SOMACLONAL VARIATION IN SUGARCANE THROUGH TISSUE CULTURE AND SUBSEQUENT SCREENING FOR MOLECULAR POLYMORPHISMS
Abstract
Sugarcane plants of cv. Ph8013, G98/28, G98/24 and GT54-C9 were used for somaclonal variation induction using the spindle leaves as ex-plants. The used varieties differed according to the vegetative traits, juice quality and their similarity index. The effect of five concentrations from 2,4-D and three concentrations of coconut water were examined for the somatic embryogenesis induction. The 2, 4-D was applied at 0, 1, 2, 3, 4 and 5 mg/l in combination with 0, 5 and 10 g/l coconut water. Also, the effect of using NAA at five concentrations i.e. 0, 1, 3, 5 and 7 mg/l in combination with five concentrations of sucrose i.e. 20, 30, 40 50 and 60 g/l were examined to obtain the highest root and shoot elongation. The obtained results indicated the different responses of studied varieties with the different treatments of 2,4-D and coconut water. The varieties gave the highest callus induction percentage and somatic embryogenesis callus induction percentage with different concentrations of 2, 4-D in range between 2 - 4 mg/l. Lowering 2, 4-D to 1 mg/l enhanced the embryos development. On the other hand, all the varieties responded to 7 mg/l NAA but differ with the sucrose concentrations, they ranged between 40, 50 and 60 g/l. Molecular genetic studies were done to assess the somaclonal variation induced through tissue culture compared to the parental varieties. Five out of six primers succeeded in amplifying DNA fragments. Tissue culture was thus responsible for the generation of new variability, since an increase in the rate of molecular polymorphism was observed. Twenty eight out of 46 loci were found to be polymorphic in the regenerated genotypes. Similarity index (as percentage) based on RAPD-PCR analysis using UPGMA computer analysis between the regenerated varieties and their parental lines Phil. 8013, G98-28 and G98-24 were 84%, 85% and 88% respectively, which indicated the induction of mutation via somaclonal variation through tissue culture. The consensus tree was developed based on the banding patterns of the four varieties and their three regenerated genotypes using SPSS statistical analysis program to study the genetic relationships among these genotypes at the molecular level. The results confirmed that in vitro culture of the sugarcane is very efficient and has tremendous potential for producing novel and useful varieties.References
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