Induction of defense-related proteins in tomato roots treated with Pseudomonas fluorescens Pf1 and Fusarium oxysporum f. sp. lycopersici

Pseudomonas fluorescens isolate Pf1 was found to protect tomato plants from wilt disease caused by Fusarium oxysporum f. sp. lycopersici. Induction of defense proteins and chemicals by P. fluorescens isolate Pf1 against challenge inoculation with F. oxysporum f. sp. lycopersici in tomato was studied. Phenolics were found to accumulate in bacterized tomato root tissues challenged with F. oxysporum f. sp. lycopersici at one day after pathogen challenge. The accumulation of phenolics reached maximum at the 5^th day after pathogen challenge. In pathogen-inoculated plants, the accumulation started at the 2^nd day and drastically decreased 4 days after the pathogen inoculation. Activities of phenylalanine ammonia-lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) increased in bacterized tomato root tissues challenged with the pathogen at one day after pathogen challenge and activities of PAL and PO reached maximum at the 4^th day while activity of PPO reached maximum at the 5^th day after challenge inoculation. Isoform analysis revealed that a unique PPO1 isoform was induced and PO1 and PPO2 isoforms were expressed at higher levels in bacterized tomato root tissues challenge inoculated with the pathogen. Similarly, -1,3 glucanase, chitinase and thaumatin-like proteins (TLP) were induced to accumulate at higher levels at 3-5 days of challenge inoculation in bacterized plants. Western blot analysis showed that chitinase isoform Chi2 with a molecular weight of 46 kDa was newly induced due to P. fluorescens isolate Pf1 treatment challenged with the pathogen. TLP isoform with molecular weight of 33 kDa was induced not only in P. fluorescens isolate Pf1-treated root tissues challenged with the pathogen but also in roots treated with P. fluorescens isolate Pf1 alone and roots inoculated with the pathogen. These results suggest that induction of defense enzymes involved in phenylpropanoid pathway and accumulation of phenolics and PR-proteins might have contributed to restriction of invasion of F. oxysporum f. sp. lycopersici in tomato roots.     

Endophytic Pseudomonas fluorescens Endo2 and Endo35 induce resistance in black gram (Vigna mungo L. Hepper) to the pathogen Macrophomina phaseolina

Abstract

The effect of endophytic Pseudomonas fluorescens isolates Endo2 and Endo35 on induced systemic disease protection against dry root rot of black gram (Vigna mungo L. Hepper) caused by Macrophomina phaseolina was investigated under glasshouse conditions. When the bacterized black gram plants were inoculated with dry root rot pathogen, the activities of peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL) were stimulated in addition to accumulation of phenolics and lignin. Activity of phenylalanine ammonia-lyase (PAL) reached the maximum 24 h after pathogen challenge inoculation, whereas the activities of PO and PPO reached the maximum at 72 h and 48 h, respectively. Isoform analysis revealed that a unique PPO3 isozyme was induced in bacterized black gram tissues inoculated with the pathogen. Phenolics were found to accumulate in bacterized black gram tissues challenged with M. phaseolina one day after pathogen challenge. The accumulation of phenolics reached maximum at the third day after pathogen inoculation. Similar observation was found in the lignin content of black gram plants. In untreated control plants, the accumulation of defence enzymes and chemicals started at the first day and drastically decreased 3 days after pathogen inoculation. These results suggest that induction of defense enzymes involved in phenylpropanoid pathway and accumulation of phenolics and PR-proteins might have contributed to restricting invasion of Macrophomina phaseolina in black gram roots.     

Influence of phylloplane colonizing biocontrol agents on the black spot of rose caused by Diplocarpon rosae

Abstract

An attempt was made to study the biocontrol efficacy of antagonistic microorganisms in phylloplane of rose cv. Edward to manage the black spot (Diplocarpon rosae) disease. Eight antagonistic microorganisms were tested in vivo against the black spot pathogen. Among these, Trichoderma viride and Pseudomonas fluorescens pf1 reduces the mycelial growth significantly. These two biocontrol agents were evaluated for their ability to induce defense-related enzymes and chemicals in plants. Increased activity of phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenoloxidase (PPO) and total phenolics were recorded in all the biocontrol agents treated leaves. P. fluorescens Pf1 recorded early and increased synthesis of the entire defense-related enzymes and total phenol within 6 days. The application of biocontrol agents induced the defense-related enzymes involved in phenyl propanoid pathway in addition to direct antagonism, which collectively contribute for enhanced resistance against invasion of Diplocarpon rosae in rose.     

Determination and quantification of sinigrin glucosinolates in Alternaria solani susceptible tomato (Solanum lycopersicum) leaves treated with moringa (Moringa oleifera) leaf extract

Abstract

The study investigates the presence and quantity of antimicrobial sinigrin glucosinolates in tomato leaves after spraying them with moringa (Moringa oleifera) leaf extract (MLAE). Moringa concentrates (0.5, 0.75, 1.00 and 1.5?kg?L^?1 (w v^?1)) were prepared. Distilled water was the control. Sampled tomato leaves were air-dried, freeze-dried and extracted firstly using pure methanol in a hot water bath and then pellet re-extracted using 5?mL of hot aqueous methanol (70% v v^?1). An ion exchange column, and sulphatase was used to achieve glucosiodesulphonation. High performance liquid chromatography (HPLC) was employed in the identification and quantitative analysis of the sinigrin glucosinolates. Tomato (Solanum lycopersicum) leaves treated with MLAE revealed highly significant (p?<?.001) content of sinigrin glucosinolates. The sinigrin standard and the desulphated sinigrin glucosinolates had a 7?s retention time difference; 5?kg?L^?1 (w v^?1) resulted in a superior amount of sinigrin in tomato leaves as compared to all the other MLAE concentrations. The study reveals that spraying MLAE on putatively diseased tomato leaves donates specific quantifiable glucosinolates like sinigrin, which may be involved in defense against tomato diseases and, hence, recommends use of 5?kg?L^?1 (w v^?1) for the highest sinigrin defense tag.     

Azoxystrobin induces lignification-related enzymes and phenolics in rice (Oryza sativa L.) against blast pathogen (Pyricularia grisea)

Abstract

This study was conducted to analyse the induction of lignification-related enzymes and phenolic content in rice to blast disease caused by Pyricularia grisea using azoxystrobin. The severity of rice blast was reduced (70% over control) through treatment by azoxystrobin. This reduction in disease severity was mainly associated with induction of host defense mechanisms by azoxystrobin. Increased production of secondary metabolite – phenolic and lignification – related enzymes, namely, peroxidase (POD), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) were observed in rice plants treated with azoxystrobin.     

PGPR mediates induction of pathogenesis-related (PR) proteins against the infection of blast pathogen in resistant and susceptible ragi [Eleusine coracana (L.) Gaertner] cultivars

The Pseudomonas fluorescens isolate 1 (Pf1) was found to protect the ragi [Eleusine coracana (L.) Gaertner] blast fungus, Pyricularia grisea. Induction of defense proteins viz. chitinase, β-1,3 glucanase, peroxidase (PO) and polyphenol oxidase (PPO) by the Pf1 isolate was studied against P. grisea. Chitinase in a resistant, susceptible and commonly used cultivar with and without challenge inoculation of P. grisea, revealed changes in the isoform pattern by UV illumination after staining the gel with fluorescent brightner 28. Native PAGE (polyacrylamide gel electrophoresis) of PO showed the single isoform in all the treatments including the control and a significant increase in the intensity of the band in the inoculated control and Pf1 treatment in all the varieties. Isoform analysis of PPO showed the induction of PPO in P. fluorescens treated plants challenged with P. grisea.     

Rhizobacterial bioformulation for the effective management of Macrophomina root rot in mungbean

Abstract

Fluorescent pseudomonads based bioformulation was evaluated for their ability to control Macrophomina root rot disease in mungbean (Vigna mungo). P. fluorescens isolate Pf1 showed the maximum inhibition in mycelial growth of Macrophomina phaseolina under in vitro conditions. Bioformulation of Pf1 with chitin was effective in reducing the root rot incidence in green gram both under glasshouse and field conditions. The rhizosphere colonization of P. fluorescens was observed appreciable with the green gram plants. However, Pf1 amended with chitin colonized effectively. Furthermore, the induction of defence-related enzymes and chemicals in plants by Pf1 amended with or without chitin and neem were tested. Increased accumulation of defence enzymes viz., phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO), chitinase, β-1,3-glucanse and phenolics were observed in Pf1 bioformulation amended with chitin, pre-treated plants challenge inoculated with M. phaseolina under glasshouse conditions. The present study reveals that in addition to direct antagonism and plant-growth promotion, PGPR strains amended with chitin bioformulation induced defence-related enzymes and pathogenesis related (PR) proteins which collectively enhance the resistance in green gram against the infection of M. phaseolina.     

Variations of Phenoloxidase Activities as a Consequence of Stresses that Induce Resistance to Fusarium Wilt of Tomato

The variations over 7–8 day of peroxidase (PO) and polyphenoloxidase (PPO) activity have been investigated in tomato plants the roots of which had been subjected to stresses (heat, chloroform and a non-pathogenic form of Fusarium oxysporum) that induce resistance to Fusarium oxysporum f. sp. lycopersici. All treatments induced increase of PO and PPO activity that reached a maximum 3 days after the treatments in leaves, 4 days in stem and roots and were higher in leaves than in other parts. Activity decreased to levels for the control plants after 8 days. Inoculation with Fusarium oxysporum f. sp. lycopersici further stimulated PO and PPO activity in all treated plants over that caused by the treatments alone. Again, activity of treated plants was lower than in controls 7 days after inoculation. It is concluded that 1. increased PO and PPO activity in tomato is a systemic response to cellular injury caused in the root by heat, chloroform and non-pathogenic Fusarium oxysporum, 2. these treatments do no prevent the pathogen from interacting with the plants and inducing further enzyme increase, 3. treated plants react more strongly to the challenge inoculation than untreated plants.     

The long-term variation of Cd and Zn hyperaccumulation by Noccaea spp and Arabidopsis halleri plants in both pot and field conditions

Three Cd and Zn hyperaccumulating plant species Noccaea caerulescens Noccaea praecox and Arabidopsis halleri (Brassicacceae) were cultivated in seven subsequent vegetation seasons in both pot and field conditions in soil highly contaminated with Cd, Pb, and Zn. The results confirmed the hyperaccumulation ability of both plant species, although A. halleri showed lower Cd uptake compared to N. caerulescens. Conversely, Pb phytoextraction was negligible for both species in this case. Because of the high variability in plant yield and element contents in the aboveground biomass of plants, great variation in Cd and Zn accumulation was observed during the experiment. The extraction ability in field conditions varied in the case of Cd from 0.2 to 2.9 kg ha^?1 (N. caerulescens) and up to 0.15 kg ha^?1 (A. halleri), and in the case of Zn from 0.2 to 6.4 kg ha^?1 (N. caerulescens) and up to 13.8 kg.ha^?1 (A. halleri). Taking into account the 20 cm root zone of the soil, the plants were able to extract up to 4.1% Cd and 0.2% Zn in one season. However, cropping measures should be optimized to improve and stabilize the long-term phytoextraction potential of these plants.     

Induction of peroxidase and polyphenol oxidase in Arachis hypogaea in response to treatment with Pseudomonas fluorescens and inoculation with Alternaria alternata

Abstract

The application of talc-formulation through seed, seed treatment plus foliar spray and foliar spray alone significantly reduced the leaf blight incidence both under greenhouse and field conditions. The groundnut plants treated with biocontrol agent and challenge inoculated with Alternaria alternata recorded significantly increased activity of Peroxidase isozyme (PO), Polyphenol oxidase isozymes (PPO) activity. Expression of PO2, PPO1 and PPO2 isoforms were found in all the plants treated with Pf1 while additional PO1, PPO3, PPO4 and PPO5 were observed in Pf1-treated plants followed by challenge inoculation with the pathogen.     

Trichoderma and chitin mixture based bioformulation for the management of head rot (Sclerotinia sclerotiorum (Lib.) deBary)–root-knot (Meloidogyne incognita Kofoid and White; Chitwood) complex diseases of cabbage

The effect of application of different biocontrol agents (Trichoderma spp (eight isolates), Pseudomonas fluorescens (four isolates) and Bacillus subtilis (two isolates)) was tested against head rot fungus, Sclerotinia sclerotiorum and root-knot nematode, Meloidogyne incognita complex diseases. In vitro studies showed that three Trichoderma isolates (Tvc1, Tvc2 and Thc) were effective in inhibition of mycelial growth of the fungus as well as egg hatching ability of the nematode. Application of talc based formulation of biocontrol agents (bioformulations) individually as well as in mixtures with or without chitin was tested against head rot–root-knot disease complex under greenhouse conditions. The combined application of bioformulation mixture (Tvc1, Tvc2 and Thc) along with chitin reduced the incidence of the complex diseases and induced significantly increased activities of phenylalanine ammonia-lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO) and chitinase in cabbage. Activities of PAL and chitinase reached maximum levels within 10 – 20 d, while the activity of PAL continued to be maintained up to 40 d after the application of Tvc1 + Tvc2 + Thc + chitin. Isozyme analyses observed that unique PO (PO1, PO2 PO3 and PO4) and PPO (PPO1, PPO2 and PPO3) enzymes were induced after 10 d in cabbage plants treated with Tvc1 + Tvc2 + Thc + chitin upon challenge inoculation with head rot–root-knot pathogens. Similarly, the bioformulation mixture with chitin was successful at reducing the incidence of head rot–root-knot apart from enhancing the crop yield under field conditions. The mechanism associated with reduced incidence of head rot–root-knot in cabbage may be due to induction of defense proteins (PAL, PO, PPO and chitinase) in the crop.     

Induction of phenylpropanoid metabolism by Pseudomonas fluorescens against tomato spotted wilt virus in tomato

Pseudomonas fluorescens (two native strains, one collection strain and their strain mixtures in all possible combinations) when applied through seed, seedling dip, soil and on leaf significantly reduced the tomato spotted wilt virus (TSWV) disease. InP. fluorescens-treated plants, the peroxidase and phenylalamine ammonia-lyase activity increased. Accumulation of phenolic compounds and lignin were shown to be increased in theP. fluorescens-treated plants. Isoperoxidase native PAGE indicated that the peroxidase isoforms in tomato plants induced by fluorescent pseudomonads were different from the control plants; this suggests that the general phenylpropanoid pathway is probably stimulated in tomato plants treated which in turn led to significant reduction in TSWV.     

Plant growth regulator interactions results enhancement of antioxidant enzymes in Catharanthus roseus

Abstract

The present investigation was carried out with the objectives to understand the effect of paclobutrazol, gibberellic acid and Pseudomonas fluorescens on the enzymatic antioxidants like Ascorbate peroxidase (APX, EC: 1.11.1.11), Superoxide dismutase (SOD, EC: 1.15.1.1), Catalase (CAT, EC: 1.11.1.6), Peroxidase (POX, EC 1.11.1.7) and polyphenol oxidase (PPO, Ec 1.10.3.1) activities of Catharanthus roseus plants under field conditions. 10 mg l^?1 paclobutrazol, 5 µM gibberellic acid and 1 mg P. fluorescens concentrations were used for the treatments, and control plants were irrigated with well water. The treatments were given 38, 53, 68 and 83 days after planting (DAP) by soil drenching. The plants were taken randomly 45, 60, 75 and 90 DAP and separated into root, stem, leaves and flowers and used for estimating the antioxidant enzymes. The results showed that these plant growth regulators have significant effects on antioxidant enzymes of C. roseus.     

Interaction between Pseudomonas fluorescens and Meloidogyne incognita on tomato plant as influenced by the different levels of phosphorus

A pot experiment was conducted on tomato (Solanum lycopersicum cv. Pusa Ruby) to assess the effect of different phosphorus (P) levels (0, 125, 250 and 500 mg/pot) and the plant growth promoting rhizobacterium, Pseudomonas fluorescens, on the growth of tomato and on the reproduction of Meloidogyne incognita. Maximum growth of tomato occurred at P rates of 125 mg/kg soil, irrespective of whether plants were uninoculated or inoculated with P. fluorescens or M. incognita or inoculated with both the agents. Nematodes per gram of roots, egg masses per root, eggs per egg mass and galls per root significantly increased by increasing levels of P. P. fluorescens performed better than other treatments and different P levels in improving tomato growth and reducing galling and multiplication of M. incognita.     

Carbon budget and methane and nitrous oxide emissions over the growing season in a Miscanthus sinensis grassland in Tomakomai,Hokkaido, Japan

Species in the Miscanthus genus have been proposed as biofuel crops that have potential to mitigate elevated atmospheric carbon dioxide (CO₂) levels and nitrous oxide (N₂O) and methane (CH₄) emissions. Miscanthus sinensis is widespread throughout Japan and has been used for biomass production for centuries. We assessed the carbon (C) budget and N₂O and CH₄ emissions over the growing season for 2 years in a M. sinensis‐dominated grassland that was naturally established around 1972 in Tomakomai, Hokkaido, Japan, which is near the northern limit for M. sinensis grassland establishment on Andisols. Average C budget was ?0.31 Mg C ha^?1, which indicates C was released from the grassland ecosystem to the atmosphere. Dominant components in the C budget appeared to be aboveground net primary production of plants (1.94–2.80 Mg C ha^?1) and heterotrophic respiration (2.27–3.11 Mg C ha^?1). The measurement of belowground net primary production (BNPP) of plants in the M. sinensis grassland was extremely variable, thus only an approximate value could be calculated. Mean C budget calculated with the approximated BNPP value was 1.47 and ?0.23 Mg C ha^?1 for 2008 and 2009, respectively. Given belowground biomass (9.46–9.86 Mg C ha^?1) was 3.1–6.5 times higher than that of aboveground biomass may provide additional evidence suggesting this grassland represents a C sink. Average CH₄ emissions across years of ?1.34 kg C ha^?1 would indicate this grassland acts as an atmospheric CH₄ sink. Furthermore, average N₂O emissions across years were 0.22 kg N ha^?1. While the site may contribute N₂O to the atmosphere, this value is lower compared with other grassland types. Global warming potential calculated with the approximated BNPP value was ?5.40 and 0.95 Mg CO₂ Eq ha^?1 for 2008 and 2009, respectively, and indicates this grassland could contribute to mitigation of global warming.     

CHLORIDE AND SODIUM UPTAKE POTENTIAL OVER AN ENTIRE ROTATION OF POPULUS IRRIGATED WITH LANDFILL LEACHATE

There is a need for information about the response of Populus genotypes to repeated application of high-salinity water and nutrient sources throughout an entire rotation. We have combined establishment biomass and uptake data with mid- and full-rotation growth data to project potential chloride (Cl^?) and sodium (Na⁺) uptake for 2- to 11-year-old Populus in the north central United States. Our objectives were to identify potential levels of uptake as the trees developed and stages of plantation development that are conducive to variable application rates of high-salinity irrigation. The projected cumulative uptake of Cl^? and Na⁺ during mid-rotation plantation development was stable 2 to 3 years after planting but increased steadily from year 3 to 6. Year six cumulative uptake ranged from 22 to 175 kg Cl^? ha^?1 and 8 to 74 kg Na⁺ ha^?1, while annual uptake ranged from 8 to 54 kg Cl^? ha^?1 yr^?1 and 3 to 23 kg Na⁺ ha^?1 yr^?1. Full-rotation uptake was greatest from 4 to 9 years (Cl^?) and 4 to 8 years (Na⁺), with maximum levels of Cl^? (32 kg ha^?1 yr^?1) and Na⁺ (13 kg ha^?1 yr^?1) occurring in year six. The relative uptake potential of Cl^? and Na⁺ at peak accumulation (year six) was 2.7 times greater than at the end of the rotation.     

Implications of rudd (Scardinius erythrophthalmus) herbivory on submerged macrophytes in a shallow eutrophic lake

The role of rudd (Scardinius erythrophthalmus) herbivory was studied in Kirkkojärvi, a shallow and turbid basin in Lake Hiidenvesi, Finland. The submerged species dominating in the rudd diets were Potamogeton obtusifolius, Ranunculus circinatus, Sparganium emersum, bryophytes, and filamentous algae. Plant consumption estimated with bioenergetics modelling increased with fish age, being highest in late summer concomitant with the macrophyte biomass peak. Depending on the age structure, a rudd biomass of 20 kg ha^?1 consumes 18–23 kg of macrophytes ha^?1 a^?1, while a rudd biomass of 100 kg ha^?1 results in plant consumption of 92–115 kg ha^?1 a^?1. Although, rudd seemed to feed rather unselectively on suitable-sized and edible plants, some species abundant in the littoral, such as Myriophyllum verticillatum and pleustophytic Ceratophyllum demersum, were not found in rudd guts, indicating selective plant consumption. In Kirkkojärvi, selective grazing by rudd and increased turbidity and high nutrient levels partly caused by bottom dwelling cyprinid fish, may promote the inedible and pleustophytic macrophytes, which have increased in Kirkkojärvi during the past decades.

     

Are the benefits of yield responses to nitrogen fertilizer application in the bioenergy crop Miscanthus × giganteus offset by increased soil emissions of nitrous oxide?

A field trial was carried out on a 15 year old Miscanthus stand, subject to nitrogen fertilizer treatments of 0, 63 and 125 kg‐N ha^?1, measuring N₂O emissions, as well as annual crop yield over a full year. N₂O emission intensity (N₂O emissions calculated as a function of above‐ground biomass) was significantly affected by fertilizer application, with values of 52.2 and 59.4 g N₂O‐N t^?1 observed at 63 and 125 kg‐N ha^?1, respectively, compared to 31.3 g N₂O‐N t^?1 in the zero fertilizer control. A life cycle analyses approach was applied to calculate the increase in yield required to offset N₂O emissions from Miscanthus through fossil fuel substitution in the fuel chain. For the conditions observed during the field trial yield increases of 0.33 and 0.39 t ha^?1 were found to be required to offset N₂O emissions from the 63 kg‐N ha^?1 treatment, when replacing peat and coal, respectively, while increases of 0.71 and 0.83 t ha^?1 were required for the 125 kg‐N ha^?1 treatment, for each fuel. These values are considerably less than the mean above‐ground biomass yield increases observed here of 1.57 and 2.79 t ha^?1 at fertilization rates 63 and 125 kg‐N ha^?1 respectively. Extending this analysis to include a range of fertilizer application rates and N₂O emission factors found increases in yield necessary to offset soil N₂O emissions ranging from 0.26 to 2.54 t ha^?1. These relatively low yield increase requirements indicate that where nitrogen fertilizer application improves yield, the benefits of such a response will not be offset by soil N₂O emissions.