International Journal of Horticulture, 2015, Vol.5, No.21, 1-45
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separation of flavonoids from plant materials have been column, paper and Thin Layer Chromatography (TLC).
Although good results have been obtained with these techniques, they are not easily adopted to quantization. High
Pressure Liquid Chromatography (HPLC) has been found to be applicable to the analysis of complex natural
mixtures of flavonoids for rapid screening and “fingerprinting” purposes. The use of these chemical markers as an
adjunct to the chemical methods presently used for plant identification will provide a more positive identification
of new cultivars, particularly those protected by the plant patent law.
Phenolic compounds have been found to be very useful for documentation of hybrids, detection of diploid
genomes constituting polyploids, detection of mutants and in solving taxonomic problems. The use of phenolic
compounds in taxonomic and genetic investigations has been reviewed earlier (Bose and Frost, 1967; Datta and
Basu, 1976; Datta, 1987). Different types of phenolic compounds are present in leaves, flowers, stem, root and
bark of plants. Distribution pattern of some of these compounds varies not only in different species but also in
cultivars of the same species. Bose (1968) reviewed the use of phenolic compounds in taxonomic and genetic
investigations. Thin layer chromatographic analysis of phenolic compounds in plant tissues led Smith and Levene
(1963) to detect the presence of species substances in the polyploids, indicating the presence of different diploid
genomes which constitutes the polyploids. Alston and Turner (1963) reported a number of species specific
phenolic compounds in
Baptisia
from the chromatographic pattern. F
1
hybrids with new hybrid substances were
reported in
Betula
,
Collinsia
,
Saxifraga
,
Dicentra
and
Vinca
(Clausen, 1963; Garber and Strmnaes, 1964;
Jaworska and Nybom, 1967; Fahselt and Ownbey, 1968; Mukherjee and Basu, 1973; Stebbins et al., 1963; Levin,
1966). Stebbins et al. (1963)and Levin (1966; 1967) demonstrated presence of all the parental compounds in F
1
interspecific hybrid. Basu and Mukherjee (1975) demonstrated species and variety specific spots in Indian
Spinach and such a study in addition to cytological and morphological investigations helped them in establishing
the taxonomic status of Indian Spinach. Chemotaxonomic affinities were determined among the different species
of Pennisetum by thin layer chromatographic compounds (Misra and Saran, 1964). This technique has also been
successfully used in mutation breeding. Mutants are often associated with changes in biochemical characteristics
which can be used as markers. Morphological mutants in fruit colour and shape have been induced in
Trichosanthes anguina
. Chromatographic analysis of phenolic compounds in leaves showed the presence of new
spots in mutant (Datta, 1976). Chromatograohic analysis of seed extracts of control and MMS induced green seed
coat colour mutant of
Trigonella foenum-graecum
showed quantitative differences of phenolic compounds (Laxmi
et al., 1983). Changes in phenolic compounds have been reported in somatic flower colour mutations in
chrysanthemum (Datta, 1987; Datta and Gupta, 1981b; Datta and Gupta, 1983a, 1983b, 1983c; Datta, 1986; Datta,
1999; Datta and Singh MS, 1999).
Electrophoresis of crude proteins and enzyme extracts has been successfully used as an additional tool to establish
these relationships. Protein and isozyme polymorphism has been successfully used for demonstrating genetic
variation, identifying interspecific hybrids and fingerprinting cultivars (Cardy and Kannenberg, 1982; Chaparro et
al., 1987; Ellstrand, 1984) Protein electrophoresis has provided a new approach to the problems of species
relationships. A crude protein extract when fractionated on a suitable gel medium produces a spectrum of bands,
which is diagnostic for the species. Homology among the bands of different species based on similarity in
migration velocity provides a criterion of genetic affinity from which evolutionary relationship may be inferred.
The homology of electrophoretic pattern of soluble proteins has been very extensively used in the study of intra -
and interspecific relationships in many crop species (Cardy and Kannenberg, 1982; Chaparro et al., 1987;
Ellstrand, 1984; Johnson, 1967; Cherry et al., 1971; Nainawatyee and Das, 1972).
Besides biochemical markers, DNA based markers provide powerful and reliable tools for discerning variation
within crop germplasm and to study evolutionary relationships (Swofford and Olsen, 1990; Gepts, 1993). The
term DNA fingerprinting was first used by Alec Jeffrey in 1985 to describe bar-code-like DNA fragment patterns
generated by multilocus probes after electrophoretic separation of genomic DNA fragments. Now-a-days DNA
fingerprinting is used to describe continued use of several single locus detecting systems and presently DNA
fingerprinting is used to analyze the various aspects of plant genus such as taxonomy, phylogeny, ecology,
genetics and breeding in inter or intraspecific level. These techniques are being widely used to detect molecular
