In vitro regeneration, detection of somaclonal variation and screening for mosaic virus in sugarcane (Saccharum spp.) somaclones

摘要

Three sugarcane accessions susceptible to sugarcane mosaic virus; HSF-240, S-2000-US-359, and S-2003-US-704 were evaluated for callogenesis and regeneration ability. For callogenesis, five?different concentrations of?2,4-dichlorophenoxyacetic acid (2,4-D) was used. The best callogenesis was?obtained when?Murashige and Skoog (MS)?was portified with?3 mg/L 2,4-D?and?the?highest?regeneration wasobtained on?media containing MS + kinetin 0.5 + 0.5 mg/L?naphthalene acetic acid (NAA). After?succesful regeneration?and rooting?on?half strength MS medium, with 1.5 mg/L indole-3-butyric acid supplementation, plantlets?were shifted to green house.Enzyme linked immunosorbent assay (ELISA)?test was performed?to detect the presence of?sugarcane?mosaic virus (SCMV)?in the regenerated plantlets?and?simple sequence repeat (SSR)?markers were used to evaluate the genetic variation at DNA level between?the?parent’s?plants?and?regenerated?somaclones of?the accession?HSF-240. A?total of 26 parent plants and 64 somaclones,?among the regenerated plantswere selected for?the?screening?of?virus through?double antibody sandwich (DAS?ELISA)?test.?Four (4)?parent plants out of the 26, showed negative reaction to the virus?test.?Ten (10)?somaclones showed positive reaction to the disease, 9 somaclones showed mild reaction to virus and 45?somaclones?showed negative reaction. For?the?detection of somaclonal variation, 38 primers pair were used and 15 simple sequence repeats (SSR) primer pairs were found to be polymorphic with 51.61% polymorphism. The study demonstrates?that SSR genetic markers are the best tool for?the?investigation of genetic variation in sugarcane.