5

INTRODUCTION AND LITERATURE REVIEW

6

INTRODUCTION AND LITERATURE REVIEW

A wide range of plants is cultivated for the oil they produce, and these oils may be

broadly divided in to the essential volatile oils and non volatile oils. These two types of

oil have basically different compositions and uses, and the fixed non volatile oils are

commercially the most important. Although the range of plants cultivated for their oils is

extensive, only a few are suitable for large scale commercial production, or produce oil

which is required in large quantities. Industries can be established using these by-product

and high protein meals, which may be obtained, can be used to help feed increasing urban

populations or to balance deficient diets one of these genus for commercial production is

Carthamus. The genus Carthamus L. belonging to the tribe Cynareae (thistle), sub-family

Tubifloreae of family Compositae has about 42 species with varying chromosome

number of 2n=20 to 2n=64 and has a wide range of adaptation. C. tinctorius, commonly

called safflower, is the only cultivated species of this genus with several ethnic names

like Alazor, Azafran, Beni-Bana, E'Sfer, Habb Et Quirthim, Huang Lan, Hung Hua,

Hung Lan Hua, Kasumba, Kesumba, Qurtum, Yao Hua. Pharmaceutical name of

Safflower (Carthamus- tinctorius) is Carthamini. Commercially produced safflower

seeds contain 32 to 52 percent oil. The plants of this species have been grown for

centuries in small plots over a vast area from China to the Mediterranean region and

along the Nile valley as far as Ethiopia. It is one of humanities oldest crop cultivated in

India mainly for oil from the seeds and reddish and yellow dyes for clothing and food

preparation from the flowers. Safflower (C. tinctorius) flowers have been used in

preparation of ayurvadic medicines in India. The Safflower flowers are used in China for

the treatment of many illnesses as well as in tonic tea. NARI (2002-2003) prepared the

herbal tea with the addition of aromatic herbs. Safflower provides three principle

products oil, meal and birdseed. Safflower oil is used by both food producers and by

industry. The development of this plant as an oil crop came later although it was known

for edible oil in pre-Christian times in the Mesopotamian region (Weiss, 1971). The

green safflower crop can be used as a green fodder for cattle as cattle relish it.

Safflower florets contain carthamin (C21H22O11 H2O), which is red and insoluble in

water, and safflower yellow (C16H20O11), which is soluble in water. Carthamin is found

7

in the florets to the extent of 0.3%-0.6% and imparts a bright red colour to cotton and

silk fabric. Reiff (1546) grew it in Europe for this purpose in the sixteenth century as far

north as southern Germany. In the seventieth century, safflower was extensively grown

in the Alsace region. It was also grown in Poland and Czechoslovakia (Celakovsky,

1867). Beck (1893) found safflower being cultivated on large scale in Hungary. Turks

emigrating from middle Asia introduced it into Asia Minor. Safflower was introduced in

China in the second century BC (Breifschneider, 1870). In Japan, it was introduced in

third century AD from China. Some horticultural varieties of safflower are C.lanatus

(yellow flowers), C.oxycantha (yellow) and C. tinctorius (orange).

Shostakovsky (1947), Ashri (1957), Ashri and Knowles (1960), Hanelt (1961, 1963),

Weiss (1971) and Kumar (1991) have reviewed literature on Carthamus. The genus is

distributed from Spain and North Africa across the Middle East to North India. Kupsow

(1932) and Vavilov (1949) suggested three centers, one each in India, Irano-Afghanistan

and Ethiopia, for the origin of Carthamus. De Candolle (1890) was of the opinion that

Arabia was geographically accurate as proposed site for the origin of this genus.

However, Ashri and Knowles (1960), Weiss (1971) and Ashri (1957) considered eastern

part of the Mediterranean as the center of origin of the genus.

The Safflower crop is usually grown in the rabi or winter season from

October/November to March/April generally as an intercrop with cereals such as

Sorghum and Wheat.

India is the largest producer of Safflower in the world with highest acreage (4.3-lakh

hectares) but with an average productivity of only 465 k.g. / hectares. Poor crop

management under input starved conditions is the most important reason for such low

per hectare yields. It is mainly grown in Maharashtra, Karnataka and parts of Andhra

Pradesh, M.P, Orissa, and Bihar etc. Maharashtra and Karnataka are the most important

Safflower growing states accounting for 72 and 23% of area and 69 and 35% of

production respectively. The acreage varies from year to year according to the demand

for Safflower oil, which is obtained from the crushed seed.

An edible oil is obtained from the seed; it is used in salad dressings and for frying

foods. There are two types of Safflower oil with corresponding types of Safflower

varities: those high in monounsaturated fatty acids (oleic) and those high in

8

polyunsaturated fatty acids (linoleic) Currently the predominant oil market is for those

varities that produce seed high in oleic acid and very low in saturated fatty acid. High

oleic Safflower oil is lower in saturates and higher in monosaturates than olive oil. High

oleic oil is a beneficial agent in the prevention of coronary artery disease. High linoleic

Safflower oil is used in human nutrition but in recent years market demand has

drastically shifted from the traditional high linoleic oils to high oleic oil. High linoleic

oil is valued as a drying agent in paints and varnishes because of its nonyellowing

property. A very stable oil, it is said to be healthier than many other edible oils and its

addition to the diet helps to reduce blood-cholesterol levels. It is much used in making

margarines. Seed roasted, fried and eaten in chutneys. The yellow is used as a saffron

substitute to flavor and color food. The fried seeds are used as a curdling agent for plant

milks etc.

Safflower is commonly grown as a food plant, but also has a wide range of

medicinal uses. To commercialize Safflower flowers in India efforts have been initiated

to popularize them as a herbal health tea for curing several chronic diseases. Regurlar

users of this tea have reported its usefulness in alleviating diseases like hypertention,

spondylosis, angina, arthritis, constipation, menstrual disorders and

hypercholesterolemia. Analysis of flowers for nutritional qualities was conducted

recently at the Central Food Technological Research Institute (CFTRI) at Maysore and

the results of this investigation are given in following table.

Table-1 Analysis of safflower flowers of NARI-NH-1

S. No. Parameters Value 1. Protein, % by wt 10.4 2. Total sugars, %by wt 11.8 3. Zinc, (mg %) 2.6 4. Sodium, (mg%) 17.0 5. Potassium, (mg%) 3264.0 6. Iron, (mg%) 42.5 7. Calcium, (mg%) 708.0 8. Magnesium, (mg%) 142.0 9. Copper, (mg%) 1.1 10. Cadmium,Magnesium,Lead, Negligible 11. Arsenic, (mg/kg) Nil

Safflower has been used traditionally in China to treat thrombotic disorders and

menstrual problem. Alcoholic extracts of the plant are used typically for direct

9

application to ulcers and wounds. Safflower has been used effectively as a daphoretic

and a diuretic. Taken hot Safflower tea produces strong perspiration and has thus been

used for colds and relative oilments. It has also been used at times for its soothing effect

in case of hysteria, such as that associated with chlorosis. Modern research has shown

that the flowers can reduce coronary heart disease and lower cholesterol levels. It is used

as alterative to analgesic, antibacterial, antiphlogistic and haematopoietic. Treats tumors

and stomatitis. The flowers are anticholesterolemic, diaphoretic, emmenagogue,

laxative, purgative, sedative and stimulant. They are used to treat menstrual pains and

other complications by promoting a smooth menstrual flow and were ranked third in a

survey of 250 potential anti-fertility plants. Safflower oil is suitable where high level of

stability at low temperature is required as in frozen desserts. It is also used in infant

foods and liquid nutrition formulation. In domestic practice, the flowers are used in

treating infants complaints such as measles, fevers and eruptive skin complaints.

Externally, they are applied to bruising, sprains, skin inflammations, wounds etc. The

plant is febrifuge, sedative, sudorific and vermifuge. When combined with

Ligusticumwallichii it is said to have a definite therapeutic effect upon coronary

diseases. The seed is purgative and tonic. It is used in the treatment of rheumatism. The

oil is charred and used to heal sores and treat rheumatism.

The oil is used for lighting, paint, varnish etc. It does not yellow with age. When

heated to 300°c for 2 hours and then poured into cold water, the oil solidifies to a

gelatinous mass and is then used as cement for glass, tiles, stones etc or as a substitute

for ‘plaster of paris’. If the oil is heated to 307°c for 2½ hours, it suddenly becomes a

stiff elastic solid by polymerization and can then be used in making waterproof cloth etc.

A yellow dye is obtained by steeping the flowers in water; it is used as a saffron

substitute. A red dye can be obtained by steeping the flowers in alcohol. It is used for

dyeing cloth and, mixed with talcum powder, is used as a rouge to color the cheeks.

The meal, which is about 24% protein and high in fiber, is used as a protein supplement

for livestock and poultry feed. Seeds contain 32 - 40% oil, 11-17% protein and 4-7%

moisture. Per 100 g, the seeds are reported to contain 482 calories, 4.8 g H2O, 12.6 g

protein, 27.8 g fat, 50.5 g total carbohydrate, 25.1 g fiber, 4.3 g ash, 126 mg Ca, 310 mg

P, 9.7 mg Fe, 0 ug beta-carotene equivalent, 0.59 mg thiamine, 0.14 Mg riboflavin, 0.5

10

mg niacin,, and 0 mg ascorbic acid. The oil contains 1.5% myristic (with lauric and

lower acids), 3% palmitic, 1% stearic, 0.5% arachidic (with trace of lignoceric), 33%

oleic, and 61% linoleic acids. Decorticated seed for animal feed contain 8.7% moisture,

10.0% fat, 45.4% protein, 20.1% carbohydrates, 8.3 fiber, and 7.5% ash. The cake

contains about 7.9%nitrogen, 1.9%potash and 2.2% phosphoric acid and its application

as manure is supposed to greatly improve the physical properties of heavy soil. Proteins

isolates prepared from debittered meal can be used to fortify bread, pasta and nutritional

drinks. Only lysine is limiting, while methionine and isoleucine are border line.

Phytochemical analyses showed the presence of following chemicals: Beta-

farnesene, Cadienals, Heptenols, Hexenols, Pentenals, Penenols, (Z)-3-hexenyl-

benzoate, (Z)-3-hexenyl-butyrate, (Z,z)-1,8,11-hepta-decatriene, (Z,z)-3,11-

tridecatriene-5,7,9-triyne, (Z,z,z)-1,8,11,14-heptade-catetraene, 1,2,3-trimethoxy-5-

methyl-benzene, 1-heptadecene, 1-hexadecene, 1-pentadecene , 1-tridecene, 2-

hydroxyarctiin, 3-methylbutyric-acid, Alloaromadendrene, Alpha-cedrene, Alpha-

copaene, Alpha-gurjunene, Alpha-muurolene, Alpha-phellandrene, Alpha-tocopherol,

Beta-cyclocitral, Beta-ionone, Beta-selinene, Carthamin, Carthamone, Caryophyllene,

Caryophyllene-epoxide, Chromium, Cobalt, Copper, Gamma-tocopherol, Germacrene-d

, Humulene, Iron, Isocarthamin, Kaempferol-glycoside, Limonene, Luteolin-7-beta-d-

glucoside, M-xylene, Magnesium, Manganese, Matairesinoside, Methylcinnamate,

Mucilage, Neocarthamin, Niacin, Nonanal, O-xylene, P-cymene, P-xylene, Pent-1-en-3-

ol, Pent-3-en-2-one, Pentanol, Phenol, Phenylacetaldehyde,Phosphorus, Safflower-

yellow, Safynol, Sd, Selenium, Serotobenine, Silicon, Terpinen-4 -ol, Tetracheloside,

Tetradecene, Ubiquinone-9, Verbenone, Zinc

The classification of the genus Carthamus has been a matter of dispute. Kupsow

(1932) classified the safflower collected from different parts of the world into two

groups (1) the eastern types and (2) the western types. Sabins and Pathak (1935) revised

the first classification given by Howard and Khan (1915). In this revised classification

single and distinctive characters like character of the bracts that is whether spinose or

spineless, the shape of outer bracts, color of the inner bracts, color of florets, growth

habit of plant etc. was used. Shostakovsky (1947) and Hanelt (1967) prepared a

comprehensive taxonomic treatment. The study of Shostakovsky is unpublished, but is

11

reviewed and summarized by Hanelt (1961, 1963). Kupsow (1932) made an extensive

study of the variability of domestic safflower. Hanelt (1963) divided the genus into five

sections on the basis of geographical distribution. The classification proposed by Ahsri

and Knowles (1960) and Estilai and Knowles (1976) are based mainly on chromosome

behavior. The genus possesses several species with n=10, seven with n=12, one each

with n=11 and with n=22 and two with n=32. Three basic chromosome numbers, x=10,

x=11, and x=12 are available in the genus Carthamus. Around 10,000 accessions of

Safflower have been collected throughout the world. With in the genus of Carthamus

there are more than 20 species with 10, 11, 12, 22 and 32 pair of chromosomes.

Safflower. It has been postulated that the most primitive condition is probably 2n=24.

The inter-specific cross could be made between some of the wild species and

cultivated. Chavan (1961) gave the following taxonomic characteristics of the genus

Carthamus. Thistle-like herbs. Leaves alternate, rigid, spinescent. Heads usually

homogamous; flowers all bisexual, fertile (rarely a few marginal female or neuter) and

similar, yellow white or purplish, tube slender; limb oblong, dilated at the base, 5-cleft.

Involucre ovoid or subglobos; bracts a-seriate, inner dry entire or with a short

fimbricate, outer with a foliaceous toothed or spinescent appendage (sometimes absent

in cultivated specimens). Receptacles flat, densely bristly. Filaments usually hairy in the

middle; anther-bases sagittate, auricles connate, tails short, fimbricate. Style arms short

or long. Achenes glabrous, oboviod, 4-angled or compressed, basal areole oblique or

lateral, all or the outer only without pappus, or all or the inner only with paleaceous a-

seriate pappus.

GENUS CHARECTERISTICS

General Plant Growth

The development of the plant has been recorded by a number of workers, but as their

methods and the varieties on which they carried out their trials were not always clearly

indicated, correlation of the result is difficult. The transition from rosetting to elongation

was between 48 to 55 days and branching and flowering began at about 76 days from

sowing. Seed formation commenced between 104 and 111 days, and the crop matured

between 125 and 132 days. Thirty four days after sowing the cotyledons were dying and

no longer contributing significantly to yield.

12

Plant Morphology

Safflower is a highly branched, herbaceous, thistle like annual varying in height from

30 to 150 centimeters. It has a strong, somewhat thickened tap root and numerous thin

laterals. The stem is stiff, solid, thick at the base and tempering with height, smooth and

glabrous. The plant has many branches, each terminating in a flower, and the extent of

branching within a variety depends mainly on environment. The leaves are simple,

usually dark-green, sessile and glabrous, exstipulate, deciduous, with short spines

scattered along the margin, having accuminate tips and a pronounced midrib. They are

cauline, alternate, pentastichous, with a phyllotaxy of two-fifths.

The inflorescence is a dense capitulum of flowers, surrounded with an involucre of

green ovoid bracts, the outer bracts separate, foliacious, sometimes spinesent, the inner

becoming fused, ovate, often covered with short white hairs. The involucre is conical,

with a small apical opening through which the corolla tubes of the flowers protrude. The

receptacle is broad flat or slightly curved, and densely bristled from the numerous floral

bracts. There are numerous flowers in the inflorescence, all regular, carried on the

receptacle, without pappus. The florets are tuber, sessile, regular epigynous, and grow

out through the apical opening of the involucre. The calyx is rudimentary. The ovary is

unilocular, with a simple basal ovule, which is composed of two united carpels, and is

inferior. The fruit is cypsela, glabrous, obovate with a flattened top, with four

longitudinal ribs. The pericarp is generally whitish, the pappus is normally absent. The

seed is dicotyledonous, oleaginous, and exalbuminous.

Root:

The plant has a well defined, frequently fleshy tap root and gives rise in the upper

layers of the soil to numerous thin horizontal laterals. The tap root commonly penetrates

to a depth of 2 to 3 meters but the length and general structure of the root depend on the

soil type and availability of soil moisture. When grown under sub irrigation, the tap root

is well developed and lateral branching may occur at the soil surface. The crop may use

considerable amounts of soil moisture but it cannot survive standing water for even a

few hours in warm weather.

Stem:

13

The stem is stiff, cylindrical, fairly thick at the base, becoming thinner as branching

increases, quite smooth, glabrous and a light gray or green to white in color, marked

with fine, longitudinal grooves. At soil level following emergence, the stem apex tends

to produce a number of leaves collected into a rosette, but rapidly develops into one or

more erect branches. There is no true rosette stage in cultivated safflower as there is in

other Carthamus species, some of which require a definite cold period to initiate stem

elongation. The rosette stage of safflower can be prolonged by inimical weather

condition i.e. when it is planted in the autumn in the Northern Hemisphere.

Turkestan varieties have a very short rosette period (Classen & Kiesselbach, 1945)

and only three weeks in certain Bombay varieties (Argikar & Salanki, 1958). This

rosette stage allows uncontrolled weed growth to quickly suppress the crop. Adequate

subsoil moisture should be available at the time of sowing (Seydlitz, 1962). The growth

of the plant is initially slow but grows rapidly as stem elongates. Application of

giberrellic acid increases the lodging (Yermanos & Knowles, 1960). Lodging is also

higher in the thin-hulled safflower strains.

The central (primary) stem branches to form secondary branches, which themselves

branch to form tertiary branches. The plant is thus usually well branched from about 20

centimeters above soil level and thus has a large number of ultimate branches each

terminating in a flower. The flower terminating the primary stem is the first on the plant

to bloom. In some varieties, branching may take place at a very low level, giving the

plant a fan like shape when mature; e.g. in the Indian variety ‘T46’ by Chavan (1961).

This branching may be due to damage of original stem. Close spacing will result in tall

plants with thin stems, and flowering heads close to the top of the plant, but wide

spacing produces bushy plants with many branches of varying length, and seed heads at

many levels. Stem girth of mature plants at ground level can vary from 3 to 12

centimeters (Chavan, 1961).

The amount of branching and type and position on the stem at which branches occur

are considerably affected by environment, but the method and nature of branching can

also be inherited. Varieties which normally have an open or intermediate branching

habit may become more appressed when grown under saline condition. The degree of

branching and the height above the ground at which branching occurs may also be

14

modified. The number of branches varied from 5 to 17 in Indian selections and the

height above ground of the first branch from 9 to 34 cms (Banerji, 1940). Nipping out

the central shoot first before flowering induced increased branching, number of seeds

per plant, and total seed yields in India (Subbia & Sivaram, 1965).

Height of safflower plants at maturity varies considerably; the tallest reported from

the Turkey-Afghanistan area, and the shortest from India. Height, however can be

influenced by factors as date of planting, soil fertility, soil salinity, type and distribution

of soil moisture, and less by plant density.

Leaf:

The branched stems bear spirally arranged leaves, but the leaves arise at uneven

intervals and may tend to be opposite. The leaves have a pronounced midrib, especially

near the base where it arises from the stem, the minor lateral veins arising from the

midrib. The leaves are between 2.5 and 5 cms broad, and may be up to 10 or 15 cms

long, with acuminate tips and with the mid rib projecting slightly from the tip of the leaf

as a short spine.

The leaves become shorter and stiffer up the plant until on the ultimate branches,

each of which is terminated by an inflorescence, the leaves are short, ovate to obovate

structure, not so strongly pointed. They are borne closer together along the ultimate

branch until at the tip they become super-imposed as the involucral bracts of the

inflorescence.

There is considerable varietal variation in the number of leaves per plant, but in

general those plants with large numbers of leaves tend to smaller leaf size. There is some

indication that leaf characteristics, either as maximum leaf length or maximum width,

can be correlated with oil yield (Argikar, 1957). According to Stern and Beech (1965)

temperature during the period up to flowering has a direct effect on the number of leaves

produced per plant. The lower leaves on most varieties are spineless, but the degree of

spininess on the upper leaves is a varietal characteristic and may vary from spineless to

strongly spined. The degree of spininess has been incorporated into a ‘spine-index’,

which was calculated by multiplying the number of spines on the outer involucral bract

by the length in millimeters of either the longer spines or the estimated average

(Claassen, 1952). Plant density has a major effect on the sizes and the number of leaves

15

per plant. The total number of leaves was directly related to the degree and numbers of

branches. Plant with the fewest branches, i.e. the highest plant population per hector, had

the lowest total leaf number.

Defoliation by disease or pests accelerated seed maturation, and the yield decreased

in direct proportion to the degree of defoliation. This decrease was due to death of plants

(Beech, 1962). Leaf-removal as the crop was maturing had a substantial effect on seed

and oil yield. Removal of all leaves at all stages of maturity reduced seed yield by only

one quarter, and removal of lower branches from almost mature plants caused the

greatest reduction. Removal of lower leaves had little effect on either plant growth or

seed yield. It was noted that the more profusely branched suffered a greater proportional

reduction in yield than those less well branched.

Inflorescence:

The inflorescence is typical of the compositae, and consists of numerous florets

collected closely together on a circular somewhat flattened receptacle, the whole

inflorescence itself appearing flower like. The receptacle is surrounded by several layers

of involucral bracts, which closely appressed, forming complete protection for

developing inflorescence, surround the receptacle. The individual flowers are themselves

also provides with bracts in the form of small hairs. The number of florets varies with

the variety and can be also affected by environment in the range of 20 to 180. The

flowers are regular, with five petals united to form a tube usually long and narrow. This

tube is divided into five lobes at tip and are of varying size. The five stamens lie inside

the corolla tube. The filaments are separate while five anthers are fused into a tube.

These rises above the corolla tube at maturity. Classen (1950) has described the process

of flower opening. Before flowering stigma is enclosed by the five fused anthers

attached to the tip of corolla tube. As florets elongates soon after sunrise, stigma and

style both elongates through the anther tube, stigma gets covered with the florets own

pollen. The stigma may remain receptive for several days. Despite of close contact

between stigma and pollen of the same flower there is every chance of cross pollination

by insects. In normal-hulled types, the anther flaps open abruptly, thus bringing pollen

into contact with the elongating style. While anthers of the mutant collapse in situ, thus

preventing release of pollen (Ebert and Knowles, 1968; Knowles, 1968). Pollen grains

16

have a prickly rough exine and their size differs between different species, as follows

(Ashri, 1957) 52 to 57 m in C. tinctorius, 54 m in C. oxycantha, 53 m in C.

palaestinus, 53 to 54 m in C. glaucus and 54 m in C. tenius.

Scales or hairs arising around the flower from the edge of the ovary represent the

calyx of the flower. It persists in the fruit as pappus or parachute. The lower bracts of the

involucre vary in number and shape from arbiculate and elliptical to lanceolate. The

inner bracts are elongate, imbricated and usually have a spinescent tip. White hairs may

be present on the outer sides of the inner bracts. Certain varieties have been classified on

the basis of smooth and green, or felted and white bracts (Sabins and Phatak, 1935). The

capitula or flower `head', are borne of the ends of the stems or branches. Each main

branches terminates in a head, which blooms before those on the ends of the secondary

branches. When the secondary branches have completed their growth and produce

heads, these begin to bloom. Thus flowering begins in the inflorescence which

terminates the main axis of the plant, followed by the most mature of the main branches.

The secondary and tertiary branches continue the process in regular order. When

secondary heads were formed on the primary stem, they flowered last of all (Beech and

Norman, 1963). The flower head can vary in diameter from 1.25 to 4.00 centimeters and

the number from 5 to 50. Row spacing between plants and heads are negatively

correlated (Williams, 1926).

Flowering normally begins at the margin of the head, and proceeds centripetal, some

three to five days being required to complete flowering. Depending on the growth habit

of the plant and the number of heads produced, total flowering may extend over a period

of ten to forty days and tends to be prolonged at low plant densities, when the number of

secondary and tertiary heads is high. The greatest proportion of flowering occurs in the

early morning, and this is accompanied by the elongation of the florets with the

projection of the style beyond the anther tube. Buds normally develop into flowering

heads after some twenty-five days. Abnormal types of inflorescence have been recorded.

Sterile heads occurred in breeding fields in India (Deshpande, 1940) and in the U.S.A.

(Claassen, 1952). Morabad and Jagannarth (1967) observed a multi-capitular type in

India. High temperature during early growth accelerates flowering and when it

occurrence is due to sowing after the optimum date, the time from emergence to

17

flowering can be reduced by more than a month when compared to plants sown at the

optimum period.

The most usual flower colors in the species are yellow and orange, sometime white or

ivory, with deep red. Both yellow and orange flowers are initially yellow, but the yellow

flower wilt to a brownish-yellow and the orange to an orange-red-shade. It has been

determined that the inheritance of flower color is due to four independently inherited

pairs of gene. Safflower is basically self-pollinated, not wind pollinated, but bees or

other insects are required for optimum fertilization and maximum yields (Kadam and

Patranker, 1942; Levin and Butler, 1966). Large numbers of honeybees are normally

attracted to safflower fields during the flowering period. The importance of the

relationship between pollinator activity and the type of safflower can be correlated

directly with the degree of self-pollination on the latter.

Fruit:

The fruit of safflower is an achene, but is normally referred to as safflower seed. The

seed generally resembles a small, slightly rectangular sunflower seed, but with a thicker

and fibrous hull. Pappus is normally absent, but it may occur on some seeds, and these

are usually from the center of the flower head. It may also be present on a majority of

seeds in a particular strain, and one major gene with some modifiers appears to

determine the degree of its presence or absence, Claassen (1950). Color is generally

cream or white, but gray or off-colored seeds also occur. Crosses of cultivated safflower

and the black-seeded wild safflower of India (C. oxycantha) produced seed colors

ranging from black, through various shades of brown and with various amounts of

mottling, to white. Testa color may vary from dark or light-brown, through various

shades of yellow to ivory or white (Kursell, 1932). It appears that several genes are

involved in hull color. It was determined that the absence of the melanin layer in the

pericarp of the seed was due to a single recessive gene (Rubis, 1966).

Seed size is variable, but frequently a varietal characteristic. Chavan (1961) has

recorded a double seeded type from the Annigeri District of India. Gila which is a

commonly grown variety has average size 6 to 7 mm in length and 100 seed weight of

about 4 gms. Twenty-six Indian selections gave a range of 3.54 to 7.55 gms per 100

seeds (Argikar et al., 1957) and one variety, Bagewadi-1, a weight of 9.92 gms (Chavan,

18

1961). The seed weights of 100 seeds from different countries were 4.4 to 5.3 gms

(Russia), 3.7 to 4.4 gms (Caucasus), 4.0 to 5.0 gms (Somalia), 3.14 gms (Kenya), 5.0

gms (Sudan), 4.8 gms (Australia), and 3.0 to 5.0 gms (Turkey). The relationship

between seed color and viability, germination and seedling growth of wild safflower

ecotypes was determined by Bassiri et al. (1976). Gray seeds were found in more

proportions in high salinity locations. White seeds weighed significantly less than

colored seeds due to their thinner coat and smaller size, while black and gray seeds were

similar in seed weight. Seed color in general, had a strong effect on germination. The

number of seeds, their germination percentage and seedling length varied between heads

of the same plant of wild safflower (Kheradnam and Bassiri, 1978). Very little

variability was found among plants of the same ecotype. The number of seeds and seed

dry weight yield per plant were considerably decreased by soil water deficit whereas the

number of florets percentage of ripened seeds, number of seeds and seed dry weight per

head were not affected (Hayashi and Handa, 1985).

Seeds from heads of different positions on the plant vary considerably in several

important characteristics. The variation in seed composition can easily be demonstrated

by selecting heads of different positions on a number of plants and comparing the oil

yields and other seed characteristics obtained from them. Seed composition varies

between the various heads from a single plant, but also between plants. Primary,

secondary and tertiary heads were harvested separately and analyzed (Yermanos, 1963;

Francois, 1963). It was found that primary heads had the highest seed weight and hull-

content, and the lowest oil-content. Salinity causes a decrease in oil percentage mainly

by increasing the hull percentage of seeds (Francois and Bernstein, 1964). Growth

regulators influence the rate of development of the plant and seed but seed yields and

composition are adversely affected. Time of planting, amount of water available and

temperature at flowering affect the seed weight, hull and oil-content. Delayed planting

reduced the seed weight and hull-content (Luebs, 1965). Seed size affects the rate and

percentage of emergence, and therefore the subsequent rate of growth (Saeed, 1966).

Large seeds emerge faster and, more regularly than small. Seedlings with three

cotyledons and multiple first leaves have been recorded (Rao and Rao, 1934). Ghorashy

et al. (1972) observed reduction in germination as salinity increased from zero to one

19

percent NaCl. Safflower appears to be only half as salt-tolerant during germination as

during later stages of growth (Francois and Bernstein, 1964). Yields of safflower were

reduced by 10% per cent at 7 milliohms per centimeter (Ece) and by 20 to 25% at 11

Ece.

The taxonomic position of Caduncellus-Carthamus complex has been a matter of

dispute Infect many workers have conducted numerous attempts to resolve this problem

but none of these have been accepted

Carthamus-lanatus L.is hybrid in origin and Carthamus-leucocaulous Sibth&Sm are

found in nature in the form of weed in the western Mediterranean region as well as in

Mediterranean climatic regions of Argentina, Australia, California, and South Africa

(Ashri&Knowles, 1960; Hanelt, 1963; Estilai&Knowles, 1978). C.tinctorius is widely

grown as an important source of oil in subtropical countries (Hanelt, 1963, 1976).

Safflower (C.tinctorius) is a substitute for saffron.

The sub tribe position of this complex with in cardueae is problematic and many

genera are not yet clearly delineated. The existing confusion was compounded by the

recent incorporation of a new genus Femeniasia Susanna (Susanna et al, 1995;

Susanna&Vilatersana, 1996; Wagenitz&Hellwig, 1996) in to the complex. Cassini

(1819) positioned the Carthamus in the sub tribe carduinae and Carthamus sensustricto

in to the centaureinaeDecandolle (1838) and Nyman (1879-1890) proposed a new sub

tribe Carthaminae for Carduncellus and Carthamus positioned between two sub tribes of

the Cardueae and Centaureinae.Subsequently Godron (1852) and Battendier (1890)

turned to the classification of Cassini (1819).

Bentham (1873) & Hoffman (1894) placed both genera in the sub tribe Cantaureinae

Femeniasia was described and classified in Carduinae (Susanna, 1988) and was

latter transferred to Centaurinae (Bremer, 1994; Susanna et al., 1995, Sussana and

Villatersana, 1996, Wageinitz and Hellwing, 1996). Two peculiar morphological

characteristics are responsible for this fluctuating subtribal placement, which were spiny

leaves and single pappus. Whereas spiny leaves were frequent characterstics of

Carduinae but unusual in Centaureinae. Single pappus found in some species of

Carduncellus (Cassini, 1819 and Dittrich, 1969) and Femeniasia (Susanna, 1988) in

Carduinae.

20

These two characteristics provide minimal systematic value (Dittrich, 1968, 1969;

Wagenitz and Hellwing, 1996). For this reason the entire complex placed in

Centaureinae (Dittrich, 1977; Bremer, 1994; Susanna et al, 1995; Wagenitz and

Hellwing, 1996).At the generic level the systematic position is not clear for two genera,

which are split from Carthamus (Kentrophyllum Necker and Phonus Hill) and another

genus (Lomottea Pomel) segregated from Carcuncellus. Hanelt, (1963) proposed four

different genera (Carduncellus, Carthamus, Femeniasia, and Phonus).

Table 2. The detailed description of the conflictory generic classifications is as follows.

Cassini (1819) Decandolle (1838) Pomel (1874) Lopez Gonzalez (1990) Kentrophyllum Kentrophyllum Kentrophyllum Carthamus Sect. Atraxyle Onobroma Sect. Atractylis Sect.

Odontognathia Sect. Euonobroma Sect. Carthamus

Sect. Thamnacantha

Sect. Cirsiastrum Sect. Odontognathus

Carduncellus Carduncellus Carduncellus Phonus Onobroma Lamottea Carthamus Carthamus Durandoa Carduncellus Bentham (1873) Battandier (1890) Henelt (1963) Boisser (1875) Hoffman (1894) Kentrophyllum Carthamus Sect. Durandoa Sect. Atractylis Carduncellus Sect. Atraxyle Sect. Carthamus Sect. Lepidopappus Carthamus Carthamus Sect. Odontognathus Carduncellus Sect.

Thamnacanthamus

Sect. Cyanoidei Sect. Cirsiastri Carduncellus Sect. Phalolepides Sect. Carduncellus

Carthamus fruticosus and Carduncellus mareoticus both were described as

Carthamus but Hanelt, (1963) transferred them to Carduncellus because they have

rudimentary appendiculate middle bracts But Lopez Gonzalez, (1990) classified them as

Phonus because they also have a semi deciduous pappus an undifferianted pericarp.

Dittrich (1969) moved to the old classification by Linne (1753). He suggested that all

the species of both genera grouped in a single genus Carthamus.Lopez Gonzalez (1990)

splitted the Carthamus and Carduncellus in to four group Carthamus, Carduncellus,

21

Lamottea and Phonus.Hanelt (1963), Dittrich (1969) and LopezGonzalez (1990)

observed that both of these are different genera For a comprehensive resolution of the

relationship with in this group R.Villatersane et al (1999) used the ITS (Internal

transcribed spacers ) sequences of nuclear ribosomal DNA. These sequences supports

the intend of LopezGonzalez (1990).ITS sequences effectively differentiated between

Carthamus and the other genera with in the complex (Caruncellus, Phonus, and

Feniasia) but the ITS phylogeny did not resolve the relationship between the two major

groups of complex

For Carthamus ITS sequences supported that the Carthamus sensustricto include

only group Carthamus and other section of Hanelt (1963), Decandolle (1838) and

Cassini (1819) classified these group as separate genera Carthamus and Kentrophyllum.

Hanelt (1963) these three Carthamus, Carduncellus and Kentrophyllum are very closely

related and Carthamus would show more clear the relationship of the two groups.

R.Villatersana (1999) proposed four different genera Carduncellus, Carthamus,

Femeniasia and Phonus. But the relationship between the western group

(carduncellus,Femeniasia and Phonus ) and the eastern genus Carthamus are not

resolved by analysis of ITS sequences, but the two groups are not close relatives. Since,

the genus Carthamus has a number of species with varying chromosome numbers

(2n=20-64) and a wide adaptation, thus provides an interesting material for cytogenetic

research, an area completely neglected in the past

The interspecific cross could be made between some of the wild species and cultivated

species. Some workers have conducted cytotaxonomic studies involving interspecific

hybridization (Ashri and Knowles, 1960; Estilai and Knowles, 1976, 1978; Khidir and

Knowles, 1970b). Both intra and interspecific relationships among different Carthamus

species were studied by workers including Khidir and Knowles (1970 a, b) and Estilai

and Knowles (1976, 1978). Schank and Knowles (1964) studied the Cytogenetics of

hybrids of Carthamus species with ten pairs of chromosome. Ashri and Knowles (1960)

studied the Cytogenetics of Safflower (C. tinctorius) species and their hybrids. The

Cytogenetics study was undertaken in order to understand better the nature of the genus,

to study the interrelationship of the wild species with each other and with the cultivated

22

species and finally to assay the possibility of transferring desirable characters from the

wild species to the cultivated one.

NARI was also a pioneer in starting Safflower hybrid development in India. The first

nonspiny hybrid in India NARI-NH-1 (PH-6) was developed at NARI. In addition to

2000 to 2500 kg seed, the Safflower hybrids were found to yield 200 to 250 kg flowers

per hectare.NARI developed high yielding and high oil containing varieties for minimal

irrigation.

It has been frequently mentioned in the literature that the somatic karyotype in safflower

is difficult due to poor stain-ability, stickiness, tendency to overlap at metaphase and

diffuse appearance of primary and secondary constrictions of the chromosomes.

Karyotype analyses of some species of Carthamus have been conducted (Knowles and

Schank, 1964; Chatterji and Rathore, 1972-73 and Pillai et al., 1981a). Srivastava and

Kalara (1996) conducted the 3-D analysis of karyotype in Carthamus and found

accessions were differing from each other not only in size and morphology but also in

the thickness of chromosome. Subramanyam (1952) carried out karyotype analysis of

the somatic chromosome complement, in C. tinctorius, C. caeruleus and C. lanatus (two

ecotypes, one from Australia and the other from Portugal). Kishore (1951) and

Deshpande (1952) observed that wild safflower has chromosome number 2n=24.

Knowles and Schank (1964) compared the somatic chromosomes of C. nitidus with

those of C. tinctorius and reported that ideograms of the two species were almost

similar, except that the satellite (SAT) chromosomes of C. nitidus appeared to be longer.

Chatterji and Rathore (1972-73) and Pillai et al. (1981a) analyzed the karyotype of C.

tinctorius. The absolute length of the individual chromosomes ranged between 1.96 m

and 4.00 m. Most of the chromosomes had median to sub median centromeres. The

total length and arm ratios of various chromosomes of the complement did not differ

very much, suggesting that the karyotype of safflower was symmetrical. Knowles and

Schank (1964), Chatterji and Rathore (1972-73) found one pair of SAT-chromosomes

each in C.nitidus, C.tinctorius and C.oxycantha, only one collection of C. tinctorius was

reported to have two pairs of SAT-chromosomes. However, Estilai and Knowles (1976,

1978) noted two pairs of chromosomes attached to the nucleolus at diakinesis in C.

divaricatus, C. leucocaulos (in some cases only) and in the F1 hybrid of C.leucocaulos x

23

C.dentatus. It appears that safflower probably has more than one pair of SAT-

chromosomes (possibly three). Villatersana (2000) analysed the karyotype of

Carthamus, Carduncellus and Phonus. The main conclusion of Villatersana is that the

descending dysploidy is the main mechanism of karyological evolution in the complex.

Jayaramu and Chatterji (1986) analyzed the karyotype for the first time on the wild

species(C. oxycantha). These species reveals the somatic complement 2n=24.

Srivastava and Gupta (1970) found irregular meiosis in one collection of

C.oxycantha; partial asyndesis was observed in 80.36% PMCs, while restitution nuclei

were found in 14.58% PMCs. Polyspory was noted in 80.23% of the microspore tetrads.

Carapetian and Rupert (1977) studied the meiotic irregularities caused by interacting

sterlity genes in cultivated Safflower. A structure tentatively designated as a ‘nuclear

body’ was observed in microsporocytes of some collections of C.baeticus and

C.turkestanicus, as well as, in their interspecific and intraspecific hybrids (Khidir, 1969).

No more than one body was noted. The body was not detected in diploid on

allotetraploids and their hybrids. The body could be observed at any stage of mitosis in

root-tip cells.

The majority of commercial varieties of the safflower are known, somewhat loosely,

as pure lines. In fact many of them are selected in the early generations following a

cross. The selection of lines or strains within the most suitable local type has proved a

successful in improving the oil percentage of seeds. Kumar and Veena (2000) developed

high yielding genotype through hybridization and mutation. Krijthe (1942) was probably

the first to use colchicine on the anthers of a cultivated safflower to induce polyploidy.

Schank and Knowles (1961) induced autotetraploidy in several varieties of cultivated

safflower and reported that most successful treatment was a 0.1% aqueous solution of

colchicine applied four times daily to a cotton swab wedged between the cotyledons of

young seedlings for a period of 3 days. Pillai (1978), following the same method found

0.5-0.1% colchicine most efficient. Khidir and Knowles (1970b) recommended the

application of colchicine on the growing tip of the seedling of Carthamus species with

n=32, by mixing 0.1% solution with 1% tragacanth gum instead of cotton swab. The

morphological changes due to polyploidy varied with the optimal level of ploidy in

relation to genotype. Schank and Knowles (1961) and Pillai (1978) noted vigorous

24

growth, longer leaves, stomata and pollen grains in diverse cultivars of C.tinctorius.

They also noted an increase of about 13- 106% in the seed index of autotetraploids, over

the diploid check. An increase in oil content to autotetraploid in relation to diploids

(Pillai, 1978) was in contrast to the earlier observation of decline in oil content (Schank

and Knowles, 1961).

Autotetraploid are characterized by the increase of varying frequency of

quadrivalents, trivalents and univalents and reduced seed fertility (Pillai, 1978; Schank

and Knowles, 1961). Estilai and Knowles, (1980) first reported a triploid as a

morphological deviant in population of autotetraploids of cultivated safflower and

considered it to be a product of outcross to a diploid cross. Kumar et al. (1984) also

reported a triploid in the gamma-irradiated material of C. tinctorius. Harvey and

Knowles (1965) obtained colchicine induced allopolyploids from various interspecific

hybrids with 10 and 12 pairs of chromosomes and crossed them to six different types of

C.lanatus. Heaton and Knowles (1981) found genetic male sterlity in Safflower treated

with colchicine. Male sterlity was controlled by a single recessive nuclear gene ms.

Plants that were homozygous for ms were completely male sterile and were not

influenced by cytoplasmic factor. NARI(2002-2003) reported male sterile maintainer

and restorer gene for induction of cytoplasmic male sterlity in Safflower

Khidir and Knowles (1970b) artificially produced allopolyploid through

chromosome doubling of F1 plants from the cross C.leucocaulos (n=10) x C.lanatus

(n=22). Thus allopolyploid was similar to C. baeticus (n=32). Similarly, an

allopolyploid developed from a hybrid of C.glaucus (n=10) with C.lanatus (n=22)

resembled C.turkestanicus (n=32). Heaton and Klislewicz (1981) synthesized a disease

resistant allopolyploid through chromosome doubling of the hybrid C.tinctorius (n=12)

with C.lanatus (n=22).

Davis et al. (1981) evaluated the world Safflower collection for resistance to

Phytophthora.

Knowles (1958) was first to report a trisomic plant in C.tinctorius. Estilai and

Knowles (1980) found a plant with varying chromosome number in derivatives of five

suspected triploids of cultivated safflower. Translocation have been induced and studied

by some workers. Schank and Knowles (1964) analyzed 20 of the possible of 21 hybrids

25

produced from intercrossings seven Carthamus species with n=10 and found that three

C.dentatus (Turkey), C.glaucus (Iran and Syria) had naturally occurring chromosome

interchange. Estilai and Knowles (1978) evidenced that C.leucocaulos had a

chromosome arrangement similar to that of C.dentatus. Khidir and Knowles (1970a)

found a reciprocal translocation in F1 hybrid of polyploids C.baeticus x C.turkestanicus.

Pillai (1978) induced translocation in cultivated safflower. Singh et al. (1981) reported

62 translocation heterozygotes in the M1 generations of gamma irradiated plus sodium

azide treated population of Safflower. Most of the simple translocations were induced at

30 and 45 KR, whereas the high dose 60 KR produced complex interchanges, which

fails to produce seed. Pillai et al. (1981b) isolated translocation homozygotes and

identified each chromosome involved in interchange through karyotype analysis.

Pillai (1978) studied the effect of gamma rays and sodium azide on root tip

chromosomes and isolated three plants showing poor growth and male and female

sterility in M1 generations after gamma irradiation. Prasad (1983) reported a desynaptic

mutant in M2 generation following gamma irradiation of safflower. The mutation was

highly male sterile and produce only a few seeds. Kumar et al. (1984) reported three

gamma rays mutants showing unreduced gametes in safflower.

Chauhan and Singh (1975) studied the effect of gamma rays and 2, 4-D on the

morphology Both 2, 4-D and gamma rays affected the cell dimensions and cell area. The

effect is less in case of gamma rays exposure only.

Imrie and Knowles (1970) studied the inheritance of self-incompability in C.

flavescens and in C. tinctorius.

Chemotaxonomic studied using phenolics and allozymes have been also carried out

in Carthamus species. In general, chromatography of extracts from leaves and head

bracts did not give much useful information on the relationships of the various species at

the diploid level or with respect to the origin of polyploidy in the genus Carthamus

(Estilai and Knowles, 1976, 1978). Estilai and Knowles (1976) noted some differences

in the chromatograms of the three flower color types of C.divaricatus. Further, the

chromatograms of C.lanatus showed more spots than those of C.turkestanicus. Estilai

and Knowles (1978) also found similarity in chromatograms of C.leucocaulos (n=10)

and C.nitidus (n=12), probably due to close morphogenetic similarity between them.

26

Efron et al. (1973) studied the ADH allozyme pattern in seeds of 1553 varieties from the

world collections of safflower (C.tinctorius) and from 36 collections of 14 wild species

with different chromosome numbers.

Rohini et al. (2000) experimented with safflower cv. A-1 and A-300 and gave a

procedure that could successfully be used to generate whole plant transformants. The

transformants were prepared by removing and growing Agrobacterium infected embryo

axes on soilrite moistened with water. To the best of our knowledge only little

information is available about the microsporogenesis and megagametogenesis. Carman

(1995) reported the presence of agamospermy manifested by apospory and adventive

embryony in Carthamus tinctorius.

Banerji (1940) described megasporogenesis, fertilization and development of the

embryo and endosperm in Carthamus-tinctorius. NARI (2002-2003) reported

polyembryony in Safflower on the basis of twin plants originating from the

polyembryonic seeds.

Azad and Gupta (2002) described the direct embryogenesis of Safflower with the

help of SEM. The SEM revealed the normal development of embryo from globular to

heart shaped, torpedo shaped and finally cotyledonary stage embryo.

Bassiri et al. (1975) studied the effect of temperature and scarification on

germination and emergence of wild Carthamus-oxycantha. and observed that the

cultivated variety had higher germination and emergence percentage and seedling height

than the wild type. The optimum temperature of wild and cultivated strain was between

15 and 200C. Scarification of the wild seed did not improve the germination or

emergence percentages and chilling of the seed for a month at 00C reduced the

emergence of the wild seed. Bassiri et al. (1977) compared the influences of simulated

moisture stress conditions and osmotic substrates on germination and growth of

cultivated and wild Safflower. Increased Ops progressively delayed and reduced seed

germination, shoot length, and fresh and dry weight of seedlings Wild ecotypes were

apparently more sensitive to high Ops than the cultivated species.

Saito (1994) isolated the carthamin dyes from dyer,s saffron flowers. Saito (2000)

introduced the stability of carthamin and Safflower yellow B on silk powders under

27

continuous irradiation of fluorescent or UV light and also improved a new technique for

redding florets with the help of UV light irradiation.

NARI (2002-2003) evaluated the spiny and nonspiny genotypes for flower yield and

other physiological traits.

Yermanos and Francois (1963) reported the seed of the primary heads were the

largest and they become progressively smaller in the secondary and tertiary heads.

The survey of the literature illustrated the requirement of the detailed cytogenetic

investigation of the genus Carthamus.