Arthrobotrys oligospora-mediated biological control of diseases of tomato (Lycopersicon esculentum Mill.) caused by Meloidogyne incognita and Rhizoctonia solani.

AIMS: To study the biocontrol potential of nematode-trapping fungus Arthrobotrys oligospora in protecting tomato (Lycopersicon esculentum Mill.) against Meloidogyne incognita and Rhizoctonia solani under greenhouse and field conditions. METHODS AND RESULTS: Five isolates of the nematode-trapping fungus Arthrobotrys oligospora isolated from different parts of India were tested against Meloidogyne incognita and Rhizoctonia solani in tomato (Lycopersicon esculentum Mill.) plants grown under greenhouse and field conditions. Arthrobotrys oligospora-treated plants showed enhanced growth in terms of shoot and root length and biomass, chlorophyll and total phenolic content and high phenylalanine ammonia lyase activity in comparison with M. incognita- and R. solani-inoculated plants. Biochemical profiling when correlated with disease severity and intensity in A. oligospora-treated and untreated plants indicate that A. oligospora VNS-1 offered significant disease reduction in terms of number of root galls, seedling mortality, lesion length, disease index, better plant growth and fruit yield as compared to M. incognita- and R. solani-challenged plants. CONCLUSION: The result established that A. oligospora VNS-1 has the potential to provide bioprotection agents against M. incognita and R. solani. SIGNIFICANCE AND IMPACT OF THE STUDY: Arthrobotrys oligospora can be a better environment friendly option and can be incorporated in the integrated disease management module of crop protection. Application of A. oligospora not only helps in the control of nematodes but also increases plant growth and enhances nutritional value of tomato fruits. Thus, it proves to be an excellent biocontrol as well as plant growth promoting agent.

Fertile plant regeneration from cryopreserved calli of Oryza rufipogon Griff. and assessment of variation in the progeny of regenerated plants.

A protocol was developed for preservation of calli of Oryza rufipogon Griff. in liquid nitrogen (-196°C). Optimal conditions for cryoprotection, pre-freezing and post-thaw recovery of calli were identified. Cryoprotectant treatment at low temperature, pre-freezing in isopropanol bath for 1 h at -70°C, rapid thawing and proper removal of cryoprotectant were critical for post-thaw survival. Genetic fidelity of the R1 plants obtained from cryopreserved, cryoprotectant-treated and untreated calli was assessed by phenotypic and molecular characterization. Comparison of phenotypic characters with seed-derived control plants showed no significant variation in the agronomic characters, but seed physical characters showed significant reduction in all the in vitro generated plants. Molecular data generated using 26 rice simple sequence repeat markers showed 4.78-7.25% change from control. Results suggested that both callus induction and cryopreservation induced heritable variations in O. rufipogon. In addition, a combination of phenotypic and molecular characterization using an appropriate marker provided better insight into genetic fidelity of recovered plants.

Earthworm Grazed-Trichoderma harzianum Biofortified Spent Mushroom Substrates Modulate Accumulation of Natural Antioxidants and Bio-Fortification of Mineral Nutrients in Tomato.

The present investigation was aimed at evaluating the impact of earthworm grazed and Trichoderma harzianum biofortified spent mushroom substrate (SMS) on natural antioxidant and nutritional properties of tomato. Results of the investigation reveal that earthworm grazing and T. harzianum bio-fortification led to significant improvement in the physico-chemical properties of fresh SMS and its application increased the accumulation of natural antioxidants and mineral content in tomato as compared to either T. harzianum biofortified SMS or fresh SMS. In particular, the earthworm grazed, T. harzianum biofortified SMS (EGTHB-SMS) was found to inhibit lipid peroxidation and protein oxidation with significant increase in total polyphenol and flavonoid content in tomato. Further, it increased Fe2+/Fe3+ chelating activity, superoxide anion radical scavenging activity compared to other treatments. The results thus suggest an augmented elicitation of natural antioxidant properties in tomato treated with EGTHB-SMS, resulting in a higher radical scavenging activity, that is highly desirable for human health. In addition, the use of SMS to enhance the nutritional value of tomato fruits becomes an environment friendly approach in sustainable crop production.

Differential methods of inoculation of plant growth-promoting rhizobacteria induce synthesis of phenylalanine ammonia-lyase and phenolic compounds differentially in chickpea.

Foliar spray and micro-injection of plant growth-promoting rhizobacterial species, viz. Pseudomonas fluorescens and P. aeruginosa on chickpea induced synthesis of phenylalanine ammonia-lyase (PAL) when tested against Sclerotinia sclerotiorum. Induction of PAL was also associated with increased synthesis of phenolic compounds such as tannic, gallic, caffeic, chlorogenic and cinnamic acids. Treatment with P. fluorescens was found to be more effective in inducing phenolic compounds as compared to P. aeruginosa. However, persistence of PAL activity was observed more with P. aeruginosa. Although both the inoculation methods were effective, foliar application was found to be superior to micro-injection in terms of rapid PAL activity leading to the synthesis of phenolic compounds.

Bio-protective microbial agents from rhizosphere eco-systems trigger plant defense responses provide protection against sheath blight disease in rice (Oryza sativa L.).

Sheath blight of rice (Oryza sativa L.) caused by Rhizoctonia solani is a major disease and attempts are being made to develop microbe based technologies for biocontrol of this pathogen. However, the mechanisms of biocontrol are not fully understood and still require indepth study in the backdrop of emerging concepts in biological systems. The present investigation was aimed at deciphering the mechanisms of biocontrol of sheath blight of rice employing Pseudomonas fluorescens and Trichoderma harzianum as model agents for biocontrol. Initially 25, 5 and 5 strains of P. fluorescens, T. viride and T. harzianum, respectively, were screened for their biocontrol potential. Out of which, six strains with higher value of percent inhibition of fungal mycelium in dual plate assay were selected. The role of P. fluorescens, T. viride and T. harzianum were investigated in induction and bioaccumulation of natural antioxidants, defence-related biomolecules and other changes in plant which lead not only to growth promotion but also protection from pathogenic stress conditions in rice. The two most promising strains, P. fluorescens PF-08 and T. harzianum UBSTH-501 selected on the basis of in planta evaluation, when applied individually or in combination, significantly enhanced the accumulation of defence-related biomolecules, enzymes and exhibited biocontrol potential against R. solani. A modified/newly developed delivery system was applied for the first time in the experiments involving inoculation of plants with both bioagents, viz. P. fluorescens PF-08 and T. harzianum UBSTH-501. Results suggested that application of P. fluorescens PF-08 and T. harzianum UBSTH-501 alone or in combination, not only helps in control of the disease but also increases plant growth along with reduction in application of toxic chemical pesticides.

Variability in Indian isolates of Sclerotium rolfsii.

Variability among 26 isolates of Sclerotium rolfsii collected from various hosts/soil samples and localities in India is reported. The isolates varied in colony morphology, mycelial growth rate, sclerotium formation, teleomorph production and sclerotial size and color. Out of 26 isolates, only 4 produced the teleomorph stage on Cyperus rotundus rhizome meal agar medium. Mycelial incompatibility among the isolates was also seen, and out of 325 combinations, only 29 combinations (8.9%) showed compatible reactions. Based on mycelial compatibility, 13 vegetative incompatibility groups (VCG) were identified among the isolates. HPLC analysis of the ethyl acetate fraction of culture filtrates of the isolates revealed 10-22 peaks. Six peaks were identified as gallic, oxalic, ferulic, indole-3-acetic acid (IAA), chlorogenic, and cinnamic acids. Oxalic, IAA, and cinnamic acids were present in the culture filtrates of all the isolates in varying amounts. The other three phenolic acids were not detected in some of the isolates. A comparative HPLC analysis of sclerotial exudate, sclerotia, mycelia, and culture filtrates of two S. rolfsii isolates (leaf spot- and collar rot-causing) producing different symptoms on their respective hosts revealed variation in the content of phenolic acids, IAA, and oxalic acid.

Effect of calcium and calmodulin modulators on the development of Erysiphe pisi on pea leaves.

The effect of calcium and calmodulin modulators, viz., ethylene glycol bis (beta-amino ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), a calcium chelator; verapamil, a plasma membrane Ca2+ channel blocker; ruthenium red, an organelle Ca2+ channel blocker; and chlorpromazine, a calmodulin antagonist; on the development of Erysiphe pisi was studied by floating the inoculated leaves on the respective solutions of chemicals. All the modulators affected the development of E. pisi by inhibiting the colony diameter, number of secondary branches, number of hyphal cells per colony and number of haustoria. The calmodulin antagonist was more effective in reducing E. pisi development. The results suggest the possible involvement of calcium and calmodulin in the development of E. pisi on pea leaves.

A novel technique to measure the group velocity of sound in dispersive media.

A technique is proposed for measuring the group velocity of sound by a frequency-modulated (FM) continuous wave (CW) technique. Detailed calculations are presented, showing that V(g)=(deltaomega(m)/deltaphi)d , where V(g), deltaomega(m), deltaphi, and d are, respectively, the group velocity, change of modulation frequency, phase change of the received signal detected by a biphase lock-in amplifier, and the distance traveled by the sound. The approach has the advantage of high resolution compared to the conventional time-of-flight measurement technique.

Antifungal activity of venenatine, an indole alkaloid isolated from Alstonia venenata.

The indole alkaloid venenatine exhibited antifungal activity against some plant pathogenic and saprophytic fungi. Venenatine in an aqueous acetic acid solution inhibited spore germination of all the 10 tested fungi, Fusarium udum, Alternaria brassicicola, Ustilago cynodontis and Aspergillus flavus showed an especially high sensitivity towards this compound, exhibiting germination levels below 10%. The spore germination and colony development of the parasitic fungus Erysiphe pisi, which causes powdery mildew in pea (Pisum sativum), on excised leaves of pea was also significantly affected. Pre-inoculation rather than post inoculation treatment of the leaves was more inhibitory against spore germination and colony development.

Role of garlic (Allium sativum L.) in human and plant diseases.

A resurgence of interest in garlic due to recent revelations of its beneficial effects in the treatment of various human and plant diseases and also due to validation of claims made in traditional systems of medicine has resulted a plethora of publications on different aspects of garlic in recent years. Chemical constituents of garlic and their variations on the methods of isolation have been discussed in the present review. Effect of garlic and its constituents against various human and plant pathogenic and saprophytic microorganisms has also been reviewed.

Potential chemoprevention of N-nitrosodiethylamine-induced hepatocarcinogenesis by polyphenolics from Acacia nilotica bark.

Chemopreventive potential of Acacia nilotica bark extract (ANBE) against single intraperitoneal injection of N-nitrosodiethylamine (NDEA, 200mg/kg) followed by weekly subcutaneous injections of carbon tetrachloride (CCl(4), 3 ml/kg) for 6 weeks induced hepatocellular carcinoma (HCC) in rats was studied. At 45 day after administration of NDEA, 100 and 200mg/kg of ANBE were administered orally once daily for 10 weeks. The levels of liver injury and liver cancer markers such as alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (gamma-GT), total bilirubin level (TBL), alpha-feto protein (AFP) and carcinoembryonic antigen (CEA) were substantially increased following NDEA treatment. However, ANBE treatment reduced liver injury and restored liver cancer markers. ANBE also significantly prevented hepatic malondialdehyde (MDA) formation and reduced glutathione (GSH) in NDEA-treated rats which was dose dependent. Additionally, ANBE also increased the activities of antioxidant enzymes viz., catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) in the liver of NDEA-administered rats. Eventually, ANBE also significantly improved body weight and prevented increase of relative liver weight due to NDEA treatment. Histological observations of liver tissues too correlated with the biochemical observations. HPLC analysis of ANBE showed the presence of gallic, protocatechuic, caffeic and ellagic acids, and also quercetin in ANBE. The results strongly support that A. nilotica bark prevents lipid peroxidation (LPO) and promote the enzymatic and non-enzymatic antioxidant defense system during NDEA-induced hepatocarcinogenesis which might be due to activities like scavenging of oxy radicals by the phytomolecules in ANBE.

Antioxidant and anti-quorum sensing activities of green pod of Acacia nilotica L.

The antioxidant and anti-quorum sensing activities of eight extracts were studied in green pods of Acacia nilotica. The specific phenolic compositions and their quantifications were performed by HPLC and MS/MS, which showed that the HEF (pH 4) was higher in gallic acid, ellagic acid, epicatechin, rutin, and GTs. In order to find antioxidant potential of various extracts, their activities were studied for TPC, AOA, FRSA, RP, inhibition of LPO, FIC activity, HO* and O(2)(-) scavenging activities. Among them HEF (pH 4) has shown potent antioxidant activity. HEF (pH 4) was also found effective in protecting plasmid DNA and HAS protein oxidation induced by HO*. Pre-treatment of HEF (pH 4) at 75 and 150 mg/kg body weight for 6 days caused a significant increase in the levels of CAT and SOD and decrease in the level of MDA content in liver, lungs, kidneys and blood when compared to CCl(4)-intoxicated rats. Eventually, the extracts were also screened for anti-QS activity. Of these extracts two showed QS inhibition: HEF (pH 4) and HCE. The results obtained strongly indicate that green pod of A. nilotica are important source of natural antioxidants.

Microwave absorption measurements using a broad-band meanderline approach.

We describe a technique that permits broad-band, field-dependent ferromagnetic and electron paramagnetic resonance absorption measurements that is applicable to thin films and patterned micro-/nanostructured arrays and is based on a wire-wound meanderline approach. Techniques to prepare meanderlines and perform microwave measurements are described along with some demonstrations involving an electron paramagnetic resonance calibration/test material, 2,2-diphenyl-1-picryl-hydrazyl, and a ferromagnetic cobalt thin film.

Polyphenolics from various extracts/fractions of red onion (Allium cepa) peel with potent antioxidant and antimutagenic activities.

In order to determine antioxidant activity, the five extracts/fractions of red onion peel were studied for their total content of phenolics (TPC), flavonoids (TFC), antioxidant activity (AOA), free radical scavenging activity (FRSA), assayed by DPPH radical in the terms of anti-radical power (ARP) and reducing power (RP), expressed as ascorbic acid equivalents (ASE)/ml. High TPC (384.7 +/- 5.0 mg GAE/g), TFC (165.2+/- 3.2 mg QE/g), AOA (97.4 +/- 7.6%), ARP (75.3 +/-4.5) and RP (1.6 +/-0.3 ASE/ml) were found for the ethyl acetate (EA) fraction. EA fraction had markedly higher antioxidant capacity than butylated hydroxytoluene (BHT) in preventive or scavenging capacities against FeCl3-induced lipid peroxidation, protein fragmentation, hydroxyl (site-specific and non-site-specific), superoxide anion and nitric oxide radicals. EA fraction also showed dose dependent antimutagenic activity by following the inhibition of tobacco-induced mutagenicity in Salmonella typhimurium strains (TA102) and hydroxyl radical-induced nicking in plasmid pUC18 DNA. HPLC and MS/MS analysis showed the presence of ferulic, gallic, protocatechuic acids, quercetin and kaempferol. The large amount of polyphenols contained in EA fraction may cause its strong antioxidant and antimutagenic properties. This information shows that EA fraction of red onion peel can be used as natural antioxidant in nutraceutical preparations.

Control of Erysiphe pisi Causing Powdery Mildew of Pea (Pisum sativum) by Cashewnut (Anacardium occidentale) Shell Extract.

The effect of methanolic extract of cashewnut (Anacardium occidentale) shell extract was seen on conidial germination of Erysiphe pisi and powdery mildew development in pea (Pisum sativum). Maximum conidial germination inhibition of E. pisi on glass slides was observed at 300 ppm. Similar effect on floated pea leaves was observed after 48 h at the same concentration. Conidial germination on intact untreated pea leaves was also assessed on II and IV nodal leaves while IV and II nodal leaves were treated with the extract and vice versa. There was tremendous reduction in conidial germination on all the nodal leaves. The disease intensity of pea powdery mildew was significantly reduced by methanolic extract of cashewnut shells. Maximum reduction was observed with 200 ppm where 39% disease intensity was recorded in comparison to 96.53% in the control. The phenolic acid content of pea leaves following treatments with this extract varied and no definite pattern was observed. Out of several phenolic compounds, namely, gallic, ferulic, chlorogenic, and cinnamic acids, only gallic acid was found to be present consistently in all the treatments with varied amounts.

Antifungal activity of 2-hydroxy 4,4'6'trimethoxy chalcone.

Antifungal activity of 2-hydroxy 4,4'6'trimethoxy chalcone individually was tested against spore germination of ten fungi of different genera. Efficacy of the chemical was also tested against conidial germination and other growth parameters of Erysiphe pisi on excised pea leaves. 2-Hydroxy 4,4'6'trimethoxy chalcone inhibited spore germination at all the concentrations. Maximum inhibition was observed at 2000 ppm where more than 78 per cent inhibition of spore germination was observed in Ustilago cynodontis, Alternaria brassicicola, A. solani and Aspergillus flavus. It also reduced conidial germination of E. pisi significantly, when applied as pre-inoculation treatment.

Foliar Application of Plant Growth-Promoting Rhizobacteria Increases Antifungal Compounds in Pea (Pisum sativum) Against Erysiphe pisi.

Systemic effect of two plant growth-promoting rhizobacterial (PGPR) strains,viz., Pseudomonas fluorescens (Pf4) and P. aeruginosa (Pag), was evaluated on pea (Pisum sativum) against the powdery mildew pathogen Erysiphe pisi. Foliar spray of the two PGPR strains was done on specific nodal leaves of pea and conidial germination of E. pisi was observed on other nodal leaves,distal to the treated ones. Conidial germination was reduced on distant leaves and at the same time,specific as well as total phenolic compounds increased in the leaves distal to those applied with PGPR strains,thereby indicating a positive correlation. The strains induced accumulation of phenolic compounds in pea leaves and the amount increased when such leaves were get inoculated with E. pisi conidia. Between the two strains, Pag was found to be more effective than Pf4 as its effect was more persistent in pea leaves. Foliar application of PGPR strains for the control of powdery mildew of pea is demonstrated in vitro while correlating it with the increased accumulation of plant phenolics.

Variation in Biochemical Composition among Indian Isolates of Sclerotinia sclerotiorum.

Biochemical variability among 20 Indian isolates of Sclerotinia sclerotiorum collected from different hosts/soil samples from different localities in India is reported. High Performance Liquid Chromatographic (HPLC) analysis of ethyl acetate fraction of culture filtrate, mycelia, sclerotia and sclerotial exudate showed 15~23 peaks but only 11 could be identified. They were tannic, gallic, oxalic, caffeic, vanillic, ferulic, O-coumeric, chlorogenic, cinnamic, salicylic and gentisic acids. The amount of phenolic compounds varied among the culture filtrates, mycelia, sclerotia and sclerotial exudates of S. sclerotiorum.

Antifungal Activity of a Phytoterpenoid (AOS-A) Isolated from Artabotrytis odoratissimus on Spore Germination of Some Fungi.

Phytoterpenoid isolated from Artabotrytis odoratissimus inhibited spore germination of some plant pathogenic as well as saprophytic fungi e.g. Alternaria alternata, A. solani, Cercospora sp., Curvularia maculans, C. pennisetti, Fusarium udum, Helminthosporium echinochlova, H. frumentacie, H. penniseti and Ustilago cynodontis. In Curvularia maculans and H. frumentacie, spore germination was completely inhibited at 2000 ppm. However, Curvularia maculans and C. pennisetti showed considerable sensitivity to this chemical even at 500 ppm.

Induction of Sexual Stage and Colony Morphology of Some Isolates of Sclerotium rolfsii Causing Spotted Leaf Rot in Plants.

Twenty-two isolates of Sclerotium rolfsii causing spotted leaf rot from Varanasi, India were grown on 6% Cyperus rotundus rhizome meal agar (CRMA) medium for the induction of athelial stage (Athelia rolfsii). Only one isolate obtained from Sphaeranthus indicus formed basidial stage on CRMA medium while the other 21 isolates did not. Basidial stage was also produced in S. indicus isolate at different concentrations (5.5, 6.0 and 6.5% w/v) of CRMA medium. Size of basidia, sterigmata and basidiospores of this isolate was measured. Basidia clavate, hyaline and measured 10~12 × 4~5 µm in size, basidiospores hyaline, unicellular, subglobose to ellipsoid produced on sterigmata and measured 3~5 × 2~4µm in size, sterigmata hyaline and measured 4~5 × 1.5~2 µm in size. The results of the present study revealed wide variation in spotted leaf rot isolates of S. rolfsii. A reddish zone around the colony of S. rolfsii isolate from Vernonia sp. was observed on CRMA medium. HPLC analysis of the zone revealed the presence of gallic and ferulic acid which were also thought to be responsible for reduced mycelial growth of the isolate on CRMA medium.