Abstract
Since 2011, chlorosis with defined chlorotic streak symptoms, yellowing, stunting, and drying of infected clumps of 155 durum (Triticum durum L.) and two bread wheat (T. aestivum L.) genotypes (30- to 45-day-old seedlings) were regularly observed at Regional Station of ICAR - Indian Agricultural Research Institute, Indore (Madya Pradesh), India. The disease incidence was greater in durum (1.7 to 20%) compared with bread wheat (1.3 to 10.4%). Ten randomly selected symptomatic leaf samples were analyzed for the presence of virus particles using leaf dip electron microscopy, PCR, and rolling circle amplification, but no virus particles viz. Wheat streak mosaic virus, Barley yellow dwarf virus, Wheat spindle streak mosaic virus, and Soilborne wheat mosaic virus were detected, nor positive serological relationships established with the antibodies of these viruses. To detect the association of phytoplasma with symptomatic wheat samples, genomic DNA from three representative durum wheat genotypes (Is-1: H18638; Is-2: PDW245; Is-3: MACS3063) and one bread wheat genotype (Is-271: HW2004) were assayed for amplification of 16S rRNA gene using primers P1 and P7 followed by nested PCR using primers R16F2n and R16R2 (Gundersen and Lee 1996). The 16S rRNA gene of phytoplasma was detected in all symptomatic samples, and no amplification was observed in asymptomatic wheat samples. The obtained PCR fragments of 1.25 kb were sequenced directly using primers 343R, 704F, 907R, and 1028F. The 1,245-kb sequences were deposited in GenBank (LT594113–16), which showed 100% sequence identity with each other and 98.16% identity with reference phytoplasma strain BGWL-C1 (Candidatus Phytoplasma cynodontis, AJ550984) when analyzed using EzTaxon database. The MEGA6 constructed neighbor joining phylogenetic analysis and virtual RFLP analysis using 17 restriction enzymes of 16SrRNA gene sequences allowed affiliating the wheat phytoplasma strains with 16SrXI-B subgroup strain with a similarity coefficient of 1 (Zhao et al. 2009). The association with the 16SrXI-B group was further established by amplifying
Published Version
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