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Salient
findings:
Extensive
DNA typing analysis was carried out for a set of 48 selected elite
genotyepes using many types of DNA markers namely , RAPD, ISSR, f-AFLP
and in-house developed novel mulberry specific microsatellite markers.
All the genotypes were also analysed for nucleotide diversity across
nuclear as well as organelle genomic domains to obtain more reliable
information on their systemic/taxonomic status.
- DNA
fingerprinting of varieties is completed with 28 RAPD primers and
9 AFLP primer combiniations. Out of around 350 RAPD markers 80%
were polymorphic , RAPD allowed scoring of around four thousand
bands for 22 genotypes with an average of 174.4 bands per genotype
.
- AFLP
revealed less polymorphism with in varieties (47.6%), out of 535
markers scored. Around eight thousand bands were scored in 22 genotypes
with an average of 407.7 bands per genotype using 9 primer combinations.
- 17
RAPD primers and 9 AFLP primer combinations were used for 26 selected
mulberry species. out of 457 RAPD markers, 453 were found to be
polymorphic (99%) where as in case of AFLP only 86.1% of total scorable
markers were polymorphic.
- All
the genotypes were also typed using 6 newly developed microsatellite
markers.
- The
RAPD , AFLP and microsatellite data were analyzed to derive genetic
affinities among varieties and species,and all the marker types
could resolve the relationships among the samples anlyzed. more
importantly,the results obtained with all the three approaches were
in good support of each other.
- More
than 110 Kb sequence data were genrated by sequencing ITS1-5SrDNA-ITS2
nuclear domain, 16s rDNA domain of mitochondrial genome, and intergenic
trn and IGS domain of the chloroplast genome.
- The
sequence data showed very little variation across all the varieties
and species that in comparision showed reasonbly high variation
with RAPD,AFLP and microsatellites. Except for few very well established
wild species like Paper mulberry, Serrata and Black mulberry the
sequence analysis could not resolve relative genetic affinities
among various varities and species.
| The
germplasm characterization studies were carried out using various
marker systems such as RAPD, SSR, AFLP and further extended directly
at the level of nucleotide variations by sequencing the phylogenetically
informative conserved genomic domains such as, internal transcribed
spacer regions ITS1-5SrDNA-ITS2 region of the nuclear ribosomal
DNA, inter genic spacer sequence of 16s rDNA and 'trn' region
of chloroplast. |
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A
representative RAPD profile of mulberry varieties and species
showing the nature of variation
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A
representative AFLP profile of mulberry varieties and species
showing the nature of variation
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A
representativeSSR profile of mulberry varieties and species
showing the nature of variation
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