International Journal of Horticulture, 2015, Vol.5, No.21, 1-45
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A RAPD-based identification system for calla lily cultivars (
Zuntedeschia
spp.) was constructed after screening 60
arbitrary IO-mer primers. Using this method, two or three sequential PCR reactions enabled clear identification of
12 cultivars within several days (Hamada and Hagimori, 1996).
In
Chrysanthemum
, cultivars are identified in flowering trials, and breeders’ rights are presented by cultivar
characteristics including flower, leaf and growth morphology. The application of isozyme technology could
largely improve the identification of
Chrysanthemum
cultivars (Roxas et al., 1993). However, the level of
polymorphism obtained is often insufficient to distinguish cultivars, and the growth conditions may influence the
quality and quantity of isozymes (Wolff et al., 2005).
Chrysanthemum
cultivars are propagated vegetatively by
cuttings. The cultivars that are propagated vegetatively must have the same DNA pattern, even after many years of
cultivation (Wolff et al., 2005). Morphological, cytological, anatomical, biochemical and RAPD work were
carried out extensively by the authors on original chrysanthemum varieties and new varieties developed through
conventional breeding and gamma ray induced mutation (Chatterjee et al., 2005, 2006a, 2006b; Datta, 1987, 1993;
Datta and Chatterjee, 2006; Datta and Datta, 1998; Datta and Gupta, 1981a, 1981b, 1983; Datta and Shome, 1994;
Gupta and Datta, 2005) and on rose (Datta and Gupta, 1983; Datta and Singh, 1995, 1999, 2001, 2002, 2003, 2004,
2005, 2006; Datta, 1997).
Genetic variation of 18 chrysanthemum cultivars was first studied by Wolff and Peter-Van Rijn (1993) in 1993
using RAPD. It is known that different conditions of the RAPD reaction (thermal cycler brand, annealing
temperature, primer, enzyme brand, magnesium concentration, pH, gelatin concentration etc.) influence the results.
Wolff et al. (1993)made extensive studies of these factors on chrysanthemum and standardized the optimum
combination of each factor to obtain reproducible patterns. Wolff et al. (1994) constructed the Pst I and Hind III
genomic library for
Dendranthema grandiflora
Tzvelev. Probes from both libraries were tested for the presence or
absence of restriction fragment length polymorphism (RFLP). The RFLP probes and primers development was
very useful for further marker assisted selection in this polyploidy crop. Scott et al. (1996) used DNA
amplification fingerprinting to study genetic relationships between 21 closely related chrysanthemum cultivars
using 11 arbitrary octamer primers. Series of new cultivars have developed in Chrysanthemum through sporting is
a well-known phenomenon. In chrysanthemum there are three different cell layers that form different parts of the
plant. The samples were taken from the leaf (layers I, II and III), from florets (layers I and 11), and the epidermis
of a leaf (layer I) differences at the DNA level among cultivars within one family and among the layers of one
cultivar were studied by RAPD analysis. Polymorphisms between cultivars as well as between different cell layers
have been observed (Wolff, 1996). Trigiano et al. (1998) studied the relation between induced mutants of
chrysanthemum ‘Charm’ family. The cultivars differ only in flower colour and were very difficult to distinguish
by DAF but could be easily distinguished by ASAP (Arbitrary Signatures from Amplification Profiles). Genetic
analysis of three hybrid combination of chrysanthemum was done by Huang et al. (2000). Forty five random
primers were screened, of which twenty two primers were selected to detect the molecular marker in three hybrids
combination of chrysanthemum by using RAPD. Jackson et al. (2000) studied the microsatellite profiling for DUS
testing in chrysanthemum. DNA fingerprinting techniques based on PCR amplification of microsatellite and Copia
like sequences were investigated for their utility in the chrysanthemum. Lema-Ruminska et al. (2004)
characterized 10 radiomutants of chrysanthemum using RAPD. They studied and confirmed the utility of RAPD
markers to identify chrysanthemum cultivars as well as to distinguish the radiomutants from the parents.
Somaclonal variation of chrysanthemum was detected by RAPD markers (Martin et al., 2002). Huang et al. (2000)
used RAPDs to detect different molecular markers in three hybrid populations of
Chrysanthemum
cultivars. They
classified the patterns of molecular markers into seven types on the basis of band sharing between parents and
hybrids. Kumar et al. (2006)applied RAPD technique to characterize eleven gamma ray induced radiomutants of
two chrysanthemum cultivars.Their study revealed that RAPD molecular markers can be used to assess
polymorphism among the radiomutants and can be a useful tool to supplement the distinctness, uniformity and
stability analysis for plant variety protection in future. Minano et al. (2009) used RAPD analysis for molecular
characterization of eight cut flowers and two pot plant cultivars of chrysanthemum. The aim of this study was to
analyze the genetic stability of micropropagated chrysanthemum shoots derived from selected commercial
