Coffee Breeding UpdateCoffee Breeding Updatewith new toolswith new tools
HCA Conference 2003Chifumi Nagai
Hawaii Agriculture Research Center
New coffee varieties with high yielding and superior quality
with rust and nematode resistance having cupping quality
Coffee Breeding and SelectionCoffee Breeding and Selection
Quality evaluation (Cupping)
Flavor chemistry
New tools
DNA markers
Genetic Engineering
Traditional tools
Existing varieties
Foreign germplasm
Hybridization
Visual selection
Data collection
Backcross
HARC Coffee Program • Breeding and Selection
HCGA/State,DOA 1997-
• Molecular Markers
USDA grant (CMBRU)
Ray Ming, Heather Pearl, C. Nagai
• Transgenic coffee for nematode resistance
USDA grant ( T-STAR )
Brent Sipes, D. Schmit. H. Atkinson, Roxana Cabos, C.Nagai
• Green Bean Chemistry
USDA grant (CMBRU) Mel Jackson, Cathy Cavaletto, Shawn Steiner, C. Nagai
Coffee Breeding and Selection
HARC- teamC. Nagai, B. Osgood, R. Ming, M. Jackson, L. Poland, K. Wiever, J. Buenafe, J. Clayton
UHC. Cavaletto, N.Nagai and S. Bittenbender,
Hawaii Coffee Growers’ Association (HCGA)
Kauaiyellow Catuaired Catuai
Oahumokka hyb.Promecafe
Molokaired Catuaired Caturra
Kona‘Guatemalan’ typica‘Old Hawaiian’Hawaiian
Mauimokkared Catuai
Common FieldKunia, HARCplanted April,1998
Crosses (spring 1999)Group # of crosses
mokka and other arabica varieties 66 with Promecafe (Catimor) 53with Icatu 8mokka and other selfs 40
Total 167
H99 - series Progeny
New New CultivarCultivar Development Development using using MokkaMokka HybridsHybrids
•Mokka was introduced to the UH coffee germplasm collection in the mid 1950s
•Commercially cultivated on Maui (Kaanapali Coffee Co). The accession is now considered to be a Mokka hybrid.
•The cupping quality of Mokka hybrid is considered excellent but bean size is very small ( about 30-40% of Red Catuai beans).
•In 1999, we made crosses focused on increasing mokka seed size while maintaining its excellent cupping quality.
Goal:The introgression of Mokka flavor traits into high yielding and larger bean size cultivars
• Tree heightRed Catuai x Mokka hyb Red catuai dwarf x mokka
hybRed Catuai x typicaYel Caturra x mokka
• Cupping Quality• Increase bean Size
Yellow catuai x MargogipeMokka hyb. SelfMokka x Red CatuaiRed Catuai x SL28Mokka hyb selfRed Catuai x 6661
Example of Progenies from 1999 CrossExample of Progenies from 1999 Cross
Cherry and green bean) size Cherry and green bean) size of families with of families with mokkamokka andand
MargogipeMargogipe crosses crosses Fall 2002Fall 2002
Cross# Parentage n 100 fruit wt Green beangram gram
H99-34 KA17-5 x Margog 3 241 ± 3 41.7 ± 0.2(Y. catuai)
H99-36 MA1-12 x Margog 5 258 ± 17 42.9 ± 4.3(mokka)
H99-22A MA1-12 self 5 179± 9 29.2 ± 1.4(mokka select )
H99-101 KA16-4 self 6 148 ± 1 25.5 ± 1.7(Y. catuai)
H99-36:MA1-12 x Margogipe(mokka)
Potential New cultivarPotential New cultivar
Cupping of 3 genotypes of coffee Cupping of 3 genotypes of coffee harvested at harvested at KuniaKunia in 2002in 2002--03 Season03 Season
By Cathy Cavaletto’s Panel ( April 2003)
Cupping Panel: 7 trained cuppers (Judge)
Genotypes ( 2 hybrids + 1 parent)H97-1700: Yellow Catuai (KA17C)H99-34: KA17 x MargogipeH99-36: MA1-12(mokka) x Margogipe
Replication: 3 days
Cupping ResultsCupping Results
_________________________________________________________Source df Dry Wet Acidity Flavor Body
aroma________________________________________________Judge 6 ** ns *** ns **Genotype 2 ns ns ns ns nsRep 2 ns ns ns ** nsResidual 43_________________________________________
**: significant at P = 0.01***: significant at P= = 0.001ns= not significant
ANOVA
Propagation of Selected Hybrids by cloning
Preparation of treesInduction of vertical shoots
Kunia H99- fields
Cloning1. Shoot tip culture
2. Root induction of cuttingsUsing misting/ bottom heat
3. Bioreactor of somatic embryos(root stock production)
Summary and plan ‘03-’04• Field Trial at Kauai Coffee- “FT7” design
Germinated Seeds from selected hybrids at MWüShip seedlings to Kauai (9/03)Plant seedlings at 2 fields, Kauai Coffee(12/03)Data collection (5/04-)
• Field planting of 2002 hybrids ( 7/03) ü
• Harvest and data collection of H99- series at Kunia (10-12/03)
• Crosses using new germplasm and selected genotypes (12/03-3/04)
• Send selected individuals for rust resistance tests.• Make new crosses of selected F1 plants
DNA Marker Study 2002-2003
Ray Ming, Heather Pearl and C. Nagai
• Genetic evaluation of Fukunaga and newly introduced coffee
• DNA analyses of green beans
• Development of coffee populations for QTL mapping
• Construction of a genetic map of Arabica coffee
DNACoffee leaves
Genetic Similarity (Cophenetic coefficient = .99)0.41 0.56 0.71 0.85 1.00
Bourbon1Bourbon3Typica-H1Typica-H2Typica-K6Typica-H3Typica-K2Typica-H4Typica-H5Typica-K10Typica-K7Typica-K9Typica-K3Typica-H8Typica-H9Typica-H7Typica-K1Typica-K11Typica-K32Typica-K33Typica-K8Catuai8Typica-K4Typica-K34MH3MH2MH5 MH6MH7Catuai1Catuai2Catuai5Caturra1Caturra2Catuai3 Catuai4Typica-K12Typica-K13MH8Catuai6Typica-K5Catimor4Catimor5Catimor7 Catimor8Catuai9MH4Catuai10Bourbon4Catimor6Bourbon2MH1Typica-H6Catuai7Catimor1Catimor3Catimor2C-canephoraC-libericaDeweveri
Genetic Relationships Within and Genetic Relationships Within and Between Between CoffeaCoffea SpeciesSpecies
Brown- Bourbon
Green- Typica
Blue- Catuai/Caturra
Lt. Blue- Mokka hybrid
Pink- CatimorBlack- Diploid Species
Application of Fingerprinting:
– Determine the genetic relationship of Fukunaga (Coffea liberica, deweveri) trees from various sources in Konaand other trees showing liberica traits
– Examine the differences between newly introduced cultivars and long established Hawaiian cultivars such as Kona typica
SamplesArabica (2n=44) 30 smples– tetraploid and self fertile– Blue Mountain(Jamaica), Blue Mountain (HARC)
Kona Typica, Mokka, Laurina, tall Mokka Hybrid, Bourbon, Mundo Nova
New Brazilian cultivars : Icatu, Obata, and Tupi
Liberica “Fukunaga “ (2n=22) 20 samples– diploid and self-sterile – torlelet nematodes , used as root stock for Kona typica
Canephora (robusta) 2 samples– diploid and self-sterile
List of List of Fukunaga Fukunaga samples from samples from KonaKona(collected by Mario (collected by Mario SarracinSarracin 11/20/01)11/20/01)
Sample Name Source Additional InformationKES1 UH Kainaliu H-Field, Marc Meinsner's selec. From P.Ito's original treesKES2 UH Kainaliu H-Field, Marc Meinsner's selec. From P.Ito's original trees
KES (Original) UH KainaliuKES 16 KraigLeeKES 6658 See MareM., Next to Arnold Diama 6658CTS 2F TwigSmith Farm Flower PlantedCTS 2S TwigSmith Farm Seed PlantedCTS 3 TwigSmith FarmCTS 4 TwigSmith FarmCTS 5 TwigSmith FarmCTS 6 TwigSmith FarmCTS 7 TwigSmith FarmCTS 9 TwigSmith FarmCTS 10 TwigSmith FarmCTS 11 TwigSmith FarmCTS 12 TwigSmith FarmCTS 13 TwigSmith Farm Reddish leaves, pointed, fruit not clustered, multiple verticalsCTS 14 TwigSmith FarmCTS 15 TwigSmith Farm Seeds Taken
C. liberica dewevrei (Fukunaga)Kraig Lee C. liberica dewevrei (Fukunaga)
Looks like dewevrei type, not fruiting
Genetic Similarity Tree
Genetic Similarity Coefficient0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00
MWBlueMountain1
BlueMountain3 KonaTypica4 KonaTypica3 KonaTypica2 MokkaHybrid4 Mokka3 Luurina4 Luurina2 Luurina3 KonaTypica1 MundoNova1 MO2-7-2 MundoNova2 Bourbon1 Bourbon2 MokkaHybrid3 Tupi1 Mokka1 BlueMountain6 MO2-7-1 YellowBourbon1 BlueMountain4 BlueMountain5 5175-1 5175-2 MWBlueMountain1 Icatu2 Obata1 Obata2 Tupi2 Canephora1 Canephora2 KESLiberica1 KESLiberica2 KESlibericaOrig KESLiberica16 KESLiberica6658 CTSLiberica12 CTSLiberica2S CTSLiberica2F CTSLiberica4 CTSLiberica13 CTSLiberica3 CTSLiberica8 CTSLiberica11 CTSLiberica14 CTSLiberica7 CTSLiberica6 CTSLiberica15 CTSLiberica5 CTSLiberica9 CTSLiberica10
Easily distinguishable groups were seen between the three species:
Arabica
Canephora
Liberica (Fukunaga)
Species Specific Marker Bands:ArabicaMarkerBand
LibericaMarkerBand
Genomic DNA analyses of green beans
• Isolated DNA from commercially processed green beans
• Tested DNA quality -- partially degraded
Polymorphic DNA Markers
Linkage Groups
Mapping of arabica coffee Development of a mapping population
P1: Promecafe1(C): X P2: Mokka (M) (1999)T5175-1 MA2-7
large bean size, small bean sizelow cupping quality, high cupping qualityRust resistance rust susceptible
F1(Pseudo F2)130 progeny
F2300 progeny(will be planted 9/03)
QTL (Quantitative Trait Locus) for marker assisted selection
Find correlation between DNA markers and coffee traits such as Tree Characters
leaf shapestree heightbranching angles
Bean Charactersbean sizeyieldcupping quality
• Developed a true F2 mapping population with 323 progeny
• Constructed the first Linkage map of Arabica coffee( Pearl et al. 2003 submitted)
• Mapped QTLs for five traits• Evaluated the possibility of applying DNA
markers on coffee green beans• Fingerprinted a new set of coffee
germplasm including Fukunaga
Summary 2002-2003
Genetic Transformation of CoffeeGenetic Transformation of Coffeefor Nematode Resistancefor Nematode Resistance
Team: Roxana Cabos(UH), Brent Sipes(UH), Don Schmitt(UH), Howard Atkinson(Leeds U)and C. Nagai
Damage to M. konaensis Infested Coffee Trees
Galls
Serracin et al ,2001
Cystatin
• From Rice Flower
• Inhibitor for Cysteine Proteinases
• Reduces Nematode Growth & Fecundity
• Tested for Safety
• Field testing in potato,rice, banana by Atkinson’s group
Atkinson et al, 2001
Coffee Tissue CultureCoffee Tissue Culture
Germination
Plants with roots Nursery
Somatic embryosCallus from young leaves
Production of Nematode Resistant Coffee Plants by Genetic Engineering
Callus Induction of Coffee Leaves
Somatic Embryos
Selection & Regeneration
Insertion of Cystatin GeneCystatin OcID86
Gene bombardment
Agrobacterium-mediated transformation
Selection of Transgenic Somatic Embryos and production of transgenic plants
Coffee Plantlets with cystatin gene
435 Plantlets Transformed w/Cystatin Constructs
Acknowledgements
hHawaii Coffee Growers Association
hUniversity of Hawaii, Kainaliu Exp. Station
hDr. Loren Gautz, University of Hawaii
hDr. Mario Serracin
hDr. H. Medina-Filho, Instituto Agronomico de Campinas
hJamaica Coffee Industry Board
hSusan Ancheta
hLaetitia Fournier, E.S.I.P.A