molecular breeding for enhancing tolerance to low …...molecular breeding for enhancing tolerance...
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Molecular breeding for Molecular breeding for enhancing tolerance to enhancing tolerance to
low iron availability low iron availability in in CalcariousCalcarious Soils Soils
Naoko K. Naoko K. NishizawaNishizawaThe University of Tokyo, JapanThe University of Tokyo, Japan
March 14, 2008, GIES 2008
順序かえた連
続
Environment
C
FeZn
Cu
Mn
MoN
Ni
PK
S
Cl
B
MgCa
H
O
Food
Crop production is critical for our life Crop production is critical for our life
Distribution of calcareous soils (high soil pH) Distribution of calcareous soils (high soil pH) Distribution of calcareous soils (high soil pH)
Distribution of CALCISOLSBased on WRB and the FAO/Unesco Soil Map of the World
- Fe
Iron is not available in Iron is not available in alkaline soilsalkaline soils
Fe
FeFe
Fe
Fe Plants diePlants die in alkalinein alkalinesoilssoils due to iron deficiencydue to iron deficiency
Iron is the essential elementIron is the essential elementfor plant growthfor plant growth
Mugineic Mugineic AcidAcid
細胞膜 細胞壁 土壌
細胞質
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
FeFe
Fe
Fe
Fe
Fe
FeFe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
Fe
FeFeFe
Fe
FeFe
Fe
Soil
FeFe
FeFe
Fe
Fe
Fe
Fe Fe
Fe
Fe
Fe
Fe
Fe
CO2H
OH
CO2HCO2H
OH
N NH
Fe YS1
Cell WallCell MembraneCell Membrane
CytoplasmCytoplasm
Iron Acquisition in Graminaceous plantsIron Acquisition in Iron Acquisition in Graminaceous Graminaceous plantsplants
NH
NH2
COOH COOH COOH
N
Nicotianamine
S-adenosyl Methionine (SAM)
COOH
H3CS NH2
Methionine
NH
O
COOH COOH COOH
N
3”-oxo acidNH
OH
COOH COOH COOH
N
2’-deoxymugineic acid
NH
OH
COOH COOH COOH
N
Mugineic acid
NH
OH
COOH COOH COOH
N
3-hydroxymugineic acid
NH
OH
COOH COOH COOH
N
3-epi-hydroxy-2’-deoxymugineic acid
NH
OH
COOH COOH COOH
N
3-epi-hydroxymugineic acid
OH
OH OH
HO HO
HO
IDS3
IDS3IDS2
IDS2
Deoxymugineic acid synthase
SAM synthetase
Nicotianamine synthase
Nicotianamine aminotransferase
Mugineic Mugineic acid familyacid family phytosiderophoresphytosiderophores
Biosynthetic Pathway of MAs Biosynthetic Pathway of MAs Biosynthetic Pathway of MAs
high
low0 1 2 3 4 5 6 (h) -Fe +Fe
DC
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5252FeFe3+3+--DMA absorption from the barley roots DMA absorption from the barley roots
Rice Suffered From Iron Deficiency Rice Suffered From Iron Deficiency Rice Suffered From Iron Deficiency
Rice Plants Uptake Iron as Rice Plants Uptake Iron as FeFe3+3+--phytosiderophore phytosiderophore and Feand Fe2+2+
OsYSLOsYSL OsIRTOsIRT
FeFe3+3+--DMADMA
FeFe3+3+
DMADMA
FeFe2+2+
FeFe2+2+
cytoplasm soil cytoplasm
Ishimaru et al, Plant J, 2006
soil
Takahashi et al,Takahashi et al,Nature Biotech, Nature Biotech, 20012001
Transgenic rice Transgenic rice tolerant to Fe tolerant to Fe deficiencydeficiency
TransformantNT
OsIRTOsIRT
FeFe2+2+
FeFe2+2+
FeFe3+3+
FeFe3+3+
Introduction ofIntroduction ofFeFe3+ 3+ Reduction Activity intoReduction Activity into Rice Rice
cytoplasm soilFeFe2+2+
Refre1-372
Mutational reconstructedMutational reconstructedferric ferric chelate reductasechelate reductase
Refre1-372Vector control Vector control
Transgenic riceTransgenic rice shows enhancedshows enhanced tolerancetolerance totolow Fe availability in a calcareous soillow Fe availability in a calcareous soil
Alkaline soilNormal soil
CACGTGG OsNAS1OsFDHOsAPT1
MTN
CACGTGG NACprotein
NAC
unknowncis-element
Fedeficiency
CACGTGG AP2 domainprotein
AP2
unknowncis-element
Fedeficiency
Fe deficiency stress
IDE-like
IDEs binding transcription factor
OsIRO2
OsIRO2
Model of gene regulation, including OsIRO2 Model of gene regulation, including OsIRO2 Model of gene regulation, including OsIRO2 Ogo et al, Plant J, 2007
Field evaluation of transgenic rice with enhanced tolerance to alkaline soils
Field evaluation of transgenic rice with Field evaluation of transgenic rice with enhanced tolerance to alkaline soils enhanced tolerance to alkaline soils
Wild typeWild typeTransgenic riceTransgenic rice
Suzuki et alSuzuki et al, Soil, Soil Sci Sci Plant Plant NutrNutr,, 20082008
Nakanishi et al, Japanese Patent 2005-9600 (2005)
New System forMarker Free Transgenic Crops
New System forNew System forMarker Free TransgenicMarker Free Transgenic CropsCrops
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