In Vitro Micropropagation and DNA Fingerprinting of Sugarcane (Saccharum officinarum L.) Varieties

Abstract

Sugarcane (Saccharum officinarum) has primary importance among commercial crop in India. It is one of the important industrial crops for sugar production grown under both tropical and subtropical conditions. It is relatively planted both as autumn and spring season crop. This crop remain in the field for 2-3 years as a plant which is a high time consuming process can be because of a large blockage in effective breading program. Plant tissue culture is an assemblage of procedures employed to sustain or culture plant groups, tissues under sterilized situations in a nutrient medium. Today, the technique of plant tissue culture has become a potent tool to study, solve basic problems and applied in plant biotechnology conducted initial trials to regenerate plants through in vitro techniques. The growth of tissue culture technology for the rapid generation of disease- free planting material has been an important step towards sufficient seed production, faithfully to the character and quality of sugarcane. Thus the application of plant tissue culture techniques provides an alternative method for the propagation and improvement of sugarcane. Plant tissue culture offers the best methodology through the micropropagation of sugarcane for planting quality material in a short period of time. Therefore it is important to focus on in-vitro culture of the plant part at an early stage for maintaining true-to-type of germplasm for future use. The present thesis focus on two research aims by establishing protocols for in-vitro micropropagation in three sugarcane genotypes viz., Co-0238, Cose-3234, Co-8272 and their DNA fingerprinting with some other sugarcane varieties. The study includes the treatment of combination of sterilants in following sequence (2.0% Labolene for 10 min followed by 0.1% Bavistin for 5 min, 0.1% HgCl2 for 5 min and 70.0% Ethanol for 45 sec) was found to be very effective for the surface sterilization. All the three genotypes showed maximum shoot initiation on MS media supplemented with BAP+ Kinetin (0.5 mg/L each). The MS media consisting of BAP +Kinetin (2.0 mg/Leach) was best for obtaining high frequency shoot multiplication in all the three genotypes. The second objective was carried out to analyse the genetic diversity of 12 sugarcane varieties which was found to be polymorphic in nature when amplified with 5 primers of SSR sequence and the Primer named UGSuM2 shows maximum number of polymorphic band and Highest PIC value was shown by UGSuM74 i.e.,0.8205. So the findings reported herein could be used for large scale commercial production of the sugarcane plant in a very less time space and efforts.