Carotenoid Pathway in Capsicum Annuum Laura Rodriguez-Uribe Department of Plant and Environmental...
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![Page 1: Carotenoid Pathway in Capsicum Annuum Laura Rodriguez-Uribe Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM.](https://reader036.fdocuments.us/reader036/viewer/2022062304/56649ced5503460f949bae5f/html5/thumbnails/1.jpg)
Carotenoid Pathway in Capsicum Annuum
Laura Rodriguez-Uribe
Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM USA
![Page 2: Carotenoid Pathway in Capsicum Annuum Laura Rodriguez-Uribe Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM.](https://reader036.fdocuments.us/reader036/viewer/2022062304/56649ced5503460f949bae5f/html5/thumbnails/2.jpg)
Carotenoids
•Most abundant natural pigments, more than 700 carotenoids have been identified in photosynthetic organisms such as plants, algae, and bacteria
Carrot Corn Tomato Pepper
Algae Bacteria β-carotene Deinoxanthin
β-carotene Lutein/Zeaxanthin Lycopene Capsanthin/Capsorubin
![Page 3: Carotenoid Pathway in Capsicum Annuum Laura Rodriguez-Uribe Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM.](https://reader036.fdocuments.us/reader036/viewer/2022062304/56649ced5503460f949bae5f/html5/thumbnails/3.jpg)
Carotenoid biosynthetic pathway in Capsicum
Peppers possess the most evolved carotenoid pathway
![Page 4: Carotenoid Pathway in Capsicum Annuum Laura Rodriguez-Uribe Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM.](https://reader036.fdocuments.us/reader036/viewer/2022062304/56649ced5503460f949bae5f/html5/thumbnails/4.jpg)
Carotenoid biosynthesis and accumulation
Fruit wall
Chromoplasts
Carotenoid composition changes duringfruit ripening
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Biological Antioxidants, Vitamin A precursorCarotenoids quench free radicals, prevent protein oxidation and DNA damage
Sources of Free Radicals: Effects:
Cigarette smoke Cancers
Environmental pollutants Atherosclerosis
Radiation Damaged DNA
Ultraviolet light Heart disease
Pesticides Rheumatoid arthritis
Nutritional supplements
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New Mexico red pepper
Pigments (natural dyes) Cosmetics
Industry
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Species Cultiv ar Phenot ype -car o tene Capsanthin Total C arotenoid
C. annuu m Hung a rian Apple Red 0.137 ± 0 0.310 ± 0.05 1.524 ± 0.18 Andy Red 0.415 ± 0.01 0.747 ± 0.04 5.558 ± 0.52 Indian PC -1 Red 1.361 ± 0.11 0.443 ± 0.12 3.756 ± 0.83 Big Red Cayenne Red 0.405 ± 0.07 0.230 ± 0 1.597 ± 0.15 Blackbird Red 0.093 ± 0.02 0.755 ± 0.27 2.105 ± 0.42 Giant Thai Red 1.319 ± 0.26 1.151 ± 0.05 6.551 ± 0.34 NuMex Garnet Red 1.158 ± 0.05 0.655 ± 0.45 5.342 ±1.38 Pimiento Red 0.175 ± 0.03 2.9 76 ± 0.77 5.675 ± 0.46 Sandia Red 0.391 ± 0.05 0.558 ± 0.03 3.348 ± 0.56 Sweet Banana Red 0.315 ± 0.09 0.094 ± 0.04 0.693 ± 0.39 NuMex Centennial Red 0.121 ± 0.10 0.036 ± 0.02 0.213 ± 0.08 Nambe Red 0.577 ± 0.22 3.028 ± 0.64 10.755 ± 2.54 NuMex Nematador Red 0.563 ± 0.40 4.014 ± 2.86 9.073 ± 2 Alba Orange 0.014 ± 0 0.047 ± 0 0.151 ± 0.01 Blushing Beauty Orange 0.000 ± 0 0.142 ± 0.05 0.311 ± 0.01 Costeno Amarillo Orange 1.239 ± 0.04 0.303 ± 0.04 3.515 ± 0.50 Early Sunsation Orange 0.116 ± 0.06 0.132 ± 0.07 0.461 ± 0.23 Orange Thai Orange 0.822 ± 0.07 0.322 ± 0.07 2.446 ± 0.43 Oriole Orange 0.277 ± 0.10 0.601 ± 0.12 1.442 ± 0.42 Gourmet Rainbow Orange 0.503 ± 0 0.954 ± 0 3.666 ± 0 Mandarin Orange 0.037 ± 0.01 0.075 ± 0.02 0.130 ± 0.01 NuMex Thanksgiving Orange 0.007 ± 0 0.028 ± 0.01 0.048 ± 0.017 . St Patri ’cks Day Orange 0 ± 0 0.011 ± 0.01 0.011 ± 0.01 NuMex Sunglo Yellow 0 ± 0 0.119 ± 0 0.301 ± 0 Yellow Cheese Pimiento Yellow 0.039 ± 0.04 0.097 ± 0 0.313 ± 0 Dove White 0.000 ± 0 0.007 ± 0 0.011 ± 0
. C baccatum Bolivian Yellow Orange 0.204 ± 0.18 0.924 ± 0.86 2.497 ± 1.20 . C chinense Aj i Dulce Red 0.228 ± 0.04 0.417 ± 0.22 1.103 ± 0.24
NuMex Suave Orange Orange 0 ± 0 0 ± 0 0.011 ± 0 J amaican Hot Chocolate Brown 0.204 ± 0.18 0.102 ± 0.04 0.891 ± 0.43 J amaican Yellow Yellow 0.280 ± 0.13 0.207 ± 0.09 1.478 ± 0.70
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Carotenoid Quantities
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Carotenoids in Orange Peppers
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Carotenoid profile in ripe orange peppers
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Thin layer chromatography with the carotenoid composition at different ripening stages
Valencia Fogo NuMex Standards Sunset
G GO O G GO O G GO O Cs Cr V
Green
Green/Orange
Orange
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Objectives
• Investigate the β-carotene content of orange fruited capsicum varieties to correlate the relationship between fruit color and β-carotene content
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Carotenoid biosynthetic genes in orange colored peppers
• Carotenoid pathway genes phytoene synthase (Psy), lycopene-β cyclase (Lcyb), β-carotenoid hydrolase (CrtZ-2), and casanthin-capsorubin synthase (Ccs), from six orange fruited peppers were cloned and sequenced.
• Mutations in the Lcyb of the six orange peppers, and a new version of the Ccs gene from Fogo pepper genes.
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Multiple alignment of the capsanthin- capsorubin synthase protein
Multiple alignment with the Ccs amino acid sequences from Yolo wonder, Valencia, and Fogo.
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Expression of carotenoid biosynthetic genes during ripening in orange peppers
EtBr
Ccs
CrtZ-2
Psy
Lcyb
G GO O G GO O G GO O
Valencia Fogo NM Sunset
Green
Green/Orange
Orange
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Approaches to test the functionality of the CCS mutant
• Biotechnological: Expression of the wild type and mutant Capsanthin-Capsorubin
synthases as recombinant proteins in an heterologous system such as Escerichia. Coli (E.coli) and assay its activity.
• Biochemical: Purification and characterization of the Capsanthin-Capsorubin
from Valencia and Fogo peppers chromoplasts
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Expression of Capsanthin capsorubin synthase in E. coli as a recombinant protein
kD 1 2 3 4 5 6 7
755037
25
Ccs
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Purification of CCS from pepper fruit wall chromoplasts
Fruit wallPercoll density gradient
Chromoplast
Isolation and Purification of CCS protein
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Carotenoid composition differs among peppers
The six orange peppers in this study have a new version of the Lcyb gene A novel version of the Ccs gene was found in the Fogo cultivar
Differences in gene expression in the carotenoid pathway might determine the carotenoid composition in mature peppers .
Conclusions
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Overall SignificanceNMSU Research
Metabolomics / Breeding
New MexicoBreeders
Nutrient + Pigment Rich Peppers↑ β-Carotene
↑ Capsanthin/Capsorubin↑ Quality in New Mexico
Improvement of other agronomically important crops
World Nutrition
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Thank You!
Dr. Mary A. O’Connell Dr. Paul BoslandDr. Richard D. Richins Chile Pepper InstituteLab members
Supported in part by NM Agricultural Experiment Station and
NIH grant R25 GM61222
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Carotenoid Chemistry
Two kinds of carotenoids
» Non Pro-Vitamin A
lycopene
» Pro-Vitamin A
-carotene