Identification of RAPD Marker Linked to Mungbean Yellow Mosaic Virus Resistance in French Bean (Phaseolus vulgaris L.)

Mungbean yellow mosaic virus (MYMV) causes yield loss up to 80 % and is becoming problematic in French bean growing areas. Molecular marker linked selection to MYMV resistance is helpful in rapid identification of genotypes carrying resistant genes. Hence, the present study was undertaken to identify the RAPD marker associated with MYMV resistance in French bean (Phaseolus vulgaris L.). Bulk segregant analysis (BSA) was used to identify RAPD marker linked to MYMV resistance. More than 140 random decamers were surveyed for identification of polymorphic markers between the DNA bulks of resistant and susceptible F2 individuals and their contrasting parents. Ninety eight per cent of these primers amplified DNA in both parents and bulks. Twenty two primers produced specific bands for resistant parent which was absent in the susceptible parent. Out of 22 primers, four primers produced specific fragments viz., OPG 13458, OPX 5670, OPW 17380 and OPP 07730, respectively in resistant parent and bulk, which were absent in susceptible parent and bulk. Amplification of individual DNA samples of segregating F2 resistant individuals using putative marker, OPP 07730, a decamer revealed polymorphism in all four resistant and four susceptible F2 segregants, indicating that the marker OPP 07730 was associated with MYMV resistance in IC-525260, a resistant genotype

The clinical relevance of oliguria in the critically ill patient : Analysis of a large observational database

Funding Information: Marc Leone reports receiving consulting fees from Amomed and Aguettant; lecture fees from MSD, Pfizer, Octapharma, 3 M, Aspen, Orion; travel support from LFB; and grant support from PHRC IR and his institution. JLV is the Editor-in-Chief of Critical Care. The other authors declare that they have no relevant financial interests. Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: Urine output is widely used as one of the criteria for the diagnosis and staging of acute renal failure, but few studies have specifically assessed the role of oliguria as a marker of acute renal failure or outcomes in general intensive care unit (ICU) patients. Using a large multinational database, we therefore evaluated the occurrence of oliguria (defined as a urine output 16 years) patients in the ICON audit who had a urine output measurement on the day of admission were included. To investigate the association between oliguria and mortality, we used a multilevel analysis. Results: Of the 8292 patients included, 2050 (24.7%) were oliguric during the first 24 h of admission. Patients with oliguria on admission who had at least one additional 24-h urine output recorded during their ICU stay (n = 1349) were divided into three groups: transient - oliguria resolved within 48 h after the admission day (n = 390 [28.9%]), prolonged - oliguria resolved > 48 h after the admission day (n = 141 [10.5%]), and permanent - oliguria persisting for the whole ICU stay or again present at the end of the ICU stay (n = 818 [60.6%]). ICU and hospital mortality rates were higher in patients with oliguria than in those without, except for patients with transient oliguria who had significantly lower mortality rates than non-oliguric patients. In multilevel analysis, the need for RRT was associated with a significantly higher risk of death (OR = 1.51 [95% CI 1.19-1.91], p = 0.001), but the presence of oliguria on admission was not (OR = 1.14 [95% CI 0.97-1.34], p = 0.103). Conclusions: Oliguria is common in ICU patients and may have a relatively benign nature if only transient. The duration of oliguria and need for RRT are associated with worse outcome.publishersversionPeer reviewe

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Not AvailablePapaya, a major fruit crop in India and worldwide, is affected by many fungal and viral diseases. A mixed infection of Papaya ring spot virus (PRSV), a linear single-stranded ( + ) RNA genome of approx 10 kb size, and Papaya leaf curl virus (PaLCV), a bipartite Gemini virus (component A & B) having circular single-stranded DNA ( + ) genome of about 5.2 kb, has hampered the production and productivity of papaya in many parts of world. Rapid detection techniques are important in prevention of spread of the disease in field conditions. In the present study, a rapid and reliable PCR based detection protocol has been standardized. Sets of primers were designed, based on the respective virus isolate sequence data available in GenBank, to obtain anticipated products of calculated size.Not Availabl

Duplex PCR to detect both Papaya ring spot virus and Papaya leaf curl virus simultaneously from naturally infected papaya (<i style="mso-bidi-font-style:normal">Carica papaya</i> L.)

269-272Papaya, a major fruit crop in India and worldwide, is affected by many fungal and viral diseases. A mixed infection of Papaya ring spot virus (PRSV), a linear single-stranded (+) RNA genome of approx 10 kb size, and Papaya leaf curl virus (PaLCV), a bipartite Gemini virus (component A &amp; B) having circular single-stranded DNA (+) genome of about 5.2 kb, has hampered the production and productivity of papaya in many parts of world. Rapid detection techniques are important in prevention of spread of the disease in field conditions. In the present study, <span style="mso-ansi-language: EN-IN;mso-fareast-language:EN-IN;mso-bidi-font-weight:bold">a rapid and reliable PCR based detection protocol has been standardized. Sets of primers were designed, based on the respective virus isolate sequence data available in GenBank, to obtain anticipated products of calculated size. </span

Identification of mixed infection caused by Badnavirus and CMV in Jasmine (<em>Jasminum multiflorum</em> Roth)

437-439The symptoms of chlorotic ring spots and irregular chlorotic patches were recently observed on leaves of landscape-grown jasmine (Jasminum multiflorum Roth) in southern parts of India. The causal agent was mechanically transmitted from symptomatic leaves of jasmine to Nicotiana glutinosa, Capsicum annum, Solanum lycopersicum, Cucurbita pepo and Cucumis sativus. Leaf dip preparations from virus-infected plants in electron microscopy revealed the presence of isometric particles similar to Cucumber mosaic virus (CMV) and bacilliform virus particles of badnavirus. The virus reacted specifically with IgG for CMV and Banana streak virus (BSV) in direct antigen coating, enzyme-linked immunosorbent assay. No reaction was observed with ilar-, poty- and tospo-viruses specific IgG. Reverse transcription polymerase chain reaction with total RNA isolated from symptomatic jasmine leaves and infected N. tabacum 'Xanthi' using CMV coat protein (CMV CP) specific primers amplified the expected product. The association of badna virus with jasmine was confirmed by PCR amplification, cloning and sequencing of 560 bp amplicon corresponding to the reverse transcriptase and ribonuclease H coding region in open reading frame III. The sequence analysis revealed maximum identity to badna virus group Diascorea bacilliform virus (88.5% at nt level) and CMV group IB (96% at nt level). To our knowledge, this is the first report of CMV and badna mixed infection in jasmine in India

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Not AvailableThe symptoms of chlorotic ring spots and irregular chlorotic patches were recently observed on leaves of landscape-grown jasmine (Jasminum multiflorum Roth) in southern parts of India. The causal agent was mechanically transmitted from symptomatic leaves of jasmine to Nicotiana glutinosa, Capsicum annum, Solanum lycopersicum, Cucurbita pepo and Cucumis sativus. Leaf dip preparations from virus-infected plants in electron microscopy revealed the presence of isometric particles similar to Cucumber mosaic virus (CMV) and bacilliform virus particles of badnavirus. The virus reacted specifically with IgG for CMV and Banana streak virus (BSV) in direct antigen coating, enzyme-linked immunosorbent assay. No reaction was observed with ilar-, poty- and tospo-viruses specific IgG. Reverse transcription polymerase chain reaction with total RNA isolated from symptomatic jasmine leaves and infected N. tabacum 'Xanthi' using CMV coat protein (CMV CP) specific primers amplified the expected product. The association of badna virus with jasmine was confirmed by PCR amplification, cloning and sequencing of 560 bp amplicon corresponding to the reverse transcriptase and ribonuclease H coding region in open reading frame III. The sequence analysis revealed maximum identity to badna virus group Diascorea bacilliform virus (88.5% at nt level) and CMV group IB (96% at nt level). To our knowledge, this is the first report of CMV and badna mixed infection in jasmine in India.Not Availabl

Monoclonal antibodies to the recombinant nucleocapsid protein of a groundnut bud necrosis virus infecting tomato in Karnataka and their use in profiling the epitopes of Indian tospovirus isolates

A tospovirus infecting tomato in the fields of Karnataka, India, was propagated in greenhouse-grown Nicotiana benthamiana plants by mechanical inoculation. The viral RNA was extracted from purified virus and used for amplification of N and NSs genes by RT– PCR using appropriate primers. The N and NSs PCR products were cloned into a pRSET vector and sequenced. The N gene of tomato tospovirus showed 98% identity with that of Groundnut bud necrosis virus (GBNV), alternate name Peanut bud necrosis virus (PBNV). Interestingly, though the virus was isolated from tomato plants, it showed only 82% identity with the N gene of GBNV-To isolate from Taiwan. The NSs gene of the virus under study showed 98% identity with GBNV. These results suggest that the tomato tospovirus in Karnataka is a strain of GBNV and is henceforth designated as GBNV-To (K). The N gene was overexpressed in Escherichia coli and the recombinant N protein was purified using Ni–NTA agarose affinity chromatography. The purified protein was used for the generation of poly- and monoclonal antibodies (mAbs). The polyclonal antiserum thus obtained had a dilution end-point >1 : 32,000 and nine unique mAbs were also obtained. These mAbs were used for epitope profiling of the tospovirus isolates from South India and for developing detection methods. The results showed that there are distinct GBNV strains in South India. A simple dot-blot assay was developed for detection of GBNV from infected field samples

Monoclonal antibodies to the recombinant nucleocapsid protein of a groundnut bud necrosis virus infecting tomato in Karnataka and their use in profiling the epitopes of Indian tospovirus isolates

A tospovirus infecting tomato in the fields of Karnataka, India, was propagated in greenhouse-grown Nicotiana benthamiana plants by mechanical inoculation. The viral RNA was extracted from purified virus and used for amplification of N and NSs genes by RT- PCR using appropriate primers. The N and NSs PCR products were cloned into a pRSET vector and sequenced. The N gene of tomato tospovirus showed 98% identity with that of Groundnut bud necrosis virus (GBNV), alternate name Peanut bud necrosis virus (PBNV). Interestingly, though the virus was isolated from tomato plants, it showed only 82% identity with the N gene of GBNV-To isolate from Taiwan. The NSs gene of the virus under study showed 98% identity with GBNV. These results suggest that the tomato tospovirus in Karnataka is a strain of GBNV and is henceforth designated as GBNV-To (K). The N gene was overexpressed in Escherichia coli and the recombinant N protein was purified using Ni-NTA agarose affinity chromatography. The purified protein was used for the generation of poly- and monoclonal antibodies (mAbs). The polyclonal antiserum thus obtained had a dilution end-point > 1: 32,000 and nine unique mAbs were also obtained. These mAbs were used for epitope profiling of the tospovirus isolates from South India and for developing detection methods. The results showed that there are distinct GBNV strains in South India. A simple dot-blot assay was developed for detection of GBNV from infected field samples

Capsicum chlorosis virus (Genus Tospovirus) Infecting Chili Pepper (Capsicum annuum) in India

Chili pepper (Capsicum annuum L.) is one of the most important spice crops in India. In October of 2006, symptoms indicative of tospovirus infection were noticed in several commercial fields of chili pepper near Bangalore in Karnataka State. Chlorotic and necrotic spots and rings on leaves, apical necrosis, and leaf distortion were observed. Disease incidence was more than 20%. Mechanical inoculation with sap extracts from these symptomatic plants showed that the host range and symptomatology of the virus was similar to those described for Capsicum chlorosis virus (CaCV) (1,3). The virus reacted with antisera specific to Groundnut bud necrosis virus (GBNV) and Watermelon silver mottle virus of serogroup IV tospoviruses in antigen-coated plate ELISA. It did not react with antisera specific to Tomato spotted wilt virus, Impatiens necrotic spot virus, or Iris yellow spot virus in double-antibody sandwich-ELISA. Immunosorbent electron microscopy of infected sap using GBNV antiserum revealed the presence of strongly decorated quasi-spherical virus particles. Reverse transcription (RT)-PCR was conducted to further identify the virus. No amplification was observed from extracts of symptomatic plants (n = 10) by RT-PCR using GBNV-specific primers (4), indicating that the diseased chili was not infected with GBNV. However, a DNA fragment of approximately 850 bp was amplified by using primers specific to the nucleocapsid (N) gene of CaCV (CaCF 5′-CTATAGAWGTACTAGGCTTTGAGC-3′ and CaCR 5′-CATGTCTAACGTCAGGCAACTTAC-3′). Direct sequencing of the amplicon (GenBank Accession No. EF625227) revealed a nucleotide sequence identity ranging from 85.5% with isolates from Thailand (GenBank Accession Nos. AY846366, AY647437, and AF134400) and China (GenBank Accession No. DQ355974) to 98.1% with isolates from Australia (GenBank Accession Nos. AY036057 and AY036058). Phylogenetic analysis of the N protein sequences showed that the chili pepper isolate from India formed a cluster with those from Australia. This cluster was distinct from the one formed by the peanut isolates from Thailand and a tomato isolate from India (2). To our knowledge, this is the first report of CaCV infection of chili pepper in India. The potential impact of CaCV on tomato and chili pepper production in India remains to be seen. References: (1) D. Knierim et al. Arch. Virol. 90:377, 2006. (2) S. Kunkalikar et al. Online publication. doi:10.1094/PHP-2007-1204-01-BR. Plant Health Progress, 2007. (3) L. A. Mc Michael et al. Aust. Plant Pathol. 31:231, 2002. (4) K. Umamaheswaran et al. Indian Phytopathol. 56:168, 2003

A Novel Curcumin-Galactomannoside Complex Delivery System Improves Hepatic Function Markers in Chronic Alcoholics: A Double-Blinded, randomized, Placebo-Controlled Study

Considering the recent interest in free (unconjugated) curcuminoids delivery, the present study investigated the efficacy of a novel food-grade free-curcuminoids delivery system (curcumin-galactomannoside complex; CGM) in improving the hepatic function markers (inflammation and oxidative stress) in chronic alcoholics. The double-blinded, placebo-controlled study randomized 48 subjects with elevated serum transaminases and gamma-glutamyl transferase (GGT) levels, who were allocated to two groups (n=24) and to receive either placebo or CGM at (250 mg × 2)/day for 8 weeks. While liver function markers (transaminases and GGT) in the placebo group showed an increase (~ 9.5%), CGM group indicated a significant decrease in transaminases (31%) and GGT (29%) from the baseline levels. The beneficial effect of CGM was also clear from the significant increase (p <0.001) in endogenous antioxidants (GSH, SOD, and GPx) and decrease in inflammatory markers (IL-6 and CRP) levels (p <0.001) as compared to both the baseline and placebo group. To summarize, the nutritional intervention of CGM-curcumin was found to offer a significant hepatoprotective effect to attenuate the alcohol induced alterations to hepatic function markers. The Indian Medical Council and Drug Controller General of India approved Clinical Trial Registry No. CTRI/2018/03/012385