Pathogenesis of barley leaves by Helminthosporium teres I: Green island formation and the possible involvement of cytokinins

Infection sites/green islands were formed in host leaf tissue infected with drops of H. teres. They exhibited higher cytokinin-like activity, sugar and starch than their surrounding tissue and tissue under water drops. The cytokinin-like activity at the infection sites increased from 24 to 72 h of incubation. However, the cytokinin-like activity of the tissue surrounding the infection drops and the tissue under water drops fell from 24 to 72 h incubation. The culture filtrate extracts of the fungus also produced cytokinin-like activity which increased from 1 to 10 days incubation. Application of this culture filtrate extract evoked green island formation. Application of kinetin to host leaves duplicated the green island effect. Thin-layer chromatographic fractions of the tissue extracts and the culture filtrate extracts revealed that a major portion of cytokinin-like activity corresponded to zeatin and zeatin riboside. The presence of zeatin and zeatin riboside (both in tissue and culture filtrate extracts) was confirmed by high performance liquid chromatography. Increases in the amounts of cytokinin-like substances, particularly zeatin and zeatin riboside, attributed to pathogen influence are suggested to be involved in infection and pathogenicity of H. teres.     

Bioassay to detect <Emphasis Type="Italic">Ascophyllum nodosum</Emphasis> extract-induced cytokinin-like activity in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Ascophyllum nodosum, a brown macroalga, is the most widely used seaweed in agriculture. We report a rapid method for the detection of cytokinin-like activity in plants treated with a commercial A. nodosum liquid concentrate (Stimplex®) using a transgenic line of Arabidopsis carrying the ARR5 promoter fused to ß-glucuronidase (GUS) reporter gene. Based on GUS activity assay, an increase in cytokinin-like activity was detected in plants grown in vitro treated with 3 mL L^?1 Stimplex®, whereas foliar spray treatments showed similar cytokinin-like activity at a concentration of 5 mL L^?1. Histochemical staining showed Stimplex®-induced GUS activity in leaf as well as in the root tissues. Taken together, our results suggest that Stimplex® contains compounds that may elicit endogenous cytokinin-like activity. Furthermore, it is shown that this bioassay can be used for rapid screening of extracts that can stimulate cytokinin-like activities using Arabidopsis AAR5::GUS reporter transgenic plants.     

Involvement of carbohydrates and cytokinins in pathogenicity ofHelminthosporium carbonum

Significant stimulation of the number of appressoria, penetration and colonization by conidia ofHelminthosporium carbonum occurred on decolorized maize leaves when exogenous carbohydrates and leaf leachates were added. Germination and germ tube length, however, did not exhibit appreciable differences on decolorized or non-decolorized maize leaves. Lower germination was recorded by leached conidia on decolorized leaves; while appressoria, penetration and colonization were absent. Addition of exogenous nutrients (sucrose>leaf leachates>yeast extract>glucose) enabled conidia to accomplish appressoria, penetration and colonization. Optimum levels for various nutrients observed were 2% (w/v) sucrose/glucose or 0.1% (w/v) yeast extract. Higher concentrations inhibited the infection stages of the pathogen. Depletion of host carbohydrates from green islands/infection sites adversely affect appressoria formation, penetration and colonization; and the loss of carbohydrates from the spore affects germination. Cytokinin-like activity at the infection site/green islands increased with the period of incubation of the host as compared to the surrounding tissue or tissue under water drops. The culture filtrate extracts ofH. carbonum recorded cytokinin-like activity which increased with growth of the fungus. TLC (thin layer chromatography) of cytokinin-like substances (tissue extract and culture filtrate) revealed major activity was confined to Rf zones 0.6 to 0.8 which co-chromatographed with zeatin and zeatin riboside. These substances increase at infection sites by virtue of which carbohydrates accumulate at these sites ensuring a continuous supply to the growing pathogen.     

Polarity of the stem and ontogenetic changes in sunflow (Helianthus annuus L.) leaves in relation to endogenou cytokinins and ethylene

The content of endogenous cytokinin-like substances and the release of ethylene were determined in leaves of different insertion of sunflower plants during their ontogeny. The content of cytokinin-like substances was highest in the leaves on the middle part of the stem (that is in leaves just before full expansion), with a decrease occurring both towards the base and the apex of the stem, when followed at four growth phases (vegetative plants, plants with inflorescence diameter up to 0.5 cm, plants with inflorescence diameter up to 3 cm, and plants in flower). Changes in the content of cytokinin-like substances during the ontogeny of the leaf also corresponded to this pattern. Data obtained with the leaf at the third node from the basis of the stem showed that the level of cytokinin-like substances first sharply increased, and then after reaching maximal value (at the time when leaf blade area reached approximately 70 % of the final value) slowly and continuously decreased. The highest amount of ethylene released from the leaves was recorded in basal leaves and then also in apical leaves, whereas the leaves with the largest blade area situated at the central part of the stem released the lowest amount of ethylene. This pattern was repeatedly found at all four selected growth phases of sunflower plants.     

Screening of endophytic bacteria isolated from leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] for cytokinin-like compounds

Endophytic bacteria are harmless in most plant species; and known to boost the growth and development of the host plants probably by secreting growth hormones. The isolation, identification and screening of endophytic bacteria for the plant growth regulators like cytokinin are needed to get the leads for their applications in agriculture sector. We describe the isolation and identification of the bacterial endophytes from the leaves of Sambung Nyawa [Gynura procumbens (Lour.) Merr.] and their screening for cytokinin-like compounds. We isolated three endophytic bacteria from the leaves of G. procumbens collected from the forest research institute of Malaysia (FRIM). They were further identified using amplified 16S rRNA gene sequence based method of bacterial identification. The ethyl acetate extracts of the isolates-broth were analyzed using cucumber cotyledon greening bioassay (CCGB) to determine the presence of cytokinin-like compounds. Consequently, the bacterial putative endophytes were identified as Psuedomonas resinovorans, Paenibacillus polymaxa, and Acenitobacter calcoaceticus. Broth-extracts from two (Psuedomonas resinovorans and Paenibacillus polymaxa) of the three putative bacterial endophytes show the positive results in their screening for cytokinin-like compounds using CCGB. Thus, we hypothesize that the bacterial putative endophytes of G. procumbens that produce cytokinin-like compounds might have a role in the growth and development of G. procumbens. ABBREVIATIONS: CCGB - Cucumber cotyledon greening bioassay, rDNA - Ribosomal DNA, K12, BAP - 6-Benzylaminopurine, Db1, MSA - Multiple sequence alignment. 8081.     

Effect of preparations exhibiting cytokinin-like activity on the specific density of leaf in grasses

The effects of synthetic preparations exhibiting cytokinin-like activity (6-benzylaminopurine, Thidiazuron, and kartolin-2) on the specific leaf area (SLA) were studied in plants of the family Gramineae (wheat, Triticum aestivum L.; meadow fescue, Festuca pratensis Huds.; and reed fescue, F. arindinacea Schreb.). At the early stages of ontogeny (until the leaf area reached 50-60% of the maximum value), treatment of plants of the three species with cytokinin-like preparations caused an increase in SLA. The SLA value in these plants was correlated with the rate of photosynthetic assimilation of carbon dioxide and activities of carbon metabolism enzymes: ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), NAD-malate dehydrogenase (EC 1.1.1.37), and NADP-glyceraldehydrophosphate dehydrogenase complex, which includes phosphoglycerate kinase (EC 2.7.2.3) and glyceraldehydrophosphate dehydrogenase (EC 1.2.1.13). However, there was no correlation of SLA with the activity of phospho(enol)pyruvate carboxylase (EC 4.1.1.31), an anaplerotic carboxylation enzyme of grasses. SLA is suggested to reflect the state and activity of the photosynthetic apparatus and can be recommended as a characteristic of photosynthesis variability (e.g., caused by cytokinin-like preparations).     

Phytotoxicity analysis of extracts from compost and their ability to inhibit soil-borne pathogenic fungi and reduce root-knot nematodes

Compost extracts are novel organic amendments, typically applied to suppress soil-borne diseases. This research evaluated the phytotoxicity of compost extracts and analyzed their ability to inhibit pathogenic fungal growth and reduce root-knot nematodes. The physical, chemical and biological characteristics of extracts from a pig manure and straw compost were analyzed. Three types of extracts were tested: direct extracts of compost (DEC), aerated fermentation extracts of compost (AFEC) and non-aerated fermentation extracts of compost (NAFEC). All compost extracts showed low phytotoxicity against lettuce and cress, but AFEC and NAFEC were more phytotoxic than DEC. All compost extracts significantly inhibited pathogenic fungal growth except for the fungus Rhizoctonia solania AG4. For two seasons, tomato root biomass of three compost extracts was 1.25–5.67 times greater than CK (water control), and AFEC and NAFEC showed the best tomato root growth promotion. The reduction ratio of root egg mass and density of soil nematodes were 34.51–87.77% and 30.92–51.37%, when applied with three compost extracts. The microbial population in compost extracts was considered to be the most significant factor of inhibition pathogenic fungal growth. No markedly correlations among bacterial community diversity, the inhibition of pathogenic fungal growth and the reduction of root-knot nematodes were observed. This information adds to the understanding of the growth-promoting and suppression effects of compost extracts and will help to enhance crop production.     

Characterization of different compost extracts using Fourier-transform infrared spectroscopy (FTIR) and thermal analysis

Compost extract or “compost tea” is a liquid extract of compost obtained by mixing compost and water for a defined period of time. Compost tea contains nutrients and a range of different organisms and is applied to the soil or directly to plants with the principal aim of suppressing certain plant diseases. In addition, the application of compost tea supplies nutrients and organic matter to the soil. Thermal analysis and Fourier transform infrared spectroscopy (FTIR) are two widely applied analytical techniques for establishing the stability of compost, and although numerous studies have evaluated the capacity of compost tea to suppress plant diseases, there are no studies employing these techniques to characterize compost-tea. For the present study, 12 compost extracts were produced under varying conditions in a purpose-built reactor. Two different composts, an stable compost produced from manure and an unstable compost produced from municipal solid waste, respectively, two aeration systems (aerated and non-aerated extracts) and three temperatures (10, 20 and 30°C) were used in these experiments. The extracts were freeze-dried and subsequently analysed, together with the two composts, by means of FTIR and thermal analysis. Extracts produced from high stability compost, independently of the conditions of aeration and temperature, showed very similar results. In contrast, differences among extracts produced from the unstable compost were more noticeable. However, the different conditions of aeration and temperature during the production of the extracts only explained partially these differences, since the transformations undergone by compost over the 3 months that the experiments lasted were also reflected in the composition of the extracts. In spite of everything, extraction process favoured the degradation of easily oxidizable organic matter, which was more abundant in unstable compost. This degradation was more intense for non-aerated processes, probably due to the longer duration of these (10 days) with respect to aerated extractions (2 days). The effect of temperature was not clear in these experiments, although high temperatures could increase micro organism activity and consequently favour the degradation of easily oxidizable organic matter.     

Comparison of cytokinin- and auxin-like activity in some commercially used seaweed extracts

Six commercially used seaweed extracts were tested for cytokinin- and auxin-like activity using the soybean callus bioassay and the mung bean rooting bioassay respectively. All the seaweed extracts tested showed cytokinin-like activity. Seamac caused the most callus growth with activity being concentrated at R _f 0.9 which co-chromatographed withiso-pentenyladenine and its riboside. The other five seaweed extracts yielded activity at R _f 0.7–0.8. This co-chromatographed with zeatin and zeatin riboside. All the seaweed extracts tested improved rooting of mung beans. Kelpak made fromEcklonia maxima (Osbeck) Papenf. gave the best rooting response.     

Effect of Preparations Exhibiting Cytokinin-like Activity on the Specific Density of Leaf in Grasses

The effects of synthetic preparations exhibiting cytokinin-like activity (6-benzylaminopurine, Thidiazuron, and kartolin-2) on the specific leaf area (SLA) were studied in plants of the family Gramineae (wheat, Triticum aestivum L.; meadow fescue, Festuca pratensis Huds.; and reed fescue, F. arindinacea Schreb.). At the early stages of ontogeny (until the leaf area reached 50–60% of the maximum value), treatment of plants of the three species with cytokinin-like preparations caused an increase in SLA. The SLA value in these plants was correlated with the rate of photosynthetic assimilation of carbon dioxide and activities of carbon metabolism enzymes: ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39), NAD-malate dehydrogenase (EC 1.1.1.37), and NADP-glyceraldehydephosphate dehydrogenase complex, which includes phosphoglycerate kinase (EC 2.7.2.3) and glyceraldehydephosphate dehydrogenase (EC 1.2.1.13). However, there was no correlation of SLA with the activity of phospho(enol)pyruvate carboxylase (EC 4.1.1.31), an anaplerotic carboxylation enzyme of grasses. SLA is suggested to reflect the state and activity of the photosynthetic apparatus and can be recommended as a characteristic of photosynthesis variability (e.g., caused by cytokinin-like preparations).     

The effects of organic amendments on mineral element uptake and fruit quality of raspberries

The organic produce industry is gaining popularity with consumers because of the perception for healthier foods and the environmental benefits of this agricultural practice. Common amendments in organic agriculture include compost and compost tea, the latter being a relatively new product in North America. The main objective of this study was to assess the effectiveness of ruminant and municipal solid waste compost and compost teas made from these composts on selected soil, leaf and fruit parameters of raspberries. Generally, foliar compost tea application was as effective as compost addition in raspberry production. Concentrations of K in leaf and fruits were significantly lower (p?=?0.05) in compost tea treated raspberries. Furthermore, the compost tea increased leaf Na compared to compost amendments which suggested that raspberries preferentially take up Na via foliar applications compared to root Na applications. The yield, total antioxidant capacity of fruit, and vitamin C content of fruit were not affected by treatment but differed greatly among years. Precipitation also varied greatly among years as did soil nutrient concentrations and may have influenced yield, total antioxidant capacity, and vitamin C content.     

Acclimation of brown seaweeds in an outdoor cultivation system and their cytokinin-like activity

Four of the most abundant intertidal eastern Mediterranean brown seaweeds were repeatedly inoculated in an outdoor cultivation system for acclimatization. Diciyopteris membranacea was best acclimated and survived for nine months in the system. D. membranacea growth rate showed a positive relationship with irradiance and temperature during nine months of cultivation. Cystoseira sp., Sargassum vulgare and Padina pavonia were not acclimated and survived for shorter times, presumably because of inadequate conditions. All four species showed a cytokinin-like activity assayed with the barley leaf senescence, and promoted radish seed germination. The cytokinin-like activity in Diciyopteris membranacea was relatively stable throughout the cultivation time, as shown by the barley leaf senescence assay, and it acted as a growth promoter of bean and lettuce seedlings. Partial fractionation of the crude seaweed extract showed an ash active component and a natural cytokinin one, which was presumably composed of minor polar and major nonpolar fractions.     

Regional distribution and in vivo release of tachykinin-like immunoreactivities in rat brain: evidence for regional differences in relative proportions of tachykinins

The regional distribution of various forms of tachykinin-like immunoreactivity (TKLI) was studied in rat brain using radioimmunoassay. TKLI was measured with two different tachykinin-antisera (K12 and E7), which react with neurokinin A (NKA) and neurokinin B (NKB) but not with substance P (SP) and with a specific SP-antiserum. TKLI-K12 and TKLI-E7 were found to have similar regional distributions which were, however, significantly different from that of the substance P-like immunoreactivity (SPLI). Thus, the ratio of the tissue concentrations of TKLI-K12 or TKLI-E7 to that of SPLI was higher in frontal cortex and hippocampus and lower in pons/medulla oblongata than in the other regions studied. Cation-exchange chromatography of neutral water extracts of brain tissue revealed two major immunoreactive components of TKLI-K12 and TKLI-E7, one of which co-eluted with synthetic NKB while the other appeared in the same region as synthetic NKA. The relative quantities of these components varied depending on the brain region studied. No TKLI-K12 or TKLI-E7 co-eluted with synthetic SP. Almost all of the SPLI in acetic acid or water extracts of brain tissue eluted as a single chromatographic component in the same position as synthetic SP. Potassium-stimulated in vivo release of TKLI-K12, TKLI-E7 and SPLI in striatum of rat brain could be demonstrated using intracerebral dialysis. The present results imply that tachykinins, which may serve as neurotransmitters or neuromodulators, are present in different proportions in different regions of rat brain.     

How to safely compost Cameraria ohridella-infested horse chestnut leaf litter on private compost heaps

Abstract:  The horse chestnut leafminer Cameraria ohridella (Lep., Gracillariidae) is a recently introduced invasive pest, which frequently defoliates horse chestnut trees Aesculus hippocastanum already in summer. Currently, the removal of leaf litter, in which pupae of C. ohridella hibernate, is the only effective short-term control measure available. In the past the deposition of pest-infested litter on private compost heaps was dissuaded because of the risk of leafminer emergence in the following spring. Thus, the aim of this study was to test safe ways to compost pest-infested horse chestnut leaf litter on small heaps so that private gardeners can save the costs for leaf disposal. The number of C. ohridella adults emerging from heaps covered with either soil or uninfested foliage was reduced by 96% compared with controls. Only single leafminers emerged from the top covered heaps and there was no difference between the use of soil or uninfested foliage as cover. However, cover did not prevent the emergence of a few leafminers over the sides of the heaps. Overall, we conclude that it is relatively safe to compost C. ohridella -infested A. hippocastanum leaf litter on small compost heaps provided they are properly covered.     

Effects of kelp (Macrocystis integrifolia andEcklonia maxima) foliar applications on bean crop growth and nitrogen nutrition under varying soil moisture regimes

A greenhouse experiment was designed to test the effects of two kelp(Macrocystis integrifolia andEcklonia maxima) concentrates, when prepared as foliar sprays, upon bean (Phaseolus vulgaris) growth and N nutrition under three soil moisture regimes. Plant growth and developmental responses in this greenhouse experiment have demonstrated the effectiveness of the two kelp foliar sprays as plant growth regulating substances. Bean growth and developmental responses to the kelp foliar spray treatments were dependent upon the soil moisture regime to which they have been subjected. Although the two kelp foliar sprays had varying and sometimes contrasting effects on bean growth and N nutrition, which were dependent on the soil moisture treatment, their developmental effects upon the number of nodes, shoot/root ratio, leaf area ratio and specific leaf area were quite similar. A soybean callus bioassay demonstrated the presence of cytokinin-like substances and a callus growth antagonist in the kelp concentrate. Increasing dilution of the kelp concentrate disproportionately reduced the callus growth antagonist relative to the growth promoting or cytokinin-like activity.     

Genotoxicity of the extracts from the compost of the organic and the total municipal garbage using three plant bioassays.

The production of compost is one of the alternatives for the disposal of non-hazardous solid wastes. Compost is used in agriculture and gardening as fertilizer. In the State of Queretaro, Mexico, there is a project to produce compost from the municipal garbage which could be used as a fertilizer. The presence of mutagenic compounds in the compost could be a major disadvantage for the selection of this alternative. For the above reason, this study was initiated as a pilot project to determine the potential mutagenic activity in the compost using three plant bioassays: Tradescantia-micronucleus (Trad-MCN), Tradescantia stamen hair mutations (Trad-SHM) and Allium root anaphase aberrations (AL-RAA). Compost was produced using both aerobic and anaerobic processes from either organic waste (from the residential area) or from the total components of the municipal garbage. Extractions from the compost were done using distilled water and organic solvents and shaking the sample for about 12 h under relatively low temperatures (15-20 degrees C). Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the extracts. Three replicates of each sample were analyzed in each one of the three bioassays. As expected the samples of compost from the total garbage showed a higher genetoxicity than those from organic waste. In conclusion, there are some substances present in the compost capable of inducing genotoxicity in the plant assays and therefore there must be some restrictions for its use as a fertilizer.     

Salmonella regrowth in compost as influenced by substrate (salmonella regrowth in compost)

Composting can eliminate pathogenic organisms, including salmonellae, from sewage sludge. However, if salmonellae are present in the compost at undetectable levels or are inoculated into the compost by infected animals or from other sources, they may regrow presenting a health hazard for certain uses of compost. In this study, we examined dilute mineral-salt extracts of three composts from widely separate composting sites in the United States and found that they supported growth ofSalmonella typhimurium. From kinetic studies of the growth of the organism on these extracts, we concluded that each compost produced on extraction a single water-soluble substrate and that the substrates from the different composts were very similar, if not identical.We thank J. Robert Burge, Statistical Consulting and Analysis, ARS, USDA for invaluable help with the statistical analyses, and Alice V. Gibson of our laboratory for technical help.     

Citrus compost and its water extract for cultivation of melon plants in greenhouse nurseries. Evaluation of nutriactive and biocontrol effects

Two different types of citrus composts, and their water extracts, were tested with regard to their utilisations as partial substitutes for peat in growing media for melon seedlings in greenhouse nurseries. Both compost showed higher plant growth than peat. Compost composed by citrus waste and green residue (C2) showed greater plant growth than compost obtained from the same organic matrices mentioned above further the addition of sludge obtained from citrus industry (C1). Compost C2 showed a greater auxinic effect than C1 and it was the only one that showed cytokinic effect. Both composts also demonstrated a biocontrol effect against Fusarium oxysporum for melon plants: the effects were also higher in C2 than in C1. Higher number of isolated fungi was active against F. oxysporum in compost C2, than compost C1. No different bacterial biocontrol efficacy was observed between both composts. The water extracts of both composts gave lower plant yields than their solid matrices, their relative effects being similar to those of the solid composts (C2 extract gave higher plant yields than the extract from C1). The biocontrol effects of compost water extracts followed the same trend.     

Estimation of maturity of compost from food wastes and agro-residues by multiple regression analysis

The composting process of food wastes and tree cuttings was examined on four composting types composed from two kinds of systems and added mixture of microorganisms. The time courses of 32 parameters in each composting type were observed. The efficient composting system was found to be the static aerated reactor system in comparison with the turning pile one. Using the multiple regression analysis of all the data (159 samples) obtained from this study, some parameters were selected to predict the germination index (GI) value, which was adopted as a marker of compost maturity. For example, using the regression model generated from pH, NH(4)(+) concentration, acid phosphatase activity, and esterase activity of water extracts of the compost, GI value was expressed by the multi-linear regression equation (p<0.0001). High correlations between the measured GI value and the predicted one were made in each type of compost. As a result of these observations, the compost maturity might be predicted by only sensing of the water extract at the composting site without any requirements for a large-size equipment and skill, and this prediction system could contribute to the production of a stable compost in wide-spread use for the recycling market.     

The Role of Uncomposted Materials,Composts, Manures,and Compost Extracts in Reducing Pest and Disease Incidence and Severity in Sustainable Temperate Agricultural and Horticultural Crop Production—A Review

This review evaluates evidence of the impactof uncomposted plant residues, composts, manures, and liquid preparations made from composts (compost extracts and teas) on pest and disease incidence and severity in agricultural and horticultural crop production. Most reports on pest control using such organic amendments relate to tropical or arid climates. The majority of recent work on the use of organic amendments for prevention and control of diseases relates to container-produced plants, particularly ornamentals. However, there is growing interest in the potential for using composts to prevent and control diseases in temperate agricultural and horticultural field crops and information concerning their use and effectiveness is slowly increasing. The impact of uncomposted plant residues, composts, manures, and compost extracts/teas on pests and diseases is discussed in relation to sustainable temperate field and protected cropping systems. The factors affecting efficacy of such organic amendments in preventing and controlling pests and disease are examined and the mechanisms through which control is achieved are described.