Journal of Herbs, Spices & Medicinal Plants

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Ocimum sp. (Basil): Botany, Cultivation,

Pharmaceutical Properties, and Biotechnology

Olga Makri & Spiridon Kintzios

To cite this article: Olga Makri & Spiridon Kintzios (2008) Ocimum sp. (Basil): Botany,
Cultivation, Pharmaceutical Properties, and Biotechnology, Journal of Herbs, Spices &
Medicinal Plants, 13:3, 123-150, DOI: 10.1300/J044v13n03_10

To link to this article: https://doi.org/10.1300/J044v13n03_10

Published online: 11 Oct 2008.

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 REVIEW

Ocimum sp. (Basil):

Botany, Cultivation, Pharmaceutical
Properties, and Biotechnology
Olga Makri
Spiridon Kintzios

ABSTRACT. The present review focuses on the various Ocimum spe-

cies, often referred to as the “king of the herbs.” The botany of more than
50 species of herbs and shrubs belonging to this genus is thoroughly re-
ported, along with traditional uses and cultivation techniques. Since ba-
sil is a rich source of natural compounds, details on the several chemical
constituents of essential oil, plant parts and derived food and medical
products, such as monoterpenes, sesquiterpenes, phenylpropanoids, an-
thocyanins, and phenolic acids, as well as their effect on sensory qualities
are included. Furthermore, particular emphasis is given to the applica-
tion of biotechnology for the clonal micropropagation of basil lines
with improved traits and the use of basil tissue culture for the derivation of

Olga Makri is Postgraduate Researcher and Spiridon Kintzios is Associate Professor,

Faculty of Agricultural Biotechnology, Laboratory of Plant Physiology, Agricultural
University of Athens, Iera Odos 75, 11855 Athens, Greece.
Address correspondence to: Spiridon Kintzios, Faculty of Agricultural Biotechnol-
ogy, Laboratory of Plant Physiology, Agricultural University of Athens, Iera Odos 75,
11855 Athens, Greece (E-mail: skin@aua.gr, info@embio.org).
The authors wish to express their warmest thanks to Mrs. Angela-Lu Petrou-Gini
(Greek Ministry of Foreign Affairs) for proofreading the text in English.
Received January 9, 2005.
Journal of Herbs, Spices & Medicinal Plants, Vol. 13(3) 2007
Available online at http://jhsmp.haworthpress.com
© 2007 by The Haworth Press. All rights reserved.
doi:10.1300/J044v13n03_10 123
124 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

valuable compounds, such as antioxidant phenolics and essential oil

components. doi:10.1300/J044v13n03_10 [Article copies available for a fee
from The Haworth Document Delivery Service: 1-800-HAWORTH. E-mail ad-
dress: <docdelivery@haworthpress.com> Website: <http://www.HaworthPress.
com> © 2007 by The Haworth Press. All rights reserved.]

KEYWORDS. Aromatic plant, culinary herb, medicinal plant, secondary

metabolites

DISTRIBUTION, HISTORY,
AND TRADITIONAL USES OF THE SPECIES

Basil, one of the most popular herbs grown in the world, is native to
Asia (India, Pakistan, Iran, Thailand, and other countries) and can be
observed growing wild in tropical and sub-tropical regions. Because of
the popularity of basil, the plant is often referred to as the “king of the
herbs.” Several name derivations and beliefs are associated with basil,
but the common name basil is probably derived from the Greek words
basileus meaning “king” or basilikon meaning “royal.” A Latin word,
basiliscus, refers to “basilisk” a mythical fire-breathing dragon so repul-
sive that a glance could kill. According to a Roman legend, basil was the
antidote to the venom of the basilisk (1). The botanical name Ocimum is
derived from the Greek meaning “to be fragrant.” In the 1600s, the Eng-
lish used basil as a food flavoring and insecticide. The plant was hung in
doorways to ward-off flies and other unwanted pests (evil spirits).
Italians used basil as the sign of love and a pot of basil placed on the
balcony meant that a woman was ready for her suitor to arrive. In turn, if
he brought a sprig of basil, he would signal to her his serious intentions
(12). Hindus believed that being buried with a leaf of basil meant they
would get into heaven. According to the ancient Greeks, basil repre-
sented hate and misfortune. Poverty was painted as a ragged woman
with basil at her side with the thought the plant would not grow unless
railing and abuse were poured forth at the time of sowing. The Romans,
in like manner, believed that the more basil was abused, the better the
plant would prosper.
The physicians of old were quite unable to agree as to the medicinal
value of basil, although some declared the plant was a poison and others
said the plant was a precious sample (a medicinal plant) (22). Basil was
sacred to the gods in India, Krishna, and Vishnu. In America where basil
has been grown for over 200 years, the plant was originally air-dried or
preserved in layers of salt and kept in earthenware crocks (75).
 Review 125

Basil has many uses, but the most common is for culinary purposes.
As a fresh herb, basil is used to flavor foods such as vegetables, poultry,
and fish. The herb is famous for use in Italian dishes, such as pesto. Basil
is commonly preserved in vinegar or olive oil and adds a delightful fla-
vor to both for salad dressings. The plant is also used as a flavoring in
jelly, honey, tea, and liquor. Basil can also be used dried.
The flowers of basil are edible and can make an attractive addition to
salads and other dishes (22). Besides being edible, basil as an aromatic
herb is often used in potpourri and sachets. The cosmetic industry uses
basil oil in lotions, shampoos, perfumes, and soaps. As an ornamental in
the flower garden, basil has attractive foliage and flowers.
Basil is a source of essential oils and aroma compounds (74, 76), a
culinary herb, and an attractive, fragrant ornamental (46, 47). The seeds
contain edible oils and a drying oil similar to linseed oil (2). Extracts of
the plant are used in traditional medicines, and have been shown to con-
tain biologically active constituents that are insecticidal, nematicidal,
fungistatic, and antimicrobial (1, 51, 74). For example, the essential oils
of O. forskolei Benth are mosquito-repellent (18, 29), while methanol ex-
tracts of O. gratissimum have been used for the control of Leishmania,
Candida, and Cryptococcus species (4, 82). The trypanocidal activity of
essential oils from O. basilicum has also been demonstrated (67). Ac-
cording to a recent report (42), sweet basil oil gave the highest anti-
proliferative activity among 17 different Thai medicinal plant species
against the murine leukemia P388 cell line. In addition, aqueous extracts
of O. sanctum has demonstrated significant mammary immunostimulant
activity (50).
In contrast, Chaha et al. (5), has observed no inhibitory activity of met-
hanolic extracts of O. gratissimum against 11 wound isolates, including
Staphylococcus aureus (four strains), E. coli (two strains), Pseudomonas
aeruginosa (one strain), Proteus spp. (three strains), and Shigella spp. (one
strain). The essential oil of O. basilicum has also been demonstrated to be
ineffective against Propionibacterium acnes, a gram-positive bacterium
linked to the inflammation of acne leading to scar formation (39).
In common with other members of the Lamiaceae family, basil, both
wild and sweet, furnishes an aromatic, volatile, and camphoraceous oil
that is much employed in France for flavoring soups, especially turtle
soup. The French also use basil in ragouts and sauces. Leafy tops of
basil are thought to be a great improvement to salads and cups. Al-
though not commonly used in England for culinary purposes, this basil
was a favorite pot-herb in the past and gave rise to the distinctive flavor
that made Fetter Lane sausages famous (22).
126 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

For the fresh market, basil should have a sweet flavor and dark green
foliage. Commercially, three major types of basil with essential oil or
dried leaves as the end product are used. French basil, reputed to be
sweetest in flavor and darkest in color, is the most valued. American
basil, noted for rich color, sweet flavor, cleanliness, and uniformity of
particle size, is considered to be of high quality. Egyptian basil, also
known as “Reunion” or “African” basil, has a somewhat camphoraceous
fragrance and unusual flavor to some palates and is thus considerably
less valued in the market (12).
While basil seed is plentiful, care must be observed in obtaining a type
that has desirable characteristics. Because several basil types may be mi-
xed together by a seed house or processor to achieve a desired blend, any
collected seeds that are later sown may vary in growth, development, and
aromatic properties.
Though generally employed in cooking for flavoring, basil has been
occasionally used for mild nervous disorders and for the alleviation of
wandering rheumatic pains. The dried leaves, in the form of snuff, are
said to be a cure for nervous headaches (84) and, indeed, research has
provided some evidence for a direct effect of basil constituents on the
central neural system. For example, leaf-derived ethanolic extracts of
O. sanctum have been shown to cancel the noise stress-induced reduction
in total acetylcholine content and increase the activity of acetylcholin-
esterase in the cerebral cortex, corpus striatum, hypothalamus, and
hippocampus of brain (70), possibly through the exertion of antioxidant
activity (66). These properties have been partly attributed to volatile com-
pounds in the plant, such as the sesquiterpenes eugenol and 1,8-cineole in
O. gratissimum, that are subject to seasonal variation (49). An infusion
made with the fresh herb is good for obstructions of the internal organs
and arrests vomiting and allays nausea. The seeds are thought to be effi-
cacious against the poison of serpents when taken internally or laid upon
the wound. Seeds are also said to cure warts.
Tohti et al. (81) observed a dose-dependent, inhibition of platelet agre-
gation in rats initiated by ADP and thrombin using basil and that 75 mg/
kg/day of an aqueous basil extract had approximately the same effect as
8.8 mg/kg/day aspirin.

BOTANY

The genus Ocimum (family Lamiaceae), collectively known as basil,

has long been acclaimed for the genetic diversity of the species within
 Review 127

the genus. Ocimum comprises at least 65 species (55), but more than 150
species according to some sources (65), of herbs and shrubs from the
tropical and sub-tropical regions of Asia, Africa, and Central and South
America. The main center of diversity appears to be Africa (56). Differ-
ent species and forms of Ocimum spp. vary in growth habit, color, and
aromatic composition, making the true botanical identity of basil diffi-
cult. Ornamental types of basil are commercially available and include
types with purple foliage, such as Dark Opal, or varying growth habi-
tats, such as bush basil.
Most commercial basil cultivars available in the market belong to the
species O. basilicum. Darrah (13,14) classified the O. basilicum cultivars
into seven types: tall slender types (including the sweet basil group)
(Figure 1A), large leafed, robust types (including ‘Lettuce Leaf’, also
known as ‘Italian’ basil) (Figure1B), dwarf types (including the short
and small leafed, such as ‘Bush’ basil), compact types (including O.
basilicum var. thyrsiflora, commonly known as ‘Thai’ basil) (Figure 1C),
purple-colored basil (which includes var. purpurascens, a purple-col-
ored basil with a traditional sweet basil flavor) (Figure 1D), purple
types (including ‘Dark Opal’, a possible hybrid between O. basilicum
and O. forskolei, with lobed-leaves and a sweet basil plus clove-like
aroma) and lemon-flavored basils (which includes the citriodorum
types (48) (Table 1).
In addition to the traditional types of basil, other Ocimum species in-
troduced into the North American horticultural trade have culinary and
ornamental uses and may be potential sources of new aroma compounds.
Interspecific hybridization and polyploidy, however, are common occur-
rences within this genus (23) and have created taxonomic confusion,
making any understanding of the genetic relationship, with the excep-
tion of the preferentially autogamous propagation of O. basilicum, be-
tween the many basil species difficult (20). In addition, the taxonomy of
basil is complicated by the existence of numerous botanical varieties, cul-
tivator names, and chemotypes within the species that may not differ
significantly in morphology (76).
A system of standardized descriptors, which include volatile oil, has
recently been proposed by Paton and Putievsky (54) and this should per-
mit easy communication and identification of the different forms of O.
basilicum. Investigations to revise the genus are underway at the Royal
Botanical Garden, Kew, London (56) and at Delaware State University
in Dover, Delaware, U.S.A. Grayer et al. (21) attempted a classification
of 15 cultivars, belonging to four different basil species (O. americanum,
O. basilicum, O. × citriodorum and O. minimum) by analyzing exudate
128 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

FIGURE 1. Basil plants. A = Sweet basil, B = Italian basil, C = Compact basil

var. thyrsiflora, D = Purple basil (var. purpurascens).

flavonoids. The highest number of flavonoids and largest amounts were

detected in specimens of O. americanum var. americanum, and the low-
est number and smallest amounts in specimens of O. minimum. Recently,
classification of different O. basilicum cultivars has been achieved by
means of DNA genotyping (36).
The perfume, pharmacy, and food industries (76) use aromatic essen-
tial oils, extracted from the leaves and flowers of basil. In chemotypes
of basil, several aroma compounds can be found, such as citral, eugenol,
linalool, methylchavicol, and methyl cinnamate that are traded in the in-
ternational essential oil market (83). These chemotypes are commonly
known by names based on geographical origins such as Egyptian,
French, European, or Reunion basil (43, 76, 78). The European type, a
sweet basil, is considered to have the highest quality aroma, containing
linalool and methylchavicol as the major constituents (76). The Egyptian
basil is very similar to the European, but contains a higher percentage of
methylchavicol. The Reunion type, from the Comoro Islands, and more
 Review 129

TABLE 1. Brief description of the most common species and intraspecific

forms of basil.

Common Name Latin Name Description1

Sweet Basil Ocimum basilicum Most commonly grown type. Erect habit.
White flowers. Bright green, one cm long
leaves. Clove-like scent.
‘Genovese’ Basil Ocimum basilicum An Italian strain, regarded as the best
‘Genovese’ variety for pesto and garlic dishes. Erect
habit. Dark green leaves up to 2 inches
long. Slow to bolt.
Bush or Greek Basil Ocimum basilicum Dwarf varieties with very small (less than
minimum one cm long), pungent leaves. White
flowers. Plants are excellent for edging or
containers. Widely available varieties
include ‘Fine Green’, ‘Green Bouquet’,
and ‘Spicy Globe’.
Purple Basil Ocimum basilicum Grown for their ornamental foliage as well
purpurascens for culinary purposes. Lavender like
flowers. Same shape and size leaf as
sweet basil. Varieties include ‘Opal’,
‘Purple Ruffles’, and ‘Red Rubin’.
Lettuce-leaf Basil Ocimum basilicum Flavor is less pronounced than other green
crispum basils, sometimes preferred for salads or
sauces. Large, wide leaves. Common
varieties include ‘Mammoth’, ‘Napoletano’,
and ‘Green Ruffles’.
Scented Basil Ocimum basilicum Cinnamon, lemon, and licorice or anise
odoratum basils possess flavors reminiscent of other
plants. Often used in fruit preserves or in
custards and sorbets.
Holy Basil Ocimum canum or Leaves are small with a clove-like
Ocimum sanctum fragrance. Violet or white flowers.
Used in some religious ceremonies.
Camphor Basil Ocimum The species is characterized by a strong,
kilimandscharicum medicinal scent. Gray-green foliage. Not
used for culinary purposes.
Peruvian Basil Ocimum Used for culinary applications, in spite of
micranthemum its medicinal, sweet flavor.
Thrysiflora Basil Ocimum thrysiflora Grown for its ornamental seed head, which
is a mound of purple flowers.
1Adapted from (3, 12, 13, 14, 15, 43, 48).

recently from Madagascar, Thailand, and Vietnam, is characterised by

high concentrations of methylchavicol (43). Methyl cinnamate-rich basil
has been commercially produced in Bulgaria (76), India, Guatemala, and
Pakistan (43). Basil from Java (76), and Russia and North Africa (43) is
rich in eugenol.
130 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

Basil, French basil, or sweet basil (Ocimum basilicum L.), a popular,

tender, annual herb is native to India and Asia (Figure 1). Although also
grown as an ornamental plant, basil is cultivated commercially for the
green, aromatic leaves that are used fresh or dried as a flavoring or spice.
The essential oil and oleoresin extracted from the leaves and flowering
tops via steam distillation is used in place of the dried leaves for flavoring
purposes. The plant is pubescent, grows about one meter high, and has
an obtusely quadrangular, stem. The labiate flowers are white, in whorls
in the axils of the leaves and the calyx with the upper lobe rounded and
spreading. The leaves, which have a greyish-green on the bottom and
dotted with dark oil cells, are opposite (1, 3, 13, 47).
Bush basil (Ocimum minumum), usually propagated by seed sown
annually, is a low, bushy plant, seldom above 6 inches in height, much
smaller than sweet basil. The leaves are ovate, quite entire, the white
flowers in whorls towards the top of the branches and smaller than those
of sweet basil. Of the two varieties of bush basil, one with black-purple
leaves and the other has variable leaves. Both bush and garden basil
are natives of India where bush basil may occasionally live through the
winter, although sweet basil never does. Both varieties flower in July and
August (12, 44, 47). The leafy tops of bush basil are used the same as
sweet basil, for seasoning and in salads.
The leaves of O. viride, a native of Western Africa, possess febri-
fugal properties and at Sierra Leone, where the plant bears the name
‘Fever-plant’, a decoction of the leaves is drunk as tea as a remedy for
the fevers. The leaves of the sweet-scented varieties O. canum, and
O. gratissimum in India, and of O. crispum in Japan, are prescribed as
a remedy for colds. O. teniflorum is regarded as an aromatic stimulant in
Java and O. guineense is employed as a medicine in cases of bilious fe-
ver. These plants are fragrant and aromatic and hence are valuable as
kitchen herbs.
Calamintha clinopodium, widely known as wild basil or hedge basil
and hedge calamint, is widely distributed throughout the North Temper-
ate Zone, and is common in the dry hedges and the copse borders
(mostly in higher elevations) of England and Scotland. In Ireland, wild
basil is somewhat rare.
Wild basil is a straggling plant with somewhat weak-looking, though
erect, stems and thickly covered with soft hairs (12, 22, 62, 74, 76).
Pairs of the opposite, short-stalked, egg-shaped leaves on the four-an-
gled stem are some distance apart. The flowers bloom from July to Sep-
tember. The entire herb is aromatic and fragrant with a faint thyme-like
 Review 131

aroma, and has been used to make an infusion for complaints similar to
calamint.

CULTIVATING BASIL
AS AN EDIBLE ORNAMENTAL HERB

Commercial cultivars. Several basil varieties, differing in the size,

shape, aroma, and color of the leaves exist. Commercial basil cultivars
also display a wide diversity in growth habit, flower, leaf, and stem col-
ors, and aromas. Many of the cultivars evaluated belong to the “sweet”
basil group with ‘Genovese’, ‘Italian large leaf’, ‘Mammoth’, ‘Napole-
tano’, and ‘Sweet’ dominating the American fresh and dry culinary herb
markets. Several other basils like ‘Sweet Fine’ appear similar to ‘Sweet’
basil though the leaves tend to be smaller (75). The lemon-scented culti-
vars (‘Lemon’ and ‘Lemon Mrs. Burns’) differ from each other in days
to flower, and total oil content, but not in citral content. The ‘Maenglak
Thai Lemon’ basil, which varies in appearance from the other lemon
basils, is an attractive ornamental. Among the purple basils, ‘Osmin Pur-
ple’ and ‘Red Rubin Purple Leaf’ are the most attractive and best retain
their purple leaf colour (75). Anthocyanins in purple basils are geneti-
cally unstable, leading to an undesirable random green sectoring and re-
version over the growing season (60). Several basils with dwarf growth
habit were developed as ornamental border plants including ‘Bush’,
‘Green Globe’, ‘Dwarf Bush’, ‘Spicy Globe’, and ‘Purple Bush’. A
group of ornamental basils were selected and named for their characteris-
tic aroma including ‘Anise’ (methyl chavical), ‘Cinnamon’ (methyl cinn-
amate), ‘Licorice’ (methylchavicol), and ‘Spice’ (-bisabolene) (1, 12, 13,
44, 47, 75).
Sensoric quality. Fresh basil leaves have a strong and characteristic
aroma, not comparable to any other spice, although a traceable hint of
cloves exists. In addition to the “Mediterranean type” most common in
the West, a plethora of other varieties or cultivars with different flavors,
many of which are hybrids, are available. India has “Sacred Basil”
(O. sanctum = O. tenuifolium) with an intensive, somewhat pungent
smell and Thailand has a sweet basil with a licorice aroma. Varieties
sold to gardeners in the West include cinnamon basil, camphor basil, an-
ise basil, and spice basil; the latter has a very pleasant, complex, and
warm flavor (1, 3, 13, 44, 47, 60, 75).
A last group of cultivars, characterized by citrus aroma, are ‘Thai Le-
mon basil’ (O. citriodorum) which has a distinct balm-like flavor and
132 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

lime basil and another lemon basil (O. americanum) which has an ex-
traordinarily pure and fresh lemon aroma (48).
Perennial basil species from Africa (O. kilimandscharicum) and Asia
(O. canum) have recently been introduced to the European herb and gar-
dening market. These species have a strong, but less pleasant flavor and
hybrids between them and Mediterranean basil are a recent innovation
with a novel appearance and flavor that are enjoying a growing popular-
ity (12, 22, 31). All basil varieties have in common that dried leaves are
much less aromatic than fresh ones; deep-freezing the herb is the best
method of preservation.
Ecology. Sweet basil is cultivated in agro climes between 7 to 27⬚C,
with 0.6 to 4.2 m annual precipitation and soil pH 4.3 to 8.2. While sus-
ceptible to frost and cold-temperature injury, the species develops best
under long days, in full sun and on well-drained soils.
Planting. Basil can be directly seeded or transplanted to the field in
late spring, after the frost period. The germination rate of the seed is 80-
95%, and seeds should not be planted if the germination percentage is
less than 70% (75). The seed is relatively small, and a good friable,
well-tilled and uniform seedbed is required for optimum plant establish-
ment. Seeds should be planted only 0.3-0.6 cm deep. Plant emergence
should occur between 8 and 14 days. To encourage lateral branching and
growth, the tops of transplants can be trimmed prior to field planting,
when they are about 15 cm tall (11).
Population and spacing. While the optimum population density is
dependent upon the end use, planting in rows 10-15 cm apart with plants
spaced every 2 cm in each row is recommended. Large variations in
growth and yield may occur due to climatic conditions, plant type, and
cultural and management practices (11-15).
Fertilization. For basic fertilization, an N-P-K ratio of 1-1-1 is sug-
gested. A side dress application with nitrogen at a rate of 250-500 kg N
ha⫺1 is recommended shortly after the first harvest. An investigation of
different nitrogen rates on the growth and development of three Italian
cultivars of O. basilicum by Sifola and Barbieria (73) indicated that nitro-
gen fertilization to 300 kg ha⫺1 increased yield of above ground, leaf fresh
biomass and leaf essential oil yield, but did not affect leaf-to-stem ra-
tio, plant height, or the number of branches per plant. The increase in es-
sential oil yield induced by nitrogen fertilization depended on an increase
in both leaf essential oil concentration and leaf biomass. The effect of ni-
trogen fertilization on essential oil composition was strongly cultivar-
dependent.
 Review 133

Irrigation. Basil is not tolerant to water stress. A regular and even sup-
ply of moisture via trickle or overhead irrigation is necessary. If trickle ir-
rigation is employed, care must be observed during harvest so no damage
is done to the irrigation line (3, 15, 75).
Weed control. Presently, high plant populations of basil coupled with
mechanical cultivation are recommended. Commercially labelled herbi-
cides for weed control in basil fields are not available in most countries.
The presence of weeds in fresh or dried basil leaves decreases the qual-
ity of the finished product (11, 15).
Insects and diseases. Several insects and diseases may infest basil,
but no pesticides are currently available for use on basil. While basil is
susceptible to at least 30 viruses, including alfalfa mosaic alfamovirus,
cucumber leaf spot carmovirus, and tobacco mild green mosaic dian-
thovirus, the plant is resistant to more than 40 viruses (3, 15).
Harvesting. Generally, basil is harvested for the leaves that are sold
fresh or dried. The foliage should be harvested, but only above the bot-
tom two to four sets of true leaves. The plant part harvested depends
upon the projected use. Where basil is grown for the dried leaves and the
extraction of essential oil, the plant is cut just prior to the appearance of
flowers. In the Mediterranean area and in other countries with similar
climates, basil is grown as a short-lived perennial and three to five cut-
tings per year are possible (11, 75). In the more temperate zones, basil
may be cut only one to three times where the first harvest is usually
quite low. The second harvest occurs just prior to open bloom.
The effect of different harvesting methods on essential oil yield and
quality of methyl eugenol-rich O. tenuiflorum has been investigated by
Kothari et al. (35). Secondary branch harvest gave the least biomass
yield, but higher total essential oil yield over primary branches and shoot
biomass cut at 30 cm above ground harvesting. Harvesting shoot bio-
mass at 30 cm above ground produced oil containing the highest amount
of methyl eugenol. The content of methyl eugenol decreased from cut-
ting at 30 cm above ground with primary branch lower and secondary
branch lowest in methyl eugenol. A reverse trend was observed, how-
ever, for oil constituents (E)-cinnamyl acetate, eugenol, and ␤-elemene.
Post-harvest handling. The quality of basil is determined by color and
aroma retention. Prior to milling or distillation, the leaves and/or flower-
ing tops should be dried at low temperatures (< 40⬚C) to retain maximum
color (3, 15). Basil can be oven dried at 50⬚C to 17.31% moisture content
in 15 h and can be sun dried to 23.79% moisture content in 28 h. Özcan
et al. (52) observed that the mineral (K, P, Ca, S, Mg, Fe, and Al) content
of oven-dried herbs was higher than sun-dried herbs.
134 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

BASIL AS A SOURCE
OF NATURAL PRODUCTS

Main constituents. The essential oil (less than 1%) is of complex and
variable composition. Within the species, several different chemical
groups exist, and furthermore soil and time of harvest influence the quan-
tity and the composition of the essential oil. Although essential oils in
different basil cultivars are variable, prevalent components are mono-
therpenes and phenylpropanoids (36). The most important aroma com-
ponents are 1,8 cineole, linalool, methyl chavicol (estragole) and, to a
lesser degree, eugenol (8, 43, 80). Further monoterpenes (ocimene, gera-
niol, camphor), sesquiterpenes (bisabolene, caryophyllene) and phenyl-
propanoids (methyl cinnamate, methyl eugenol) are present in varying
amounts and strongly influence the flavor (8, 44, 83). Sweet basil,
O. basilicum, contains primarily phenol derivatives, such as eugenol,
methyl eugenol, chavicol, estragole, and methyl cinnamate, often com-
bined with various amounts of linalool (45, 85).
The quality traded in Europe, Western Asia, and North America (Med-
iterranean type) is characterized by 1,8 cineole and linalool, plus smaller
amounts of estragole and eugenol. This description holds for both green
leaved and red-leaved (anthocyanin containing) strains. A couple of
chemotypes are dominated by methyl chavicol (estragole) and these
stand apart by their sweet anise (licorice) fragrance (anise basil, sweet
Thai basil), while lemon-scented varieties contain mostly citral. The Afri-
can species, O. kilimandscharicum, is characterized by much camphor
besides 1,8 cineole; camphor is also found, albeit in lesser quantities, in
kilimandscharicum hybrids with O. basilicum. The scent of cinnamon
basil comes from methyl cinnamate, a chemical also found in cinnamon.
Last, a very pleasantly scented cultivar called ‘spice basil’ was found to
contain beta-bisabolene, 1,8 cineole und estragole (5, 7, 9, 24, 75, 76, 80).
Indian ‘sacred basil’ (O. sanctum = O. tenuifolium) owes a stronger,
somewhat pungent taste than other basils to a sesquiterpenoid, beta-
caryophyllene, and a phenylpropanoid, methyl eugenol. Chemotypes of
O. gratissimum centering on eugenol are rare; their aroma resembles
cloves closely (8, 44).
Anthocyanins. The dark red foliage of some basil varieties is caused
by pigments, anthocyanin type, commonly found in reddish leaves. Some
basil varieties contain 200 µg anthocyanins/g fresh weight in their leaves.
Phippen and Simon (60) analyzed the anthocyanins present in purple
basils utilizing high performance liquid chromatography, spectral data,
 Review 135

and plasma-desorption mass spectrometry. A total of 14 different antho-

cyanins were identified of which 11 were cyanidin based with cyanidin-
3-(di-p-coumarylglucoside)-5-glucoside as the major pigment. Also
identified were three minor pigments based on peonidin. Purple basils
can be an abundant source of acylated and glycosylated anthocyanins
and could provide a unique source of stable red pigments to the food in-
dustry. The large-leafed cultivars ‘Purple Ruffles’, ‘Rubin’, and ‘Dark
Opal’ had an average extractable total anthocyanin content ranging from
16.6-18.8 mg per 100 g fresh tissue, while the ornamental small-leafed
cultivar ‘Purple Bush’ had only 6.5 mg per 100 g fresh tissue.
Antioxidant compounds. Basil extracts display potent antioxidant prop-
erties, which are stable over an extended pH and temperature range (to
80⬚C in neutral or acidic solutions). These properties are expressed as
strong superoxide anion scavenging activity, Fe2⫹ chelating activity,
and reducing power in a concentration-dependent manner (30). Among
the compounds (terpenoids, flavonoids, and tocopherols) thought to
have the strongest antioxidant activity are some phenolic acids ob-
served in relatively large amounts in the essential oil of species of the
Lamiaceae family, especially in oregano (Origanum spp.), sage (Salvia
spp.), rosemary (Rosmarinus officinalis), thyme (Thymus spp.), and ba-
sil (17). Some representative phenolic acids in these plants are caffeic,
p-coumaric, ferulic, syringic, and vanillic acids (71). Inatani et al. (27)
has isolated and characterized rosmanol, a phenolic diterpene with anti-
oxidant activity, from rosemary. The metabolite has also been found in
other species of Lamiaceae (37). Other compounds isolated from rose-
mary that show antioxidant efficiency are rosmarindiphenol (25), and
rosmariquinone (26). The antioxidant properties of Origanum species,
such as O. sanctum, have been associated with antidiabetic activity (6).
Rosmarinic acid (␣-o-caffeoyl-3,4-dihydroxyphenylactic acid) (Fig-
ure 2) is an antioxidant phenylpropanoid whose chemical structure was
identified in rosemary extracts (68). Biosynthesis of rosmarinic acid in-
volves the phenylpropanoid pathway. Razzaque and Ellis (64) confirmed
the biosynthesis pathway with cell suspensions of Coleus blumei. The
two precursors involved in this biosynthesis procedure are tyrosine and
phenylalanine. In 1989, the last enzyme in the chain, rosmarinic acid
synthase, was isolated and described by Petersen (59).
A phenylalanine ammonia-lyase (PAL) has been purified from leaves
of O. basilikum chemotype methyl cinamate. The enzyme catalyses the
conversion of L-phenylalanine-d8 into trans-cinnamic acid-d7. Compar-
ing the Ki values between trans-cinamic acid and trans-methyl cinamate
for L-phenylalanine indicated that the regulation of PAL is related to
136 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

FIGURE 2. Rosmarinic acid.

synthesis mechanisms besides the mechanism of feedback inhibition in

the biosynthesis of trans-methyl cinamate (87).
The essential oils of basil also have potent antioxidants properties.
According to a recent investigation by Leea et al. (38), eugenol, thymol,
carvacrol and 4-allylphenol had stronger antioxidant activities than other
volatile components of basil and these antioxidant activities were compa-
rable to those of the known antioxidants, ␣-tocopherol and butylated
hydroxy toluene (BHT).

BIOTECHNOLOGY

An increasing interest in the use of efficient protocols for the tissue

culture and micropropagation of basil for in vitro production of second-
ary metabolites and for clonal multiplication of elite genotypes has de-
veloped.
Callus induction and culture. Callus cultures from basil leaf and
shoot explants can be established on solid Murashige and Skoog (MS)
medium supplemented with 3 percent (w/v) sucrose and a combination
of a cytokinin, such as kinetin (Kn), and an auxin, such as indoleacetic
acid (IAA), as the growth regulators (41). Callus cultures usually grow
continuously and demonstrate a significant increase in both fresh and
dry weight after the first or second month of incubation.
Dalton (11) investigated the use of gas analysis during continuous
plant cell cultures in O. basilicum, in particular in the culture vessel inlet
and outlet streams under normal light conditions and during brief peri-
ods of darkness. The amounts of O2 and CO2 were measured and three
 Review 137

equations were derived relating yield to the rate of CO2 production

(RCO2), the rate of O2 consumption (RO2), and the ratio of RCO2/RO2
referred to as the gaseous exchange quotient (GEQ) or respiratory quo-
tient (RQ). Results indicated that gas analysis could be applied to plant
cell cultures with utility, particularly for the measurement of in situ pho-
tosynthesis and yield. Dalton (10) also reported on the effect of the
sugar supply rate on the photosynthetic development of sweet basil cells
in continuous culture. O. basilicum cells were cultured continuously in
a fermentor. The medium used for each steady state differed only in the
concentration of fructose. When fructose concentration was decreased
while maintaining a constant dilution rate, the rate of chlorophyll pro-
duction increased. The photosynthetic rate measured in the steady state
condition also increased. When fructose supply rate was increased by
increasing the dilution rate, both chlorophyll production and photosyn-
thetic rate increased.
Micropropagation. Basil micropropagation is usually achieved by
axillary shoot proliferation from node explants (more seldom leaf or in-
florescence explants) (Table 2). Culture media (solid) commonly con-
sist of MS salts, 3% (w/v) sucrose and benzyl adenine (BA) (alone or
with gibberellic acid, GA3) as growth regulators (Figure 3). Rooting of
the adventitious shoots takes place in half-strength MS medium supple-
ment with BA and indolebutyric acid (IBA).
Shetty (72) developed and patented a method for identifying a clonal
sweet basil line containing an elevated level of a secondary metabolite
that involved contacting propagules from at least one clonal line with
mucoid, non-pathogenic bacteria. Researchers working at Kanebo Ltd.
(31) presented a process for the multiplication of basil seedlings involv-
ing induction of multiple buds from apical, lateral or adventitious buds
of sacred basil (O. sanctum), sweet basil (O. basilicum), lemon basil (O.
citriodorum), and camphor basil (O. kilimandscharicum) in bud-induc-
tion culture medium; multiplication of the buds in a multiplication me-
dium; and regeneration of the plant body in a regeneration medium. Bud
induction, multiplication and regeneration culture media are also claimed,
containing 5-50 ␮M, 1-10 ␮M, and 0-0.1 ␮M cytokinin, respectively, and
0-10 ␮M, 0-0.1 ␮M and 0-0.1 ␮M auxin, respectively. Basil buds were
cut and cultured successively in MS, Linsmaier, B5, and Nitsch-Nitsch
or White media for induction, multiplication, and regeneration, at 15 to
35⬚C in the light (1,000-20,000 Lux) for 10 to 40, 30, and 10 to 20 days,
respectively. Cytokinins used included BA, Kn, and zeatin, and auxins
used included naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic
138
TABLE 2. Summary of micropropagation protocols of basil.

Species Explant Medium Plant growth regulator(s) Growth response Reference

O. basilicum apical, lateral MS, Linsmaier 0-10 µM BA ⫹ 5-50 µM NAA Bud induction Kanebo Ltd. (31)
O. sanctum or adventitious B5, Nitsch- 0-0.1 Mm Kin ⫹ 0-10 µM 2,4-D Bud multiplication
O. citriodorum buds Nitsch White 0-0.1 µM Zea ⫹ 0-0.1 µM IAA Plant regeneration
O. kilimandscharicum
O. basilicum Adventitious MS 0.5 mgL-1 BA ⫹ 0.3 mgL−1 GA3 Shoot induction Pattnaik et al.
O. sanctum buds 1.0 mgL−1 BA ⫹ 0.5 mgL−1 GA3 Shoot induction (58)
O. americanum 0.25 mgL−1 BA ⫹ 0.5 mgL-1 GA3 Shoot induction Patnaik and
O. gratissimum 0.5 mgL-1 BA ⫹ 0.3 mgL−1 GA3 Shoot induction Chand (57)
1.0 mgl−1 IBA/NAA Root induction
O. basilicum Adventitious MS 1 mgL-1 BA ⫹ 0.4 mgL−1 GA3 Shoot induction Sahoo et al.
buds ½ MS 0.25 mgl−1 BA Shoot proliferation, (65)
1 mgl−1 IBA in vitro flowering
Root induction
O. sanctum Young MS 1 mgL−1 BA + 0.05 mgL−1 IAA Shoot induction + Singh and
inflorescences proliferation Sehgal (76)
Root induction
O. basilicum Leaves, MS 1.5 mgl−1 BA Shoot induction + Makri and
internodal proliferation Kintzios (41)
stem ½ MS 0,5 mgL−1 BA ⫹ 1 mgL−1 IBA Root induction
segments
 Review 139

FIGURE 3. Micropropagated sweet basil plants on half-strength MS medium

supplemented with 0.5 mgL⫺1 BA and 1 mgL⫺1 IBA.

acid (2,4-D), and IAA. A stable production of basil plants of the same
quality and with a high essential oil content was achieved quickly.
Pattnaik et al. (58) and Pattnaik and Chand (57) reported the clonal
propagation of four basil species: O. americanum (hoary basil), O. basi-
licum (sweet basil), O. gratissimum (shrubby basil) and O. sanctum L.
(sacred basil). Healthy axillary vegetative buds were collected from the
four species were cultured on MS medium with 0.25-2.0 mgL⫺1 BA
and 0.25-0.5 mgL⫺1 GA3 either individually or in combinations. In
O. americanum, sprouting of the axillary buds and development of multi-
ple shoots was best observed on MS with 0.25 mgL⫺1 BA and 0.5
mgL⫺1 GA3. For O. basilicum and O. gratissimum, the optimal level of
BA and GA in the culture medium was recorded as 0.5 mgL⫺1 and 0.3
mgL⫺1, respectively. In O. sanctum, high frequency sprouting of the
axillary buds and development of multiple shoot was best elicited on
MS with 1.0 mgL⫺1 BA and 0.5 mg L⫺1 GA3. The shoots formed in vitro
140 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

were excised and rooted on 1/2-strength MS medium with 1.0 mgL⫺1

IBA/NAA. Indolebutyric acid (IBA) (1.0 mgL⫺1) was most effective
for O. americanum, O. basilicum, and O. gratissimum. In O. sanctum,
rhizogenesis was best induced with an NAA (1.0 mgL⫺1) supplement.
Root induction rates varied between 94.6 and 98 percent. The rooted
plantlets were acclimatized on artificial soil and successfully trans-
ferred to field conditions with a 75 to 80 percent survival rate. In a sub-
sequent transfer to the greenhouse, 80 to 85 percent of the plants
survived.
Sahoo et al. (65) developed an efficient method for in vitro propagat-
ing sweet basil by axillary shoot proliferation from node explants carry-
ing a single axillary bud, collected from field-grown plants. High
frequency bud break and maximum axillary shoot formation was in-
duced in node explants on MS medium with BA. The node explants re-
quired the presence of BA at a higher level (1 mgL⫺1) at the 1st stage of
bud break, but further growth and proliferation required transfer to a
medium with BA at 0.25 mgL⫺1. Gibberellin at 0.4 mgL⫺1 added to the
medium with BA (1 mgL⫺1) enhanced the frequency of bud break. The
shoot clumps that were maintained on proliferating medium for longer
durations developed inflorescences and flowered in vitro. The shoots
formed in vitro could be rooted on half-strength MS plus 1 mgL⫺1 IBA.
Rooted plantlets were acclimated in vermi-compost inside a growth
chamber and eventually established in soil. All regenerated plants were
identical to the donor plants with respect to vegetative and floral mor-
phology.
Singh and Sehgal (77) have accomplished the micropropagation of
holy basil on MS medium utilizing young inflorescence explants. MS
supplemented with 2,4-D or thidiazuron (TDZ) produced only non-
morphogenetic callus. Direct multiple shoots differentiated within 2 to
3 weeks when explants were cultured on MS containing BA. Of the var-
ious levels of BA tested, MS ⫹ BA (1.0 mgL⫺1) produced the maxi-
mum number of shoots. Incorporation of IAA (0.05 mgL⫺1) along with
BA (1.0 mgL⫺1) in the culture medium showed a marked increase in the
number of shoots. About 92 percent of the in vitro regenerated shoots
rooted on MS hormone-free medium within two to three weeks of culture
and 85 percent of the micropropagated plantlets could be successfully es-
tablished in soil and grew normally.
Makri and Kintzios (41) identified a number of preliminary protein
markers for the association of distinct in vitro developmental phases of
sweet basil. Proteins, found at 63 and 92 kD, appeared in both donor
plants, leaf explants, and leaf-derived callus cultures. In contrast, two
 Review 141

other proteins, each smaller than 20 kD, were expressed in stem explants,
stem-derived callus cultures and micropropagated plants, indicating a de-
pendence of the expressed protein pattern on the original explant used
for culture initiation. Associating quantitative protein changes with cer-
tain traits of basil callus cultures, such as growth and total phenolics ac-
cumulation was, however, not possible. These markers could serve as
reliable indicators for the identification of distinct clones and the detec-
tion of somaclonal variants.
Somatic embryogenesis. Gopi and Ponmurugan (19) recently de-
scribed a protocol for plant regeneration via somatic embryogenesis
from leaf explants of O. basilicum. Globular embryos were induced on
MS medium supplemented with 2,4-D (0.5 mgL⫺1) and BA (1 mgL⫺1)
and further embryo development and maturation took place on MS me-
dium supplemented with NAA (1 mgL⫺1), BA (1 mgL⫺1) and Kn (0.5
mgL⫺1). The maximum induction rate was 75 percent, corresponding to
36.5 somatic embryos/culture.
Protoplast isolation and culture. Iseki (28) described a method for the
preparation of protoplasts by treating the callus from a leaf explant with
an enzyme solution containing cellulase (EC-3.2.1.4), polygalacturo-
nase (EC-3.2.1.15), calcium chloride, and mannitol to purify and sepa-
rate the protoplasts that were cultured on 8p culture medium containing
auxin. Protoplasts were obtained in a high yield.
Kao (32) presented another protocol for basil protoplast culture that
involved culture of basil protoplasts in the presence of a basil cell sus-
pension, resulting in efficient colony formation. In an example, 5 to 15
mm wide sweet basil leaves were cultured on solid Linsmaier-Skoog
(LS) culture medium to produce callus culture, and calli were subcul-
tured in LS liquid medium weekly to produce a suspension cell cul-
ture. The suspension was treated with enzyme solution containing 1
percent cellulase-Onozuka-RS, 0.5 percent pectinase-SE-150, 0.1 per-
cent Pectolyase-Y-23, 0.5 M mannitol and 5 mM MES at 26⬚C for 2 h.
Protoplasts were obtained by filtration and centrifugation, followed by
suspension in Nagata-Takebe (NT) medium. Gelrite (0.24%) was added
and 1 ml medium and 1 ml suspension were mixed in a 35 mm diameter
dish. Gel containing protoplasts was formed into 4 to 5 mm beads, and
0.5 ml of beads were mixed with 3 ml NT medium in 6-well plates. A
cell-culture insert with 3 ml basil suspension cell culture was added and
cultured at 26⬚C for 25 days under 100 lux light. The colony formation
rate was 19.6 percent, compared with 0.1 percent for a single culture
control.
142 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

In vitro production of secondary metabolites. Tada et al. (79) cul-

tured five clones of O. basilicum hairy roots, A-1 and A-2 (induced by
Agrobacterium rhizogenes ATCC 15834) and J-1, J-2 and J-3 (induced
by A. rhizogenes MAFF 03-01724) in three plant growth regulator-free
liquid media (MS) containing 30 gL⫺1 sucrose, Gamborg-B5 (B5) con-
taining 20 gL⫺1 sucrose and woody plant (WP) media containing
20 gL⫺1 sucrose). After inoculation into these media, all clones had
rapid proliferation at the early stages of culture particularly in the WP
medium. The highest amount of biomass recorded for any culture was
702.8 mg dry weight/flask for A-2 grown in MS medium for six weeks.
Although in all media the hairy root cultures produced substantial
amounts of rosmarinic acid (RA), particularly high levels were ob-
served in MS medium (14.1% for J-1 at wk 8) and B5 medium (14.0%
for A-2 at wk 6). These levels were almost 3.5-fold higher than those of
an intact plant. The maximum yield of RA in all cultures examined was
73.5 mg/flask produced by J-1 in MS medium at wk 5. Small amounts
of the related phenolics, lithospermic acid, and lithospermic acid-B
were also obtained.
Madhavi et al. (40) established callus cultures from sweet basil ‘Pur-
ple Ruffles’ which accumulated anthocyanins in the dark. Plant growth
regulators were found to have a distinct effect on callus induction and
pigmentation. A reverse phase high pressure liquid chromatography
(HPLC) analysis of anthocyanins revealed 6 major peaks in the leaf ex-
tract. The profile of anthocyanins from callus cultures was comparable
to the in vivo extract. Analysis of the acid hydrolysate of the total extract
indicated that the major aglycone was cyanidin.
Zeldin et al. (86) described a system for recovering essential oils from
in vitro grown lemon basil plants. Shoot cultures, grown in one-liter fl-
asks, were outfitted with an apparatus that supplied humidified air to the
culture. A column, fitted with Amberlite XAD-2 and attached to the air
outlet, was used to collect the volatile compounds emitted. These com-
pounds were eluted from the collector with methylene chloride and ana-
lyzed by thin layer chromatography (TLC) and HPLC. The method may
potentially be used for large-scale flavorings and fragrances production.
The advantages of this method over the present destructive agronomic
methods include biomass conservation, increased productivity (to 100-
fold), and recovery of highly purified compounds.
Purohit and Khanna (61) investigated the production of essential oil
from 24-month old callus tissues of O. basilicum. The cultures were
grown as static and in suspension cultures for the production of essential
oil. Both cultures were maintained by frequent subculturing every 4-6
 Review 143

weeks in fresh RT medium. The tissues were kept in the dark and culti-
vated at 26⬚C with a relative humidity of 55 percent. An individual lot of
static cultures was transferred to continuous light (1600 lux). The maxi-
mum oil yield with dark-grown and light grown static cultures was
0.035 and 0.043 percent of fresh weight, respectively. Maximum oil
yield for dark-grown suspension cultures was 0.026 percent. The cul-
tures grown under illumination showed a significant increase in the con-
stituent level, particularly linalool, and total oil yield.
Callus cultures of sweet basil (incubated on solid MS ⫹ 2 mgL⫺1 Kn ⫹
2 mgL⫺1 IAA) have been demonstrated to accumulate various phenolic
compounds with antioxidant activity (41). Callus tissues were extracted
in 80 percent (v/v) methanol and analyzed by HPLC with an APEX ODS
analytical column. Rosmarinic acid usually accounted for approximately
90 percent of the total phenolic content. The pattern of this accumulation
varied with time with production maxima (expressed on a callus fresh-
weight basis) observed during the 3rd, 4th, and 7th week of culture.
When results were expressed on callus dry weight, maximum produc-
tion was observed only during the 4th week of culture. The observed
RA levels were almost 200 percent higher than that of the intact plant.
Panagiotopoulos (53) investigated the RA accumulation in cell suspen-
sions of sweet basil cultured in MS supplemented with 50 gL⫺1 sucrose,
0.5 mgL⫺1 L-Phe, 2 mgL⫺1 2,4-D, and 2 mgL⫺1 Kn during a total period
of five weeks. Maximum RA production, approximately 2 mg RA g⫺1
fresh weight (1.2 mg RA g⫺1 dry weight), was achieved during the second
week of culture. Culture growth (in terms of fresh and dry weight), oxygen
consumption, and antioxidant activity of the extracts demonstrated a sim-
ilar pattern, dependent on the pH of the medium. Increasing the volume of
culture medium from 25 to 50 ml significantly increased RA accumula-
tion. Kintzios et al. (34) investigated the accumulation of RA in different
types of sweet basil cell cultures (callus, cell suspension, and immobilized
cell). Leaf-derived suspension cultures grown in a liquid Murashige and
Skoog (MS) medium were able to accumulate RA at a maximum con-
centration of 10 mg g⫺1 dry weight, a value 8.5 to 11 times higher than
for callus cultures and donor plants. These results may have been due to
feeding culture media with the RA precursor L-phenylalaline (at a con-
centration of 0.5 g L⫺1). Immobilization of basil cells in 2-3 percent (w/v)
calcium alginate, however, failed to increase production (< 15 µg g⫺1)
over non-immobilized cell suspensions (69).
144 JOURNAL OF HERBS, SPICES & MEDICINAL PLANTS

Increased RA accumulation has been associated with an advanced state

of differentiation in vitro. For example, Rady and Nazif (63) achieved the
synthesis of 3 mg g⫺1 d.w. RA in O. americanum plantlets regenerated
from shoot cultures grown on MS medium supplemented with BA (1
mgL⫺1) and NAA (0.25 mgL⫺1). By comparing RA accumulation and
the results of an RAPD analysis of in vitro regenerated plants, molecu-
lar differences between the donor and the regenerants could be identi-
fied, suggesting the existence of genetic variation that might have
affected the biosynthetic pathway of RA in vitro.
Cell suspensions accumulating RA tend to increase their productivity
with the volume of the culture vessel. When cultured in 5-liter airlift
bioreactors for three weeks (33), the fresh weight of sweet basil suspen-
sion cultures increased by 241 percent while RA was accumulated at 29
µg g⫺1 dry weight. During the same period, RA accumulation in biore-
actor-micropropagated basil plants (Figure 4) reached 178 µg g⫺1 dry
weight, which is 44 times higher than in suspension cultures in a 250-ml
flask. Since product formation was growth associated, a minimum in-
oculum was required for culture initiation and satisfactory RA accumula-
tion was achieved within a short culture period.
Other studies. Deufel et al. (16) suggested the use of phototropic ba-
sil cell cultures for labeling proteins expressed by cells (recombinant
proteins).

FIGURE 4. Micropropagated sweet basil plants in five liter airlift bioreactors.

Picture adapted from Kintzios et al. (33).
 Review 145

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doi:10.1300/J044v13n03_10