H. S. Prakash

Improvement of seed quality and field emergence of Fusarium moniliforme infected sorghum seeds using biological agents

Five different cultivars of sorghum seeds infected with a varied degree of Fusarium moniliforme were treated with biocontrol agents. Pure cultures of Pseudomonas fluorescens, Trichoderma harzianum and Chaetomium globosum at the rate of 1 x 10(8) cfu g(-1) and talcum based formulations of (28 x 10(7) cfu g(-1)), (19 x 10(7) cfu g(-1)) and (4 x 10(6) cfu g(-1)) at the rate of 6 g kg(-1) and 10 g kg(-1) of seeds were used, respectively. The treated seeds were evaluated for per cent reduction of F moniliforme, seed germination, vigour index and field emergence. It was found that the pure culture of P fluorescens was more effective in reducing the F moniliforme infection followed by T harzianum and C globosum than the Bavistin treated and untreated seeds. Formulations of P fluorescens were effective in reducing the F moniliforme infection and also in increasing the seed germination, vigour index and field emergence, followed by T harzianum and C globosum treatments in comparison with control

A network map of FGF-1/FGFR signaling system

Fibroblast growth factor-1 (FGF-1) is a well characterized growth factor among the 22 members of the FGF superfamily in humans. It binds to all the four known FGF receptors and regulates a plethora of functions including cell growth, proliferation, migration, differentiation, and survival in different cell types. FGF-1 is involved in the regulation of diverse physiological processes such as development, angiogenesis, wound healing, adipogenesis, and neurogenesis. Deregulation of FGF-1 signaling is not only implicated in tumorigenesis but also is associated with tumor invasion and metastasis. Given the biomedical significance of FGFs and the fact that individual FGFs have different roles in diverse physiological processes, the analysis of signaling pathways induced by the binding of specific FGFs to their cognate receptors demands more focused efforts. Currently, there are no resources in the public domain that facilitate the analysis of signaling pathways induced by individual FGFs in the FGF/FGFR signaling system. Towards this, we have developed a resource of signaling reactions triggered by FGF-1/FGFR system in various cell types/tissues. The pathway data and the reaction map are made available for download in different community standard data exchange formats through NetPath and NetSlim signaling pathway resources.

Zearalenone induced toxicity in SHSY-5Y cells: The role of oxidative stress evidenced by N-acetyl cysteine.

Zearalenone (ZEN) is a mycotoxin from Fusarium species commonly found in many food commodities and are known to cause reproductive disorders, genotoxic and immunosuppressive effects. Although many studies have demonstrated the cytotoxic effects of ZEN, the mechanisms by which ZEN mediates its cytotoxic effects appear to differ according to cell type and route of exposure. Meantime, the available information on the neurotoxic effects of ZEN is very much limited. In the present study we evaluated the role of oxidative stress in ZEN mediated neurotoxicity in SH-SY5Y cells and investigated the possible underlying mechanism. ZEN induced ROS formation and elevated levels of MDA, loss of mitochondrial membrane potential (MMP) and increase in DNA damage in a dose dependent manner as assessed by COMET assay and agarose gel electrophoresis. However, there was no DNA damage by plasmid breakage assay at 6, 12 and 24h time points. DAPI staining showed apoptotic nuclei at 12 and 24h. Further, ZEN treated SH-SY5Y cells showed a marked suppressive effect on the neuronal gene expression. Use of an antioxidant N-acetylcysteine (NAC) reversed the toxin-induced generation of ROS and also attenuated loss of MMP. Collectively, these results suggest that ROS is the main upstream signal leading to increased ZEN mediated neurotoxicity in SH-SY5Y cells.

Beauveria bassiana - A potential mycopesticide for the efficient control of coffee berry borer, Hypothenemus hampei (Ferrari) in India

The Coffee Berry Borer (CBB), Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) has been a serious insect pest of coffee cultivars C. robusta and C. catimor in India since 1991, causing 40-80% coffee bean loss. To combat this important pest, an indigenous entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin was isolated from dead and moribund coffee berry borers from the wild. The fungus was cultured on yeast extractpeptone supplemented liquid medium. The mycelial mat was harvested from 12-day old cultures and lyophilized. A suspension of the most virulent isolate (Bb2) was prepared in sterile water and used as a mycopesticide. The laboratory studies were conducted on coffee berry borers by applying conidial suspensions at a dosage rate of 1 2 10 6 conidia ml -1 . Pest mortality with the Bb2 isolate increased from 69.3% to 95.3% with an increase in relative humidity (RH) at 25 - 2°C. Field experiments were conducted in a coffee plantation area in the Kodagu district of Karnataka, and the results showed that, under favourable environmental conditions (27- 29°C; 82-91% RH; 10-15 inches rainfall per year), the fungus required only eight days to colonize and kill the target pest. A maximum of 75.6% insect mortality was recorded 24 days after spraying. Large-scale field trials conducted in five plantation plots between September 1995 and September 1998 showed significant insect mortality both in C. robusta and C. catimor cultivars of coffee. The potential use of this indigenous fungal strain of B. bassiana as a mycopesticide for management of CBB in India is discussed.

Rosmarinic acid mediated neuroprotective effects against H2O2-induced neuronal cell damage in N2A cells.

AIMS Oxidative stress plays a key role in several ailments including neurodegenerative conditions. The aim of the study was to demonstrate the effect of rosmarinic acid (RA) in preventing oxidative stress related death of neuronal cell lines. MAIN METHODS In the present study, we demonstrated direct neuroprotective effect of RA using H2O2-induced oxidative challenge in N2A mouse neuroblastoma cells. The mechanism of neutralization of H2O2-induced toxicity by RA was evaluated using MTT, lactate dehydrogenase, mitochondrial membrane potential (MMP), intracellular ROS, and comet assays. Up-regulation of brain neuronal markers at molecular level was performed by RT-PCR. KEY FINDINGS Results presented in the paper indicate that H2O2-induced cytotoxicity in N2A cells was suppressed by treatment with RA. Moreover, RA is very effective in attenuating the disruption of lactate dehydrogenase, mitochondrial membrane potential and intracellular ROS. Pretreatment with RA significantly prevents genotoxicity (3.7-fold, p<0.01) and promotes the up-regulation of tyrosine hydroxylase (TH) (4.5-fold, p<0.01), and brain-derived neurotrophic factor (BDNF) genes (5.4-fold, p<0.01) against H2O2-induced cytotoxicity in N2A cells. SIGNIFICANCE Our results revealed that N2A cells are suitable cellular models to evaluate neuroprotective effects of RA, and suggest that RA may potentially serve as an agent for prevention of several human neurodegenerative diseases caused by oxidative stress.

Prospects of molecular markers in Fusarium species diversity

Recent developments in genomics have opened up for newer opportunities to study the diversity and classification of fungi. The genus Fusarium contains many plant pathogens that attack diverse agricultural crops. Fusarium spp. are not only pathogenic to plants but are also known as toxin producers that negatively affect animal and human health. The identification of Fusarium species still remains one of the most critical issues in fungal taxonomy, given that the number of species recognized in the genus has been constantly changing in the last century due to the different taxonomic systems. This review focuses of various molecular-based techniques employed to study the diversity of Fusarium species causing diseases in major food crops. An introduction of fusarial diseases and their mycotoxins and molecular-marker-based methods for detection introduce the concept of marker application. Various well-known molecular techniques such as random amplified polymorphic DNA, amplification fragment length polymorphism, etc. to more modern ones such as DNA microarrays, DNA barcoding, and pyrosequencing and their application form the core of the review. Target regions in the genome which can be potential candidates for generation of probes and their use in phylogeny of Fusarium spp. are also presented. The concluding part emphasizes the value of molecular markers for assessing genetic variability and reveals that molecular tools are indispensable for providing information not only of one Fusarium species but on whole fungal community. This will be of extreme value for diagnosticians and researchers concerned with fungal biology, ecology, and genetics.

Detection of Tobacco mosaic virus and Tomato mosaic virus in pepper and tomato by multiplex RT–PCR

Aims:  To develop a highly sensitive and rapid protocol for simultaneous detection and differentiation of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in pepper and tomato. In this study, we use the multiplex PCR technique to detect dual infection of these two viruses.

Seed biopriming with novel strain of Trichoderma harzianum for the control of toxigenic Fusarium verticillioides and fumonisins in maize

Fusarium verticillioides is one of the most important fungal pathogens in maize causing both pre- and post-harvest losses and also capable of producing Fumonisins. In the present study attempts have been made for screening potential T. harzianum from native rhizosphere and to study its effect on Fusarium ear rot disease, fumonisin accumulation in different maize cultivars grown in India. Eight isolates of T. harzianum were isolated and T. harzianum isolate Th-8 exhibited better antifungal activity than carbendizim. Th-8 was formulated in different solid substrates like wheat bran, paddy husk, talcum powder and cornstarch. Maize seeds of kanchan (moderately resistant), pioneer (resistant) and sweet corn (susceptible) were selected for laboratory and field studies and these seeds were treated with a conidial suspension of T. harzianum at the rate of 1 × 108 spore/ml and formulation at the rate of 10 g/kg. Treated seeds were subjected to evaluate F. verticillioides incidence, seed germination, seedling vigour and field emergence, yield, thousand seed weight and fumonisin production. It was found that the pure culture of T. harzianum was more effective in reducing the F. verticillioides and fumonisin incidence followed by Talc formulation than the carbendizim treated and untreated control. Formulations of T. harzianum were effective at reducing the F. verticillioides and Fumonisin infection and also increasing the seed germination, vigour index, field emergence, yield, and thousand seed weight in comparison with the control.

Purification and properties of lipoxygenase induced in downy mildew resistant pearl millet seedlings due to infection with Sclerospora graminicola

Induction of lipoxygenase (LOX) was studied in pearl millet seedlings upon inoculation with Sclerospora graminicola. Resistant pearl millet seedlings exhibited a 2.4-fold increase in LOX activity after inoculation with the downy mildew pathogen S. graminicola. This increase was mainly due to the synthesis of a new LOX isozyme designated as LOX-6. Three of the six isozymes of lipoxygenase designated LOX-1, -3 and -6, were purified from inoculated downy mildew resistant pearl millet seedlings as electrophoretically homogeneous proteins. The isozymes were purified stepwise by ammonium sulfate fractionation, DEAE-sephadex A-50 and sephadex G-200 gel-filtration. Purification factor for LOX-1, -3 and -6 were 46.5, 73.6 and 115.7, respectively. The purified LOX-1, -3 and -6 had molecular weight of 83, 77 and 73 kDa with a pI of 5.5. 5.8 and 6.2, respectively. The results indicate that the LOX isozymes were dimers composed of two unequal subunits of 43 and 40 for LOX-1. 40 and 37 for LOX-3, 38 and 35 for LOX-6. The optimum pH for LOX 6 was 6.5 being stable from pH 4.5 to 8.0. The LOX-1 and -3 had a similar optimum pH of 9.0. LOX isozymes had a different thermal stability ranging from 0 to 30degreesC. Esculetin, NDGA, SHAM, N-propylgallate and copper sulfate were found to be the potent inhibitors of all three isozymes of LOX. All the isozymes of LOX revealed similar reaction with the metal ions like Ca2+, Ba2+, Mn2+, Cu2+, Fe2+ and Mg2+ with very few exceptions. Butyl hydroxyanisole caused the strongest inhibition of the pearl millet LOX isozymes among the various antioxidants tested. The LOX isozymes showed preferential activity towards linoleic acid followed by linolenic acid as substrate

Control of Fusarium verticillioides, cause of ear rot of maize, by Pseudomonas fluorescens

BACKGROUND Maize is one of the staple food crops grown in India. Fusarium verticillioides (Sacc.) Nirenberg is the most important fungal pathogen of maize, associated with diseases such as ear rot and kernel rot. Apart from the disease, it is capable of producing fumonisins, which have elicited considerable attention over the past decade owing to their association with animal disease syndromes. Hence, the present study was conducted to evaluate ecofriendly approaches by using a maize rhizosphere isolate of Pseudomonas fluorescens (Trev.) Mig. and its formulation to control ear rot disease and fumonisin accumulation, and also to study the capacity to promote growth and yield of maize. In vitro assays were conducted to test the efficacy of P. fluorescens as a seed treatment on seed germination, seedling vigour and also the incidence of F. verticillioides in different maize cultivars. The field trials included both seed treatment and foliar spray. For all the experiments, P. fluorescens was formulated using corn starch, wheat bran and talc powder. In each case there were three different treatments of P. fluorescens, a non-treated control and chemical control. RESULTS Pure culture and the formulations, in comparison with the control, increased plant growth and vigour as measured by seed germination, seedling vigour, plant height, 1000 seed weight and yield. P. fluorescens pure culture used as seed treatment and as spray treatment enhanced the growth parameters and reduced the incidence of F. verticillioides and the level of fumonisins to a maximum extent compared with the other treatments. CONCLUSION The study demonstrates the potential role of P. fluorescens and its formulations in ear rot disease management. The biocontrol potential of this isolate is more suited for fumonisin reduction in maize kernels intended for human and animal feed.