Epidemiological Studies and Molecular Characterization of Phytoplasma Associated With Sugarcane

Abstract

Gladiolus (Gladiolus hybridus Hort.) belongs to the family Iridaceae is one of the most important bulbous commercial ornamental plant grown for cut flowers all over the world as well as in India. In this study, the effect of various concentration of plant growth regulators (NAA and 2,4-D for in-vitro callus induction of gladiolus were used with shoot bud of cormels as explants. The higest rate of 76% explants for callus induction was observed on MS medium supplemented with 4mg/l NAA. In-vitro genetic transformation of Gladiolus cultivar White prosperity was achieved using callus. The calli were co-cultivated with Agrobacterium strain EHA-105 (pCAMBIA 1301), in the presence of 100μM of acetosyringone. The calli were found GUS positive which were grown in SIM for shoot induction. The calli were screened by PCR also for presence of selectable marker in the transformed callus. A method of transgenic development called “In-planta” transformation method, where Agrobacterium is used to infect the plantlets but the step of in-vitro regeneration of plants is completely avoided. In this step we have reported a simple in-planta method for efficient transformation of gladiolus cv. White Prosperity using Agrobacterium tumefaciens EHA-105 harbouring binary vector plasmid pCAMBIA 1302 whose T-DNA contains the GFP gene as well as hygromycin phosphotransferase (hpt) gene under the control of cauliflower mosaic virus 35S constitutive promoter. The culture is injected with 1ml niddle in cormels and infect the meristematic tissue. The transgenic event obtained, were not positive in further PCR screening using gene specific 35S primer.