The role of pigment carotenoids in flower crops

2. THE role of Carotenoids IN Flower crops
M. Sanghamitra
Presented by,

3. Contents:
 Introduction to pigments
 Carotenoids and its Distribution
 Classification
 Structure
 Biosynthetic pathway
 Extraction
 Functions
 Carotenoids in flower crops
 Factors influencing the Carotenoid content in flowers
 Conclusion

4. • Pigment is an organic molecule that selectively absorbs light of specific
wavelengths.
• They produce the colors that we observe at each step of our lives,
because pigments are present in each one of the organisms in the world,
and plants are the principal producers.
•They are present in leaves, fruits, vegetables, and flowers , and also
present in skin, eyes, and other animal structures and in bacteria and
fungi.
INTRODUCTION

5. Types of pigments:
Photosynthetic organs of plants always contain an assortment of
pigments
• Chlorophylls
• Carotenoids
• Phycobilins
• Anthocyanins
• Betalains
• Flower color is predominantly due to three pigments i.e., Betalains,
Carotenoids, and Anthocyanins.
• Betalains are least abundant of all the above two pigments and
contribute to various hues of ivory, yellow, orange, red and violet.
• Anthocyanins contribute to various hues of red, orange, brown and
bronze.

6. CAROTENOIDS
•This is a class of accessory pigments that occur in all
photosynthetic organisms, they are especially abundant in yellow-
orange Fruits, Vegetables and Flowers.
• Carotenoids absorb light maximally between 460 nm and 550 nm
and appear as red, orange, or yellow colors.
• More than 700 naturally occurring
Carotenoids have been identified in
all plants. In flowers more than 40
Carotenoid pigments have been
identified.

7. Distribution:
• Carotenoids are widely distributed in nature.
• They are completely hydrophobic (fat soluble) and exist in lipid
membranes. They are therefore not located in the vacuoles but in the
membrane structures of plastids (exclusively in the chromoplasts of
flower petals).
• It is a common pigment in the many yellow flowers of the Asteraceae
family.
• Carotenoids differ from Anthocyanins and betalains in that they play
essential roles in plant life, for example photo protective function
during photosynthesis and provision of substrates for bio-synthesis of
the plant growth regulator ABA.

8. TYPES OF CAROTENOIDS

9. Carotenoids are classified by their chemical structure as:
(1) Carotenes that are constituted by carbon and hydrogen.
(α -Carotene, β- Carotene, γ -Carotene, Lycopene)
(2) Oxycarotenoids or Xanthophylls that have carbon, hydrogen,
and, additionally, oxygen.
(Lutein, Zeaxanthin, Violoxanthin, β- Cryptoxanthin, Astaxanthin)
Also, carotenoids have been classified as primary or secondary
(1)Primary carotenoids: Those compounds required by plants in
photosynthesis (β-carotene, violaxanthin, and neoxanthin)
(2) Secondary carotenoids: These are localized in fruits and flowers
(-carotene, β-cryptoxanthin, zeaxanthin, antheraxanthin,
capsanthin,capsorubin)
Classification

10. Structure: In general, Carotenoids are compounds comprised of
eight Isoprenoid units (ip) with polyene chains that may contain 15
conjugated double bonds whose order is inverted at molecular centre.
At one or both ends of the isoprene chain Carotenoids carry an ionone
ring.

11. β-Carotene(C40H56) , Xanthophyll(C40H56O2)
 Xanthophylls contain their oxygen either as hydroxyl groups
and/or as pairs of hydrogen atoms that are substituted by oxygen
atoms acting as a bridge (epoxide).

12. Typeof
carotenoids
Occurrence
I. Carotenes
-Carotene In many leaves
β-Carotene Major carotene of all plants (carrot, sweet potato)
γ-Carotene Traces in some plants
Lycopene Tomato, Rose hips
II. Xanthophylls
Lutein Major form of Xanthophyll (Marigold, Chrysanthemum)
Zeaxanthin Found in many plants (Safron)
Neoxanthin Found in some plants
Violoxanthin Second major Xanthophyll
β-Cryptoxanthin Found in few species
Astaxanthin Bacteria, Fungi
Flavoxanthin and
Auroxanthin
Calendula officinalis

13. Name of carotenoids Characteristic color
Lycopene Red
- Carotene Yellow
β- Carotene Orange
γ- Carotene Red- orange
Lutein Yellow
Violoxathin Yellow
β- Cryptoxanthin Orange
Zeaxanthin Yellow- orange
Astaxanthin Red
Source: Carotenoids and food preparation
Washington, DC, 1997
Carotenoids and its Characteristic color:

14. Biosynthesis of Carotenoids:
All Carotenoids can be considered as Lycopene derivatives by
reactions. The basic skeleton can be modified in many ways
including cyclization at one or both ends of the molecule to give
different end groups, changes in hydrogenation level, de-
hydrogenation and introduction of oxygen containing functional
groups, double bond migration, methyl migration, chain elongation,
chain shortening or combination of these processes resulting in a
great diversity of structures.

15. Glyceraldehyde 3-Phosphate+ Pyruvate
1-deoxyxylulose
5-Phosphate
synthase
1-Deoxyxylulose 5-Phosphate
(3) Isopentenyl- diphosphate+ (1) Dimethylallyl-diphosphate
Geranylgeranyldiphosphate
synthase
(2) Geranylgeranyldiphosphate(20 C)
Phytoene synthase
Phytoene (40 C)
1- hydroxy-2-methyl-2-butenyl
4-diphosphate reductase

16. Phytoene
Phytoene desaturase
Zeta-carotene isomerase
Lycopene
Carotene isomerase
Zeta-carotene desaturase
ε-Cyclase
β-Cyclase
β-Cyclase
β-Cyclase
-Carotene
β-ring
Hydroxylase
ε-ring
Hydroxylase
β-Carotene
β-carotene hydroxylase
Lutein Zeaxanthin
Violaxanthin
Zeaxanthin epoxidase
Astaxanthin

17. Evaporate the combined ether layers which contain carotenoids in a hot
water bath
Pool the filtrates & partition thrice with equal volumes of peroxide free
ether using a separatory funnel
Repeat the extraction until the tissue is free from pigments
Filter on a buchner funnel through whatman No.42 filter paper
Cut the fresh plant material and grind (2g) in a mortar with 20 ml distilled
acetone or methanol
Extraction of total carotenoids:

18. Measure the absorbance of this solution at 450 nm & calculate the
Carotenoid content (mg/100g) in the sample using a calibration curve
Evaporate the combined ether layers as before and dissolve the residue in
minimum volume of ethanol
Add equal volume of water & partition twice with ether
Keep the mixture overnight at room temperature or boil it for 5-10 min
Add 60% aqueous KOH at the rate of 1 ml for every 10 ml of the ethanol
extract
Dissolve the residue in minimum quantity of ethanol

19. a.Color: Carotenoids provide colors to flowers, seeds & fruits. Color has
an important role in reproduction. Coloration attracts animals that
disperse pollen, seeds or spores.
b.The function of carotenoids in photosynthesis: The red, orange and
yellow carotenoids function as accessory pigments in photosynthesis.
They contribute to photosynthesis by absorbing and transmitting the light
energy to chlorophyll and also protect photosynthetic tissues from photo-
oxidation.
Functions

20. c.Xanthophyll cycle:
• In higher plants there are three carotenoid pigments that are active
in the xanthophyll cycle violaxanthin, antheraxanthin and zeaxanthin.
• During light stress violaxanthin is converted to zeaxanthin via the
intermediate antheraxanthin.
• This is one of the plant protection mechanisms against light
damage.
• The number of carotenoid molecules is higher in sun-exposed
leaves than darkness maintained leaves. Also, xanthophyll cycle
carotenoids (violaxanthin, antheraxanthin, and zeaxanthin) are
increased in sun-exposed leaves.

21. Contud…..
•This phenomenon is very important, sun exposed leaves in a fast-
growing stage use not more than 50% of absorbed energy during
the stage of maximum radiation (midday), and in some species only
10% is used.
• Thus, 50 to 90% of absorbed light is in excess and must be
eliminated in order to avoid cellular damage.
• Xanthophyll cycle is a process that makes the energy dissipation
easy and protects the photosynthetic apparatus.

22. d. Anti-oxidant:
• Carotenoids are also a popular source of antioxidants. Antioxidants
protect our body from free radicals.
• Free radicals are unstable oxygen molecule, which damage our body
cells through destructive oxidation process.
• According to studies, several carotenoids are capable of preventing
growth of malignant cells.
• They can be fighter of lung cancer, breast cancer, prostate cancer and
skin cancer.
• Researches have shown that if beta-carotene and lycopene is consumed
with antioxidant rich vitamins C and E, the adverse side effects of
radiation therapy and chemotherapy, used in cancer treatment, can be
significantly reduced.
• Among the known carotenoids, lycopene is the most powerful
antioxidant. Canthaxanthin and astaxanthin show better antioxidant
activity than β-carotene or zeaxanthin.

23. e. Provitamin A:
 Carotenoids also play an important role in human nutrition and
health, providing provitamin A. Beta-carotene is known as
provitamin A, since it is converted into vitamin A in the body. Nearly
ten percent of carotenoid present in our food can be converted into
vitamin A in the liver and intestine. Besides beta-carotene, alpha
carotene and nearly 50 other types of carotenoids can be converted
into vitamin A.
f. Macular degeneration:
 Lutein and other carotenoids may protect photoreceptors in the
retina of the eye from wavelength blue light and help prevent age-
related disorders to the eye such as macular degeneration.

24. • Natural and synthetic carotenoid pigments are used in medicines,
clothes, furniture, cosmetics, and in other products.
• A lot of pigments have a well-known pharmacological activity in
sickness such as cancer and cardiovascular diseases, and this has
stressed pigment importance for human beings.
USES

25. Additives, colourants:
• β-Carotene and other synthetic or natural carotenoids or carotenoid-
rich extracts are widely used as additives to colour processed food,
drinks, confectionery, ice-cream etc.

26. •The carotenoids extracted from flower petals are added to poultry
feed for intensification of yellow color of egg yolk.
• They are used as dyes for coloring the textiles.
• The carotenes promote the renewal of skin tissue + antibacterial
properties prevent infections.

27. • Carotenoid formation in flowers is associated with the conversion of
chloroplasts into chromoplasts.
• Chromoplast formation commences with the disruption of grana
thylokoids and disappearance of the chlorophylls.
• In many chloroplasts compartments are formed in place of the grana in
which carotenoids are synthesized.
PIGMENT FORMATION

28. Carotenoids in flower crops:
Flowers have been identified that synthesize:
 Highly oxygenated carotenoids
 Principally β carotenes
 Carotenoids that are species specific ( eg: Eschscholzxanthin in
Poppies)
California golden poppy (Eschscholzia californica)

29. Common name: Marigold
Botanical name: Tagetus erecta
Family : Asteraceae
•Xanthophylls are the major carotenoid fraction in the flower petals.
• Lutein accounts for 80 – 90% of total Xanthophylls content.
• Used for poultry feed to intensify the yellow colour of the egg yolk
and broiler skin and also for fish.
• Also used for colouring food stuffs.
Source:
Archives of Biochemistry
and Biophysics(2010)

30. Screening of African marigold (Tagetus erecta L.) cultivars
for flower yield and carotenoid pigments.
Chandrashekar Rao et al, (2005)
Case study: 1

31. Cultivar Days to first
flowering
Flower
duration(days)
Flower
diameter(cm)
Number of
flowers
Total
caroteniods(mg/g)
Percent
petal
weight
Pusa Narangi
Gainda
97 35 7.7 30.0 2.69 66.4
Orange Double 95 44 13.4 18.3 2.66 87.8
Hyd. Local sel-1 99 23 7.4 32.3 1.07 43.8
Pusa Basanti
Gainda
110 38 8.2 17.0 1.37 64.7
African Tall
Double Orange
100 31 6.5 25.0 1.87 58.3
Hyd. Local sel-2 102 20 7.1 13.3 0.20 68.6
Hyd. Local sel-3 98 18 5.9 19.0 0.64 48.6
Hyd. Local sel-4 103 21 8.1 7.0 0.32 71.8
Yellow double 109 29 8.5 15.0 0.87 75.3
Lemon yellow 105 25 6.9 11.0 0.55 68.8
Table: Variability in qualitative characters in African marigold cultivars

32. Common name: Chrysanthemum
Botanical name: Chrysanthemum morifolium
Family: Asteraceae
• Chrysanthemum is a commercially
valuable ornamental plant with bright
yellow petals, mainly reflecting the
accumulation of lutein.
Source: Archives of Biochemistry and Biophysics
(2010)

33. Common name: Pot marigold
Botanical name: Calendula officinalis
Family : Asteraceae
• The petals and pollen of Calendula officinalis contain the carotenoids
flavoxanthin and auroxanthin as antioxidants, & source of their yellow-
orange coloration
• As a rinse for the hair, pot marigold gives a golden tinge to fair hair.
• Petals used to add color to salads , extract is common food additive to
produce darker egg yolks.
( Bako et al., 2002)

34. Common name: Summer pheasant’s eye
Botanical name : Adonis aestivalis
Family : Ranunculaceae
• It is an ornamental plant native to Europe.
•The petals of Adonis aestivalis and A. annua anomalously accumulate a
large amount of astaxanthin, resulting in their blood-red color.
• Astaxanthin is a ketocarotenoid that is produced in a number of
bacteria, fungi and algae. Only a few plant species are known to produce
Astaxanthin.
Source: Archives of
Biochemistry and
Biophysics(2010)

35. Botanical name :Osmanthus fragrans
Family: Oleaceae
• It is a flower native to China that is valued for its delicate fruity-
floral apricot aroma.
• The flowers of osmanthus range from silver-white (Osmanthus
fragrans Lour. var. latifolius Mak.) to gold-orange (Osmanthus
fragrans Lour. var. thunbergii Mak.) to reddish (Osmanthus fragrans
Lour. var. aurantiacus Mak.).
• Various workers have examined the different colored varieties and
find that the gold-orange variety (e.g., Osmanthus fragrans Lour. Var
thunbergii ) tends to have more of the desirous notes and tend to be
higher in carotenoid derived materials. Among the carotenoids of
Osmanthus accumulate beta-Carotene in higher amounts.
•The flowers are used to flavour jam, sauce and sweets.

36. Osmanthus fragrans

37. Common name: Weld
Botanical name: Reseda luteola
Family: Resedeceae
Dyer's Rocket or dyer’s broom, is an European plant with long
spikes of small, yellowish-green flowers contain carotenoids cultivated
as a source of yellow dye.
Source: Archives of Biochemistry and Biophysics(2010)

38. Common name : Dandelion
Botanical name: Taraxacum officinale
Family: Asteraceae
• The flower heads are yellow to orange color.
• Analysis have detected the presence of beta- carotene,
cryptoxanthin, flavoxanthin and lutein.
• Mixed carotenoids can act as cancer preventing anti-oxidants.
• With this dandelion flowers we can make a tea that helps muscles
to relax.

39. Common name: Lilies
Botanical name: Lilium candidum
Family: Liliaceae
• The Asiatic hybrid lily (Lilium spp.) is another commercially
valuable ornamental plant with flower colors ranging from red,
orange and yellow (carotenoids).Carotenoid profiling has shown
that most of the carotenoids in yellow petals are antheraxanthin,
violaxanthin and lutein .
Source: Archives of Biochemistry and
Biophysics(2010)

40. Common name: Christamas bells
Botanical name: Sandersonia aurentiaca
Family: Colchicaceae
 The pigments responsible for
the golden orange flower color of
S. aurantiaca have been
characterized and the major
carotenoids were zeaxanthin and
β-cryptoxanthin
(Neilsen et al., 2003)

41. Carotenoids in some other flower crops:
Narcissus (β- Carotene, Lutein)
Coreopsis
Viola tricolor
(Yellow cultivars of
Coreopsis and Viola
contain carotenoids)
Source:
Liedvogel et al., 1976.,
Hansmann and Kleinig, 1982

42. Golden rod
Tansy
Source: Archives of
Biochemistry and
Biophysics(2010)
The flower heads of Golden rod and Tansy contains small
amounts of Carotenoids.

43. Tulips(β –Carotene, Lutein) Oncidium
(Violoxanthin)
Source: Archives of Biochemistry and Biophysics (2010)

44. Effects of Environmental Conditions and Cultivation Practices on
carotenoids:
• Environmental conditions and cultivation practices influence
Carotenoid content, since it is related to photosynthetic efficiency and
density of chloroplasts.
• Heat stress, drought stress, and stress by pollution or salt are
detrimental to Carotenoid content, as they effect the plant growth and
health in general.
• The age and maturity of plant tissues at harvest is a significant factor
leading to apparent variations in Carotenoid concentration.

45. Hot Weather Increases Carotenoids:
• Carotenoids increase production in response to heat and lots of sun.
•The hotter and sunnier the weather gets, the more carotenoids a plant
produces, and flower color becomes brighter orange and red.
•When weather cools off, carotenoid production decreases, and colors
become softer oranges and yellows.

46. • It works with Hibiscus. In hot, sunny, summertime heat, Hibiscus
is bright red with vivid yellow markings. In cool cloudy weather,
hibiscus bloom with soft orange color. Banga and Bruyn noticed
synthesis of more carotenes at higher temperature (23°C) than at the
low temperature (17° or 10°C).
Bright red with vivid yellow
markings Soft orange color

47. Fertilization enhances the carotenoid content of flowers:
• By application of potassium increases the carotenoid content of flower
because ‘K’ play a special role in the process of carotenoid biosynthesis
by activating several enzymes participating in biosynthetic pathway as
well as the precursors of isopentanyl diphosphate. (Pyruvate and
Glyceraldehyde 3 phosphate)
• By application of ‘K’ in the form of sulphate of potash also increases
the carotenoid content through supply of sulphur along with potassium
from sulphate of potash.(Ananthi et al., 2004)

48. Fertilization enhances growth, yield, and xanthophyll contents of
marigold
Iftikhar et al,(2010)
Case study:2

50. Light Absorption and Carotenoid Synthesis of Pot Marigold
(Calendula officinalis L.) in Response to Phosphorous and Potassium
Varying Levels.
Sedhgi et al, (2011)
Case study:3

51. P2O5 (kg/ha) K2O (kg/ha) β-Carotene(mg/g) Biological
yield(g/m2)
0 0
50
100
150
2.5
2.8
3.1
2.9
360.4
385.3
489.1
495.6
40 0
50
100
150
2.6
3.2
3.4
3.9
368.9
392.4
498.3
507.6
80 0
50
100
150
2.7
3.5
4.1
4.3
376.1
402.1
524.6
534.2
120 0
50
100
150
2.5
3.5
3.4
3.4
411.5
510.7
548.3
552.1
Table: Compaision of means of carotenoids and yield of pot marigold as effected by interaction of K and P

52. Improvement of the Yield and Essential Oils Quantitative in
Calendula (Calendula officinalis L.) by Using Different Planting
Arrangement and Potassium Fertilizer
Hashemabadi et al, (2012)
Case study:4

53. Treatment Carotenoid( mg/g)
A1 (Square arrangement) 6.68
A2 (Lozeng arrangement) 6.58
A3 (Rectangular arrangement) 6.20
K1 (Control) 5.79
K2 (100 kg /ha) 6.45
K3 (200 kg/ha) 6.31
K4 (300 kg/ha) 7.38
Table: Effect of plant arrangement , K2SO4 and interaction between plant
arrangement and K2SO4 on traits

54. • Recovery of carotenoids depends on the stage of harvest.
• Larger flowers have more carotenoids compared to small and over
mature flowers.
• As the age of the flower increases there will be degradation of
carotenoids.
Recovery of Carotenoids

55. Recovery of carotenoids and its fractions from marigold flowers as
influenced by genotype, grading and stage of harvest.
Kanwar et al,( 2008)
Case study: 5

56. Geno
type
Total carotenoid
content(mg/100g)
Carotene content (mg/100g)
Harvesting
stage
Flower grade Harvesting
stage
Flower grade
Half
bloom
Full
bloom
Small
flower
Large
flower
Half
bloom
Full
bloom
Small
flower
Large
flower
Pusa
Narangi
Gainda
415.84 385.48 355.77 378.41 25.01 20.72 18.21 23.14
Sel-8 405.19 347.15 308.77 321.72 25.06 20.36 18.45 20.78
Sel-19 397.05 326.75 302.58 319.28 47.13 45.09 41.64 44.37
Sel-20 380.95 312.56 291.43 301.41 21.01 17.45 15.14 19.09
Sel-22 369.83 318.78 295.99 304.97 17.67 14.59 13.73 15.69
Sel-29 366.99 331.05 298.04 319.86 46.77 44.07 42.24 43.89
Table: Total carotenoid and carotene content in different marigold genotypes at different
harvesting stages and flower grades

57. Flavonoid and carotenoid pigments in flower tissue of
Sandersonia aurantiaca
David et al, (1997)
Case study: 6

58. Stage of development Zeaxanthin Cryptoxanthin
1:An immature green
bud
0.34 Not detected
4:Flower open but still
has green tips
0.79 0.52
7:Mature flower
completely orange
1.69 2.54
10:Petals wilting and
orange color fading
1.01 1.52
Table: Concentration and content per flower for individual carotenoids in a
saponified extract from flowers of Sandersonia at different stages of development.

59. Conclusion
 Flowers accumulates large amount of different
carotenoids.
 Considering the importance of carotenoids for
industry, human health and plant development, emphasis
has been given to targeted manipulation of carotenoid
biosynthesis in flowers.
 Diet rich in carotenoids represents lower risk for
several diseases.
By the addition of natural carotenoids to healthy
foods, the food products containing carotenoids may
become superfluous in the future.
 Carotenoid engineering is expected to contribute to
human health, as carotenoids are important pigments as
well as nutrients.

60. Future trends:
• More studies must be carried out to have complete vision of the
biosynthesis and regulation of Carotenoids.
• The researchers should put an interest in two areas ie., generation
of crops with improved characteristics and pigment production at the
industrial level and under controlled conditions.
•Food technologists should continue affronting the problems of
availability and stability of natural pigments in order to replace the
synthetic ones.
• Carotenoid production by Yeasts, Bacteria and fungi require the
development of better biotechnological approaches.