Evaluation of Tissue Culture Seedlings for Their Genetic Fidelity and Virus Indexing in Sugarcane

Abstract

Plant tissue culture technique is a powerful tool for rapid and large scale multiplication of virus free seed material. Clonal fidelity of the in vitro raised plants was carried out using RAPD. A total of fifteen primers were used for detecting polymorphism by RAPD analysis and the results revealed that all the tissue culture plants were grouped into one cluster and indicated that all the tissue culture developed plantlets are true-to-type. There are no somaclonal variations and are genetically identical with the mother plant. Virus indexing through ELISA indicated that the tissue culture developed seedlings does not contain the protein particles of ScMV and are free from the viral disease. The wells coated with leaf samples with suspected ScMV infection (97 A 85, C3) gave visual confirmation by yellow colour change, with OD 405 values of 0.526. As the reading is 2 times more than the negative control (OD 0.215) we can presume mild infection of mosaic in the test sample with lower titre of ScMV. All the ScMV +ve controls showed significant higher OD values of more than 2.515 confirming the earlier observations. Thus, viral detection using DAS-ELISA method can be used for detection and indexation in large scale sugarcane crop especially for in vitro regenerated plants with relatively cheaper cost and faster for supplying virus free seedlings to farming community in a large scale.