高オレイン酸ダイズ（ GmFad2-1, Glycine max (L.) Merr.）(260-05, OECD UI :DD- Ø26ØØ5-3) 申請書等の概要
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1 GmFad2-1, Glycine max (L.) Merr. (260-05, OECD UI : DD- Ø26ØØ5-3) ( ) ( ) ( ) ( ) ( ) ( ) ( ) i
2 ( ) 23 ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) GmFad2-1, Glycine max (L.) Merr. (26Ø-Ø5, OECD UI : DD- Ø26Ø5-3) ii
3 (GmFad2-1, Glycine max (L.) Merr.) (260-05, OECD UI DD-Ø26ØØ5-3) 1
4 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) ( ) II A2396 ( ) Glycine soja Sieb. & Zucc., 1994 OECD, ,000 11, 1994 OECD, ,900 2,000, 2002,
5 50 40, 2002, FAO Statistical Database, , 80%, , ( ) 18 28, 1994 ph 6 7,
6 2002 OECD, 2000, %, 2002, 1994 G. soja Sieb. & Zucc., 1994 OECD, 2000, 2002, 2002 Kasahara, m m 0 1 4
7 Optimum Quality Grains, L.L.C G. max -12 GmFad2-1 1 Fad2-1 80% ( )
8 GmFad2-1 pbs43 dapa pml dapa pbs43 pml pbs43 pml GmFad2-1 dapa GUS amp r (Kinny, 1994) Fad2-12 (C18: ) -12 n-6 (C18:2) (Okuley et al., 1994 Heppard et al., 1996) Fad (n-3) C18:3 (Yadav et al., 1993) 12 1 Fad2-12 6
9 Fad2 GmFad2-1 (Hepperd et al., 1996) 19 GmFad2-2 GmFad2-1 RNA GmFad2-2 RNA (Hepperd et al., 1996) Fad2 GmFad GmFad2-1, 1996 DNAP GmFad GmFad % A α α A2 B1A 4 - α α GmFad Nordlee, 1995 Takahashi et al., 1994 Kitamura, α α A2 B1A α α
10 % 8
11 1 pbs43 (kbp) GmFad Glycine max - α GmFad Glycine max δ-12 cdna Phaseolus vulgaris 3 GUS 35S (CaMV) (Harpster et al., 1988) Cab 22L untranslated leader Petunia hybrida var. Mitchell 22L a/b 5 untranslated leader GUS E. coli uida - (uida ) NOS Agrobacterium tumefaciens 3 lac Lac l Plac -D- lacza ori E. coli puc19 amp r E. coli δ-12 n-6 untranslated leader mrna 3 D- E. coli E. coli E. coli 9
12 f1 ori f1 f1 DNA f1 lacza lacza 3 10
13 2 pml102 (kbp) dapa ptz18r Escherichia coli pbr322 Kti Glycine max Kunitz ssu CTC Glycine max Rubisco N- dapa Corynebacterium glutamicum Kti Glycine max Kunitz 3 3 lac Lac l Plac -D- lacza ori E. coli puc19 amp r E. coli - f1 ori f1 E. coli 2 E. coli E. coli E. coli f1 DNA f1 lacza'3' lacza 3 11
14 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 Fad dapa 2,3- - N ,6-2 12
15 mrna Northern Probe SEED DAF L R St GmFad 2-2 GmFad GmFad2-1 RNA GmFad2-2 RNA L R St DAF Days after flowering (Hepperd et al., 1996) 13
16 ( ) pbs43 4 pml102 5 Escherichia coli pbr pbs43 10,303 bp pml102 6,247 bp 2 pbr322 ( ) amp r amp r ( ) ( 5 16 ) 14
17 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
18 [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
19 II A2396 pbs43 pml102 GUS R0 GUS GmFad2-1 PCR R1 R R 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 R0 1 17
20 ( ) 18
21 R GmFad2-1 R1 GmFad2-1 G168 R1 GmFad2-1 G94 G168 R2 R2 GmFad2-1 G94-1 G94-19 GmFad2-1 R3 (R4 ) R0 GmFad2-1 ( ) dapa ( ) GUS R0 R1 GmFad2-1 ( ) dapa ( )GUS G168 R1 R2 GmFad2-1 ( ) dapa ( )GUS G94-1 G GmFad2-1 1 GUS 1 GUS G168 G94-1 G94-19 GmFad2-1 ( ) dapa ( )
22 R2 R6 R R1 R2 R6 R4 R5 R6 R7 GmFad2-1 GUS dapa amp r GUS R R4 R5 8 R8 9 20
23 ( ) R A GmFad2-1 dapa GmFad2-1 2 GmFad2-1 1 GUS 1 GUS 2 amp r 1 amp r dapa dapa GmFad2-1 BamHI Phaseolin R1 R R A2369 GmFad R A R6 21
24 A2396 GmFad2-1 dapa GUS amp r 2005 R R10 GmFad2-1 GmFad2-1 2 dapa R6 GmFad2-1 dapa ( ) GmFad2-1 pbs R4 R5 R6 R7 A % % 6 22
25 ( ) DNA 2 18bp 19bp PCR 7 DNA 2 18bp PCR PCR ng DNA ( ) 2 GmFad2-1 GmFad A α α A2 B1A 4 GmFad (Julie Nordlee, 1995)Takahashi et al., 1994 Kitamura,
26 A A A , A2396 (12 ) A 24
27 A2396 A A A A % A A
28 0% 0.5 1%(, 2002, 1994) % (Nakayama Y. and Yamaguchi H., 2002) ( ) 26
29 ( ) ( ) ( ) ( ) ( ) /
30 ( ) 28
31 2 (Glycine max (L.) Merr.) IP A ( ) A
32 % ( ) ( ) ( ) 2 ( )
33 ( ) ( ) ( ) 3 ( ) ( )
34 0.7% (Nakayama Y. and Yamaguchi H., 2002) 2.3% Kiang % Fujita 1997 ( ) IP ) 2.3% Kiang et al., Fujita et al., % 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) cm 30cm 0% 0.5 1%,
35 , A ) DNA DNA 3 I II III I III G.gracilis Abe 1999 DNA 330 DNA III I I 4 dedomestication IP
36 DNA DNA
37 GmFad2-1 GmFad IP 10 IP A
38 A A % (Nakayama Y. and Yamaguchi H., 2002) %
39 Abe, J., Hasegawa, A., Fukushi, H., Mikami, T., Ohara, M. and Shimamoto, Y Introgression between wild and cultivated soybeans of Japan revealed by RFLP analysis for chloroplast DNAs. Economic Botany 53(3), FAO Statistical Database ( Harpster et al Mol. Gen. Genet. 212: Heppard, E.P., Kinney, A. J., Stecca, K.L., Miao, G-H Plant Physiol. 110: Nordlee, J. A 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 In: Geobotany 2; Biology and Ecology of Weeds. (Holzner, W., Numata, M. eds.) pp W. Junk, Netherlands. Kinney, A.J. (1994) Curr. Opin. Biotechnol. 5: Kitamura, K JARQ 29:1-8., pp Nakayama, Y. and Yamaguchi, H Weed Biology and Management 2: ,. 1994: OECD Series on Harmonisation of Regulatory Oversight in Biotechnology, No. 15: Consensus Document on the Biology of Glycine max (L.) Merr. (Soybean). ( Okuley, J., Lightner, J., Feldman, K., Yadav, N., Lark, E., Browse, J Plant 37
40 Cell 6: : Takahashi, K., Banba, H., Kikuchi, A., Ito, M., Nakamura, S Breeding Science 44: The International Plant Names Index ( Yadav, N.S., Wierzibicki, A., Aegerter, M., Caster, C.S., Perez-Grau, L., Kinney, A.J., Hitz, W.D. et al Plant Physiol. 103: ( 38
41 ( ) (GmFad2-1, Glycine max (L.) Merr.) (260-05, OECD UI DD-Ø26ØØ5-3) ( )
42
43 GmFad2-1, Glycine max (L.) Merr. (26Ø-Ø5, OECD UI : DD- Ø26Ø5-3) pbs43 pml102 3 ampr GmFad2-1 dapa GUS
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4) H. Takayama et al.: J. Med. Chem., 45, 1949 (2002). 5) H. Takayama et al.: J. Am. Chem. Soc., 112, 8635 (2000). 6) H. Takayama et al.: Tetrahedron Lett., 42, 2995 (2001). 9) M. Kitajima et al: Chem.
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