, CBG100 W SA BD, Towson, MD) to pass through a 150-lm sieve (W.S. Tyler, Nentor, OH). Ground soybean samples were stored in hermetic bottles at 4 C before analysis. 2.2. Chemical composition analysis The moisture content of ground soybean was determined according to AACC Approved Method 44-15A (AACC, 2000). Duplicate samples of 2 g ground soybean our were accurately weighed into a weighing dish, and dried in an oven at 130 C for 2 h. Crude protein content was determined using the combustion method (Campbell, Kraught, Yackel, & Yang, 1985) by Elementar Variomax CN (Elementar Americas, NJ). A conversion factor of 6.25 was used to convert total nitrogen to crude protein content. Lipid content was determined by accurately weighing 1 g of sample into a 15ml centrifuge tube, and then adding 10 ml hexane to extract lipid. The tubes were rotated end-over-end at room temperature for 2 h, and then centrifuged at 1700 g for 10 min. The supernatant containing oil was transferred into a pre-weighted dish, and hexane

was evaporated by heating on a hot plate (Lam & Proctor, 2001). Total starch content was determined according to AACC Approved Method 79-13 (AACC, 2000) using an amyloglucosidase/alphaamylase assay method (Megazyme, Wicklow, Ireland). Soluble saccharides in 100 mg ground soybean were extracted with 5 ml 80% ethanol at 75 C for 10 min with magnetic stirring and centrifuged at 1500 g for 10 min. The extraction was done with two replicates of each ground sample. The phenolsulphuric acid method (Dubois, Gilles, Hamilton, Smith, & Rebers, 1956) was used to estimate the amount of dissolved saccharides in the supernatant.

2.3. Extraction and quantication of saccharides Ground soybean (0.15 g) was extracted with 1.5 ml of double distilled water in a 2-ml centrifuge tube. The tube was shaken horizontally (Barnstead 1314, Melrose Park, IL) at 200 rpm and ambient temperature for 20 min, and then centrifuged at 20,000 g for 10 min. Supernatant (500 ll) was removed and mixed with 0.7 ml acetonitrile in a 1.5-ml tube, and then kept at room temperature for 30 min. The mixture was centrifuged at 20,000 g for 10 min, and the supernatant was ltered through a 0.2-lm membrane. Filtrate (24 ll) was dissolved in 576 ll double distilled water prior to HPLC analysis. A high-performance anion-exchange chromatography with pulsed amperometric detection (HPAECPAD) system was used for separation and quantication of each saccharide. Samples were injected via a Dionex autosampler equipped with a 25-ll sample loop. Sugars were separated on a Dionex CarboPac PA 10 pellicular anion-exchange resin column (250 mm 4 mm i.d.), preceeded by a Dionex CarboPac PA 10 guard column (50 mm 4 mm i.d.) and a Dionex AminoTrap column (30 mm 3 mm i.d.) at a ow rate of 1 mL/min (Dionex, Sunnyvale, CA). The mobile phase consisted of 90 mM NaOH solution prepared by dilution of carbonate-free 50% (w/w) NaOH solution in distilled water, which was previously ltered through a 0.45-lm membrane. The detection was accomplished by an ED40 electrochemical detector. Commercial saccharides, including glucose, fructose, sucrose, rafnose, stachyose and verbascose, purchased from SigmaAldrich (St. Louis, MO), were used as external standards to identify and quantify each saccharide based on their retention times and peak areas. All chemical composition analyses were performed in duplicate, and results were expressed as percentage on a dry-weight basis.

2.4. Statistical analysis The measurement of soluble sugars at different growth stages qualies as a repeated measures experimental design. Because repeated measures analysis is subject to the violation of the homogeneity of variance assumption in the ANOVA model, the existence of a specic variancecovariance structure of the repeated measurements, also known as the HuynhFeldt condition, has to be tested to determine whether the interpretation of the ANOVA will be performed using F statistics with adjusted degrees of freedom. We tested the HuynhFeldt condition using the Maulchys sphericity test (Huynh, 1978; Kuehl, 2000; Mauchly, 1940) in SAS (SAS Software Institute, Cary, NC). As none of the variables met the HuynhFeldt condition (data not shown), the factor week and its interaction with genotype in the ANOVA had only an approximate F distribution with reduced degrees of freedom, so unbiased estimation of F statistics and p values was not possible. Therefore, we opted to use the Greenhouse-Geisser (Kuehl, 2000) and Huynh Feldt (Kuehl, 2000) F statistics with adjusted degrees of freedom for the interpretation of the factor week and its interaction with genotype in the ANOVA.

Table 1 List of soybean genotypes used in this study and their specialty traits and uses. Genotype Jack Ozark 03-CB14 R03-144 R95-1705 V94-3168T V97-3000 Source Illinois Arkansas Virginia Arkansas Arkansas Virginia Virginia Specialty traits and uses Commodity soybean, early maturing, high in glucose and fructose, low in sucrose Commodity soybean, late maturing, high yield Food soybean, large seeded, low in oligosaccharides, edamame, tofu, soymilk Commodity soybean, high in oil, vegetable oil Commodity soybean, high in protein, feed Food soybean, late maturing, large seeded, edamame, tofu, soymilk Food soybean, small seeded, natto

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3. Results and discussion The results of 2007 followed a similar trend as those obtained from the preliminary experiment conducted in 2006; therefore, the individual data of 2006 are not presented here because 2007 data were more complete and representative. Analysis of the pooled data showed no signicant difference between these two experiments in terms of genotype and growth periods for seed chemical compositions, but signicant interactions between growth periods and genotypes were detected (p < 0.05) for soybean seed chemical compositions.

3.2. Change in protein content during seed development and maturation The changes in protein content of the seven soybean genotypes during seed development and maturation in 2007 are shown in Fig. 2. The protein levels in mature soybean seeds varied from 43.4 to 51.3% (dry basis, db), with R95-1705 and V94-3168T having over 50% protein. Levels of protein decreased 26% during the rst 35 weeks after owering and gradually increased thereafter until maturity, which was attributed to the rapid synthesis of oil and starch in early seed development. Protein content in most genotypes increased from 5 to 10 11 weeks after owering (R6R8 stages). The high protein genotype, V94-3168T, accumulated protein drastically in the last three weeks before maturation, whereas low protein genotypes, such as R03-144 and Ozark, exhibited a slower increase in protein accumulation during the same time period. On the other hand, protein accumulated rapidly in Jack and R95-1705 between 4 to 8 weeks (R5R7 stages), and then their protein contents remained unchanged until fully maturity. The pattern of protein accumulation during seed development is important for breeding vegetable soybean varieties because vegetable soybean is harvested at R6 stage, around 67 weeks after owering. Although Jack, R95-1705 and V94-3168T had similar levels of protein at maturity, Jack and R95-1705 had signicantly higher levels of protein than V94-3168T at R6 stage, which is a desired trait for vegetable soybean genotypes.

3.1. Seed size change during seed development and maturation Soybean reproductive growth and development can be divided into 8 stages according to morphological features (Table 2): R1 and R2 describing owering; R3 and R4 describing pod development; R5 and R6 describing seed development; R7 and R8 describing plant maturation (McWilliams, Berglund, & Endres, 1999). This study used seed size (length) and colour to establish time of maturity. Seed size increased to a peak value at R6R7 stages and decreased afterwards (Fig. 1). Soybean seeds remained green at R5 and R6 stages and began to turn yellow at R7 stage (TeKrony, Egli, Balles, Pfeiffer, & Fellows, 1979). Jack, an early maturing genotype, reached R6 stage at 5 weeks after owering. In contrast, Ozark and V94-3168T, both late maturing genotypes, reached R6 stage at 78 weeks after owering. The other genotypes reached R6 stage at 6 weeks after owering. The average length of mature soybean seeds was 8.08.5 mm. Large seeded genotypes Ozark and V94-3168T were approximately 9.0 mm in length, whereas the small seeded genotype V97-3000 was 6.0 mm in length (Fig. 1).

3.3. Change in oil content during seed development and maturation Unlike protein synthesis, the seven soybean genotypes displayed a similar pattern of oil accumulation (Fig. 3). Oil was accumulated rapidly at 37 weeks after owering (R5R6 stage) and remained at a similar level afterwards. R03-144 had the highest oil content during seed development, whereas V94-3168T had signicantly lower oil content at the beginning of seed development between 4 and 8 weeks after owering. When the protein contents were correlated with the oil contents (Figs. 2 and 3), a negative correlation was found between protein and oil among the seven genotypes, which agrees with Hymowitz et al. (1972). They reported that protein and oil in mature soybean were negatively correlated between different genotypes. Genotypes V94-3168T and R95-1705 had low oil contents and high protein contents, whereas R03-144

Table 2 Stages of soybean development. Stages R1 R2 R3 R4 R5 R6 R7 R8 Description Beginning bloom, rst ower Full bloom, ower in top two nodes Beginning pod, 3/16 inch pod in top four nodes Full pod, 3/4 inch pod in top four nodes Beginning seed, 1/8 inch seed in top four nodes Full size seed in top four nodes Physiological maturity Full maturity, 95% of pods on the plant are mature

18

16

14 Seed length (mm)

R6 R6

Jack Ozark
R6

12
R6

R6 R6

03-CB14 R03-144

10
R6

R95-1705 V94-3168T V97-3000

4 3 4 5 6 7 8 Weeks after Flowering 9 10 11

Fig. 1. Change in seed length of developing soybean seeds over time. R6 stage of each genotype was marked as R6 in gure.

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54 52 Protein (g/100g dry weight) 50 48 46 44 42 40 38 36 3 4 5 6 7 8 9 10 11 Weeks after Flowering

Fig. 2. Change in protein content in soybean during seed development and maturation. The average coefcient of variation is 1.65%.

Jack Ozark 03-CB14 R03-144 R95-1705 V94-3168T V97-3000

24 22 20 Oil (g/100g dry weight) 18 16 14 12 10 8 6 4 2 0 3 4 5 6 7 8 9 10 11 Weeks after Flowering

Fig. 3. Change in oil content in soybean during seed development and maturation. The average coefcient of variation is 4.25%.

Jack Ozark 03-CB14 R03-144 R95-1705 V94-3168T V97-3000

12 Soluble Saccharides (g/100g dry weight) 11 10 9 8 7 6 5 4 3 2 3 4 5 6 7 8 9 10 11 Weeks after flowering

Fig. 4. Change in soluble saccharides content in soybean during seed development and maturation. The average coefcient of variation is 4.79%.

Jack Ozark 03-CB14 R03-144 R95-1705 V94-3168T V97-3000

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had the highest oil content and the lowest protein content among the seven genotypes. 3.4. Change in soluble saccharides during seed development and maturation The changes of total soluble saccharides as measured by the phenolsulphuric acid method during soybean maturation are shown in Fig. 4. In general, levels of soluble saccharides decreased at the beginning of seed formation, remained relatively unchanged from 5 to 8 weeks, and increased toward the end of seed maturation. The decrease in the level of soluble saccharides in the beginning, similar to the decrease in protein, was attributed to the rapid increase of oil and starch at early stages. A wide range of saccharides, including monosaccharides of glucose and fructose, disaccharide of sucrose, and oligosaccharides of rafnose, stachyose and verbascose, were present in soybean seeds as analysed by the HPAECPAD system (Table 3). In general, mono-

saccharides and disaccharide decreased, whereas oligosaccharides increased in all the soybean genotypes during seed development and maturation. Among the seven soybean genotypes, the average glucose content decreased from 7.2 to 1.6 mg/g, and the average fructose content decreased from 6.4 to 0.4 mg/g dry seeds. Yazdi-Samadi et al. (1977) and Dornbos and McDonald (1986) reported similar trends for glucose and fructose during soybean development. At maturity, Jack had the highest content of monosaccharides of 1.9 mg/g glucose and 0.6 mg/g fructose; 03-CB14 had the lowest of 1.1 mg/g glucose and 0.2 mg/g fructose. Sucrose was the most abundant soluble saccharide. The average sucrose content in mature seeds was 56.4 mg/g on a dry-weight basis, with 03-CB14 having the highest sucrose content of 94.7 mg/g. Jack showed the most signicant decrease in sucrose during soybean development, followed by V94-3168, whereas Ozark and R95-1705 had only slight decreases. The decrease in sucrose level in these genotypes was consistent with the results of

Table 3 Changes in soluble saccharides (mg/g dry seeds) in soybean during seed development and maturation.* Genotype Jack Weeks 5 (R6 ) 6 7 8 9 10 4 5 6 7 8 (R6) 9 10 3 4 5 6 (R6) 7 8 9 10 5 6 (R6) 7 8 9 10 3 4 5 6 (R6) 7 8 9 10 4 5 6 7 (R6) 8 9 10 11 4 5 6 (R6) 7 8 9 10

Glucose 1.4 1.1a 1.9a 1.4a 1.8a 1.9a 4.6a* 1.9b 1.5bc 1.1c 1.1c 1.6b 1.8b 11.8a 9.7ab 8.4abc 4.5bcd 3.3cd 3.0cd 1.1d 2.1d 1.7a 1.4a 1.4a 1.4a 1.5a 1.2a 4.4a 2.6b 1.1d 1.2cd 1.1cd 1.9c 1.3cd 1.3cd 14.8a 4.2b 2.8bc 1.5c 1.5c 1.5c 2.1bc 1.8c 4.1a 1.9bc 1.8bc 1.6c 1.6c 2.7b 1.7bc
a

Fructose 1.0 0.8a 1.0a 0.6a 0.4a 0.6a 4.0a 0.9b 0.7bc 0.8bc 0.5bc 0.4c 0.4c 12.6ab 20.4a 8.6ab 2.6b 3.0b 3.2b 0.2b 0.2b 1.0a 0.4b 0.4b 0.4b 0.3bc 0.2c 4.0a 1.0b 0.4c 0.4c 0.3c 0.5c 0.2c 0.3c 14.7a 3.9b 2.7b 1.2c 1.2c 0.5c 0.9c 0.5c 3.0a 1.1bc 0.9bc 0.7c 0.7c 1.4b 0.6c
a

Sucrose 88.4 75.0b 53.7c 40.2d 41.3d 32.4d 67.9a 57.1b 54.5b 58.8b 61.2b 54.1b 58.5b 105.4a 70.2c 65.4c 65.7c 57.8c 70.3c 90.0b 94.7ab 59.1ab 59.6ab 64.2a 56.6ab 45.5bc 51.4c 66.5a 71.8a 50.3b 50.4b 50.8b 48.1b 46.4b 45.0b 85.9a 83.0a 80.2a 79.4a 75.3a 59.8b 53.8b 49.4b 86.1a 93.3a 96.0a 92.5a 81.1a 61.2b 58.7b
a

Rafnose 0.0 0.2c 0.6c 2.6b 8.7a 9.2a 0.0c 0.0c 0.0c 0.0c 0.5c 3.6b 8.9a 0.0c 0.0c 0.0c 0.1bc 0.1bc 0.5b 3.3a 2.9a 0.2c 0.3c 0.8c 2.2b 5.5a 5.7a 0.0c 0.1c 0.2c 0.4c 1.2c 6.4b 8.9a 9.3a 0.0b 0.0b 0.1b 0.4b 0.3b 1.9b 7.3a 8.4a 0.0c 0.0c 0.2c 0.3bc 0.9b 4.6a 4.6a
c

Stachyose 0.0 0.0c 0.8c 6.5b 30.9a 29.6a 0.0c 0.0c 0.0c 0.0c 0.7c 13.0b 42.2a 0.0c 0.0c 0.0c 0.0c 0.1c 0.2c 2.9a 2.2b 0.0d 0.1d 0.7d 3.2c 35.3b 39.6a 0.0c 0.0c 0.0c 0.0c 1.4c 21.1b 38.0a 39.3a 0.0b 0.0b 0.0b 0.1b 0.3b 4.7b 37.8a 40.1a 0.0c 0.0c 0.2c 0.3c 1.5c 30.6b 39.9a
c

Verbascose 0.0c 0.0c 0.0c 0.0c 1.1b 1.6a 0.0b 0.0b 0.0b 0.0b 0.0b 0.0b 1.4a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0a 0.0b 0.0b 0.0b 0.0b 0.9ab 1.8a 0.0b 0.0b 0.0b 0.0b 0.0b 0.1b 1.3a 1.2a 0.0b 0.0b 0.0b 0.0b 0.0b 0.0b 0.6a 1.0a 0.0c 0.0c 0.1c 0.0c 0.0c 0.3b 1.9a

Ozark

03-CB14

R03-144

R95-1705

V94-3168T

V97-3000

Means with different letters in the same column within a genotype are signicantly different (p < 0.05). R6 Stage.

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Yazdi-Samadi et al. (1977). Genotype 03-CB14 had the highest initial sucrose content, which decreased rapidly from 3 to 7 weeks after owering and then increased afterwards. The sucrose contents in V97-3000 and R03-144, slightly increased from 46 weeks after owering (R5R6 stages) and then decreased until maturity. Dornbos and McDonald (1986) reported that genotype Williams 79 showed an increase in sucrose at R5-R6 stages and then unchanged until full maturity. These results suggest that the changes of sucrose level during soybean development vary among different soybean genotypes. Oligosaccharides were not detected until 67 weeks after owering, around R6 stage, in the seven soybean genotypes, and accumulated rapidly during the last 3 weeks before full maturity. Stachyose was the predominant oligosaccharide in soybean, followed by rafnose, and verbascose. The average contents of rafnose and stachyose in mature soybean were 6.8 and 33.0 mg/g, respectively, with 03-CB14 having signicantly lower values of 2.9 and 2.3 mg/g, respectively. Oligosaccharides are accumulated in legume seeds during maturation as energy sources for germination (Kuo, VanMiddlesworth, & Wolf, 1988). Because they are non-digestible, their presence has been linked to abdominal discomfort when soy food and other legume products are consumed (Calloway, Colasito, & Mathews, 1966). Because of its low oligosaccharide content at maturity, 03-CB14 has an advantage as a food or feed soybean genotype. The formation of rafnose oligosaccharides requires sucrose and galactinol. Lowell and Kuo (1989) reported that at R6 stage the decrease in sucrose and high galactinol synthase activity coincided with initial accumulation of rafnose and stachyose. The changes in the amounts of sucrose and oligosaccharides during soybean seed development demonstrate the advantages of vegetable soybean in terms of sensory and nutritional attributes over mature soybean. Vegetable soybean offers a sweeter taste and less digestive problems than mature soybean because it is harvested at R6 stage when sucrose content is high and oligosaccharides are minimally accumulated (Rackis, Hoing, Sessa, & Moser, 1972). The amount of total soluble saccharides in soybean seeds as measured by the phenolsulphuric acid method was generally lower than the sum of the six saccharides analysed by HPAEC PAD. The discrepancy was attributed to the standard curve for quantifying soluble saccharides in the phenolsulphuric acid method, which was prepared by D-glucose and not optimised for other saccharides.

3.5. Change in starch content during seed development and maturation Soybean has the distinction of oilseed legumes that they lack considerable levels of starch in seeds at maturity (Isleib, Pattee, & Giesbrecht, 2004; Wilson, Bermingham, Moon, & Snyder, 1978). Levels of starch in the seven soybean genotypes were 5.69.3% at the initial stage of seed development, and drastically decreased to 0.21.0% at maturity (Fig. 5). Among the genotypes, the starch level in V94-3168T signicantly increased from 4 to 7 weeks after owering and then rapidly decreased. Ozark and V94-3168T, both late maturing genotypes, reached R6 stage at 78 weeks after owering and at the same time their starch levels began to decrease. Jack, an early maturing genotype, reached R6 stage at 5 weeks after owering and concurrently its starch level began to decrease. These results suggest the initiation of starch degradation at the commencement of soybean maturation around R6 stage of soybean seed development. In addition, the degradation of starch from 7 8 weeks in most genotypes coincided with the increase in soluble saccharides, suggesting that starch was consumed for the production of soluble saccharides, mostly oligosaccharides. All the traits measured showed a signicant genotype week interaction (Table 4), suggesting that the trends over time of seed component accumulation or degradation were signicantly different among genotypes. This is partially attributed to the differences in maturity.

Table 4 Effects of genotype, week and genotype week on soybean seed compositional traits. Trait Genotype Pr > F Week Adj Pr > F GG Protein Oil Starch Glucose Fructose Sucrose Rafnose Stachyose Verbascose <.0001 0.0001 <.0001 0.0122 0.0011 0.0001 0.0027 0.0001 0.0277 <.0001 <.0001 <.0001 <0.001 <0.001 <0.001 <0.001 <0.001 0.0002 HF <.0001 <.0001 <.0001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 Genotype Week Adj Pr > F GG 0.0973 <.0001 0.0001 0.009 0.002 <0.001 <0.001 <0.001 0.0522 HF 0.0188 <.0001 <.0001 0.0002 <0.001 <0.001 <0.001 <0.001 0.0033

GG: GreenhouseGeisser adjusted p-value; HF: HuynhFeldt adjusted p-value; Pr: Probability; F: F-value.

16 14

Starch (g/100g dry weight)

12 10 8 6 4 2 0 3 4 5 6 7 8 9 10 11

Jack Ozark 03-CB14 R03-144 R95-1705 V94-3168T V97-3000

Weeks after flowering

Fig. 5. Change in starch content in soybean during seed development and maturation. The average coefcient of variation is 11.23%.

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4. Conclusions Food grade soybeans are either consumed directly or processed into soy foods and may be harvested at immature stage. During soybean seed development, oil accumulation was very rapid at early stages, whereas protein was accumulated at later stages. Levels of monosaccharides and disaccharides decreased over the course of seed development and maturation, while oligosaccharides were accumulated during the last 3 weeks before seed full maturity. Starch was present in a signicant amount in early development of soybean seeds, but was degraded to less than 1% at maturity. The information from this research will be helpful for soyfood processors in determining the optimum harvest time for the crop and for breeders to develop specialty soybean varieties for specic food applications. Acknowledgement The authors wish to thank Dr. Andronikos Mauromoustakos and Luciano Jaureguy for their assistance with statistical analysis of the data. References
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