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Improving Phosphorus Efficiency in Sorghum by the Identification and
Validation of Sorghum Homologs for Pup1, a Major QTL Underlying
Phosphorus Uptake in Rice (SorghumPup1)
ICRISAT - Mali
Eva Weltzien, Fred Rattunde, Willmar Leiser
IRRI/JIRCAS
Sigrid Heuer (IRRI)
Rico Gamuyao (IRRI)
Matthias Wissuwa (JIRCAS)
GCP – GRM 2013, Lisbon - Portugal
Moi Unversity
Sam Gudu
INRAN - Niger
Soumana Souley
Cornell University/USDA-ARS
Leon Kochian (USDA/CU)
Randy T Clark (USDA/CU)
Brittany Green (USDA/CU)
Jon Shaff (USDA/CU)
Sharon Mitchell (IGD/CU)
Sylvia de Sousa
Robert Schaffert
Claudia Guimarães
Lidiane Braga (PhD S)
Gabriel Corradi (PhD S)
Barbara Negri (Und)
Fabiano Matos (Und)
ESALQ/USP
Antonio Augusto F. Garcia
Rodrigo Gazaffi
ICRISAT - Niamey
Bettina Haussmann and Tom Hash
Jurandir Magalhaes
Barbara H Maciel (PhD student)
A Molecular Tool Kit for Highly
Weathered Tropical Soils
Al toxicity
High P fixation
Low activity
clay
Variable charge
> 70 % Kaolinate/
Fe and Al oxidesEquator
Capricorn
Low P soils Acid soils (Al tox)
NH
COOH
Cytoplasm
3
*
AltSB
Pstol1
Plant Phys 156-1041
Fe-O
Citrate
PO4
--
Band A
Band B
Band C
10 ppm (P)
5 ppm (P)
0 ppm (P)
10 ppm (P)
5 ppm (P)
0 ppm (P)
0 - 20 cm
- Spatial models (r/c)
- Environmental covariates
- Phenological covariates
Phenotyping for Phosphorus Efficiency
- Soil moisture strongly
impaires P diffusion
- PAE x PUTIL
Phenotyping for Root Morphology (2D)
- Importance for PAE
- Shallow roots?
PSTOL1 increases grain yield
PSTOL1 increases early root growth
Comparative Strategy to Validate Pstol1 Homologs
as Genes Conferring Enhanced P acquisiton
Sorghum:
association mapping
US (converted) Panel
Contrasting parental linesPstol1 - Rice
Maize:
QTL mapping L22
Grain yield:
low P soils
Root morphology: nutrient solution
Wide Diversity Panel
Parentoni et al. (2010) Maydica
Polymorphism Discovery for Linkage and Association Mapping
• Primer design for amplicon sequencing
• SNP detection
• Conversion to the Kaspar system
• Genotyping in the association panel
Primer-BLAST
Conserved Regions
Paralog-specific primers
Step 2:
Producing
F1: Ms3ms3
Step 3:
Self-pollinate
F1s
Step 4:
Random Matting
Cycle 1
Step 5:
Random Matting
Cycle 2
Step 1:
Base population
development
Step 6:
- Random
Matting Cycle 3
- Pup1/AltSB
validation
Selection for AltSB based on
allele mining with tag markers
Genotyping
Genotyping S1s to produce S2s
Phenotype S2s and relate genotypic information
on AltSB/Pstol1 with the phenotype for validation
- Provides means to validate Pup1/AltSB (higher LD)
- Effect estimate
- Amenable to gene pyramidation
- Contingency plan based on a familiar breeding
scheme (high risk project)
- Entry points for new material (SorghumCI)
- Cultivar release, regardless
Sorghum RMPs: pyramiding AltSB and multiple Pstol1 homologs
4 cycles of random matting
Products:
- Agronomic performance under low P (SAP)
- Root morphology and architecture (SAP)
- Root morphology QTL
- Functional markers

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GRM 2013: Improving phosphorus efficiency in sorghum by the identification and validation of sorghum homologs for Pup1, a major QTL underlying phosphorus iptake in rice (SorghumPup1) -- J Magalhaes (redacted)

  • 1. Improving Phosphorus Efficiency in Sorghum by the Identification and Validation of Sorghum Homologs for Pup1, a Major QTL Underlying Phosphorus Uptake in Rice (SorghumPup1) ICRISAT - Mali Eva Weltzien, Fred Rattunde, Willmar Leiser IRRI/JIRCAS Sigrid Heuer (IRRI) Rico Gamuyao (IRRI) Matthias Wissuwa (JIRCAS) GCP – GRM 2013, Lisbon - Portugal Moi Unversity Sam Gudu INRAN - Niger Soumana Souley Cornell University/USDA-ARS Leon Kochian (USDA/CU) Randy T Clark (USDA/CU) Brittany Green (USDA/CU) Jon Shaff (USDA/CU) Sharon Mitchell (IGD/CU) Sylvia de Sousa Robert Schaffert Claudia Guimarães Lidiane Braga (PhD S) Gabriel Corradi (PhD S) Barbara Negri (Und) Fabiano Matos (Und) ESALQ/USP Antonio Augusto F. Garcia Rodrigo Gazaffi ICRISAT - Niamey Bettina Haussmann and Tom Hash Jurandir Magalhaes Barbara H Maciel (PhD student)
  • 2. A Molecular Tool Kit for Highly Weathered Tropical Soils Al toxicity High P fixation Low activity clay Variable charge > 70 % Kaolinate/ Fe and Al oxidesEquator Capricorn Low P soils Acid soils (Al tox) NH COOH Cytoplasm 3 * AltSB Pstol1 Plant Phys 156-1041
  • 3. Fe-O Citrate PO4 -- Band A Band B Band C 10 ppm (P) 5 ppm (P) 0 ppm (P) 10 ppm (P) 5 ppm (P) 0 ppm (P) 0 - 20 cm - Spatial models (r/c) - Environmental covariates - Phenological covariates Phenotyping for Phosphorus Efficiency - Soil moisture strongly impaires P diffusion - PAE x PUTIL
  • 4. Phenotyping for Root Morphology (2D) - Importance for PAE - Shallow roots?
  • 5. PSTOL1 increases grain yield PSTOL1 increases early root growth
  • 6. Comparative Strategy to Validate Pstol1 Homologs as Genes Conferring Enhanced P acquisiton Sorghum: association mapping US (converted) Panel Contrasting parental linesPstol1 - Rice Maize: QTL mapping L22 Grain yield: low P soils Root morphology: nutrient solution Wide Diversity Panel Parentoni et al. (2010) Maydica
  • 7. Polymorphism Discovery for Linkage and Association Mapping • Primer design for amplicon sequencing • SNP detection • Conversion to the Kaspar system • Genotyping in the association panel Primer-BLAST Conserved Regions Paralog-specific primers
  • 8. Step 2: Producing F1: Ms3ms3 Step 3: Self-pollinate F1s Step 4: Random Matting Cycle 1 Step 5: Random Matting Cycle 2 Step 1: Base population development Step 6: - Random Matting Cycle 3 - Pup1/AltSB validation Selection for AltSB based on allele mining with tag markers Genotyping Genotyping S1s to produce S2s Phenotype S2s and relate genotypic information on AltSB/Pstol1 with the phenotype for validation - Provides means to validate Pup1/AltSB (higher LD) - Effect estimate - Amenable to gene pyramidation - Contingency plan based on a familiar breeding scheme (high risk project) - Entry points for new material (SorghumCI) - Cultivar release, regardless Sorghum RMPs: pyramiding AltSB and multiple Pstol1 homologs 4 cycles of random matting Products: - Agronomic performance under low P (SAP) - Root morphology and architecture (SAP) - Root morphology QTL - Functional markers