Analysis of Genetic Distance among Four Glycine

.林學報
50(3) : 55-65(2001)
- 55-
Analysis of Genetic Distance among Four Glycine Species
Collected from Taiwan : Revealed by DNA Polymorphisms
Tzu-Kai Lin 1)
Shu-Tu Wu2) Tzer-Kaun Hu3)
Fu-Sheng Thseng2 )
(Accepted for publication: June. 28 , 2001)
ABSTRACT: Subgenus Glycine are generally distributed in Australia, Micronesia, and southem
Chi悶，
but G.
tomente /l a and G. tabacina can all be found in Taiwan , which is the northem most distribution area of subgenus
叫ycine.
G. formosana of subgenus Soja are also found in Taiwan. However, these three wild species of soybeans
are very complícated in the variation of morphological characters. In the past , their relationships in taxonomy were
rather
unclear 甘le
researcher have investigated the
mo巾hological
characters of Taiwanese wild soybean and
re- c1 assified these three species mentioned above into four species , G. tomente /l a, G. tabacina , G. dolichocarpa ,
and G.
m叫 ssp.
formosana (Hosokawa) Tateishi et Ohashi. In this report , four species and G. soja was analyzed
their interspecies relations by random amplified polymorphic DNA (RAPD).
The results of RAPD analysis showed that G.
s句a
and G. max ssp. formosana are the most c1 osely-related
among the five species , and that G. tabacina is the least related to the rest of the species. As a result , they can be
classified into two groups: A) G. soja and ‘ G.
m囚 ssp.
formosana ; B) G. tomentella , G. tabacina , and G
dolichocarpa . In group A , the similarity coefficients are between 0.50 and 0.65 , while being between 0.09 and 0.17
in group B.
Key words: Coefficients of genetic similarity; Glycine species; Random
ampli 釘 ed
polymorphic DNA (RAPD).
The subgenus Glycine contains at least 17 species ,
all being perennial , which are mainly distributed in
INTRODUCTIO
Australia, Philippine , Southem Pacific archipelago ,
southem
Chi 悶，
and Taiwan (Singh and Hymowitz
1) Department of Agronomy, Chung Hsing University. (Present address: Department of Horticulture, Taiwan
Agricultural Institute , W ufe嗯， Taichung 413 , Taiwan.)
2) Professor of Department of Agronomy , Chung-Hsing University , respectively
3) Associate professor of Department of Agronomy , Chung Hsing University. (Co汀esponding author).
56 -
Anal:ysis of Genetic Distance among Four G扯ineSpeó的 CoUected from Taiwan : Revealed by DNA Pol)'mo帆sms
1985; Hymowitz and Singh 1987; Tindale 1984, 1986
investigate the relationships among wild soybean
a, b; Tindale and Craven 1988 , 1993; Tateishi and
species ,
Ohashi 1992). Th ree subgenus Glycine species , G
and G. formosana collected from Taiwan
G一 tabacina.
G. tomentella, G. dolichocarpa.
tabacina. G. tomentella. and G. dolichocarpa. are
known to be native in Taiwan. G. tabacina is only
MATERIALS AND
found in Pong-Fu archipelago. According to the
investigation on G. tabacina populations in eight areas
METHODS
of Pong-Fu by Yiu and Th seng (1 997b), continuous
variations in stems, leaves , and flowers exists among
different populations. However, further investigation
on isozymes showed little variation among populatíons.
A.
Plant material
G. tomentella are distributed in Taichung , Pingtong,
With the exception of nine accessions of G. soja
and Taitung ( Tang and Lin 1962; Tateishi and Ohashi
introduced from China, Japan , and Korea , the other
1992). Yiu and Th seng (1 997 a , c) found significant
wild species , G. formosana. G. dolichocarpa. G
variations of plant characteristics among Taichung ,
tomentella and G. tabacina were all collected 介om
Kenting (Pingtong) , and Taitung populations. Hayata
Taiwan and nearby islands. A total of twenty seeds
(1917) named a wild soybean found in Taitung as G.
from accession were grown to the flowcring stage in
tabacina , however, Chuang and Huang (1966)
greenhouse
renamed this wild soybean found in south-eastem
Taiwan as G. tomentosa. Later, Huang and Ohashi
(1 977) and Huang and Huang (1987) named it as G
B.
DNA extraction
DNA was extracted using a modified version of
G
Doyle et al (1990). Leaf materials (1 g fresh weight)
dolichocarpa (Tateishi and Ohashi 1992; Thseng et al
were ground to fine powder in liquid nitrogen. The
1997).
powdered leaf tissue was transferred to a
tomentella.
At
present ,
it
is
classi 日 ed
as
Doyle and Beachy (1 985) analyzed variation in
beak 汀，
and
5ml of pre-heated extraction buffer (2% CT AB; 1.4 M
2-mercraptoethanol 、 20
18S. 25S rDNA repeat length and restriction enzyme
NaCI , 0.2%
site locations among the Glycine subspecies. They
mM Tris-HCI , pH
found that G. tabacina from Taiwan had repeat length
at 的'(， 5
of 9.2 kbp , being significantly different from 8.0 kbp
was added , and the supe tJ1 atant was collected by
of those from Australia , and that G. tabacina from
centrifugation at 10000 xg for 5 minutes at 4'C. Th en ,
Taiwan had one more EcoR 1 enzyme site than those
5 ml of Chloroform-isoam ly alcohol was added , and
from Australia. Differences in seed proteins and DNA
the cell debris was removed by another 10 minutes of
tomentella
centrifugation at 4'C. The DNA was prccipitated by
were also found
among Taitung G.
population and other populations (Hsing et al. 1995)
G. tomentella and G. dolichocarpa had been classifíed
into different groups by RAPD analysis
(甘lseng
et al
1997). In this experiment , we used RAPD analysis to
mM EDTA , 100
= 8.0) was added. After 30 minutes
ml of Chloroform-isoamyl alcoho l. (24: 1 v/v)
the addition of 3.3 ml of isopropanol and recovered by
centrifugation
for
5
min utes
incubation in a
freezer(一 20'C)
at
10000xg
after
for 3 hours. Thc pellet
農林學報
50(3) :
55~65(2001)
-
57 一
was dried and re-dissolved in 2 ml TE buffer (1 0 mM
estimate Jaccard similarity coefficients
Tris-HCl , ImM EDTA , pH = 7 .4), 0.2 ml 2M NaCl ,
where a = band number of positive coincidences in
and 5 ml a1c ohol(95%) for 1 hour at -20'C. Then the
OTUj (1
precipitate was collected by centrifugation at 10000 xg
number of positive incidence only
for 5 minutes at 4 'C. Th e pellet was re-dissolved in 5
band number of positive incidence only in OTUj
centrifugation 剖
(Dunn and Everitt 1982). Th e similarity coefficients
12000 xg for 5 minutes at 4 'C. The final pellet was
calculated by SAS computer package were used to
ml alcohol (95%) and re-collected by
dried and dissolved in 0.5 ml TE buffer. Th e DNA
concentration was determined using a fluorometer
following the procedures supplied by the
j; i, j were accessions codes) , b = band
in OTUi , and c =
construct a dendrogram using UPGMA (Unweighted
Pair-Group Method with Arithmetic mean).
manufact叮叮.
The extracted DNA was stored at 4'C in a cooler.
C.
'Î'
aJ(a+b+c) ,
RESULTS AND DISCUSSION
DNA amplification
A set of 20 random 10-mer primers (Operon
Technologi 的 Inc.
USA) was used in single-primed
A set of 20 random 10-mer primers was used in
the RAPD analysis performed on 18 accessions of
PCR reactions to generate polymorphisms. A 25μr 1
rcaction was sct up as follows: 0.2mM dNTPs , 0.2μM ，
3ngμ1
1
Genomic DNA and 0.04Uμr l template
subgenus Soja and Glycine species. In 246 amplified
polymo中hism
bands , a total of 240 bands showed
(98
%) (Table 1). In all 20 primers, OPB-19 produced the
highest number of bands. Figure I-A shows the results
DNA in IX buffer Th e reaction was ovcrlaid with
mineral oil ,
and the mix was heated to 94'C for 5
of RAPD analysis on OPB-19 primer: A total of 18
bands were produced , and
pol 戶norphisms
were
Ampli 日 CatIOn
detected among all the accessions tested. Figure 1-8
proceeded on a DNA Thcrmolcycler (Perkin-Elmer)
shows the results ofRAPD analysis on OPB-16 primer:
42'C 、 3
only the accessions of subgenus of Soja species
minutes to denature the template DNA.
for 43 cycles of 1 minute at 94'C , 2 minutes at
minutes at 72'C , and the tìnal step of 10 minutes at 72
℃
ηle
total volume of the reaction was loaded onto a
showed the presence of 1700 bp bands.
Th e similarity coefficients among accessions are
listed in Table 2 , and they can be classified into two
1.5% agarose gcl containing 0.3μg mr l ethidium
groups by cluster analysis (Fig. 2). Group A being the
bromide in 0.5X TBE buffer (pH = 8.3 , with 45 mM
12 accessions of subgenus Soja
speci 的，
Tris-base , 45 mM boric acid and 1 mM EDTA). The
similarity coefficients between 0.5
~
samples wcre separated by electrophoresis at 11 OV for
being the 6 accessions of subgenus
3.5 hours , then the gels were photographed under UV.
have similarity coeftïcients between 0.18
all have
0.94. Group B
G砂cine
species , all
~
0.95. Th e
similarity coeftïcient between these two groups is 0.12 ,
D.
Data analyses
Reproducible DNA bands
which indicates that the two groups, subgenus Soja
were scored
for
and Glycine species are , the most
distant妙-related.
G
presence (1) or absence (0) of amplitìcation products.
lomentella is the c1 0sest one to subgenus Soja in
Palr-Wlse comparisons of sample were used to
subgenus
G今czne specles 甘le
similarity coeftïcients
一 58
Analys的 of Genetic Dis恤.ce
-
among Four Glycine Species CoUected from Taiwan : Revealed by DNA Polymorp地m
between G. formosana and G. tomentella are 0.16 -
between 0.12 - 0.15 , which shows that G. formosana
。 18 ，
and G.
and those of G. s句。 and G. tomentella are
tomentella 訂e
closer than other species are
Table 1. Th e total numbers of amplified DNA fragment (band) and the numbers of polymorphic band for each
primer used in RAPD analysis of 18 accessions of five wild species of Glycine
Operon
Sequence (5 to 3 )
Total bands'
Operon
pnmers
Sequence (5 to 3 )
Total bands'
pnmers
OPB-OI
GTTTCGCTCC
11 (11)
OPB-II
GTAGACCCGT
11 (11)
OPB-02
TGATCCCTGG
11 (1 0)
OPB-12
CCTTGACGCA
11 (1 0)
OPB-03
CATCCCCCTG
12 (1 2)
OPB-13
TTCCCCCGCT
10 (9)
OPB-04
GGATCGGATG
14 (1 4)
OPB-14
TCCGCTCTGG
15 (1 4)
OPB-05
TGCGCCCTTC
10 (9)
OPB-15
GGAGGGTGTT
14 (1 4)
OPB-06
TGCTCTGCCC
15 (1 5)
OPB-16
TTTGCCCGGA
10 (10)
OPB-07
GGTGACGCAG
17 (1 7)
OPB-17
AGGGAACGAG
7 (7)
OPB-08
GTCCACACGG
12 (12)
OPB-18
CCACAGCAGT
16 (1 6)
OPB-09
TGGGGGACTC
9 (9)
OPB-19
ACCCCCGAAG
18 (1 8)
OPB- lO
CTGCTGGGAC
12 (11)
OPB-20
GGACCCTTAC
12 (12)
Total
246 (240)
a) Number within the parenthesis indicates polymorphic band
E.I.-'
，且，
1
，1
EA
， IEI
1
，1
1
， l'1
17
， B且，自且，
可缸，
， IBI'l
過
Y
、3
d
司，'，3
令
治斗『
‘
而yh 司'，但情
J
島、 3
aa
冉、d 令、d
弓，.司4.l'IBI--zJ
，
Iz--a7B
缸，
2
、3/OOO
『 d5
A斗
且仇 υAUIE3111000J4
且，
， galz--z1
弓，
-7
可 &7.
，且，
司且
1.I'ihy
，
、
J44
， 11111
， ISIt-
、
34
咀 SA--2.Ba--a
、3
U 白UAU
勻，
uQFQJt
il﹒仇
GE-'l'1.ItIAυAυ'Ig1
勻，
f-'Iz--a1JnUEIl
-E且，自且，
弓，缸司3
3
「J
弓r
-AVAUAυAOAOAUAUAVAUAMVAUnvnUAVAU
-qb
-勻，
'F、-2ga--且
團
句/呵，可
， It--a'121
ll-qdAdTA 啥?.，
IElv--IAU
"、
，益，
1
MFEE--11
， 121
l
21
、3
J
仇"/利pb 仇M/ 呵，，
1dny/011
-AHUAHVAHUAHVAUAUAHVAυAυAHVAHVAHVAHVAU
A判，
--oo7
勻，但?但勻，-、 J
令、
J'EaAHV
勻，
『 u
ai
--
-AU 仇。 AUAOAUAVAUnUAOAυAVAVAV
，A
且可 AUTfo74
JhvAUQ/Q/
司
I--LVLUζUAaT
dA
戶，EAOXUOXU'
-『
qd
2-6544777767rb
'I
d 呵，
仇。
Aυ' 、 d
， UtozO/OFAU
句/勻
『 /O/O/0
A
-AHVAUAHVAHVAHυAHU仇HVAHVAOAυ
吟
，
31ν
-AHVAHUAUAHVAHUAHunHvnυGV
-『
-nynyA
，
。一 555777667
1
句32
，力 66677b
-7 缸，
9-6
。OOO
-AVAUAUAυAUAUnv
-ζυ
且可司3
-AUAVAUAυAOnυAU
-AUAVAUAUA
7τ355777
JAY
弓
『 d 勻，
-fOAaYAaT/O/ 。
『 d
-A且可司‘d
-AυnunVAUAυ
-AHvnHUAHυAHV
仇。
7 G. s句。
8 G. soja
9 G. soja
10 G. s句。a
11 G. s句。
12 G. s句G
13 G. tomentella
14 G. tomentella
的 G. dolichocarpa
16 G. labacina
17 G. labacina
18 G. tabacina
qd一『 d
-AUAυ
， Aυ'l
-A斗
勻，但 or
Table 2. Similarity index between each pair of 18 accessions offive wild species of G砂cme.
4-555 6-55577
l
l
3-90
8-5557766
1
l- --oooooooooaa
l-oaoo
:.ill-llli--l
Accession
-AV
-AUTAVMnynyhy
-AVAHVAHUAOAOAHVQVAHVAHVAHVAV
-AHVAHVAHVAHUAHUAHVAHVA
-AUAUnHvnununv •.
.
.
1 G. formosana
2 G. formosana
3 G.formosana
4 G. soja
5 G. s.句a
6 G. so)，。
0.84 0.80
。 .89
農林學報
50(3) :
-59-
55~65(2001)
拉紛前
、企 \t.....
ψ".~C)..~().i:>~-$-v勻。
q
~、:.s:
~t"""
.A
MAdcd::34叭叭-抖抖〈咚WV
(bp)
2072-
600-
車站﹒
A
'。
。抖。。1/.otfDC丸。.'I'<>_\'I'~
M
(坤)
2072-
600軍沁.
B
Fig. 1.
RAPD profiJes in five GJycine species.
A: primer OPB‘ 19
B: primer OPB-16
M: JOObp DNA Ladder
一 60
-
0.00
An alysis of Genetic Distance among F，ωr Glycine Species CoUected from Taiwan : Revealed by DNA Polym呵地問
0.25
0.50
0.75
1.00
G.formosanα
G.formosαm
G.formosαm
G.s吋α
G.
s吋α
G. s吋α
G.
s吋α
G.
s吋α
G.
s吋α
G. s吋α
G.
s吋α
G.
s吋α
G. tomentella
G. tomentella
G. dolichocarpa
G. tαbα cina
G.
tabαcin α
G. tabacina
Fig. 2.
Dendrogram of G. formosana , G. soja , G. fomentella , G. dolichocarpa and G. tabacina based
on Jaccard 's genetic similarity coeffïcients by using UPGMA method.
農林學報
50(3) : 55-65(2001)
一 61 一
1n the group B , two accessions of G. tomentella
indicated that G. formosana is close to G. tomentella
show high similarity and are closely related. High
both genetically and geographically.
similarity coefficients are also found among three
bands produced in the experiment , as many as 240
accessions of G.
bands showed polymorphism (98%) , which means that
tabacina , which shows small
Howev缸，
of 246
variations among these three species. However, the
the accessions tested so far are h_i ghly genetically
RADP analysis of OPB-07 showed several distinct
divergen t. Moreover, from the similarity coefficient of
polymorphic bands among three species G. tomentel旬，
0.12 and distantly genetic re1ationship between these
G. dolichocarpa , and G. tabacina in the group
two subgenera, it is not clear whether G. tomentella is
B(Figure 3-A).
1t indicates that 750 bp, 510 bp, 420
the ancestor of G.
s句a
and G. formosana or no t. 1n
bp , 350 bp, and 310 bp are exclusive to G. tomentella.
fact , it had been discovered that G. lomen/ella is c1 0ser
650 bp , 600 bp , 450 bp , and 200 bp are exclusive to G
to Soja genetically than G. tabacina is (Hui and
dolichocarpa , and 1500 bp , 900坤， 850 bp , and 530 bp
Zhuang 1996). It was also suggested that a G. curvata
are exclusive to G. tabacina.
in subgenus
Among these three species , G. tomenlella is
closer to G. dolichocarpa than to G.
tabacin日.
G秒'CÍne
species was c1 0ser to subgenus
Soja species. However, because they have different
Besides,
genome (Kolipara et al. 1997), further investigation is
the similarity coefficient between G. dolichocarpa and
necessary to clarify the relationship between those two
G. lomentella and the similarity coeffícient between G.
subgenera.
dolichocarpa and G. tabacina are higer than that
For the subgenus Glycine species native in
betwecn G. /omen/ella and G. /abacina , which shows
Taiwan , the classification of Taitung-originating G.
that the parentage of G. dolichocarpa Ii es between G
dolichocarpa has been wi1dly disputed. As early as
/omenlella and G. tabacina. Figure 3-B shows the
1917, Hayata had determined the wild soybean in
result of RAPD analysis on OPB-14 primer. It is found
Taitung as G. tabacina. Later, It was determined as G
that 350 bp bands only appears in G. /omentella and G
tomentella by Huang and Ohashi (1977) and Huang
dolichocarpa , and 800 bp and 850 bp bands appear in
and Huang (1987). Nowadays, it has been identified as
G. dolichocarpa and G. labacina , which shows G
a new species and is named G. dolichocarpa (Tateishi
dolichocarpa is similar to both G. /omentella and G
and Ohashi 1992). 1n our experiment, G. dolichocarpa
/abacina. It had been proposed that G. /omentella or G
indeed showed significant difference to G. tomenlella
tabacina could be the ancestor of G. Soja ( Hardley
and G. tacacina in DNA sequences. 1n addition , it is
and Hymowitz 1973). However, the results of this
closely rclated to G. /omenlella than to G. labacina,
cxperiment revcal that G. /omentella is closer to
which is agreed with previous research using RFLP
subgenus Soja species than G. tabacina. Moreover ‘ the
analysis (Hsing et al. 1995)
crossing
of Soja
and
G.
lomen/ella had
been
Although
many
bands
exc1usive
to
G
successfully achieved (Newell and Hymowitz 1982)
dolichocarpa were discovered in this experiment,
As a result, it is suggested that G. /omentella rather
bands
than G. tabacina is the ancestor of G. soja. Sincc G
but absent in G. labacina were also discovered
formosana and ci. /omentella have higher similarity
Moreover, bands common to G. dolichocarpa and G
coefficient than the rest of wild Soja subspecies, it
/abacina but absent ìn G. /omen/ella were a1so
coηlITI on
to G. dolichocarpa and G. lomenlella
- 62 -
Analysis of Geneti c Distance among F，∞r Glycine Species CoIIαted from Taiwan : Reveal叫 byDNAPoly油α前1Ís間
observed. In addition , considering the fact that the
tabacina was sterile , and G. tabacina had not been
parentage of G. dolichocarpa Ii es between the other
found
two , we therefore suspect it being the introgressive
investigation is essential on the relationship among
hybridization of G. tomentella and G. tabacina or the
these three and other subgenus Glycine species, in
intermediate type in evolution. However, research on
order to understand the genetic characteristics and
cross breeding (Singh and Hymowitz 1985) had
c1 assification of G. dolichocarpa
on Taiwan
island.
As a result , further
indicated that the crossing of G. tomentella and G
呻吟恭、、、
鼠，主舟、忠心念念
呻吟恭、、、
可 L" ,."c,'" ~'-~念﹒令 ~V
MAdgydJf
SjJSY
().' ().' ().'" ().' ():本
().'JFd、三
M ().'- ().' JSJ
().'"J().,.
G"- G.
(句~
2仰'2-
(句)
_
• •
••
2的'2-
倒﹒
軍沁﹒
•
E三1
因
單恥 ﹒
A
Fig.3.
B
RAPD profiles in G. tomentella, G. dolichocarpa and G. tabacina.
A: primer OPB-07
B: primer OPB-14
h
M: 100bp DNA Ladder

一窩棚輔騙
ω- ---于一-品品置圓圓
鐘，品也回
a
扭轉趣聽
•
•
-j"酋﹒叫
W璽ERE
w
.-ac
Ewwwzama
農林學報
50(3) : 55-65(2001)
- 63-
ACKNOWLEDGEM ENT
This work was supported by a grant of the National Science Council (N SC 88-2311-B-005-043 , NSC
89-23 13"B005-009).
LITERATURE CITED
1. Chuang , C. c. , and C. Huang. 1966. Glycine. In The Leguminosae of Taiwan for.pasture and soil improvement.
P.54-57.
2. Doyle, J. J. , and R. N. Beachy. 1985 Ribosoma1 gene variation in soybean
(G秒'cine)
and its re1atives. Th eor.
App l. Genet. 77 :768-776
3. Doyle ,J. D. , 1. L. Doyle, and L. H. Bailey. 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13-15
4. Dunn , G. , and B. S. Everitt. 1982. An introduction to mathematical taxonomy. Cambridge University Press,
Cambridge
5. Hardley , H.
H., and T.
Soybean: Impro \i eme肘，
H戶nowitz.
Production
1973 . Speciation and cytogenetics. Jn : Caldwell B. E.(ed.)
and Uses. p.97-116. Amer. Soc. Agro. Madison , Wis.
6. HayataB. 1917. G徊 ne. in Icones Plantarum Formosanarum 6 (supp l. ):20.
7. Hsing Y. 1., K. L.
Hsi 帥，
Y. C. Huang , and J. S. Hsieh.
1995 . 而 e
relationships of cu It ivated soybeans and their
wild relatives collected from Taiwan : revealed by seed proteins. Bot. Bul l. Acad. Sin . 36:65-72.
8. Huang S. F., and Huang T. C. 1987. Glycine. In Taxonomic treatment of the Papilionoideae (Leguminosae) of
Taiwan. Taiwania 32:63-65
9. Huang , T. c. , and H. Ohashi. 1977. Glycine. In : Flora ofTaiwan 3:293-298.
10. Hui , D. W. and 8. C. Zhuang 1996. Phylogeny of genus Glycine reconstructed by RAPD fingerprinting. Acta
Genetica Sinica 23(6):460-468
1 1. Hymowi 眩， T., and R. J. Singh . 1987. Taxonomy and speciation. In:Caldwell , B. E. (ed) Soybeans:
Improvement , Production , and Uses. 2nd.ed . p.23-48 . Amer. Soc. Agro., Madison , Wis.
12. Kol 1i para , K. P., R. J. Singh , and T. Hymowitz 1997. Phylogenetic and genomic rel ationships in the genus
Glycine Wild. 8ased on sequences from the Its region ofnuclear rDNA. Genome 40:57-68.
13 . Newell , c. , and T. Hymowitz. 1982. Successful wide hybridization between the soybean and a wild perennial
relati \i e, G. tomentella Hayata. Crop. Sci. 22:1062-1065.
14. Singh , R. J. , and T. Hymowitz. 1985 . The geomic relationships among six wild perennial species of the genus
Glycine subgenus Glycine Wi l1 d. Theo r. App l. Gene t. 7 1: 221-230.
一 64
Analysis of Genetic Dis恤ce amoog Four Glycíne S阿ies CoUected from Taiwan : Rev，臼led by DNA Polymoφhisms
-
的 Tang ，
W. T. and C. C. Lin 1962. Studies on the characteristics of some Glycine spp. Found in Taiwan. 1. Agric
Assoc. China37:15-19.
16. Tateishi , Y. , and H. Ohashi. 1992. Taxonomic studies on Glycine ofTaiwan , 1. Jpn. 80t. 67:127-147.
17 甘lseng，
F. S. , T. H. Tsai and S. T. Wu 1997. Population variation of wild soybean in Taiwan V. Glycine
tomentella Hayata and G. dolichocarpa Tateishi et Ohashi: leaf1 et and seed
m。中hology
and DNA
polymorphism. Chinese Agron. J. 7:107-114.
18. Tindale , M. D. 1984. Two new Eastem Australian species of G砂cine Willd. (Fabaceae). 8runonia 7:207-213.
19. Tindale , M. D. 1986a. A new North Queensland species of Glycine Willd. (Fabaceae). 8runonia 9:99-103 .
20. Tindale , M. D. 1986b. Taxonomic noted on three Australian and Norfols :Island species of Glycine Willd
(Fabaceae: Phaseolae) including the choice of a neotype for G. c1andestina Wend l. 8runonia 9: 179-191.
2 1. Tindale , M. 0. , and L. A. Craven. 1988. Three new
speci的，
of Glycine (Fabaceae:Phaseolae) from
North-westem Australia, with noths on amphicarpy in the genus. Aus1. Sys1. 801. 1:399-410.
22. Tindale , M. D. and L. A. Craven 1993 . Glycine pindanica (Fabaceae: Phaseolae) , a new species from West
Kimberley, Westem Australia. Aus 1. Sys 1. 801. 6:371-376
23 . Yiu , T. J. and F. S. Th seng 1997a. Population variation of wild soybean in Taiwan. I. Plant characteristics in
natural populations of Glycine formosana , G. tabacina and G. tomentel/.ι
24. Yiu , T. J. and F.
S. 甘lseng
1.
Agric. Assoc. China. 177:287-40.
1997b. Population variation ofwild soybean in Taiwan . II. Intraspecific variation of
plant characteristics and isozymes of Glycine tabacina (l abil l.) 8enth. in Pongfu islets. 1. Agric . Fors t.
46(1):111-127.
25. Yiu , T. J. and F. S. Th seng 1997c. Population variation of wild soybean in Taiwan . I1I. lntraspecific variation
of plant characteristics and isozymes of Glycine dolichocarpa Tateishi et Ohashi and G. tomentel!a Hayata in
Taiwan. 1. Agric. Forest. 46(2)59-74.
ρ宮、
農林學報
- 65-
50(3) : 55-65(2001)
利用逢機增痘多型性 DNA(RAPD)探討
台灣四個野生種大豆間之關係
林子凱 1)
吳詩都 2)
(接受刊載日期
胡澤寬 3)
曾富生 2)
中華民國 90 年 6 月 28 日)
摘要:台灣雖然是 G伊ine 亞屬最北之分佈地區，有常見於澳洲 Micronesia 、南中國等地之 G. tomente l/a 及 C
tabacina 之自生種。但也有固有種 Soja 亞屬 G. formosana 之分佈。然而在台灣這三種植物之變異及分佈複
雜，過去在分類學上較不一致。目前台灣自生之野生大豆依形態特徵由過去之三個種被重新區分為四個種:
G. tomente/(a , G. tabaci間. G. dofichocarpa , G. max ssp. formosana • 因此本試驗以此分額之四個野生種加上
G. s句a 共五個為材料，利用逢機增殖多型性 DNA (RAPD)進行系統間關係之分析。
經 RAPD 分析的結果，此五個野生種大豆中最近緣的種為 G. SO)。與 G.
為 G.
G.
tabacina
• 而此五個種可區分為 A)
G. 5句α 、 G. max
dolichocarpa 等兩群。其額似{系數在 A 群為 0.50~0.65
max ssp. formosa間，而最連者
SSp. formosana 及 B)
. B 群為
G. tomentella • G. tabacina •
0.18-0 .3 8 :而 A 與 B 群問為 0.09~0.17 。
關鍵詞:類似係數、大豆屬、逢機增撞多型性核酸。
1)國立中興大學農藝系研究生(現任職行政院農委會農業試驗所圍藝系，台中縣霧峰鄉
2) 國立中興大學農藝系教授
3) 國立中興大學副教授(通訊作者)
413 '台灣)

Download Report