Modifying DNA methylation pattern in papaya embryos to harness useful and stable variants

Abstract

The purpose of this study was to investigate the effect of various DNA methylation modifying agents on genome wide global DNA methylation status and to develop papaya cv. Surya plants with useful and stable horticulturally important variations. DNA methylation-status modifying agents (Zebularine, 5-azacytidine, 5-aza deoxycytidine (DNA hypomethylating agents), 3’-azido-deoxythymidine (DNA hypermethylating agent) were used at two concentrations (0.3mM and 0.5mM) on papaya cv. Surya zygotic embryos for incubation periods ranging from 0-120 hrs. The technique of MS-RAPD-PCR which resolves global DNA methylation pattern was employed. A total of 87 primers were tested on the control samples, 55 primers showed PCR amplification, of which 11 showed polymorphic bands. Six primers showing clear and reproducible bands were selected further for analysing methylation-modifying-agent treated samples. This technique was found to be reliable and efficient for detecting changes in DNA methylation at both the scale and pattern in a genome wide sta ndpoint.