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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