Post on 13-Mar-2020
鉄欠乏耐性イネ説明資料
資料 2
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Distribution of CALCISOLS Based on WRB and the FAO/Unesco Soil Map of the World
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Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
FeFe
Fe
Fe
Fe
FeFe
Fe
Fe
FeFe
Fe
FeFe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
FeFe
FeFeFe
Fe
FeFe
Fe
Fe
FeFeFeFe
Fe
FeFe
Fe Fe
Fe
Fe
Fe
Fe
CO2H
OH
CO2HCO2H
OH
Mugineic acidN N
Fe
H
YS1
Strategy-II
㊪
NH
NH2
COOH COOH COOH
N
(NAS)
S- -L- (SAM)
SAM(SAMS)COOH
H3C S NH2
㌹(NAAT)
NH
O
COOH COOH COOH
N
3”-oxo acid
(DMAS)NH
OH
COOH COOH COOH
N
2’-
NH
OH
COOH COOH COOH
N
NH
OH
COOH COOH COOH
N
3-
NH
OH
COOH COOH COOH
N
NH
OH
COOH COOH COOH
N
3- -2’-
NH
OH
COOH COOH COOH
N
3-
OH
OH OHHO HO
HO
IDS3
IDS3IDS2
IDS2
HO
naat-B
-Fe+Fe
naat-A
Shoot
Root
Shoot
Root
naat-A
naat-B
B C
A
NP NPT II NT MCSLacZ 35P iGUS NT 35P HPT NT
RB LB
Ri ori RK2 ori
Hin dIII, Spe I, Not I (multicloning site)
NAAT genomic fragment 11kbNot I Not I
naat-Anaat-B
0
10
20
0
2
4
6
8
10(g) (g)Shoot Grain
C1 C6 C8 N1 N3 N8 N9 N17
Daysaftertransfertoanalkalinesoil(days)
(cm)
N1N3N8N9N17C1C6C8
85756555453520
40
60
80
100
A
B
C1 C6 C8 N1 N3 N8 N9 N17
NAAT(Takahashi et al., Nature Biotechnology 19:466-469, 2001)
A B
NAAT(Takahashi et al., Nature Biotechnology 19:466-469, 2001)
NAAT
NAATcontrol
control
NH OH
COOH COOHCOOH
N
DMA
OH
NH OH
COOH COOHCOOH
N
MAepiHMA
OH
NH OH
COOH COOHCOOH
NHO
NH OH
COOH COOHCOOH
NHO
epiHDMA
MAs
AdeAMP
ATPPRPPPPi
CH3 S CH2 CH2 COO-
CH2CHNH3+
Ade
R COO-CHNH3+
OHCH2
O
OH
CH2 AdeS
OPO3H-
CH2CH3 S
ATP
R COO-CO
ATP
ADP
H2O
HCOO-
O2
HPO42-O
OH OH
O
OH OH
O
OH OH
methionine
SAM
OH
CO2
NAD+NADH
NH NH2
COOH COOHCOOH
N
NA
NH O
COOH COOHCOOH
N
keto-form
IDI1
NAS
APRT
NAAT
IDS3 IDS3
Ids2
Ids2
SAMsynthetase
S CH2 OPO3H-CH2CHCH3 CH COOH
OH
Yang Cycle(Methionine Cycle)
CH3 S
CH3CH3 S CH2 CH2 COO-CO
S CH2 C=CH-O-CH2CH3 CO COOH
CH3 S CH2 CH2 COO-CHNH3+
OHS CH2 OPO3H-CHCH2CH3 CO C=
S CH2 OPO3H-CH2CH2CH3 CO CO
FDH
ムギネ酸類の生合成経路2
Ids3 genome 20.0 kb
HvNAS1 cDNANaat-A cDNAHvAprt cDNA
Ids3 5' 2.2kIds3 5' 2.2kIds3 5' 2.2k
Ids3 5' Ids3 5'
Ids3 5'Ids3 5'Ids3 5'
HvNAS1 cDNA Naat-A cDNA
Naat-A cDNAHvNAS1 cDNA HvAprt cDNA
HvNaat genome 11.0 kbHvNAS1 genome 13.0 kb1
6
5
4
3
2
7
8
Ids3 cDNANaat-A cDNA
HvNAS1 cDNA9
11
10
35Spro 0.8k
Ids6 genome 20.0 kb
12
HvNAS genome 13 kb HvNaat genome 11 kb14
鉄欠乏耐性イネ作成に使用したコンストラクト
(赤字は今回申請したもの)
35Spro 0.8k35Spro 0.8k
gHvNAS1-gHvNNAT
I3pNas-Naat-Aprt
gHvIDS3
I3pAPRT
gHvNAS1 gHvNAATvector control
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0
1
2
3
4
5
0
10
20
30
40
50
60
70
80
90
5月24日 6月21日 7月19日 8月16日
gHvNAS1
gHvNAS1-gHvNAAT
gHvIDS3
gHvNAAT
I3pAPRTI3pNasNaatAprt
vector control
㈻ モ