Commercial formulations of strobilurins (azoxystrobin, kresoxim-methyl, trifloxystrobin and pyraclostrobin) were evaluated for their efficacy against Bean common mosaic virus (BCMV) in screenhouse and field conditions. Highest seed germination and seedling vigour were recorded with 20 lg/ml pyraclostrobin seed treatment in comparison with the control. In screenhouse studies, 76% protection against BCMV was recorded with pyraclostrobin seed treatment at 10 lg/ ml. Under field conditions with natural BCMV inoculum, pyraclostrobin seed treatment resulted in 65% protection against BCMV. The protection offered by strobilurins against BCMV was evaluated by ELISA, with lowest immunoreactive values recorded in common bean seedlings raised from seeds treated with pyraclostrobin and kresoxim-methyl. Strobilurins in addition to exerting a direct positive physiological effect on common bean plants also protect bean plants against BCMV infection in screen house and field conditions. Thus, it is proposed that these reducedrisk pesticides are potential inducers against BCMV and growth enhancers and could be a beneficial component of integrated disease management of common bean.
In different legume-growing regions of India, a total of 136 seed samples of cowpea (Vigna unguiculata (L.) Walp.) were collected and tested for the presence of the blackeye cowpea mosaic strain of Bean common mosaic virus, Cowpea aphid-borne mosaic virus, Cowpea mosaic virus, Cucumber mosaic virus and Bean yellow mosaic virus using growing-on test, enzyme linked immunosorbent assay, differential host test, electron microscopy and immuno-capture reverse transcription polymerase chain reaction. Among the 136 seedlots tested, 43 cowpea seedlots were found to be infected with BCMV-BlCM. The identity of three of these isolates as BCMV-BlCM was further supported by nucleotide sequencing in the 3′ region of the genome. The incidence of seeds carrying transmissible virus ranged from 0.67% to 13.49%. In most cases, only symptomatic seedlings in a growingon test were found infected with the virus.
Pearl millet (Cenchrus americanus L.) field-grown plants of cv. 7042S shown unusual water-soaked lesions on leaf tips spreading towards the leaf base from Manasagangothri region (12.31°N 76.61°E), Karnataka, a southern Indian state during March 2020. Later those infected plants showed extensive necrosis and typical leaf blight symptoms with 70% disease incidence and 59% severity. Surface sterilized (3 x 3 mm) infected leaf tissues were crushed in 1mL sterile distilled water and streaked onto nutrient agar media. Bright-yellowish, circular, mucoid single bacterial colonies (PPi-M1) with regular margin were recovered after 24 hours of incubation at 28oC, and the same bacterial colonies were used for further biochemical and molecular characterization. The isolate, PPi-M1 found as gram-negative rods, gelatin, starch hydrolysis negative, and catalase, indole production positive. The partial sequence of 16S rRNA gene (primers: 27F/1492R) of the isolate PPi-M1 was amplified, sequenced, and curated sequence submitted to NCBI GenBank (accession number: MN808555). In nucleotide BLAST search for homologous sequences, 99.5% nucleotide matching similarity (1410bp) was observed with other Pantoea stewartii subspecies indologenes strains (MF163274; NR_104928) at NCBI database indicating that our isolate PPi-M1 belongs to this species. In Phylogenetic analysis using the Maximum Likelihood method and Tamura Nei model (1993), PPi-M1 formed a distinct cluster with other Pantoea stewartii strains with bootstrap value >95 and it was distant from P. allii, P. ananatis, P. agglomerans, and P. dispera. Besides, the subspecies-specific PCR assay and subsequent sequencing of galE and recA genes (primers: 3614galE/3614galEc; 3614recA/3614recAc; 372 and 223 bp) also confirmed the identity of the isolate as Pantoea stewartii subspecies indologenes. Further, the pathogenicity test was performed in-planta on 21 days old seedlings of pearl millet cv. CO-10. The bacterial suspension of isolate PPi-M1 (1x108 CFU/ml) was used for inoculation by leaf clipping method (Ke et al. 2017). All the inoculated plants (n=4 leaves per plant; 15 plants) maintained under greenhouse conditions (Temp: 27-29oC; RH: 80-85%) except mock (sterile water inoculation) shown similar water-soaked lesions from the cut end of the leaf, with a definite spreading margin and a typical leaf blight symptom in 8 dpi, as observed in the field. Re-isolated bacterial colonies from infected leaves shared similar morphological characters and molecular identity with inoculated culture, thus proving Koch’s postulates. This pearl millet leaf blight causing bacterial strain PPi-M1 was deposited in the National Agriculturally Important Microbial Culture Collection, Mau, India (accession no.: NAIMCC-B-02508). Previously, P. stewartii was reported to cause leaf blight and rot diseases on rice and maize (Kini et al. 2016; Roper et al. 2011), also the international seed federation has instigated the phytosanitary measures highlighting its true seed transmission ability (Pataky et al. 2003). This study will supplement future pearl millet breeding programs, and to our knowledge, this is the first report of P. s. subsp. indologenes inciting pearl millet leaf blight disease in India.
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