Co-inoculation of Urea and DAP Tolerant <Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> and <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> as Integrated Approach for Growth Enhancement of <Emphasis Type="Italic">Brassica juncea</Emphasis>

Two plant growth promoting rhizobacteria––Sinorhizobium meliloti RMP1 and Pseudomonas aeruginosa GRC₂ were studied for integrated nutrient management to obtain improved yield of Brassica juncea. Low concentrations of urea and diammonium phosphate (DAP) stimulated the growth of both S. meliloti RMP1 and P. aeruginosa GRC₂. 1 M of urea and 0.35 M of DAP was found lethal for RMP1, while 1.3 M and 0.37 M concentrations of urea and DAP proved to be toxic for GRC₂. Lc₅₀ was observed as 0.49 M of urea and 0.15 M of DAP for RMP1, and 0.66 M urea and 0.18 M of DAP for GRC₂. Urea and DAP adaptive variants of RMP1 and GRC₂ was isolated. Adaptive bacterial variants had better growth rates at sub-lethal (Lc₅₀) concentrations of urea and DAP as compared to non-adaptive variants. They also retained plant growth promoting attributes similar to non adaptive variants. GRC₂ and RMP1 did not affect the growth of each other and were chemotactically active for DAP, urea as well as root exudates of B. juncea. Both the isolates colonized well in the rhizosphere of B. juncea, as their populations were recorded ≈5 log₁₀ cfu g⁻¹ after 120 days. Interestingly, the colonization ability was found even better when both strains were co-inoculated, as their population was recorded in the range of ≈6 log₁₀ cfu g⁻¹ after 120 days. In field trials, application of RMP1 and GRC₂ resulted in significant increase in biomass and yield of B. juncea as compared to control. However, yield was better with application of half dose and full dose of recommended fertilizers. Interestingly, the biomass as well as yield improved further when both isolates were applied together along with half dose of recommended fertilizers.     

Biological Control of Soil Pests by Mixed Application of Entomopathogenic and Fungivorous Nematodes

In greenhouse experiments, massive application of the fungivorous nematode, Aphelenchus avenae, in summer at 26-33 C (1 x l0⁵ nematodes/500 cm³ autoclaved soil) or in autumn at 18-23 C (5 x 10⁴ nematodes/500 cm³ autoclaved soil) suppressed pre-emergence damping-off of cucumber seedlings due to Rhizoctonia solani AG-4 by 67% or 87%, respectively. Application of 2 x l0⁵ A. avenae to sterilized soil infested with R. solani caused leafminer-like symptom on the cotyledons, which did not occur in mixed inoculations with the entomopathogenic nematode, Steinernema carpocapsae. When 1 x 10⁶ A. avenae were applied 3 days before inoculation with 100 Meloidogyne incognita juveniles, gall numbers on tomato roots were reduced to 50% of controls. Gall numbers also were suppressed by S. carpocapsae (str. All). Reduction in gall numbers was no greater with mixed application of A. avenae and S. carpocapsae than with application of single species, even though twice the number of nematodes were added in the former case. These nematodes were positively attracted to tomato root tips. Aphelenchus avenae suppressed infection of the turnip moth, Agrotis segetum, but not the common cutworm, Spodoptera litura, by S. carpocapsae.     

Evaluation of the adverse effects of two commonly used fertilizers,DAP and urea,on motility and orientation of the green flagellate Euglena gracilis

The effect of two commonly used fertilizers, DAP (diammonium phosphate) and urea was studied on the freshwater flagellate Euglena gracilis using the automatic biotest ECOTOX. NOEC and EC₅₀ values for various parameters like motility, velocity, cell shape and gravitaxis were calculated. The NOEC and EC₅₀ values obtained for DAP were much lower than those for urea; i.e. DAP showed a stronger inhibitory effect as compared to urea. The inhibition caused by DAP increased with increasing exposure time over 24 h but urea showed no augmentation with increasing exposure time. Application of DAP resulted in an increased pH and high concentrations of ammonia but urea did neither affect the pH nor affect the ammonia concentration. Recovery experiments in deionized water after urea application showed a reconstitution of motility after 72 h. After an application of 1.35 g L⁻¹ (24 h EC₅₀ for motility) DAP motility recovered after 72 h but motility did not recover when the concentration was doubled (2.7 g L⁻¹). The EC₅₀ values obtained were compared with the EC₅₀/LC₅₀ values reported for other aquatic organisms and were found to be comparable with the reported values.     

Effect of urea and certain NPK fertilizers on the cereal cyst nematode (Heterodera avenae) on wheat

Two outdoor pot experiments were conducted in two consecutive years under outdoor conditions during the wheat growing season in Saudi Arabia to determine the effects of urea and certain compound fertilizers (NPK), compared to the effects of the nematicide fenamiphos on the cereal cyst nematode (CCN), Heterodera avenae, and wheat growth. The results showed that all of the treatments, except the fertilizer diammonium phosphate (DAP), reduced the number of nematode cysts/root system and increased (P ⩽ 0.05) the dry weight of nematode-infected wheat plants. Fenamiphos and urea resulted in the best control, followed by the NPK fertilizers. The combined application of urea and fenamiphos resulted in the most significant effect in decreasing (P ⩽ 0.05) the number of cysts/root system and increasing (P ⩽ 0.05) the growth of nematode-infected wheat plants.     

Evaluation of Phosphate Fertilizers for the Immobilization of Cd in Contaminated Soils

A laboratory investigation was conducted to evaluate the efficiency of four phosphate fertilizers, including diammonium phosphate (DAP), potassium phosphate monobasic (MPP), calcium superphosphateon (SSP), and calcium phosphate tribasic (TCP), in terms of the toxicity and bioavailability of Cd in contaminated soils. The efficiency of immobilization was evaluated on the basis of two criteria: (a) the reduction of extractable Cd concentration below the TCLP regulatory level and (b) the Cd changes associated with specific operational soil fractions on the basis of sequential extraction data. Results showed that after 50 d immobilization, the extractable concentrations of Cd in DAP, MPP, SSP, and TCP treated soils decreased from 42.64 mg/kg (in the control) to 23.86, 21.86, 33.89, and 35.59 mg/kg, respectively, with immobilization efficiency in the order of MPP > DAP > SSP > TCP. Results from the assessment of Cd speciation via the sequential extraction procedure revealed that the soluble exchangeable fraction of Cd in soils treated with phosphate fertilizers, especially TCP, was considerably reduced. In addition, the reduction was correspondingly related to the increase in the more stable forms of Cd, that is, the metal bound to manganese oxides and the metal bound to crystalline iron oxides. Treatment efficiency increased as the phosphate dose (according to the molar ratio of PO₄/Cd) increased. Immobilization was the most effective under the molar ratio of PO₄/Cd at 4:1.     

Sustainable phosphorous management in two different soil series of Pakistan by evaluating dynamics of phosphatic fertilizer source

Phosphorous (P) plays the prominent role to promote the plants storage functions and structural roles, as it is recognized as a vital component of ADP, ATP, Cell wall as well as a part of DNA. Soils acts as the sink to supply P to plants because soil pH and its physical condition are the main factor which regulate the solubility and availability P element. Phosphorus is not deficient in Pakistani soils but its availability to plants is the serious matter of concern. A pot experiment was conducted to evaluate P dynamics in two different soil series of Pakistan (Bahawalpur and Lyallpur) using Maize as test crop. The treatments applied were T0: Control (without any fertilizer), T1: Recommended DAP @648 mg pot^?1, T2: Half dose DAP @324 mg pot^?1, T3: Recommended rate of TSP @900 mg pot^?1, T4: Half dose TSP @450 mg pot^?1. Soil analysis showed that Bahawalpur soil has sandy clay loam texture with 33% clay and Lyallpur series has sandy loam texture with 15.5% clay; furthermore, these soil contain 4.6 and 2.12% CaCO₃ respectively. Results showed an increase in P concentration in roots (23 mg kg^?1) with the application of half dose of TSP in Lyallpur series and lowest in Bahawalpur series (14.6 mg kg^?1) at recommended dose of DAP. Concentration of P in shoots responded the same; increase at half dose of TSP (16.7 mg kg^?1) and lowest at full dose of DAP in Bahawalpur series as (15.58 mg kg^?1). Adsorbed P (17 mg kg^?1) was recorded highest in Bahawalpur soil with more clay amount in pot with DAP application but lower in Lyallpur soil series (14 mg kg^?1) with the application of applied TSP. The PUE was recorded highest in Lyallpur series with the application of half dose of TSP and it was 61% more than control and was Highest in Bahawalpur series was with the application of recommended dose of DAP is 72% more than control treatment. On estimation; results showed that applied sources made an increase in P availability than control, but TSP gave better P uptake than DAP unless of rates applied. Soil of Lyallpur series showed better uptake of P and response to applied fertilizers than Bahawalpur series which showed more adsorption of P by high clay and CaCO₃ amount. Conclusively, the study suggested that soil series play a crucial role in choosing fertilizer source for field application.     

Correlations of Nematodes and Soil Properties in Soybean Fields

Soil samples from 40 soybean fields were collected in 1967 and 1968 and analyzed for nematodes and soil properties. Correlations o f total nematodes, non-stylet nematodes, Dorylaimoidea (excluding Xiphinema americanum), X. americanum, Helicotylenchus pseudorobustus, Tylenchus spp., Aphelenchus avenae, and other groupings of nematodes were made with pH; percentage sand, silt, and clay; percentage organic matter; cation exchange capacity; saturation percentage, and percentage saturation. Organic matter, pH, and cation exchange capacity were most consistently highly correlated with the nematodes. H. pseudorobustus had the most consistently significant correlations with the soil factors. Correlations of nematodes were with more soil factors and were stronger in a wet than in a dry year. The highest numbers of nematodes were usually found in the lighter soils, except in the loamy sand where moisture probably was limiting. In general, soil moisture levels below 20% saturation were probably limiting for most nematodes studied, except for the dorylaims which survived in large numbers in soils with less than 20% saturation.     

Influence of Aphelenchus avenae on Vesicular-arbuscular Endomycorrhizal Growth Response in Cotton

The influence of Aphelenchus avenae on the relationship between cotton (Gossypium hirsutum ''Stoneville 213'') and Gigaspora margarita or Glomus etunicatus was assessed by its effect on the mycorrhizal stimulation of plant growth and microorganism reproduction. The mycophagous nematode usually did not suppress stimulation of shoot growth resulting from mycorrhizae (G. margarita) at inoculum levels of 3,000 or 6,000 nematodes per pot, but retarded root growth at 6,000 per pot. When the nematode inoculum was increased to 10, 40, or 80 thousand, G. margarita stimulation of shoot or root growth was retarded at the two higher rates. Shoot growth enhancement by G. etunicatus was suppressed by 10 thousand A. avenae but not by 40 or 80 thousand. A. avenae reproduced better when the nematode was added 3 wk after G. margarita than with simultaneous inoculations. Sporulation of both fungi was affected little by the mycophagous nematode. The high numbers of A. avenae required for an antagonistic effect probably precludes the occurrence of any significant interaction between these two organisms under field conditions.     

Interactions between Heterodera avenae and Pratylenchus neglectus on Wheat

In a long-term field experiment, differential population densities of Heterodera avenae were produced by frequent cropping with resistant (cv. Panema) or susceptible (cv. Peniarth) oat. The two oat cultivars were equally good hosts of Pratylenchus neglectus in a glass house experiment with field soil. On wheat crops grown after oats in field experiments, P. neglectus population densities in roots were higher in plots where H. avenae had been controlled than in plots with moderate infestations (40 H. avenae eggs/g soil). The field observations indicated that the reduction in population densities of P. neglectus coincided with the development in roots of sedentary stages of the cyst nematode. Evidence for an indirect effect of H. avenae on P. neglectus was found in vitro in a split-root experiment. In the same field, grain yields of two wheat cultivars susceptible or resistant to H. avenae, but both susceptible to P. neglectus, was not reduced by P. neglectus. Alternation of H. avenae resistant and susceptible cultivars is a possible way of exploiting the inverse relationship between these nematodes, whilst controlling cyst nematode -populations in intensive cereal production systems.     

Effects of aldicarb and benomyl on nematodes, vesicular arbuscular mycorrhizas and the growth and yield of forage maize

The effects of aldicarb and benomyl on plant-parasitic nematodes, vesicular arbuscular mycorrhiza and the growth of forage maize were measured in 1980—1982 in two field experiments at Woburn, Bedfordshire and in a pot experiment using loamy sand soil from the field site. The most numerous migratory nematode, Tylenchorhynchus dubius increased three to four-fold during each season in untreated soil and was effectively controlled by aldicarb. Pratylenchus species were fewer but equally well controlled. The cereal cyst-nematode (Heterodera avenae), a serious maize pathogen in Northern France, was relatively scarce in untreated roots and was further decreased by aldicarb treatment; post-harvest H. avenae egg numbers were not affected by treatments; they declined equally because maize is such a poor host. Significant yield benefits (up to 37%) followed aldicarb treatment and were ascribed to nematode control in the absence of attribution to insect or other pests. Benomyl did not increase yields nor did it significantly affect the incidence of mycorrhiza. The results confirm that considerable losses of forage maize can be caused by nematodes in light soil and that aldicarb is an effective nematicide even at the rate of 1·7 kg a.i./ha.     

Responses of morphological, physiological, and biochemical parameters in Euglena gracilis to 7-days exposure to two commonly used fertilizers DAP and urea

Diammonium phosphate (DAP) and urea are commonly used fertilizers throughout the world. The effects of these fertilizers on the freshwater flagellate Euglena gracilis was studied after 7 days of growth using morphological, physiological and biochemical parameters as end points. NOEC and EC₅₀ values for various parameters like cell density, motility, velocity, cell shape, gravitaxis, chlorophyll a, b and total carotenoids were calculated. NOEC and EC₅₀ values of DAP varied from 0.5 to 2.5 g L⁻¹ and 3.14 to 5.96 g L⁻¹, respectively, for different parameters. NOEC and EC₅₀ values for urea ranged from 5 to 25 g L⁻¹ and 28 to 44.05 g L⁻¹, respectively, for various parameters. Photosynthetic pigments were found to be more sensitive to both fertilizers as compared to other measured end points. The NOEC and EC₅₀ values obtained for DAP were much lower than those for urea; i.e., DAP showed a stronger inhibitory effect as compared to urea. Application of DAP resulted in an increased concentration of ammonia in Euglena cultures but urea did not. The stronger inhibitory effect of DAP is attributed to release of free ammonia in the culture due to DAP decompostion. No release of ammonia by urea occurred due to the absence of the enzyme urease in E. gracilis.     

Carbamate and Organophosphorus Nematicides: Acetylcholinesterase inhibition and Effects on Dispersal

The sensitivities of acetylcholinesterases (ACHE) from the fungus-feeder Aphelenchus avenae and the plant-parasitic species Helicotylenchus dihystera and Pratylenchus penetrans and the housefly, Musca domestica, were compared using a radiometric assay which utilized H³ acetylcholine as a substrate. Nematode ACHE were generally less sensitive to inhibition by organophosphorns and carbamate pesticides than were ACHE from the housefly. ACHE from the plant-parasitic species and A. avenae were generally similar in sensitivity. In soil, carbamates were more toxic than the organophosphorus pesticides to A. avenae. All pesticides tested affected nematode movement, but fenamiphos was more inhibitory than others. The effects on dispersal of nematodes may be an important mechanism in control by some nematicides.     

Consumption of nematodes by fungivorous mites,Tyrophagus spp. (Acarina: Astigmata: Acaridae)

Summary Free-living nematodes may be attacked and eaten by soil mites which are normally considered fungivores or saprophages. Three species in the genus Tyrophagus, common inhabitants of grassland soils and also common pests in stored products, museums and laboratories, are predators of nematodes. All active stages of the mites will voraciously consume nematodes. When offered nematodes and a choice of other food (baker's yeast and algae), 11% of the Tyrophagus putrescentiae, 23% of the T. zachvatkini, and 56% of the T. similis tested fed on nematodes. Tyrophagus zechvatkini and similis were reared on a diet consisting entirely of nematodes, and developed at rates similar to a fungal diet and produced viable offspring. Agar cultures of Aphelenchus avenae which were inoculated with five mating pairs of Tyrophagus zachvatkini had populations that were one-third less than mite-free controls. Observations indicate that nematodes may be attacked by tyrophagid mites when in a dry, anhydrobiotic state.     

Metabolism of Glycogen and Neutral Lipids by Aphelenchus avenae and Caenorhabditis sp. in Aerobic,Microaerobic and Anaerobic Environments

Starving Aphelenchus avenae survived 3-4 weeks in microaerobic and anaerobic environments, but Caenorhabditis sp. survived less than 80 hr. Aerobically, both nematodes metabolize neutral lipid reserves: there was no microaerobic ( <5% O₂) or anaerobic neutral lipid catabolism. Early in anaerobiosis both nematodes utilized endogenous glycogen. Caenorhabditis sp. depleted the glycogen and died. A. avenae under oxygen stress longer than 120 hr entered cryptobiosis, during which there was neither measurable O₂ uptake nor glycogen or neutral lipid utilization, Only when re-aerated, did A. avenae recover and resume "''normal" metabolism.     

Management of <Emphasis Type="Italic">Sclerotium rolfsii</Emphasis> with integration of non-conventional chemicals,vermicompost and <Emphasis Type="Italic">Pseudomonas syringae</Emphasis>

Two non-conventional chemicals, ZnSO₄ (10⁻⁴ mM) and oxalic acid (4 mM) were tested (alone as well as in combination with seeds bacterized with Pseudomonas syringae strain PUR46 and vermicompost substitution in the potting soil), for their ability to suppress collar rot of chickpea (Cicer arietinum) caused by Sclerotium rolfsii under greenhouse conditions. ZnSO₄ and oxalic acid were applied as pre-inoculation foliar spray on chickpea and subsequently challenged with S. rolfsii. Both the chemicals provided significant protection to chickpea compared to control (100% plant mortality) when used alone as well as in combination with PUR46 and vermicompost. However, ZnSO₄ was more effective than oxalic acid against S. rolfsii. Amongst the treatments tried, plant mortality was least when ZnSO₄ was used in combination with seed bacterization with PUR46 and 25% vermicompost substitution. The findings indicate the utility of integration of the above factors in managing collar rot efficiently.     

Inhibition of Acetylcholinesterases from Aphelenchus avenae by Carbofuran and Fenamiphos

Exposure to carbofuran and fenamiphos for 72 hours reduced the numbers of active Aphelenchus avenae in aqueous suspension by > 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides.     

Nematodes in Dryland Field Crops in the Semiarid Pacific Northwest United States

Soils and roots of field crops in low-rainfall regions of the Pacific Northwest were surveyed for populations of plantparasitic and non-plant-parasitic nematodes. Lesion nematodes (Pratylenchus species) were recovered from 123 of 130 non-irrigated and 18 of 18 irrigated fields. Pratylenchus neglectus was more prevalent than P. thornei, but mixed populations were common. Population densities in soil were affected by crop frequency and rotation but not by tillage or soil type (P < 0.05). Many fields (25%) cropped more frequently than 2 of 4 years had potentially damaging populations of lesion nematodes. Pratylenchus neglectus density in winter wheat roots was inversely correlated with grain yield (r² = 0.64, P = 0.002), providing the first field-derived evidence that Pratylenchus is economically important in Pacific Northwest dryland field crops. Stunt nematodes (Tylenchorhynchus clarus and Geocenamus brevidens) were detected in 35% of fields and were occasionally present in high numbers. Few fields were infested with pin (Paratylenchus species) and root-knot (Meloidogyne naasi and M. chitwoodi) nematodes. Nematodes detected previously but not during this survey included cereal cyst (Heterodera avenae), dagger (Xiphinema species), and root-gall (Subanguina radicicola) nematodes.     

Interaction of dietary zinc and growth implants on weight gain,carcass traits and zinc in tissues of growing beef steers and heifers

In two separate studies, 60 beef heifers (379 kg BW) and 60 beef steers (348 kg BW) were randomly assigned to six treatments in 2×3 factorial arrangements. The treatments were with or without Synovex^® implants combined with either a control diet or diets supplemented with 200 ppm Zn from ZnSO₄ or zinc methionine (Zn-Met). Near the mid-point of the feeding periods, cattle were vaccinated with a modified live virus and subsequent titers and concentrations of immunoglobulin G (IgG) were measured. Liver and blood samples were obtained 1 week prior to the start of the experiments and at intervals during the experiments. In experiment 1, average daily gains of beef heifers were (P<0.05) affected by the interaction of implant and source of dietary Zn. Compared to control and ZnSO₄ treatments, supplementation with Zn-Met increased (P<0.05) the concentration of Zn in serum. Antibody titers and concentrations of IgG in serum were highest (P<0.05) in heifers fed ZnSO₄ compared to heifers fed the control or Zn-Met supplemented diets. The Synovex-H^® implant reduced the concentrations of Zn and Cu in liver. In experiment 2, Synovex-S^® implants improved (P<0.05) weight gains of steers supplemented with 200 ppm dietary Zn from ZnSO₄ compared to non-implanted steers. However, the implant had no effect when Zn-Met was the dietary Zn source. The implant increased (P<0.05) concentrations of Zn in liver of steers supplemented with 200 ppm dietary Zn and reduced Zn in liver of steers fed the control diet. Implanted steers had higher (P<0.05) Cu status and IgG concentrations in serum than non-implanted steers. Steers supplemented with either ZnSO₄ or Zn-Met had greater (P<0.05) concentrations of Zn in liver and plasma than steers fed the control diet. These results indicate both the level and source of Zn supplementation in diets of feedlot cattle affect their response to growth implants.     

The parasitic and lethal effects of Trichoderma longibrachiatum against Heterodera avenae

Heterodera avenae is a devastating plant pathogen that causes significant yield losses in many crops, but there is a lack of scientific information whether this pathogen can be controlled effectively using biocontrol agents. Here we determined the parasitic and lethal effects of Trichoderma longibrachiatum against H. avenae and the possible mechanism involved in this action. Both in vitro and greenhouse experiments were conducted. In vitro, T. longibrachiatum at the concentrations of 1.5 × 10⁴ to 1.5 × 10⁸ spores per ml had a strong parasitic and lethal effect on the cysts of H. avenae, with the concentration of 1.5 × 10⁸ spores per ml having >90% parasitism 18 days after treatments. In greenhouse, T. longibrachiatum inoculation decreased H. avenae infection in wheat (Triticum aestivum) significantly. Observations with microscopes revealed that after mutual recognition with cysts, the spore of T. longibrachiatum germinated with a large number of hyphae, and reproduced rapidly on the surface of cysts. Meanwhile, the cysts surface became uneven, with some cysts producing vacuoles, and the others splitting. Finally the cysts were dissolved by the metabolite of T. longibrachiatum. Chitinase activity increased in the culture filtrates of T. longibrachiatum and reached the maximum 4 days after inoculation in the medium supplemented with colloidal chitin (1.02 U/min per ml) and nematode cysts (0.78 U/min per ml). The parasitism and inhibition of cysts through the increased extracellular chitinase activity serves as the main mechanism with which T. longibrachiatum against H. avenae. In conclusion, T. longibrachiatum has a great potential to be used as a biocontrol agent against H. avenae.