"DIVERSITY ANALYSIS OF SUGARCANE VARIETIES USING SOLUBLE PROTEIN PROFILES, ISOZYME PROFILES AND RAPD MARKERS"

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Date
2007
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Mahatma Phule Krishi Vidyapeeth, Rahuri.
Abstract
The present investigation was undertaken to analyze the genetic diversity in seven sugarcane varieties having different sugar content viz., CoC 671, CoM 0254, Co 94012, MS 202, CoM 9908, Q 63 (high sugar content varieties) and CoM 0251 (low sugar content variety) and nine sugarcane varieties differing in cane yield potential viz., MS 6847, CoM 0261, CoM 0265, Co 740, MS 9906, Co 86032, CoM 0254 (high cane yield potential varieties) and Co 775, MS 0219 (low cane yield potential varieties) using protein profile, isozyme pattern and RAPD analysis. The Native- and SDS-PAGE of soluble proteinsexhibited the polymorphism among the high and low sugar content varieties and high and low cane yield potential varieties of sugarcane,, in respect of presence and absence of bands and differential band intensity. In relation to sugar content, seven protein bands were produced in seven sugarcane varieties when resolved by Native-PAGE, out of which, three were polymorphic. by However^SDS-PAGE, seventeen polypeptideswere resolved out of which which 10 bands were polymorphic. The variety specific band of the size of" -36.03 kDa was produced in high sugar content variety?CoM 0254 when resolved by Native-PAGE, however, with SDS-PAGE of soluble protein, it was revealed that the polypeptide of the size.- 30.85 kDa was present in all the high sugar content varieties and remained absent in a low sugar content variety. Further variety specific unique bands were observed of the size of- 28.04 and 13.37 kDa in CoM 0251, a low sugar content variety with SDS-PAGE of soluble proteins. By Native-PAGE, 10 bands and by SDS-PAGE 14 bands were observed in nine sugarcane cultivars out of these nine and six bands were polymorphic^ respectively. The variety specific unique bands were observed of the size of 40.45 kDa in MS 6847, a high cane yield potential sugarcane variety and 27.13 kDa in Co 775," low cane yield potential variety by Native-PAGE of soluble proteins. The esterase isozyme zymogram showed low polymorphism among the sugarcane varieties having varying cane yield potential, and was unable to detect the polymorphism among the sugarcane varieties having varying sugar content. The RAPD analysis of genomic DNAs by 20 RAPD primers showed polymorphism in seven sugarcane varieties w!#i varying sugar content and nine sugarcane varieties v&Hvarying cane yield potential. The primer OPB 16 produced^highest 75% polymorphism. It was observed that 12 variety-specific RAPD markers were amplified in a low sugar content variety^CoM 0251. However in high sugar content varieties,unique variety specific bands produced were 7 in CoM 9908, 6 in Q 63, 5 in MS 202, 4 in CoC 671, 3 in CoM 0254 and 1 in Co 94012. Further, it was revealed that 9 amplified products of the sizeA1847 bp, 1524 bp, 883 bp (OPB 01), 1456 bp (OPB 03) and 2001 bp (OPB 09) were consistently present only in six high sugar varieties. From the Dice similarity coefficient values and UPGMA based consensus tree generated using Win Boot computational analysis indicated that the low sugar variety was found to be^most divergent and formed an independent cluster. However, high sugar : . ., varieties viz., CoC 671, Co 94012, CoM 0254 formed one cluster and MS 202, Q 63, CoM 9908 . or, ! another cluster. In relation to yield potential, out of nine varieties variety specific RAPD markers were synthesized in eight. In low cane potential varieties,MS 0219 and Co 775, 5 and 2 unique bands were observed,respectively. In high cane yield potential varieties, unique bands produced were 11 in CoM 0254, 5 each in Co 740 and CoM 0261, 3 in MS 6847, 2 in Co 775 and 1 in MS 9906. The Dice similarity coefficient values and consensus tree constructed using Win Boot computational analysis revealed that a high cane yield variety CoM 0254 formed an independent cluster, however, low cane yield varieties Co 775 and MS 0219 formed the cluster with high cane yield variety CoM 0261. The remaining high cane yield varieties viz., MS 6847, CoM 0265, Co 740, formed a cluster and MS 9906^ Co 86032 formetlthe another cluster. Thus from the studies of protein profile, isozyme pattern Hie. and RAPD analysis, it is revealed that.RAPD markersiwejtinformative for detecting the diversity in sugarcane varieties in relation to cane yield and sugar content.
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