Effects of heavy metal salts on susceptibility of rice seedlings to brown spot disease

Heavy metal salts, known to induce phytoalexin production in different plant species, were used at dilute concentrations (10^-3 to 10^-5 M) in different treatments to control brown spot infection in rice seedlings caused by Drechslera oryzae. Foliage sprays, root-dips for 24 h at the time of transplanting, or seed soaking for 24 h provided moderate to substantial protection to rice seedlings, at different stages after treatment, against artificial inoculation with the pathogen or natural infection under field conditions and mostly caused significant reductions in symptoms. Of the three methods, greatest and most durable protection was achieved with the wet seed treatment. The number of successful infections was appreciably reduced in most treatments and some also inhibited lesion expansion. Ferric chloride and nickel nitrate were the most effective and mercuric chloride the least effective of the chemicals tested. There was no fungitoxicity in leaf diffusates from 3- and 5-wk-old seedlings that had been treated with heavy metal salts and only mild toxicity in that from untreated, inoculated seedlings. However, treated seedlings inoculated at the 3-and 5-wk stages produced leaf diffusates with significantly greater toxicities which were in proportion to their abilities to resist attack.     

3Hthymidine incorporation of rhizosphere bacteria influenced by plant N-status

The effect of plant-root N-status on bacterial growth in the rhizosphere was studied with 5-week-old wheat plants grown in soil with low N content obtained by mixing 9:1 gravel:sandy loam. As a consequence of N limitation, significant increase in³Hthymidine (Tdr) incorporation rate occured 3 days after addition of 30 mM ammonium compared to controls without ammonium. Plants were grown with split-roots to separate the effect of soil N from effect of plant root derived organic matter-N on bacterial activity. The increase in nitrate concentration from 10 mM to 30 mM at one part of the root system led to significant increased³HT dr incorporation in the rhizosphere at the other part of root system after 4 days showing that the composition of root exudates became more favourable for bacterial growth when plants were fertilized with the higher level of nitrate.     

Factors affecting plant regeneration from immature inflorescence of two winter wheat cultivars

Inflorescence explants of two winter wheat cultivars, Triticum durum cv. Kızıltan-91 and T. aestivum cv. Bezostaja-01, were used to evaluate the effects of vernalization period of donor plants, callus age and medium composition on regeneration capacity. Donor plants were grown for 7 d and they were exposed to 4 °C for 1, 2, 3, 4, and 5 weeks. The maximum inflorescence formation was observed as 79 % at 4 weeks and 73 % at 5 weeks of vernalization period for Kızıltan-91 and Bezostaja-01, respectively. Among 6 different callus induction and regeneration mediums, I₁-R₁ and I₃-R₃ have to be the best responding mediums for Kızıltan-91 and Bezostaja-01, respectively. In Kızıltan-91, calli induced from donor plants, vernalized for 3 weeks, showed a significantly lower regeneration capacity than counterparts vernalized for 4 and 5 weeks. The highest regeneration capacity of 69 % was obtained from 6-week-old calli produced from 4 weeks vernalized Kızıltan-91 donor plants. In contrast to Kızıltan-91 cultures, the effects of vernalization period and callus age on regeneration capacity were not significant in Bezostaja-01 cultures. The maximum numbers of tillers were obtained from 6-and 15-week-old calli for Bezostaja-01 and Kızıltan-91, respectively. In contrast to vernalization period of donor plants, callus age had no effect on seed number.     

Effect of salicylic acid and methyl jasmonate on phenylalanine ammonia-lyase activity and total phenol in wheat infected by Pratylenchus thornei

The effect of biochemical responses in wheat seedlings cultivar Falat against lesion nematode (Pratylenchus thornei) was investigated by two factors inducer resistance salicylic acid (SA) and methyl jasmonate (MeJA). Foliar sprays with 1?ml SA 1?mM and 1?ml MeJA 100?μM was conducted on 20-day-old wheat (cv. Falat) seedlings. About 24?h after foliar sprays, plants were inoculated by root lesion nematode. Phenylalanine ammonia-lyase (PAL) activity was increased significantly in all treatments two days after inoculation over control. However, PAL activity was decreased in all treatments five days after inoculation in compared to control. What is more, in both time points, total phenol increased in all treatments over control. The comparison of both inducers SA and MeJA on total phenol in healthy and infected wheat seedlings in both days two and five after inoculation of nematode indicated that total phenol has decreased significantly in all treatments in the fifth day.     

Wheat Resistance to Spot Blotch Potentiated by Silicon

Spot blotch, caused by the fungus Bipolaris sorokiniana, is one of the most important diseases on wheat. The effects of silicon (Si) on this wheat disease were studied. Plants of wheat cultivars BR‐18 and BRS‐208 were grown in plastic pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). The content of Si in leaf tissue was significantly increased by 90.5% for the +Si treatment. There was no significant difference between Si treatments for calcium content, so variations in Si accounted for differences in the level of resistance to spot blotch. The incubation period was significantly increased by 40% for the +Si treatment. The area under spot blotch progress curve, number of lesions per cm² of leaf area, and real disease severity significantly decreased by 62, 36 and 43.5% in +Si treatment. There was no significant effect of Si on lesion size. The role played by total soluble phenolics in the increased resistance to spot blotch of plants from both cultivars supplied with Si was not clear. Plants from cultivar BR‐18 supplied with Si showed the highest values for concentration of lignin‐thioglycolic acid derivatives during the most advanced stages of fungus infection. Chitinase activity was high at the most advanced stages of fungus infection on leaves from both cultivars supplied with Si and may have had an effect on fungus growth based on the reduction of the components of resistance evaluated. Peroxidase activity was found to be high only at 96 h after inoculation of both cultivars supplied with Si. Polyphenoloxidase activity had no apparent effect on resistance regardless of Si treatments. Results revealed that supplying Si to wheat plants can increase resistance against spot blotch.     

Reaction and survival of four types of sunflowers against Sclerotinia sclerotiorum under controlled conditions

Sclerotinia wilt of sunflower caused by Sclerotinia sclerotiorum (Lib.) De Bary is the major disease of sunflower (Helianthus annuus L.) in Iran. North-western areas of Iran demonstrate the most yield losses through the disease. Development of cultivars with adequate genetic resistance is necessary to avoid these losses. Evaluation of sunflower genotypes varies based on researchers and environmental conditions. Meanwhile, greenhouse tests of the sunflower genotypes are more reliable because of controlled conditions during the assessment activities. To study the reaction of the host plants under controlled conditions, three stem inoculation techniques, including mycelium plug (MP), oxalic acid solution (OAS) and wheat seeds infested (IWS) with Sclerotinia mycelium, were employed. Wounded and non-wounded treatments were used in the experiment to find their effect on the disease progress. Four genotypes, including Ghalami (local variety in market), Confeta, Alstar and Master, were inoculated in this study. The factor was the lesion length to evaluate the effectiveness of different inoculation procedures. The lesion length was measured after 3, 7, 10 and 14 days post inoculation. The analysis of variance demonstrated significant differences between IWS and two other methods (MP and OAS), where the IWS produced the longest lesion lengths. In contrast, the non-significant differences between MP and OAS methods might help researchers to employ the pathogenicity factor (oxalic acid) as an alternative inoculum for their studies. Master variety was the most tolerant genotype among the treatments and its viability was 100% even at 14?days after inoculation and incubation. Interestingly, there was no mortality in all cultivars before 7-day incubations, regardless of resistant or susceptible reactions.     

Fungitoxic and Phytotoxic Studies with Essential Oil of Ocimum adscendens

On screening the leaf extracts of some higher plants for their volatile antifungal activity against the test organism Aspergillus flavus, the extract of Ocimum adscendens exhibited the strongest fungitoxicity. The leaves showed maximum, fungitoxicity as compared with other plant parts. The volatile fungitoxic fraction obtained as an essential oil was standardized by various physico-chemical properties. The oil showed its fungicidal nature and broad range of activity at its minimum inhibitory concentration. The oil was thermostable and the toxicity remained unchanged even on autoclaving and on storage for up to 360 days. Moreover, the oil proved more active than some prevalent synthetic fungicides and exhibited no phytotoxic effect onVigna radiata.     

The effect of quinoid and phenolic compounds on urease and dehydrogenase activity and nitrification in soil

Summary 1,4-Naphthoquinone; 2-methyl-1,4-napthoquinone; 2,3-dichlorohydroquinone; 4,6-di-tert.butyl-o-benzoquinone; 4,6-di-tert.butylpyrocatechol and 4-tert.butylpyrocatechol at concentrations of 10 and 20 ppm inhibit mineralization of urea N by reducing urease activity and/or nitrification in soils. Coating of urea with these chemicals was more effective than direct application to the soil. There was no adverse effect of any of the chemicals either on germination or on the growth of wheat plants at concentrations of 20 and 50 ppm added to soil.     

Cross Protection Between Strains of Fusarium oxysporum f. sp. vasinfectum and its Effect on Vascular Resistance Mechanisms

The severity of symptoms induced in cotton plants by stem inoculation with Fusarium oxysporum f. sp. vasinfectum was reduced by inoculation with a non-pathogenic strain (NPS) of the fungus 3 days previously. Symptom reduction was associated with reduced populations of the pathogen in the stele tissue of the stem. This cross-protective response became effective within 24 h of inoculation with the NPS. Although both vascular occlusion and terpenoid aldehyde synthesis were induced by theNPS, the short time required for cross protection to become effective and the duration of the effect, indicated that vascular occlusion rather than fungitoxicity of the terpenoid aldehyde compounds was the mechanism of protection.     

Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection

Tomato spotted wilt virus (TSWV) is one of the most devastating plant viruses and often causes severe crop losses worldwide. Generally, mature plants become more resistant to pathogens, known as adult plant resistance. In this study, we demonstrated a new phenomenon involving developmentally regulated susceptibility of Arabidopsis thaliana to TSWV. We found that Arabidopsis plants become more susceptible to TSWV as plants mature. Most young 3-week-old Arabidopsis were not infected by TSWV. Infection of TSWV in 4-, 5-, and 6-week-old Arabidopsis increased from 9%, 21%, and 25%, respectively, to 100% in 7- to 8-week-old Arabidopsis plants. Different isolates of TSWV and different tospoviruses show a low rate of infection in young Arabidopsis but a high rate in mature plants. When Arabidopsis dcl2/3/4 or rdr1/2/6 mutant plants were inoculated with TSWV, similar results as observed for the wild-type Arabidopsis plants were obtained. A cell-to-cell movement assay showed that the intercellular movement efficiency of TSWV NSm:GFP fusion was significantly higher in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves. Moreover, the expression levels of pectin methylesterase and β-1,3-glucanase, which play critical roles in macromolecule cell-to-cell trafficking, were significantly up-regulated in 8-week-old Arabidopsis leaves compared with 4-week-old Arabidopsis leaves during TSWV infection. To date, this mature plant susceptibility to pathogen infections has rarely been investigated. Thus, the findings presented here should advance our knowledge on the developmentally regulated mature host susceptibility to plant virus infection.     

Proline antioxidant role in the common ice plant subjected to salinity and paraquat treatment inducing oxidative stress

Leaves of 4-week-old (juvenile) and 9-week-old (adult) plants of the halophyte Mesembryanthemum crystallinum L. (the common ice plant), cultured under controlled conditions in the phytotron, were treated with paraquat (0.1 μM), which produces superoxide radical, and (or) paraquat combined with introduction of NaCl (100 mM) or proline (5 mM) into nutrient medium. After a 20-h dark period (23°C), plants were transferred into light (4 h at 54.1 W/m² of photosynthetically active radiation) for stimulation of O°₂⁻ formation in plastids. Activities of antioxidant enzymes, the contents of MDA, H₂O₂, chlorophyll, and free proline were measured in leaves. Plant responses in two age groups, which differed in the type of photosynthesis (juvenile plants had C₃ type of photosynthesis, whereas adult plants were at the transition stage to Crassulacean Acid Metabolism (CAM) photosynthesis), differed in the levels of constitutive proline and proline, induced by NaCl and paraquat, as well as in activities of superoxide dismutase (SOD) and catalase. Changes in SOD activity and proline accumulation in response to paraquat treatment combined with NaCl revealed opposite dependence to accumulation of proline: the more proline accumulated in leaves, the lower activity of the enzyme. In response to paraquat treatment, the content of chlorophylls a and b most drastically declined in juvenile plants. Negative effect of salinity on the content of chlorophylls was lower than that of paraquat and was almost the same in plants of both age groups. Protective effect of exogenous proline was most profound in the case of paraquat treatment. Exogenous proline decreased the rate of lipid peroxidation, the content of superoxide radical and, consequently, SOD activity (almost fivefold), and increased the content of chlorophylls (a and b) in leaves of adult plants. The obtained data suggest that stress-induced accumulation of proline in the common ice plant has both osmoprotectory and antioxidant functions.     

Fast growth responses of barley and durum wheat plants to NaCl- and PEG-treatment: resolving the relative contributions of water deficiency and ion toxicity

Effects of adding osmotically comparable concentrations of PEG and NaCl to nutrient solution were studied in 1-week-old durum wheat (Triticum durum Desf., salt-sensitive) and barley (Hordeum vulgare L., more salt tolerant). Both treatments arrested leaf extension immediately, followed by partial recovery within 40–60 min. Similarities and differences were observed between the responses of the two species and between the two treatments. Typically, PEG-treatment increased leaf extensibility of both barley and wheat as measured by changes in leaf extension rate monitored by an electromechanical sensor to which a stretching counter-weight was added. In barley, NaCl-treatment increased extensibility in a similar way to PEG, but in wheat, extensibility did not change within 40 min of NaCl-treatment. The contrasting fast responses of wheat plants to NaCl and PEG treatments suggests that, unlike barley, wheat leaf extension was sensitive not only to osmotic effects of salinity, but also to ion toxicity. Thus, separate PEG- and NaCl-treatments enabled us to distinguish between ionic and osmotic effects and to show, for the first time, that ionic effects of salinity on leaf extension can commence after only short-term exposure (<1 h) in the case of a salt-sensitive species such as durum wheat.     

Different concentrations of Melia azedarach L. (Meliaceae) ethanolic extract on the developmental time and egg laying of Tetranychus urticae Koch

The use of plant-derived chemicals is a new recommended method to control Tetranychus urticae. According to recent studies, Melia azedarach has several compounds which could be effective on insects and mites. The effect of the extract on the fertility and pre-mature period of the mites were investigated at concentrations of 1, 3, 4 and 5?mg mL^?1 of the extract under laboratory conditions. The effect of chemicals extracted from plants was determined using the spraying bioassay method. The leaves being the location of the mites’activity, were sprayed with ethanol extract, then the mites were placed on the discs. In this experiment, the results show that all concentrations were effective, but extract concentrations 4 and 5?mg mL^?1 caused a reduction of more than 50% in the egg laying of the mites and a significant increase in their pre-mature period.     

Evaluation of fungicides applied to soil to control naturally-occurring take-all using a balanced-incomplete-block design and very small plots

Six sterol biosynthesis-inhibiting fungicides representing several combinations of properties were applied to soil to control naturally-occurring take-all (caused by Gaeumannomyces graminis var. tritici) in winter wheat in field experiments in two successive years. The average take-all severity category was never more than moderate in the different clay-loam and sandy loam sites used in each year. At each site in each year there were six treatments and an untreated control in an arrangement based on a balanced-incomplete-block design for six treatments in 10 blocks each with three treatments. Each block had three treated plots and a control plot and was paired with the complementary block of three treatments (plus control) to form a complete replicate, of which there were 30 per site. Take-all assessments in June or July showed that after incorporation into the seed bed (at 2 kg ha“¹and sometimes at 1 kg ha”¹) in autumn, two non-volatile, strongly lipophilic compounds, nuarimol and triadimenol, with good intrinsic toxicity to the take-all fungus and slow rates of degradation, partially controlled take-all. However, another compound, flutriafol, with similar properties to nuarimol and triadimenol, controlled take-all less. Two slightly volatile, strongly lipophilic compounds, flusilazole and penconazole, with good intrinsic activity, were less effective (at 2 kg ha^-1). A volatile, less lipophilic compound, PP 969, with less intrinsic activity, also partially controlled take-all, but only after application as a drench in the spring (2 kg ha^-1). The most effective treatments were generally more effective the greater the level of disease (as indicated by assessments of disease in control plots), especially in spring assessments of disease. Although flutriafol did not perform as expected, it still seems reasonable to conclude that the requirements for a soil-applied fungicide to control take-all are likely to be: (i) good intrinsic fungitoxicity, (ii) some mobility in soil water (i.e. not strongly lipophilic), and (iii) season-long persistence.     

Efficacy of Selective Gametocides on Lolium perenne L.

The efficacy of six chemicals as selective male gametocideson perennial ryegrass plants was studied by making use of agenetic marker technique. In a preliminary screening experimenteach of the chemicals was used in five concentrations and infive treatments at several stages of development at or nearear emergence. Promising treatments from within this experimentwere assessed in more detail subsequently. In most treatmentsfemale fertility was impaired more than male fertility. In thetreatments in which the desired effect was found, i.e. malefertility was depressed appreciably more than female fertility,the differential gametocidal selectivity of the chemicals couldnot be considered distinct enough to merit application on afield scale.     

Effect of <Emphasis Type="Italic">Trichoderma</Emphasis> spp. isolates for biological control of tan spot of wheat caused by <Emphasis Type="Italic">Pyrenophora tritici-repentis</Emphasis> under field conditions in Argentina

Trichoderma spp. have been used as biocontrol agents to protect plants against foliar diseases in several crops, but information from field assays is scarce. In the present work, experiments were carried out to determine the effect of six isolates of Trichoderma harzianum and one isolate of T. koningii on the incidence and severity of tan spot, caused by Pyrenophora tritici-repentis (anamorph: Drechslera tritici-repentis) under field conditions. Significant differences between years, wheat cultivars and treatments were found. In 2003, two of the isolates assayed (T5, T7) showed the best performance against the disease applied as seed treatments or sprayed onto wheat leaves at different stages. The application of six of the treatments on wheat plants significantly reduced disease severity by 16 to 35% in comparison with the control. Disease control provided by isolate T7 was similar to that provided by the fungicide treatment (56% reduction). This is the first report on the efficacy of Trichoderma spp. against tan spot under field conditions in Argentina.     

Temperature-induced Alterations in the Expression of Susceptibility of Cassava to Colletotrichum gloeosporioides f. sp. manihotis

Cassava plants (Manihot esculenta Crantz) were incubated at 15, 25 or 35 °C after inoculation of green stems with Colletotrichum gloeosporioides (Penz.) f. sp. manihotis Chev. At 25 °C, the cultivar TMS 30211 was less susceptible than the cultivar TMS 30337. At 35 °C, compared to 25 °C, the lesion diameter was reduced in both cultivars, the cassava stem extracts showed an enhanced fungitoxicity, the grem tubes were more sensitive to the stem extracts and a layer of lignified, cells developed earlier around the infected tissues. At 15 °C, the lesions in both cultivars extended more rapidly than at the other temperatures, even though 15 °C, compared to 25 °C, caused a 50% reduction of mycelial growth rate on oat meal agar.     

Reaction of sunflower varieties to Sclerotinia sclerotiorum under various inoculation methods,time of field evaluation and phylogenic stages of host

Sunflower cultivation is affected seriously by Sclerotinia sclerotiorum (Lib.) de Bary in Iran, particularly north-western areas. Because of economic and environmental harms by chemical control, it is necessary to develop cultivars with adequate genetic resistance for reduction of yield losses. The purpose of this study was to find an effective method of inoculation with S. sclerotiorum under field evaluations. Three stem-inoculation techniques including: 1 – mycelium plug, 2 – oxalic acid solution (OAS) and 3 – infested wheat seeds with Sclerotinia mycelium were employed under field conditions. Four genotypes including Ghalami (local variety in market), Confeta, Allstar and Master were used in this study. The lesion length, lesion width and lesion as up and down leading on the stem from inoculation site were measured after 3, 7, 10 and 14 days of inoculation. The analysis of variance showed significant difference between all employed techniques and incubation days after inoculation. Mycelial plug (MP) inoculation technique produced significantly more developed lesions on the treated stems. In spite of this effect, Master variety demonstrated reasonable resistance reaction against the disease. The progress of disease in wounded treatments was also faster than the non-wounded ones. And, the shortest time to obtain significant differences between varieties was 10?days after inoculation. By comparison of results of lesion length at flowering and seed-filling stages, the more obvious effectiveness of the disease was observed at the second stage. Finally, there were negative correlations between mean temperature and mean lesion length in all three inoculation methods.     

Prospects for fungicidal control of take-all of wheat

A range of fungicides and herbicides was tested against Gaeumannomyces graminis (causal agent of take-all) on agar plates, and on wheat seedlings in pots and in liquid culture. Benomyl, the standard in all tests, was consistently most effective: like iprodione, nuarimol and KWG 0599 , it diminished infection from inoculum placed just below the seeds more effectively when applied as a drench than as a soil-mix. Benomyl as a soil-mix was most effective in soils with least organic matter. Some compounds toxic to the pathogen on agar plates and in plants grown in liquid culture were ineffective as soil treatments. The practical limitations of soil treatment with conventional fungicides and application methods are discussed.     

Chitin-supplemented Foliar Application of Serratia marcescens GPS 5 Improves Control of Late Leaf Spot Disease of Groundnut by Activating Defence-related Enzymes

Chitinolytic Serratia marcescens GPS 5 and non‐chitinolytic Pseudomonas aeruginosa GSE 18, with and without supplementation of chitin, were tested for their ability to activate defence‐related enzymes in groundnut leaves. Thirty‐day‐old groundnut (cv. TMV 2) plants pretreated with GPS 5 and GSE 18 (with and without supplementation of 1% colloidal chitin) were challenge inoculated after 24 h with Phaeoisariopsis personata, the causal agent of late leaf spot (LLS) disease of groundnut. GPS 5 and GSE 18, applied as a prophylactic spray, reduced the lesion frequency by 23% and 67%, respectively, compared with control. Chitin supplementation had no effect on the control of LLS by GSE 18, unlike GPS 5, which upon chitin supplementation reduced the lesion frequency by 64%, compared with chitin alone. In a time course study the activities of chitinase, β‐1,3‐glucanase, peroxidase and phenylalanine ammonia lyase were determined for the different treatments. There was an enhanced activity of the four defence‐related enzymes with all the bacterial treatments when compared with phosphate buffer and colloidal chitin‐treated controls. In correlation to disease severity in bacterial treatments, chitin‐supplemented GSE 18 was similar to GSE 18, whereas chitin‐supplemented GPS 5 was much more effective than GPS 5, in activation of the defence‐related enzymes. The high levels of enzyme activities following chitin‐supplemented GPS 5 application continued up to the measured 13 days after pathogen inoculation.