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Agricultural Sciences in China

2007, 6(2): 150-158 ScienceDirect February 2007

Correlation Between Antioxidation and the Content of Total Phenolics and

Anthocyanin in Black Soybean Accessions

XU Jin-rui1.2,ZHANG Ming-weil, LIU Xing-hua3, LIU Zhang-xiong4, ZHANG Rui-fed, SUN Ling1 and QIU
Li-juad

J Key Laboratory of Functional Food, Ministry of Agriculture/Bio-Tech Research Institute, Guangdong Academy of Agricultural Sciences,
Guangzhou 51 064, P.R.China
2 School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510310, P.R.China

3 College of Food Science and Engineering, Northwest A&F University, Yangling 712100, P.R. China

4 National Crop Gene Resources and Genetic Improvement/Key Laboratory of Crop Germplasm and Biotechnology, Institute of Crop

Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

Abstract
The objective of this study was to investigate the correlations between antioxidations and the contents of the total
phenolics and anthocyanin in 127 accessions of black soybean. A T-test, a fast clustering procedure, and a correlation
coefficient analysis were used for experimentation. The variation ranges of the total antioxidant capacity (TAC), the
total phenolics, and anthocyanin contents in 127 black soybean accessions were 0.44-3.56, 7.05-74.82, and 0.22-1.87
mg g-1, respectively, displaying significant genotype differences. The major differences in TAC, the total phenolics,
and the anthocyanin contents existed among various types of accessions from geographical regions. The differences
between the accessions from black and yellow soybeans, spring and autumn, summer and autumn, Dongbeichun and
Nanfangchun, Dongbeichun and Nanfangxia, Beifangchun and Nanfangchun, and Beifangchun and Nanfangxia were
significant at 0.01 or 0.05 levels, respectively. The general tendency was that the TAC, the total phenolics, and the
anthocyanin contents of Beifangchun accessions were higher than that of Dongbeichun ones, while that of Nanfangchun
accessions were the worst. 127 black soybean accessions could be clustered into 6 clusters, which consisted of 3, 24,
20, 31, 37, and 12 accessions, respectively. The most significant (P<0.01) correlations existed respectively between
the TAC and the total phenolics content, and the TAC and the anthocyanin content of black soybean. The results
showed that the total phenolics and anthocyanin in black soybean seed coat were the important antioxidation
substances.

Key words: seed coat of black soybean, antioxidation, total phenolics content, anthocyanin content, correlation

anthocyanin in its seed coat. Anthocyanin remaining

INTRODUCTION
Black soybean has been used as food and medicinal
materials in China and is an important source for the
development of natural antioxidants owing to the rich
with polyphenolic substance can effectively scavenge
oxygenic free radical. For a long time, research on the
anthocyanin from black soybean seed coat has focused
on the chemical structure and properties, while research
on physiologically active of anthocyanin is few (LU and

This paper is translated from its Chinese version in Scientia AgriculfuraSinica.

XU Jin-rui, Ph D, E-mail: xujr343@sina.com; Correspondence ZHANG Ming-wei, Ph D, Professor, E-mailmwzhh@163.net
Correlation Between Antioxidation and the Content of Total Phenolics and Anthocyanin in Black Soybean Accessions
Wu 1997; Cui et al. 2004; Meng 2002; Wang et al.
1995; Wang et al. 1998). Takanori et al. (1996), Long
(1999), Xiao et al. (2000), Azevedo et al. (2003) found
that the anthocyanin from black soybean seed coat could
effectively scavenge oxygenic free radical, and inhibit
lipid peroxidation and DNA damage in vitro. Xu et al.
(2005), Huang et al. (2004) studied the extraction tech-
nology of anthocyanin from black soybean coat (ABSC)
and found that ABSC could well scavenge the hydroxy
free radical (OH.), the superoxide free radical ( 0 2 y ) ,
and l,l-dipheny1-2-picryl-hydrazyl (DPPH-) in vitro.
They also found that the most significant positive cor-
relation existed between the total antioxidant capacity
(TAC) and the anthocyanin content, by analyzing their
correlation. Owing to the limited quantities of black
soybean accessions analyzed, the diversity of the total
phenolics and anthocyanin contents and the
antioxidations among various black soybeans were not
fully shown, and the internal relation between the anti-
oxidant and the total phenolics and anthocyanin con-
tents was not fully revealed. By collecting various black
soybean accessions, the article compares the differ-
ence of TAC in viti-o of black soybean accessions in
large numbers and analyzes the relations between the
total phenolics and anthocyanin contents and the
antioxidation in vitro, in order to pick out black soy-
bean accessions with intensive antioxidant properties
and provide evidence for unveiling the antioxidant ma-
terial basis of black soybean.
MATERIALS AND METHODS
Experimental materials
The resources of 127 black soybean accessions The
101 black soybean accessions were provided by Insti-
tute of Crop Sciences, Chinese Academy of Agricul-
tural Sciences, and the remaining were provided by Bio-
tech Research Institute, Guangdong Academy of Agri-
cultural Sciences, China, of which 54 were from north
of China, 44 from central, and 29 from south, and 111
yellow cotyledon and 16 green cotyledon. 15 yellow
soybean accessions were used for comparison.
The main reagents 2,4,6-tripyridyl-s-triazine(TPTZ)
was purchased from Fluka company; the other analyti-
cal regents were domestically manufactured.
151
Experimental methods
Sample treatment Various black soybean seed coats
(60 meshes) were pulverized and 1.O g flour was accu-
rately weighted. It was then extracted with 60% etha-
nol (20 mL, pH 3.0) at 60°C for 2 h, and the filtrate
volume was re-fixed to 20 mL with then extraction
reagent; it was then shaken up and reserved for later
use.
TAC assay According to the FRAP assay (Iris and
Strain 1996; Sean and Ranjeet 2004), the reagent and
sample quantities were enlarged. 0.2 mL extraction of
black soybean seed coat was added to 0.6 mL water; 6
mL FRAP reagent was heated up to 37°C (made up of
10 mmol L-l TPTZ, 20 mmol L-' FeCl,, and 0.3 mmol
L-' sodium acetate, in the proportion 1:l:lO) in turn,
and was shaken up and allowed to stand for 4 min.
The absorption value was determined in 593 nm. At
the same time, 0.1-1 mmol L-I FeSO, was used to es-
tablish the calibration curve, the correlation coeffi-
cient r = 0.9994, and the regression equation y=0.3 117x-
0.0006. The FeS0,content contained in one gram of
extraction was used to represent TAC, and was ex-
pressed as mmol g-'.
Total phenolics assay According to the assay (Wu et
al. 2006; Djeridane et al. 2006),1 mL extraction of
black soybean seed coat was added to 1 mL Folin
Ciocaiteu's reagent and 2 mL sodium carbonate in turn,
shook up and maintained at 20-25°C for 60 min. The
absorption value was determined in 765 nm. The cali-
bration curve was established using gallic acid as the
standard sample, with correlation coefficient r= 0.9973,
and regression equation y = 0.056~-0.021. The gallic
acid content was used as the total phenolics content,
and was expressed as mg g-'.
Anthocyanin contents assay According to the assay
(Fuleki and Francis 1968; Dong et al. 2003), the ex-
traction solution was diluted with 60% ethanol (pH
3.0), and the absorption value was determined in 535
nm. The calculation formula was: MF= A,,, nm x V x
N/98.2/m, where, MF represents the anthocyanin
content in black soybean seed coat (mg g-l), AS3,nm
represents the absorption value in 535 nm, V repre-
sents the total volume (mL), N represents the dilution
multiple, 98.2 represents the extinction coefficient in
535 nm, and m represents the mass of black soybean
92007,CAAS. All rights reserved.Publishedby Elsevierltd.
 152 XU Jin-rui et al.

seed coat (8). significant genotype differences exist among

antioxidations and the total phenolics and anthocyanin
Statistical analysis contents among black soybean accessions. Further
analysis of accessions distribution (Figs. 1-3) indicated
that TAC of black soybean accessions presented nor-
According to the distribution distance of black soybean
mal unimodal distribution, with TAC value of 76.3%
TAC, the total phenolics, and anthocyanin contents
total black soybean accessions distributed between
among black soybean accessions, the bar diagram was
made by the expressions (max-min)/N, and the per-
centage that the accession numbers in each distribution 25 23
distance occupied the total sample numbers was
20
reckoned. The significant analysis of differences of
TAC, the total phenolics, and anthocyanin contents in
127 black soybean accessions adopted the t-test for
average; the clustering analysis of TAC, and the total
phenolics and anthocyanin contents of 127 black soy-
bean accessions adopted fast clustering method; cor- 047 0.78 1.09 1.43 1.72 2.03 2.34 2.66 297 3.28 3.59
relation analysis adopted the correlation coefficient
TAC (mmol g ')
methods among TAC, the total phenolics, and antho-
cyanin contents. The AND SPSS 11.5 software pack- Fig. 1 Distributionof TAC in seed coats of black soybean accessions.
age was used to set up the database in data processing
and statistical analysis.

RESULTS 3o
25 c 23 23 -
-
24

15
Antioxidations and the total phenolics and 1
anthocyanin contents in different black soybean 9

accessions
- -
The ranges and coefficients of variation (CVs) from 10.0 17.5 25.0 32.5 40.0 47.5 55.0 62.5 70.0 77.5

TAC, the total phenolics, and anthocyanin contents of Total phenolics content (mg 9')
127 black soybean accessions of different eco-regions
and sowing types (Tables 1 and 2) showed that the Fig. 2 Distribution of the total phenolics content in seed coats of
ranges and CVs were relatively large, indicating that black soybean accessions.

Table 1 The ranges and CVs of TAC, and the total phenolics and anthocyanin contents in seed coats of black soybean accessions from
different ecological regions

Total 0.44-3.55 35 7.05-74.82 35 0.22-1.87 34

Dongheichun 1.25-2.95 27 21.61-70.0 31 0.6X-1.40 21
Beifangchun I .12-3.18 29 20.89-69.02 32 0.45-1.79 34
Huanghuaichun 0.67-3.55 51 7.05-71.43 50 0.34-1.74 59
Huanghuaixia 1.46-3.50 20 33.21-74.82 17 0.55-1.87 28
Nanfangchun 0.44-2.60 48 11.34-64.64 44 0.22-1.71 43

Nanfangxia 0.72-2.56 26 17.86-57.59 27 0.36-1.47 28

Nanfangqiu I 33-2.70 19 37.23-53.04 12 1.10- 1.68 16

@2K17.CMS. All rightsresewed. PublishedLY ElsevierLM.

Correlation Between Antioxidation and the Content of Total Phenolics and Anthocvanin in Black Soybean Accessions 153

39 Antioxidation and the total phenolics and

n anthocyanin contents in different eco-regionsand

25
I I sowing types of black soybean

20
15
10
5 the max variation of TAC occurred in the Huanghuaichun
0 eco-region, followed by the Nanfangchun eco-region,
0.31 0.50 0.69 0.88 1.06 1.25
Anthocyanin content (mg g')
Fig. 3 Distribution of the total anthocyanin content in seed coats
of black soybean accessions.
1.25 and 2.81 mmol g-I. The total phenolics contents
showed the same normal unimodal distribution as TAC
among 21.3-58.8 mg g-', in which inclusive accessions
accounted for 77.9%; however, anthocyanin contents
showed normal skew unimodal distribution, which ac-
counted for 75.6% total accessions among 0.59- 1.34
mg g-l, indicating that the majority of the total pheno-
lics and anthocyanin contents of black soybean acces-
sions concentrated on 21.3-58.8 and 0.59-1.34 mg g-',
respectively. The strongest one of TAC in 127 black
soybean accessions was shuilizhan from the
Huanghuaichun region, and the highest one of the total
phenolics and anthocyanin contents was both Zaoshu
Heidou from the Huanghuaixia region.
The results showed (Table 1) that the range and CVs in
different eco-regions of black soybean were significant,
1.44 1.63 1.81
while the min occurred in the Nanfangqiu eco-region,
and the variations of the total phenolics and anthocya-
nin contents were entirely similar to the variation of
TAC. The results also showed (Table 2) that the range
and CVs in different sowing times of black soybean
were great; the max variation of TAC, the total
phenolics, and anthocyanin contents all occurred in
spring, while the min all occurred in autumn.
Further comparison of the average values of TAC,
the total phenolics, and anthocyanin contents of black
soybean/yellow soybean, yellow cotyledon/green
cotyledon, Dongbeichun/Beifangchun, Dongbeichunl
Nanfangchun, Dongbeichun/Nanfangxia, Beifangchd
Nanfangchun, BeifangchudNanfangxia, and
NanfangchudNanfangxia showed (Tables 3 and 4) that
the differences in different eco-regions and the sowing
types of black soybean accessions were great, and the
average values of TAC, the total phenolics, and antho-
cyanin contents of black soybean accessions were re-
markably higher than that of yellow soybean, reaching

Table 2 The ranges and CVs of TAC, and the total phenolics and anthocyanin contents in seed coats of black soybean accessions at different
sowing times
TAC Total phenolics Anthocyanin
Sowing time
Range (-01 g-l) . CV (46) Range (mg g-9 cv (46) Range(mgg') , CV(46)
Total 0.44-3.55 35 7.05-74.82 35 0.22-1.87 34
Spring 0.44-3.55 39 7.05-71.43 39 0.22-1.79 36
SUmmer 0.72-3.50 31 17.86-74.82 32 0.36-1.87 30
Autumn 1.58-2.70 19 37.23-53.04 12 1.10-1.68 16

Table 3 Differences of TAC. and the total ohenolics and anthocvanin contents in seed coats of black sovbean between eco-types
~~~

TAC Total phenolics Anthocyanin

Eco-type
Mean (mmol e t value Mean (mpa I ) f value Mean (ma E I ) I value
Black soybeanlyellow soybean 1.9410.03 31.56" 4 1.4213.32 28.8" 0.9610.19 32.95"
Yellow cotyledonlgreen cotyledon 2.0111.14 3.48" 42.73134.09 3.15" 0.9810.83 1.85
Springlsummer 1.8612.04 1.45 39.28143.9.5 1.75 0.9310.97 0.7
Springlautumn 1.8612.08 1.26 39.28145.49 2.20* 0.9311.34 2.94"
Summerlauhnnn 2.0412.08 0.25 43 .95145.49 0.53 0.9711.34 3.01"

'p.tO.05: "'P<0.01. The same as below.

632007,CAAS.All tights reserved. Publishedby Elsevier Ltd.

154 X U Jin-rui et al.

the most significant (P < 0.01) level, indicating that
antioxidation and active material contents of black soy-
bean are obviously superior to yellow soybean. As far
as the cotyledon type is concerned, the TAC and the
total phenolics contents of yellow cotyledon type were
both higher than that of the green cotyledon type, but
the difference of anthocyanin contents was not
significant. As far as the sowing type was concerned,
the best was the autumn sowing type, followed by the
summer sowing type, and the worst was the spring
sowing type in TAC, with no significant difference
among the three types. Regarding the total phenolics
contents, the best was the autumn sowing type, fol-
lowed by the summer sowing type, and the worst was
the spring sowing type, in which the difference be-
tween the autumn and spring type reached a significant
(Pc0.05) level. Regarding the anthocyanin contents,
the best was the autumn sowing type, followed by the
summer sowing type, and the worst was the spring
sowing type, in which differences of the autumdspring
and the autumnhummer types both reached the most
significant (Pc0.01) level. As far as the eco-region
type was concerned, the best was Beifangchun, fol-
lowed by Dongbeichun, and the worst was Nanfangchun
in TAC, in which differences of Dongbeichunl
Nanfangchun, Dongbeichun/Nanfangxia, and
Beifangchun/Nanfangchun, Beifangchuaanfangxia
reached the significant (Pc0.05) level and the most sig-
nificant ( P < 0.01) level, respectively. Regarding the
total phenolics contents, the best was Beifangchun, fol-
lowed by Dongbeichun, and the worst was Nanfangchun,
in which differencesof DongbeichudNanfangchun and
BeifangchunINanfangchun, and Beifangchunl
Nanfangxia reached the significant (P<0.05) level and
the most significant (P<O.Ol) level, respectively. Re-
garding the anthocyanin contents, the best was
Beifangchun, followed by Dongbeichun, and the worst
was Nanfangchun, in which the differences were not
significant among the three types.
In summary, as far as TAC, total phenolics, and an-
thocyanin contents are concerned, the general tendency
is that the Beifangchun is the best, followed by
Dongbeichun, and the worst is Nanfangchun. This
may be caused by the differences in the ecological con-
ditions of the habitat and the cultivation techniques, or
may be determined by the genetic differences among
the various accessions.
Cluster analysis of antioxidation and the total
phenolics and anthocyanin contents of black
soybean accessions
127 black soybean accessions were clustered into 6
clusters using the fast clustering method, each con-
taining 3,24,20,31,37, and 12 accessions, respectively.
The clusters with the majority accessions were the
fourth and fifth clusters, accounting for 24.4 and
29.1% of the total black soybean accessions,
respectively. The first and the sixth clusters contained
relatively fewer accessions. The clustering results were
not related to the habitats of accessions.
The results of analyzing the final cluster centers of
TAC, the total phenolics, and anthocyanin contents of
various clusters (Table 5 ) indicated that the TAC, the
total phenolics, and anthocyanin contents of the first
cluster were the lowest, while those of the sixth cluster
were the highest; those of the second cluster were higher
than those of the first cluster, those of the sixth cluster
were only higher than those of the third cluster, which
were higher than those of the fourth cluster, and those
of the fourth cluster were higher than those of the fifth
cluster, relatively. Comparatively speakmg, the order
of quality of the six clusters was: sixth > third > fourth
> fifth > second > first.

Table 4 Differences of TAC, and the total phenolics and anthocyanin contents in seed coats of black soybean between eco-regions
TAC Total phenolics -- Anthocyanin
Eco-region
Mean (mmol g-1) f value Mean (mgg-1) T value Mean (mg g-'1 t value
Dongbeichnn/Beifangchun 1.94/2.19 1.33 40.87/45.33 1.05 0.98/0.10 0.22
DongbeichunlNanfangchun 1.9411.46 2.60' 40.87/3 1.61 2 34' 0.9W0.83 1.74
Dongbeichun/Nanfangxia 1.9411.67 2.07' 40.87/35.25 1.77 0.98/0.88 0.67
BeifangchudNanfangchun 2.19/1.46 3.46" 45.33131.61 3.12" 0.10/0.83 1.56
BeifangchurdNanfangxia 2.19/1.67 3.37" 45.33135.25 2.93" 0.10/0.88 1.41
Nanfangchunrnanfangxia 1.4610.67 1.22 31.6U35.25 1.07 0.83/0.88 0.58
Correlation Between Antioxidation and the Content of Total Phenolics and Anthocyanin in Black Soybean Accessions 155

Table 5 Final cluster centers

Cluster 1 2 3 4 5 6
TAC 0.56 1.16 2.52 2.10 1.73 3.12
Anthocyanin 0.31 0.7 1 1.13 1.06 0.86 1.41
Total phenolics 10.18 23.57 56.62 45.43 35.35 67.92

The distances between the focus of the clusters (Table tive correlation existed between the TAC and the total
6) showed that the largest distance was between the phenolics and anthocyanin contents of black soybean,
first and the sixth, followed by the distances between respectively, with the correlation coefficient reaching
the first and the third, the second and the sixth; the the most significant level ( P < 0.01), inhcating that the
relatively less distances were between the first and the most significant correlation existed between the
second, the third and the fourth, the second and the antioxidation and the total phenolics and anthocyanin
fifth, the third and the sixth; the least distance was be- contents of black soybean, that is, the antioxidation of
tween the fourth and the fifth, while the genetic dis- black soybean is related to the total phenolics and antho-
tances among other clusters were moderate. cyanin contents of its seed coat. It can also be inferred
from Fig.6 that the most significant positive correlation
The correlations between the antioxidation and existed between the TAC and the seed coat rate.
the total phenolics and anthocyanin contents of
black soybean .v = 0.0427~+ 0.1667
r=0.9153
3.5
The correlations between the antioxidation and the total
phenolics and anthocyanin contents of black soybean - 3.0 -
-
b 2.5 -
are shown in Figs.4,5.
It can be inferred from Figs.4, 5 that significant posi-
g 2.0 -
v
V 1.5 -
2 1.0 -
0.5 -
Table 6 Distances between the final cluster centers
01
Cluster 1 2 3 4 5 0 20 40 60 80
2 13.41
3 46.49 33.08 Total phcnolics content (mg g I)
4 35.29 21.89 11.20
5 25.21 11.80 21.28 10.09
Fig. 5 The correlation between TAC and the anthocyanin content
6 57.81 44.40 11.32 22.51 32.60
in the seed coat of black soybean.

v = 1.616xt0.3848 ,v = 1.2072~+ 8.1368

4.0 r r = 0.7736 r = 0.2943
3.5 -
3.0 - 3
v
20.0
r
** +
2.5 - 0
5 15.0 -
2.0 - c
8 -
1.5 - " 10.0
B
1.0 - 8
5.0 -
0.5 -

0 0.5 1 1.5 2
, 0.0 '
0.00 I .on 2.00 -3.00 4.00
Anthocyanin content (mg g ') TAC (nimol g')

Fig. 4 The Correlation between TAC and the total phenolics content Fig. 6 The correlation between the seed coat rate and TAC in the
in the seed coat of black soybean. seed coat of black soybean.

02037. CAAS.All rightsreserved.Publshed by ElsevierLld.

156
DISCUSSION
The difference of antioxidation and the total
phenolics and anthocyanin contents of black
soybean between accessions
The great variation range and coefficient of TAC, the
total phenolics, and the anthocyanin contents of black
soybean existed among accessions and types; the gen-
eral tendency was that the TAC, the total phenolics,
and the anthocyanin contents of Beifangchun acces-
sions were higher than those of Bongbeichun ones, and
those of Nanfangchun accessions were the least, which
may either be determined by genetic genes or may be
caused by planting conditions and ecological
environments. Zhang e f al. (1994, 2005) adopted the
periodic seeding method and analyzed the relation be-
tween the anthocyanin of black rice and the ecological
environment at the heading and grain filling stages, and
the result showed that significant negative correlation
existed between the anthocyanin contents of black rice
and the average daily temperature and the average daily
sunshine hours, whereas significant positive correla-
tion existed between the anthocyanin contents of black
rice and the average daily relative humidity 30 days af-
ter heading. Namely the low average daily temperature;
short hours of sunshine and great relative humidity are
favorable to accumulate the anthocyanin of black rice.
The air temperature in the north was significantly lower
than that in the south, and the growth duration of the
former was usually longer and the grain filling rate was
slower; all these factors are favorable for the accumu-
lation of anthocyanin in black soybean. Further, the
fact that the latitude was higher in the north than in the
south can be attributed to this reason. The analysis of
the correlation between chemical components and eco-
logical distributions of soybean accessions in China
showed that significant positive correlation existed be-
tween the fattiness content and the habitat latitude of
soybean. However, significant negative correlation ex-
isted between the protein content and the habitat lati-
tude of soybean and the variation of mineral compo-
nents also displayed the geographical distribution rule,
namely, the increase or decrease from north to south
(Fei et al. 1983; Xu et al. 1993; Wang 1979; Wang et
al. 1979; Dong and Zhang 1997; Ren et al. 1987).
Xl! Jin-mi et aE.
These results were similar to the analysis of black soy-
bean active components in this study, and thus the im-
provement of black soybean accessions with the inten-
sive antioxidant property must combine the genetic im-
provement with the ecological environment control.
Breeding of antioxidation black soybean
accessions with abundant anthocyanin
The research showed that the distributions of TAC,
the total phenolics, and anthocyanin contents of 127
black soybean accessions displayed normal unimodal
or normal skew unimodal characteristics, showing that
the genetic features of quantitative characters were
controlled by micro effect multiple genes. In breed-
ing practices, accessions with large or moderate ge-
netic distances may be selected for single crossing or
multiple crossing by referring to the clustering result,
where 127 black soybean accessions were clustered
into 6 clusters in this study. With the development of
the living standard, the appraisal and selection of na-
ture resources with strong antixoidative activities has
become the topic of biology, iatrology, and food
science. This research showed that the most signifi-
cant positive correlation existed between the TAC and
the anthocyanin content of black soybean, namely,
the antioxidation of black soybean accessions with
deeper (color) anthocyanin in seed coat was stronger,
thereby providing useful information for selecting new
black soybean accession with strong antioxidation.
Breeding methods such as multiple crossing and even
recurrent selection will be adopted in breeding, the
successive multiple generation directional selection for
dark colored individual of seed coat will be conducted
with the seed coat color as the primary objective, and
antioxidation in higher generations can be evaluated,
so that new quality black soybean accessions (lines)
with higher antioxidation can be selected. This is also
helpful in accumulating more cumulative genes related
to anthocyanin and antioxidation, so as to combine the
dark colored seed coat and the high antioxidation.
CONCLUSION
The range and coefficient of variation between the TAC
02007,CAAS. All rights resewed. Publishedby Elsevier LM.
Correlation Between Antioxidation and the Content of Total Phenolics and Anthocyanin in Black Soybean Accessions
and the total phenolics and anthocyanin contents of dif-
ferent black soybean accessions are great, indicating
the existence of genotypic variance. Normal unimodal
or normal skew unimodal between accessions indicates
that genetic control is dominated by minor genes, and
exhibits genetic features of quantitative characters.
Significant differences of TAC, the total phenolics, and
anthocyanin contents of black soybean existed among
accessions and types, and the general tendency was
that the TAC, the total phenolics, and anthocyanin con-
tents of Beifangchun accessions were higher than those
of Dongbeichun ones, while those of Nanfangchun ac-
cessions were the least. The strongest one of TAC in
127 black soybean accessions was Shuilizhan from the
Huanghuaichun region, and the highest one of the total
phenolics and anthocyanin contents was both Zaoshu
Heidou from the Huanghuaixia region.
By adopting the fast clustering method, 127 black
soybean accessions could be clustered into 6 clusters
and the clusters with the majority accessions are the
fourth and fifth clusters. The order of quality of the
six clusters is: sixth > third > fourth > fifth > second >
first.
Significant correlations exist between the TAC and
the total phenolics and anthocyanin contents,
respectively, where the correlation coefficients of both
reached the most significant level (P<0.01), indicating
that the total phenolics and the anthocyanin of black
soybean seed coat is the major material basis for
antioxidation.
Acknowledgements
This work was supported by the National Natural Sci-
ence Foundation of China (30200171).
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