Research Article
Jyothi G, Chandrakala A, Cheta
Abstract
Crotalaria is a tropical Asian plant grown as a source of green manure, fodder and fiber. Phyllody disease in crotalaria caused by phytoplasma has been gaining importance in the recent years. Symptoms of phyllody infected plants appear pale green and bushy due to excessive stunting, malformation of floral parts into green leaf like structures and production of excessive spikelets in an upright fashion within inflorescence axis. In the present investigation, Fluorescent microscopy and Polymerase chain reaction (PCR) techniques were standardized for detection of crotalaria phyllody phytoplasma. Fluorescent microscopic technique using 41-61 diamidino-2-phenylindole (DAPI) stain revealed the association of phytoplasma with bright fluorescent spots in the phloem region of infected stem sections, but no such spots were observed in the phloem region of healthy stem sections. Annealing temperature of 55 and 56oC for one minute consistently amplified the phytoplasma specific products of 1800 and 1250 bp using P1/P7 and R16F2n/R16R2 universal primers respectively.