GmFad2-1, Glycine max (L.) Merr. (260-05, OECD UI : DD- Ø26ØØ5-3)...1 1... 2 1... 2 ( )... 2... 2... 2... 2 ( )... 2... 2... 2 ( )... 3... 3... 3... 3... 4... 4... 5... 5 2... 5 ( )... 5... 5... 6 ( )... 14... 14... 14 ( )... 14... 14... 17... 17 ( ).. 21... 21... 21... 22 i
... 22... 22 ( ) 23 ( )... 23... 23... 23 3... 26 ( )... 26 ( )... 27 ( )... 27 ( )... 27 ( )... 27 ( )... 27 2... 29 1... 29 ( )... 29 ( )... 30 ( )... 30 ( )... 30 2... 30 ( )... 30 ( )... 31 ( )... 31 ( )... 31 3... 31 ( )... 31 ( )... 31 3... 35... 37 ( )... 39 GmFad2-1, Glycine max (L.) Merr. (26Ø-Ø5, OECD UI : DD- Ø26Ø5-3)... 41 ii
16 17 2 11 1 4 2 (GmFad2-1, Glycine max (L.) Merr.) (260-05, OECD UI DD-Ø26ØØ5-3) 1
1 1 ( ) Soybean /Soyabean Glycine max (L.) Merr. The International Plant Names Index, 2004 (GmFad2-1, Glycine max (L.) Merr.) (260-05, OECD UI DD-Ø26ØØ5-3) ( 260-05 ) II A2396 ( ) Glycine soja Sieb. & Zucc., 1994 OECD, 2000 5,000 11, 1994 OECD, 2000 1,900 2,000, 2002, 2002 2
50 40, 2002, 2002 2002 1 8 75 41 FAO Statistical Database, http://apps.fao.org/page/collections 27 2002 5 76 380, http://www.customs.go.jp/toukei/info/index.htm 80%, 1994 2002 4, http://www.customs.go.jp/toukei/info/index.htm ( ) 18 28, 1994 ph 6 7, 1994 3
2002 OECD, 2000, 2002 0.5 1%, 2002, 1994 G. soja Sieb. & Zucc., 1994 OECD, 2000, 2002, 2002 Kasahara, 1982 1 2 2m 0.036 10m 0 1 4
2 260-05 Optimum Quality Grains, L.L.C. 260-05 G. max -12 GmFad2-1 1 Fad2-1 80% 260-05 260-05 ( ) 260-05 1 5
GmFad2-1 pbs43 dapa pml102 2 260-05 dapa 260-05 pbs43 pml102 1 2 11 pbs43 pml102 2 260-05 1 9 2 11 2 GmFad2-1 dapa GUS amp r 3 260-05 2 (Kinny, 1994) Fad2-12 (C18: ) -12 n-6 (C18:2) (Okuley et al., 1994 Heppard et al., 1996) Fad3-15 -15(n-3) C18:3 (Yadav et al., 1993) 12 1 Fad2-12 6
Fad2 GmFad2-1 (Hepperd et al., 1996) 19 GmFad2-2 GmFad2-1 RNA GmFad2-2 RNA (Hepperd et al., 1996) 13 3 260-05 2 Fad2 GmFad2-1 260-05 -12 GmFad2-1, 1996 DNAP 5034323 GmFad2-1 -12 3 GmFad2-1 17 30% 80 260-05 A2396 - α α A2 B1A 4 - α α GmFad2-1 - - Nordlee, 1995 Takahashi et al., 1994 Kitamura, 1995 - α α A2 B1A 260-05 - α α 260-05 1995 7
1996 24 1 80% 8
1 pbs43 (kbp) GmFad2-1 - 0.630 Glycine max - α GmFad 2-1 1.490 Glycine max δ-12 cdna 3 1.186 Phaseolus vulgaris 3 GUS 35S 1.380 (CaMV) (Harpster et al., 1988) Cab 22L untranslated leader 0.059 Petunia hybrida var. Mitchell 22L a/b 5 untranslated leader GUS 1.856 E. coli uida - (uida ) NOS 3 0.796 Agrobacterium tumefaciens 3 lac 0.231 Lac l Plac -D- lacza ori 0.599 E. coli puc19 amp r 1.008 E. coli - 5 3 δ-12 n-6 untranslated leader mrna 3 D- E. coli E. coli E. coli 9
f1 ori 0.461 f1 f1 DNA f1 lacza 3 0.166 lacza 3 10
2 pml102 (kbp) dapa ptz18r 0.035 Escherichia coli pbr322 Kti3 2.091 Glycine max Kunitz ssu CTC 0.167 Glycine max Rubisco N- dapa 0.923 Corynebacterium glutamicum Kti3 3 0.195 Glycine max Kunitz 3 3 lac 0.231 Lac l Plac -D- lacza ori 0.599 E. coli puc19 amp r 1.008 E. coli - f1 ori 0.461 f1 E. coli 2 E. coli E. coli E. coli f1 DNA f1 lacza'3' 0.166 lacza 3 11
C3:0-ACP C3:0-CoA C4:0-ACP KAS III C2:0-CoA ACCase KAS X KAS I KAS II C8:0-ACP C16:0-ACP C18:0-ACP C18:1-ACP II I C18:3-CoA C16:0-CoA C18:0-CoA C18:1-CoA C18:2-CoA 18:3-O OR O-PdtC Fad 3 OR 18:2-O Fad 2 O-PdtC 18:1- O OR O-PdtC 1 C18:1 C18:2 C18:3 Fad2-12 Fad3-15 + dapa 2,3- - N- - - - - - - - 1 - - 2,6-2 12
mrna Northern Probe SEED DAF 13 17 19 21 26 L R St GmFad 2-2 GmFad 2-1 3 GmFad2-1 RNA GmFad2-2 RNA L R St DAF Days after flowering (Hepperd et al., 1996) 13
( ) 260-05 pbs43 4 pml102 5 Escherichia coli pbr322 260-05 pbs43 10,303 bp pml102 6,247 bp 2 pbr322 ( ) amp r amp r 260-05 3 ( ) 5 9 5 15 10( 5 16 ) 14
amp probe BspHI (9533) HindIII (1) BspHI (384) GmFad 2-1 probe BspHI (8525) r amp β-conglycinin Promoter GmFAD2-1 pbs43 [10303 bp] 3' Phaseolin XbaI (2117) SstI (2169) SstI (2223) Phaseolin 3 probe SstI (7409) 3' NOS GUS 35S Promoter HindIII (3308) XbaI (3317) XbaI (3338) BamHI (3344) BspHI (5634) EcoRV (5536) EcoRV (5305) XhoI (4686) SstI (4689) 35S Promoter probe GUS probe 4 pbs43 15
[Vector: ptz18r] BspHI (5536) HindIII (1) BamHI (25) HindIII (404) BspHI (427) amp probe HindIII (1126) amp r pml102 [6247 bp] Kti 3 promoter BspHI (1253) BspHI (4528) CTS Kti 3 terminator Corynebacterium dapa NcoI (2126) EcoRV (2386) BamHI (3422) HindIII (3289) HindIII (2979) dapa probe 5 pml102 16
II A2396 pbs43 pml102 GUS R0 GUS GmFad2-1 PCR R1 R1 260-05 R0 6 19 R0 R1 GmFad2-1 dapa GUS GUS GUS GmFad2-1 dapa G168 G94-1 G94-19 GUS GUS 21 GmFad2-1 1 GUS 1 GUS GmFad2-1 dapa GmFad2-1 dapa 260-05 R0 1 17
3 260-05 260-05 3 20 260-05 1999 5 2000 5 ( 2001 3 2003 3 ) 18
R0 260-05 GmFad2-1 R1 GmFad2-1 G168 R1 GmFad2-1 G94 G168 R2 R2 GmFad2-1 G94-1 G94-19 GmFad2-1 R3 (R4 ) 6 260-05 260-05 R0 GmFad2-1 ( ) dapa ( ) GUS R0 R1 GmFad2-1 ( ) dapa ( )GUS G168 R1 R2 GmFad2-1 ( ) dapa ( )GUS G94-1 G94-19 20 GmFad2-1 1 GUS 1 GUS G168 G94-1 G94-19 GmFad2-1 ( ) dapa ( ) 3 260-05 19
R2 R6 R6 3 260-05 R1 R2 R6 R4 R5 R6 R7 GmFad2-1 GUS dapa amp r GUS R6 260-05 3 4 5 1 8 5 11 12 6 R4 R5 8 R8 9 20
( ) 260-05 1996 R6 260-05 A2396 260-05 GmFad2-1 dapa 5 1 8 GmFad2-1 2 GmFad2-1 1 GUS 1 GUS 2 amp r 1 amp r dapa dapa 5 9 10 GmFad2-1 BamHI Phaseolin 3 5 11 12 1993 1995 R1 R2 1996 R6 260-05 A2369 GmFad2-1 1993 1995 R2 260-05 A2396 1996 R6 21
260-05 A2396 GmFad2-1 dapa GUS amp r 2005 R10 260-05 R10 GmFad2-1 GmFad2-1 2 dapa R6 GmFad2-1 dapa ( ) 260-05 GmFad2-1 pbs43 2 5 9 1995 1997 R4 R5 R6 R7 A2396 17-30% 260-05 80% 6 22
( ) 260-05 DNA 2 18bp 19bp PCR 7 DNA 2 18bp PCR PCR 260-05 20ng DNA ( ) 2 GmFad2-1 GmFad2-1 -12 260-05 A2396 - α α A2 B1A 4 GmFad2-1 - - (Julie Nordlee, 1995)Takahashi et al., 1994 Kitamura, 1995 1995 1996 24 1 260-05 5 23
5 6 8 2 3 260-05 9 260-05 A2396 260-05 23.2 A2396 20.1 260-05 A2396 9 3 6 8 33, 2002 260-05 A2396 (12 ) 60 2 60 9 8 260-05 260-05A 24
260-05 260-05 A2396 A2396 260-05 260-05 A2396 260-05 A2396 260-05 A2396 19 260-05 91% A2396 88 260-05 A2396 260-05 260-05 25
0% 0.5 1%(, 2002, 1994) 260-05 0.7% (Nakayama Y. and Yamaguchi H., 2002) 260-05 2005 10 260-05 260-05 8 260-05 260-05 260-05 3 ( ) 26
( ) ( ) ( ) ( ) ( ) 260-05 4 4 260-05 / 1997 1997 2000 2000 2000 2000 1999 5 27
2000 5 ( 2001 3 2003 3 ) 28
2 (Glycine max (L.) Merr.) 260-05 IP 260-05 260-05 260-05 260-05 A2396 1 ( ) 10 260-05 260-05 A2396 29
260-05 80% 260-05 ( ) ( ) ( ) 2 ( ) 2005 260-05 10 260-05 8 260-05 30
260-05 260-05 260-05 ( ) ( ) ( ) 3 ( ) ( ) 260-05 260-05 31
0.7% (Nakayama Y. and Yamaguchi H., 2002) 2.3% Kiang 1992 13% Fujita 1997 ( ) 260-05 IP 260-05 260-05 1) 2.3% Kiang et al., 1992 13Fujita et al., 1997 0.5% 3% (Ahrent and Caviness, 1994; Carlson and Lersten, 1987;Caviness, 1996; Chang and Kiang, 1987; Garver and Odland 1926; Wever and Hanson, 1961) 2m 0.036% 10m 0% 1 2) Nakayama Y. and Yamaguchi H., 2002 Nakayama Gls/93-J-01 50cm 0.7% (Nakayama Y. and Yamaguchi H., 2002) 3) 10 260-05 60cm 30cm 0% 0.5 1%, 2002 32
, 1994 260-05 A2396 260-05 260-05 4) DNA DNA 3 I II III I III G.gracilis Abe 1999 DNA 330 DNA III I 1.8 6 I 4 dedomestication 260-05 260-05 IP 2 0.036 10 0 260-05 30 80 0 50 260-05 33
260-05 260-05 13 2.3 DNA DNA 260-05 260-05 34
3 260-05 GmFad2-1 GmFad2-1 -12 260-05 260-05 260-05 IP 10 IP 260-05 260-05 260-05 10 260-05 260-05 A2396 260-05 35
260-05 A2396 260-05 A 2396 260-05 0.7% (Nakayama Y. and Yamaguchi H., 2002) 260-05 0.5 1% 260-05 260-05 260-05 36
Abe, J., Hasegawa, A., Fukushi, H., Mikami, T., Ohara, M. and Shimamoto, Y. 1999. Introgression between wild and cultivated soybeans of Japan revealed by RFLP analysis for chloroplast DNAs. Economic Botany 53(3), 285-291. FAO Statistical Database. 2004. (http://apps.fao.org/page/collections) Harpster et al. 1988. Mol. Gen. Genet. 212:182-190. Heppard, E.P., Kinney, A. J., Stecca, K.L., Miao, G-H. 1996. Plant Physiol. 110:311-319. Nordlee, J. A. 1995. Screening of Soybean (Glycine max) Germplasm for Binding to Immunoglobulin E by Immunoelectroblotting. A thesis for the Degree of Master of Science. The Graduate College at the University of Nebraska. Kasahara, Y. 1982. In: Geobotany 2; Biology and Ecology of Weeds. (Holzner, W., Numata, M. eds.) pp285-297. W. Junk, Netherlands. Kinney, A.J. (1994) Curr. Opin. Biotechnol. 5:144-151. Kitamura, K. 1995. JARQ 29:1-8.,. 1996 pp 161-169 Nakayama, Y. and Yamaguchi, H. 2002. Weed Biology and Management 2: 25-30 2,. 1994: 537-541... 2002. OECD. 2000. Series on Harmonisation of Regulatory Oversight in Biotechnology, No. 15: Consensus Document on the Biology of Glycine max (L.) Merr. (Soybean). (http://www.olis.oecd.org/olis/2000doc.nsf/linkto/env-jm-mono(2000)9) Okuley, J., Lightner, J., Feldman, K., Yadav, N., Lark, E., Browse, J. 1994. Plant 37
Cell 6:147-158.. 2003: 77-95. Takahashi, K., Banba, H., Kikuchi, A., Ito, M., Nakamura, S. 1994. Breeding Science 44:65-66. The International Plant Names Index. 2004. (http://www.ipni.org/index.html). Yadav, N.S., Wierzibicki, A., Aegerter, M., Caster, C.S., Perez-Grau, L., Kinney, A.J., Hitz, W.D. et al. 1993. Plant Physiol. 103:467-476.. 2002. (http://www.customs.go.jp/toukei/info/index.htm) 38
( ) 16 8 17 2 11 1 (GmFad2-1, Glycine max (L.) Merr.) (260-05, OECD UI DD-Ø26ØØ5-3) ( 260-05 ) 1 260-05 2 260-05 39
3 260-05 260-05 4 260-05 5 260-05 40
GmFad2-1, Glycine max (L.) Merr. (26Ø-Ø5, OECD UI : DD- Ø26Ø5-3) 1 2 1 5-1. 2 pbs43 pml102 3 ampr GmFad2-1 dapa GUS 4 260-05 5 6 7 260-05 8 260-05 9 260-05 10 41