Nested PCR assay for detection of sugarcane grassy shoot phytoplasma in the leafhopper vector Deltocephalus vulgaris: a first report

Abstract

Nymphs of Deltocephalus vulgaris , the leafhopper vector of sugarcane grassy shoot (SCGS) disease, fed on SCGSinfected and healthy sugarcane leaves, and SCGS-infected and healthy plant tissue of sugarcane cv. CoLk 8102, were examined by nested PCR using phytoplasma-specific rRNA operon primers for detection of the SCGS phytoplasma. Samples of SCGS-infected plants with symptoms and SCGS-exposed D. vulgaris nymphs yielded SCGS-exclusive DNA bands when nested PCR was performed. Negative results were obtained when symptomless plant host and unexposed insect vector samples devoid of phytoplasma DNA templates were used. Such a reliable molecular tool for the precise detection of SCGS phytoplasma in the D. vulgaris population would help forecast the potential of secondary spread of SCGS in a susceptible sugarcane variety, and may facilitate control of the disease.

Detection of sugarcane grassy shoot phytoplasma infecting sugarcane in India and its phylogenetic relationships to closely related phytoplasmas

“…Up to 70% or higher incidence of SCGS has been recorded resulting in 100% per cent yield 1 3 loss (Rao and Dhumal, 2002;Rao et al, 2005). The leafhopper Deltocephalus vulgaris Dash and Viraktamath has been reported to transmit SCGS phytoplasma in India (Singh et al, 2002;Srivastava et al, 2006).…”

“…However, inoculation experiments need to be carried out in order to prove definitively the pathological role of low-titer phytoplasma infections, in sugarcane. Although there are a few reports on detection of phytoplasmal infections in SCGS-affected sugarcane plants in India with PCR-based methods (Rao et al, 2003;Viswanathan et al, 2005;Srivastava et al, 2006), the extensive survey of phytoplasmal infections in sugarcane plants collected in several sugarcane-growing areas of India using PCR assays as well as detection of phytoplasmal infections in nonsymptomatic sugarcane plants and/or in sugarcanes with nonspecific symptoms had previously not been reported. Also, the phylogenetic relationships of SCGS phytoplasma strains to each other and to other closely related phytoplasmas, at both 16S rRNA gene and 16S/23S rDNA spacer region sequence levels were examined for the first time in the present work.…”

Detection and Characterization of Phytoplasma and Sugarcane Yellow Leaf Virus Associated with Leaf Yellowing of Sugarcane

“…; Srivastava et al. ). Thus, the 16S rDNA gene and 16S‐23S rDNA intergenic spacer region have been widely used as targets to detect and identify many different types of phytoplasma (Gundersen et al.…”

Disease Scenario and Management of Major Sugarcane Diseases in India

“…Universal phytoplasma-specific primer pairs P1 and P7 were used for nested PCR assays that successfully detect the SCGS phytoplasma in sugarcane and its reported leafhopper vector Deltocephalus vulgaris Srivastava et al 2006). Since the diseased plants exhibit various phenotypic symptoms under field conditions, diagnosis of the disease becomes difficult.…”