DIVERSITY ANALYSIS IN SUGARCANE USING SOLUBLE PROTEIN PROFILES, ISOZYME PATTERNS AND RAPD MARKERS
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Date
2004
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Mahatma Phule Krishi Vidyapeeth, Rahuri.
Abstract
The genus Saccharum comprises of three cultivated
species S. officinarwn, S. barberi and S. sinense and two wild
species S. spontaneum and S. robustum. The present investigation
entitled "Diversity analysis in sugarcane using soluble protein
profiles, isozyme patterns and RAPD markers" was undertaken
with a view to analyse the diversity within cultivated varieties and
among the four species. The genetic relationship among Saccharum
species and cultivated varieties was assessed using soluble
proteins, two isozyme systems and fourteen RAPD markers. The
electrophoretic spectra of soluble proteins from 11 species, when
resolved on 10 per cent polyacrylamide gel, exhibited seventy one
bands of which sixteen were distinct polypetide bands with the relative mobility ranging from 0.1 to 0.9 and exhibiting both the
uniformity as well as diversity of pattern between species and
varieties. The similarity indices (SI), indicating evolutionary affinity
between the species, ranged from 0.15 to 0.83. Comparison of the
similarity indices between the cultivated varieties showed
significant differences with a minimum similarity index value of
0.33 between CO-95020 and CO-86032 and a maximum similarity
index value of 0.80 between CO-740 and CO-86032. Variability
among Saccharwn species and cultivated varieties was also
examined which revealed detectable variation in the number and
frequency of isoenzymes. Peroxidase diversity among seven
commercial hybrids quantified in terms of similarity indices showed
variation, which ranged from 0.20 between CO-86032 and CO
94012 to 0.66 between in CO-95020 and CO-95008. The SI values
when compared between species showed variation from 0.20
between S. officincurum and S. spontaneum to 0.66 between S.
offtcinarum and S. sinense. Among the seven cultivated varieties, a
late maturing variety, CO-740 exhibited only one band, whereas
the early maturing variety, CO-94012 showed 5 distinct bands for
esterase. RAPD analysis exhibited a primer specific amplification
profile. A minimum of 25 amplicons with the OPE-09 primer across
7 cultivated varieties of sugarcane and a maximum of 67
amplification bands with OPE-03 primer were observed. Fourteen
primers showing amplification with almost all the eleven species
were tested and the reproducibility confirmed. The average
similarity index between species, based on average of 14 primers, ranged from 0.52 to 0.82. A binary matrix of all the bands present
in each species was generated and the pairwise genetic similarities
between the species under study were determined using Win Boot
computational analysis and the dendogram was constructed. The
consensus tree constructed of varieties clearly showed three
distinct groups. The late and midlate maturing varieties, CO-740,
CO-86032 and CO-8014, formed a separate cluster; whereas three
early maturing and high sugared varieties showed another cluster.
Two varieties viz., CoC-671 and CO-94012 with common
percentage (Q-63 x CO-775) were grouped under one cluster. The
RAPD results in comparison with the protein electrophoresis
revealed more discriminatory power as evident from polymorphic
amplification profile among species. The study demonstrated
usefulness of both the soluble protein electrophoresis and RAPD
analysis for diversity studies. Use of more isozyme system (> 5) and
more number of random primers further needs to be attempted.
Use of more isozymes, RAPD markers and varieties belonging to
distinct groups economic characters will help us to know the extent
of diversity and identification of trait specific markers.
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