Biological Control of F. Oxysporum F. Sp. Lycopersici Causing Wilt of Tomato by Pseudomonas Fluorescens

Abstract- Pseudomonas fluorescens is one of the major fungal biocontrol agents found in the soil and the rhizosphere of various crop systems. Ten isolates of P.fluorescens were isolated from rhizosphere soil samples collected from various tomato-growing fields and evaluated for their efficacy in increasing seed quality variables of tomato and in inhibiting the mycelial growth of Fusarium oxysporum. Pseudomonas isolate 2 produced effective results and was selected and mass multiplied. Talc and sodium alginate formulations of mass multiplied using different agents were prepared and evaluated for their effects against fusarium wilt under greenhouse conditions. Fresh cultures of Pf2 isolate was found to increase seedling emergence and reduce fusarium wilt disease incidence when compared to the control and the formulations

Establishing inoculum threshold levels for Bean common mosaic virus strain blackeye cowpea mosaic infection in cowpea seed

Bean common mosaic virus strain blackeye cowpea mosaic (BCMV-BlCM) is an important seed-borne virus infecting cowpea and is transmitted both by seeds and aphids. Infected cowpea seeds can act as primary source of inoculum for disease epidemics. Four field experiments were conducted during 2003 - 2006 to assess the role of different amounts of seed-borne inoculum in the dissemination of BCMVBlCM virus in cowpea under field conditions. The identity of BCMV-BlCM was confirmed by ELISA and IC-RT-PCR. Plants infected at an early growth stage appeared to serve as the primary source for subsequent virus spread by aphids. The mean disease incidence during four field experiments reached88-93% in plots sown with 10% infected seed. The disease incidence in plots sown with 5% infected seed recorded 46-63% while for plants raised from 3 and 2% BCMV-BlCM seed infection, disease incidence reached 32-49% and 17-23%, respectively. Mean yield losses in terms of seed yield per plant from four field experiments were 74 and 54% for initial seed infection of 10 and 5%, respectively. Seed infection of 2% BCMV-BlCM incidence resulted in an average of 24% mean seed yield loss/plant-1. The infection appeared to decrease the seed yield in terms of number and size. The BCMV incidence in harvested seed ranged from 0.3 - 19% for the different levels of initial seed infection. The field experiments demonstrated that sowing > 1% BCMV-BlCM infected seed can lead to significant losses in grain yield, while the spread of BCMV-BlCM infection resulting from sowing 1% infected seed did not significantly decrease seed yield. The role of establishing damage or inoculum thresholds from BCMVBlCM seed-borne infections is discussed in the present study.Keywords: Cowpea, potyvirus, seed-borne virus, thresholds, yield los

Biochemical characterization of Fusarium oxysporum f. sp. cubense isolates from India

The Fusarium wilt caused by Fusarium oxyspoum f. sp. cubense (Foc) is a major biotic constraint for banana production. The characteristics of F. oxyspoum f. sp. cubense isolates were investigated using electrophoretic studies of isozyme and whole-cell protein. The morphological characteristics of the isolates were very similar to each other. All the Foc isolates were pathogenic to banana cultivar 'Nanjangud Rasabale' but they did not induce any disease symptoms on cultivar 'Cavendish'. F. oxyspoum (Isolate 6) did not induce wilt symptoms on either 'Nanjangud' or 'Cavendish' cultivar. Isozyme banding patterns showed 46 scoreable markers and cluster analysis with UPGMA using genetic distance showed that the isolates belonged to three main groups. Group 1 contained isolates 1, 2, 4, 5, 7 and isolate 3 and 6 were placed in group 2 and 3. Results indicated that the estimated intra-specific variation may be more pronounced with isozyme analysis than with protein markers. The level of isozyme variability detected within F. oxysporum f.sp. cubense suggested that it is reliable, efficient and effective in determining genetic relationships among Foc isolates

Fusarium oxysporum f. sp. lycopersici causal agent of vascular wilt disease of tomato: Biology to diversity– A review

Tomato (Lycopersicon esculentum) is one of the widely grown vegetables worldwide. Fusarium oxysporum f. sp. lycopersici (FOL) is the significant contributory pathogen of tomato vascular wilt. The initial symptoms of the disease appear in the lower leaves gradually, trail by wilting of the plants. It has been reported that FOL penetrates the tomato plant, colonizing and leaving the vascular tissue dark brown, and this discoloration extends to the apex, leading to the plants wilting, collapsing and dying. Therefore, it has been widely accepted that wilting caused by this fungus is the result of a combination of various physiological activities, including the accumulation of fungal mycelia in and around xylem, mycotoxin production, inactivation of host defense, and the production of tyloses; however, wilting symptoms are variable. Therefore, the selection of molecular markers may be a more effective means of screening tomato races. Several studies on the detection of FOL have been carried out and have suggested the potency of the technique for diagnosing FOL. This review focuses on biology and variability of FOL, understanding and presenting a holistic picture of the vascular wilt disease of tomato in relation to disease model, biology, virulence. We conclude that genomic and proteomic approachesare greater tools for identification of informative candidates involved in pathogenicity, which can be considered as one of the approaches in managing the disease

Draft genome sequence of Sclerospora graminicola, the pearl millet downy mildew pathogen:Genome sequence of pearl millet downy mildew pathogen

Sclerospora graminicola pathogen is one of the most important biotic production constraints of pearl millet worldwide. We report a de novo whole genome assembly and analysis of pathotype 1. The draft genome assembly contained 299,901,251 bp with 65,404 genes. Pearl millet [Pennisetum glaucum (L.) R. Br.], is an important crop of the semi-arid and arid regions of the world. It is capable of growing in harsh and marginal environments with highest degree of tolerance to drought and heat among cereals (1). Downy mildew is the most devastating disease of pearl millet caused by Sclerospora graminicola (sacc. Schroet), particularly on genetically uniform hybrids. Estimated annual grain yield loss due to downy mildew is approximately 10?80 % (2-7). Pathotype 1 has been reported to be the highly virulent pathotype of Sclerospora graminicola in India (8). We report a de novo whole genome assembly and analysis of Sclerospora graminicola pathotype 1 from India. A susceptible pearl millet genotype Tift 23D2B1P1-P5 was used for obtaining single-zoospore isolates from the original oosporic sample. The library for whole genome sequencing was prepared according to the instructions by NEB ultra DNA library kit for Illumina (New England Biolabs, USA). The libraries were normalised, pooled and sequenced on Illumina HiSeq 2500 (Illumina Inc., San Diego, CA, USA) platform at 2 x100 bp length. Mate pair (MP) libraries were prepared using the Nextera mate pair library preparation kit (Illumina Inc., USA). 1 ?g of Genomic DNA was subject to tagmentation and was followed by strand displacement. Size selection tagmented/strand displaced DNA was carried out using AmpureXP beads. The libraries were validated using an Agilent Bioanalyser using DNA HS chip. The libraries were normalised, pooled and sequenced on Illumina MiSeq (Illumina Inc., USA) platform at 2 x300 bp length. The whole genome sequencing was performed by sequencing of 7.38 Gb with 73,889,924 paired end reads from paired end library, and 1.15 Gb with 3,851,788 reads from mate pair library generated from Illumina HiSeq2500 and Illumina MiSeq, respectively. The sequences were assembled using various assemblers like ABySS, MaSuRCA, Velvet, SOAPdenovo2, and ALLPATHS-LG. The assembly generated by MaSuRCA (9) algorithm was observed superior over other algorithms and hence used for scaffolding using SSPACE. Assembled draft genome sequence of S. graminicola pathotype 1 was 299,901,251 bp long, with a 47.2 % GC content consisting of 26,786 scaffolds with N50 of 17,909 bp with longest scaffold size of 238,843 bp. The overall coverage was 40X. The draft genome sequence was used for gene prediction using AUGUSTUS. The completeness of the assembly was investigated using CEGMA and revealed 92.74% proteins completely present and 95.56% proteins partially present, while BUSCO fungal dataset indicated 64.9% complete, 12.4% fragmented, 22.7% missing out of 290 BUSCO groups. A total of 52,285 predicted genes were annotated using BLASTX and 38,120 genes were observed with significant BLASTX match. Repetitive element analysis in the assembly revealed 8,196 simple repeats, 1,058 low complexity repeats and 5,562 dinucleotide to hexanucleotide microsatellite repeats.publishersversionPeer reviewe

Resultative Compound Verb in Modern Chinese : A Comment on Imai(1985) and Lu(1986)

<p>A. API and DMO suppresses NF-κB DNA binding ability in HCT116 cells. HCT116 cells were treated with DMO and API at indicated doses, nuclear extracts were prepared, and 20 μg of the nuclear extract protein was used for the ELISA-based DNA-binding assay *p<0.05; **p<0.005). B & C. NF-κB responsive elements linked to a luciferase reporter gene were transfected with wild-type or dominant-negative IκB and transfected cancer cells were treated at indicated doses for 6 h and luciferase activity was measured as described in Materials and Methods section. All luciferase experiments were done in triplicate and repeated twice (*p<0.05; **p<0.005). D. API abrogates constitutive IκBα phosphorylation in dose-dependent manner in HCT116 cells. HCT116 cells were treated with different concentrations of API (0, 5, 10 and 20 μM) for 6 h and cytoplasmic extract was prepared. Lysates were resolved on SDS gel and electrotransferred to a nitrocellulose membrane and probed with anti-phospho-IκBα/IκBα. The blot was washed, exposed to HRP-conjugated secondary antibodies for 1 h, and finally examined by chemiluminescence. GAPDH was used as loading control.</p

Effect of toxigenic Aspergillus flavus and aflatoxins on seed quality parameters of Sorghum bicolor (L.) Moench.

The Aspergillus flavus and aflatoxins are known to be detrimental to plant and animals affecting their productivity and yield. This study evaluated effects of toxigenic A. flavus and aflatoxins on physical parameters like seed germination, seedling vigor, root length, shoot length and also biochemical parameters like chlorophyll content, protein, sugars and amylase activity in sorghum seeds. The sorghum seeds were treated with 100, 250 and 500 μg ml-1 concentrations of aflatoxins and likewise, A. flavus spore suspension adjusted to 1x108 spores ml-1 were also treated to seeds in different treatments. The experimental results revealed maximum inhibition of seed germination, seedling vigor, chlorophyll, proteins, total sugars and α-amylase activity in the sorghum seedlings was observed at 500 µg ml-1 followed by 250 and 100 µg ml-1. But seed treatment with toxigenic A. flavus spore suspension showed slight inhibition all the above parameters tested when compared to untreated control, but there was no significant decrease was observed. The study highlighted negative effects of the A. flavus and aflatoxins on the tested seed quality parameters tested there by necessitating need of monitoring of toxigenic fungi and their metabolites in sorghum seeds