CHEMOTAXONOMY

1.1. CHEMISTRY OF NATURAL PRODUCT: SCIENCE OF ALL TIMES

Natural products, as the term implies, are those chemical compounds derived from living
organisms and the study of natural products is the investigation of their structure,
formation, use and purpose in the organism. Drugs derived from natural products are
usually secondary metabolites and their derivatives. Today those must be pure and highly
characterized compounds. Since prehistoric times, the humans have relied on natural
products as a primary source of medicine. Plants and animals were used to bring back the
health of sick and frail. Plant were found to be beneficial as food, fodder, medicine etc. but
also harmful as being poisonous and toxic (Fuller and Hemrick, 1985). The application of
herbs for external and internal use has always been a major factor in practice of
medicine (Steiner, 1986). The experience and knowledge gained in using the traditional
medicines in different regions over the millennia resulted in the complex science of modern
medication.

The various approaches to drug discovery from nature are:

Ethnobotanical: Ethnic and traditional medicine

Random screening: Bioassay guided routes
Chemotaxonomic: Screening of relatives

1.2. TAXONOMY OF PLANTS

Biological classification or scientific classification, is a method by which
taxonomists group and categorize plants by biological type, such as genus or species.

The classification of plants by grouping data according to morphological similarities is

probably the oldest and most widely-used of all the approaches (Quinlan, 1993).

However many approaches evolved over time towards the taxonomy of plants.

1.2.1. CAUSES OF TAXONOMIC COMPLEXITY

Morphological variants of a species results because of different factors. In addition,
many species exhibit considerable genetic variation, both florally and vegetatively. This
variation may occur in different populations of the same species, or may characterize
different infraspecific categories of a species. Sometimes, the character may be genetically
controlled in one species, but phenotypically plastic in another. Quaternary climatic
changes have had a profound impact on speciation, structuring of genetic diversity and
the shaping of the present-day distributions of plant and animal taxa (Avise, 2000; Hewitt,
1996, 2000, 2004; Vuilleumier, 1971). Oscillations of population sizes, bottle necks,
founder events and other population historical events associated with climatic shifts
have further contributed to differentiation among regional population groups. As a
combined effect of range shifts and population differentiation, divergent lineages have
occasionally formed contact zones, leading to reticulate speciation by means of
hybridization and polyploidization (Grant, 1981; Stebbins, 1984). Polyploid speciation has
long been recognized as an important process in plant evolution (Mntzing, 1936;
Stebbins 1950; Grant, 1981). Recent genomic studies have made it clear that
angiosperms possess genomes with considerable gene redundancy, indicating that most
(if not all) plants have undergone one or more episodes of polyploidization (Soltis et al.,
2003).

1.3. CHEMOTAXONOMY
Chemotaxonomy is also called chemosystematics or biochemical systematics. The
science of chemical taxonomy is used on the classification of plants on the basis of their
chemical constituents which are deeply concerned with the molecular characteristics.
The method of chemical taxonomy is simple in principal and is based on the investigations
of the distribution of chemical compounds or groups of biosynthetically related

compounds in series of related plants. Different plants sometimes contain substances

which although belong to different chemical compounds appear to be biosynthetically
analogous. Such plants may contain similar enzyme systems, and the compounds produced
by such enzymes are indicative of the relationships that exist between the plants. However,
the chemotaxonomic studies include the investigation of the patterns of the compounds
existing in plant. Climatic conditions have a major influence on the distribution of plants
containing certain substances e.g. fats, volatile oils, alkaloids, flavonoids etc. It is well
known that for tropics, and perhaps for all climates, the chemical products are highly
organized. According to Reichert (1919), it is possible to identify many plants by their
starch grains. Stress has been given on the importance of -Cyanins and -Xanthins in plant
taxonomy. -Cyanins are commonly met within the families of order centrospermae. The
other chemicals are also found specifically in particular orders or families of flowering
plants e.g. Isoquinoline (Alkaloids) is found in the families of Ranales; retanone in the
families of leguminales, biflavonoils in casuarinas equisetifolia (casuarinaceae). The
presence of such chemicals in different groups of plants has great taxonomic significance
(Stuessy, 2008). Taxonomic studies for various plant taxa by using different parameters
have been successfully carried out in Pakistan including that of cereals (Ashraf et al.,
2003), Legumes (Ahmad et al., 2007) and Maize (Nawaz and Ashraf, 2007).

1.3.1. APPLICATION OF CHEMOTAXONOMY (Stuessy, 2008)

There are a few angiospermic taxa which are characterized by specific
compounds

of

general

occurrence.

For

example

leaving

aside

the

family of

caryophyllaceae, the rest of the families such as chenopodiaceae, amaranthaceae,

aizoacaceae etc. of the taxon caryophyllales (centrospermea) contain -cyanin a colored
substance

but

differs

from

anthocyanins.

It

appears

that,

with

the

exception

caryophyllaceae, these families are closely related and therefore caryophyllaceae may be
isolated. -cyanin also occurs in cactaceae and therefore, the members of caryophyllales
are phylogenetically related. There are certain other chemical connections between
cactaceae

and

members

of

caryophyllales

e.g.

common presence of isoquinole

alkaloids in Salsola of Chenopodiaceae and cactaceae. Another example that may be

cited is in the family crucifereae, where unsaturated acid erucic acid is prominent and
also in Tropaelum erucic acid is present; it indicates the relationship between Geraniales
and Rhocadales. In umbellifereae and Araliaceae petroselinic acid (a structural isomer of

Oleic acid) occurs and these two families are related and belong to the same order. The
other examples are from Magnoliales

Ranals

taxa,

where

it

is

shown

that

magnoliaceae, lauraceae, Ranulculaceae, Annoraceae, the alkaloid isoquinoline is present,

this supports that these

families

are

loosely

related.

On

the

other

hand

Asclepiadaceae and Gentianaceae are allied due to the common occurrence of pyridine. The
lilliaceae and Amarylldeceae are closely associated and this is supported by the
presence of Isoquinoline in both. A number of citations regarding chemotaxonomy
and secondary metabolites further.

1.4. FAMILY SOLANACEAE

The Solanaceae, to which the genus Solanum belongs, is a cosmopolitan family
which is widely distributed throughout tropical and temperate regions of the world, with
centers of diversity occurring in Central and South America and Australia. It is
composed of approximately 84 genera and 3000 species. The name of the family comes from
the Latin word Solanum, meaning "the nightshade plant", but the further etymology of that
word is unclear; it has been suggested it originates from the Latin verb solari meaning "to
soothe". This would presumably refer to alleged soothing pharmacological properties of
some of the psychoactive species found in the family. It is more likely, however, that
the name comes from the perceived resemblance that some of the flowers bear to the
sun and its rays, and in fact a species of S. nigrum Complex (Solanum retroflexum) is
known as the sunberry. The family is also informally known as the nightshade or potato
family (Yasin, 1985).

1.6. SOLANUM NIGRUM: TAXONOMIC COMPLICATIONS

Solanum nigrum is the most variable species of the genus Solanum. The species related
to S. nigrum have been reclassified innumerable times. Characters used by later
taxonomists to separate and describe additional taxa often differed very slightly from those
given for species by earlier workers. These Solanum species display varying amounts of
phenotypic variation, particularly in their vegetative features such as plant habit, leaf
size and form, and stem winging. In addition, senescence is often accompanied by smaller
and fewer flowers and fruits (Ganapathi and Rao, 1986).

S. nigrum was first delimited in four taxa with polynomials by Dillenius. Linnaeus
subsequently modified Dilleniuss work, describing these in six varieties under the
binomial S. nigrum (Edmonds and Chweya, 1997). Since then, the plants morphologically
related to S. nigrum have been reclassified many times. Over 300 post-Linnean specific
and infraspecific names have now been published, and synonymy is extensive within
the section. However, no satisfactory revision of the whole section has yet been devised.
The boundaries between many of the species are still ill-defined, with many of the new
taxa proving to be no more than slight morphological

variants

of those

already

described. The situation is further complicated by the researchers who either treated
different members of the section as varieties of S. nigrum or considered them as
different species on the basis of morphological
Chweya,

1997;

Schilling

differences

(e.g.

Edmonds

and

and Andersen, 1990; Stebbins and Paddock, 1949; Symon,

1970). These Solanum species display varying amounts of phenotypic variation,

particularly in their vegetative features such as plant habit, leaf size and form, and
stem winging. In addition, senescence is often accompanied by smaller and fewer flowers
and fruits than usual. Natural hybridization is probably more widespread in this section
than generally supposed. It is now named as Solanum nigrum Complex because it is
composed of a large number (about 30) of morphologically distinct taxa (Schilling and
Andersen, 1990). Only during the revision of Solanum section Solanum appear in 1979
for Flora Europaea 3, drawn turned out that in Europe two different forms of the species
coexist. The most widespread form was considered subspecies S. nigrum ssp.
nigrum, the second, rarely encountered species as S. nigrum ssp. schultesii classified
(Edmonds and Chweya, 1997).

Deadly nightshade, identified as S. nigrum, causes belladonna poisoning (Hubbs, 1947),

with symptoms including widely dilated pupils (characteristic of the atropine group, but
either unexpressed, or expressed very mildly, in poisonings by plants whose major
poisoning principle is of the solanine group). Deadly nightshade is identified as S. nigrum,
but black nightshade is botanically unidentified and gives a strong cat's-eye test for atropine
(Case, 1955). According to a research at University of Pennsylvania, unripe berries are said
to be more toxic than ripe berries. Berries are more toxic than leaves which, in turn, are
more toxic than stems or roots. Overall plant glycoalkaloid content is often higher in the
autumn than in the spring. These problems clearly state the importance of proper

identification and detailed composition analysis of each taxa of S. nigrum Complex.

The taxa of S. nigrum Complex are difficult to distinguish because
1. They are morphologically similar.
2. These species are all highly phenotypically plastic.
Three taxa belonging to S. nigrum Complex viz.: S. americanum Mill., S. nigrum L.
and S. villosum Mill. had been reported in Pakistan (Schilling and Andersen, 1990).
S. chenopodioides Lam. and S. retroflexum Dunal are two other species that were found
growing wild in and around Botanic Garden, GC University, Lahore. Morphologically S.
nigrum is different from S. villosum in the respect that the former has black matured berries
with peduncles longer than pedicels while latter has orange/orange-red matured berries and
peduncles shorter than or equal to the pedicels. Classification of S. nigrum and S.
villosum as varieties or distinct species started taxonomic controversy between Linnaeus
and Miller (Edmonds and Chweya, 1997).

Though

S.

americanum

Mill.,

S.

chenopodioides Lam. and S. retroflexum Dunal have morphological resemblance with S.

nigrum, yet no chemotaxonomic relationship has so far been established due to lack of a
comprehensive study of their chemical composition.

1.7. BOTANICAL ASPECTS OF THE INVESTIGATED TAXA

The five locally available taxa investigated were:
1. Solanum americanum
2. Solanum chenopodioides
3. Solanum nigrum
4. Solanum retroflexum
5. Solanum villosum

These Solanum species display varying amounts of phenotypic variation, particularly in

their vegetative features such as plant habit, leaf size and form, and stem winging. In
addition, senescence is often accompanied by smaller and fewer flowers and fruits than
usual, while the gene for anthocyanin pigmentation in flowers seems to be dependent on
light intensity and temperature for its expression, in some species. It is therefore often
difficult to define the limits within which such features are genetically fixed (Baylis, 1958;

Henderson, 1974; Edmonds, 1977).

Natural hybridization is probably more widespread in this section than generally

supposed. Though this is probably followed by subsequent genetic breakdown in F1
or F2 generations (Edmonds, 1977), it may also be followed by back-crossing to the
parental species. This would result in morphogenetically complex population variation:
the collection of specimens from such populations would
difficulties

encountered

in

the

explain

some

of

the

morphological differentiation of these species in the

herbarium (Edmonds, 1979).

1.11. AIMS AND OBJECTIVES

There are various approaches to the taxonomic studies of the plants based on the structural,
cytological and chemical constituents. The ancient classification of the plants was mainly
carried out on the comparative morphological and anatomical concepts of the natural
plant flora.

However with the rapid progress in the isolation, purification, identification, elucidation of
structure and the configuration of natural plant products, the phytochemists and
ethnobotanists believe that it is possible to characterize and classify the plants on the
basis of their chemical constituents. The chemical constituents are formed within the plants
by definite biosynthetic pathways aided by the specific enzymes.