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Twenty five years of azadirachtins (1986-2011)

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Azadirachtins are known as phagorepellent natural products from seeds of the neem tree Azadirachta indica A. Juss (Meliaceae), which impede the development of larval insects and sterilize adults. The labors of the past three decades ultimatively lead to a chemical structure, which has been unanimously but independently elaborated by German, British and American authors. This structure is in force ever since 1985/87 and was ultimately confirmed by Veitch & al. in Ley’s group in Cambridge, England, through total synthesis. Its 25th birthday is presently being celebrated. Marrangin (=Azadirachtin L) was discovered in Gießen and chemically identified by Kalinowski and colleagues. It occurs in seeds of the marrango tree, Azadirachta excelsa (Jack) Jacobs. In some insect species but also in mites like Tetranychus urticae, its biological activity is significantly superior to azadirachtin A. The value of these biorational compounds, with their very low vertebrate toxicity, and their low toxicity today is globally recognized. Quite recently, azadirachtin and analogues gain acceptance in veterinary and human medicine.
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Placing biopesticides on the Polish market

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Placing biological plant protection products on the Polish market undergo completely different rules depending on the group they do belong: if they are micro or macro organisms. The microorganisms and viruses undergo a very strict, costly and time consuming registration procedure more or less uniform for all EU countries. Placing products belonging to this group on the Polish market requires very significant financial and time investment. The rules regarding macro organisms registration are established in EU on the member state level. In Poland at present there are no rules at all and the macro organisms used in biological control can be imported and used in Poland without any supervision. However the list of 25 biological products registered under the old regulations can be still used as a guide informing about safety and efficacy of some products.
Many species of Trichoderma produce secondary metabolites such as volatile organic compounds (VOCs) that reduce plant diseases and promote their growth. In this work we evaluated the antagonistic effects of VOCs released by eight strains of two Trichoderma species against Pyrenophora teres Drechsler, the causal agent of barley net blotch. Antagonism was estimated based on the percentage of mycelial growth inhibition according to the confronted cultures method. VOCs extraction and identification were performed by gas chromatography and mass spectrometry, through different methodologies for VOCs emitted by antagonists and pathogens alone or when confronted. VOCs produced by all Trichoderma strains inhibited mycelial growth of the pathogen in a range of 3 to 32%, showing weak and unpigmented mycelia with vacuolization. In addition, P. teres stimulated the release of VOCs by both Trichoderma species. The major groups of VOCs detected were sesquiterpenes, followed by diterpenes, terpenoids and eight-carbon compounds. This is the first report about characterization of volatiles emitted by Trichoderma in the presence of P. teres.
Freshly prepared garlic (Allium sativum L.) juice, containing the antimicrobial allicin, was evaluated as a possible grain pro-tectant against the maize weevil, Sitophilus zeamais (Motsch.). Each experiment was set out in Completely Randomized Design (CRD) with four replications, and there was a control treatment. Adult mortality and weight loss percentage were investigated. There was an observed increase in adult mortality following days of exposure in all treatments. Statistically significant (p < 0.05) reduced grain loss was also observed in all the treatments when compared with the control. The juice samples were freshly prepared from an indigenous Nigerian garlic cultivar (GUN) and a cultivar purchased from a supermarket in Germany (GAG). These garlic juice samples exhibited lethal effects causing at least 90% adult mortality in contact toxicity tests. The amount of allicin in GUN was 1.88 mg/ml according to High Pressure Liquids Chromatography (HPLC) analysis, while the amount of allicin in GAG was 3.50 mg/ml. This study highlights the potential of A. sativum containing allicin for biorational control of maize grains against S. zeamais infestation and damage.
Biocontrol of plant diseases has emerged as an eco-friendly measure of plant protection and has experienced a lot of devotion in the last two decades. Biocontrol agents include application of microbial agents, their secretion products and natural extracts from different parts of several plants. The present study, therefore, aimed at evaluating the potency of aqueous extracts of Tagetes erecta L. (marigold) in controlling bacterial speck disease in tomato plants. The experimental design consisted of two groups of 50 plants each: group 1 – sprayed with sterile water (control); and group 2 – sprayed with marigold extract. Spraying was performed under aseptic conditions at the third node from the base of each plant. Challenge inoculation with the bacterial speck pathogen Pseudomonas syringae pv. tomato was performed to analyze the disease severity on the test plants. The parameters of study were analysis of alteration in the activity and gene expression of peroxidase (POX), phenyl ammonia lyase (PAL), and polyphenol oxidase (PPO), as well as isoform expression of POX and PPO. The results demonstrated strong inductive effects of the extract on the activity and genes of POX, PAL and PPO. De novo expression of POX and PPO isoforms following marigold extract treatment was also observed. The observations indicate that marigold extract could be a promising biopesticide.
The two-spotted spider mite (Tetranychus urticae Koch) is an important pest of many horticultural crops. A study was conducted to evaluate the effect of three fluorescent Pseudomonas isolates obtained from rhizospheric soil of tomato (Solanum lycopersicon) in Agadir, Morocco: Q110B, Q036B and Q172B, as potential biological control agents for T. urticae. Both acaricide and repellent activities were assessed on homogenous adult mites. The acaricidal activity test evaluated five concentrations of bacterial suspensions: 0 (control), 102, 104, 106, 108, and 1010 cfu ⋅ ml–1, while only the 1010 cfu ∙ ml−1 concentration of each bacterium was used for the repellent bioassay. The mortality rate and repellentindex were recorded 24, 48 and 72 h after application. Results indicated that the survival rate of T. urticae was reduced (p ≤ 0.01) by all three bacterial isolates compared to control. Within the 24–72 h time period the mortality rates ranged from 8 to 87%, 16 to 99%, and 13 to 89%, for Q110B, Q036B and Q172B isolates, respectively. The isolate Q036B (LC50 = 0.598 cfu ⋅ ml–1) provided higher mortality rates than Q172B and Q110B with LC50 values of 90,846 and 169,585 cfu ⋅ ml–1, respectively. Repellent activity was also the highest with Pseudomonas Q036B having a 71% repellence index at 48 h after application. Regarding the mechanism of action, all three isolates produced hydrogen cyanide, and exhibited protease and cellulose activities, although only Q036B and Q172B had potential chitinase action. Identification analysis showed that the isolates were either Pseudomonas putida (Q172B) or P. fluorescens (Q110B and Q036B). Our results indicate that the P. fluorescens isolate Q036B is a promising candidate for biological control of T. urticae, and has potential to contribute to an integrated pest management program to control this important pest. Then the fruits produced will be qualified as safe for consumers and the environment. The present work was customized to give support for policy decision makers as an agroecological potential meeting needs of industries and ecological balance.
Changes in activity of the grain aphid peroxidase (Px) and polyphenol oxidase (PPO) towards phenolics isolated from leaves of black currant, sour cherry and walnut were examined. Slight increase in activity of peroxidase was found within insect tissues after 24 h of walnut extract treatment, whereas black currant and sour cherry strongly inhibited activity of this enzyme. Later on, the walnut extract reduced activity of the enzyme, finally about 30%. The other extracts showed slight changes in Px activity. The grain aphid’s polyphenol oxidase was stimulated during the first 24 h of the experiment. Further treatment with the phenolics extracts reduced activity of the grain aphid polyphenol oxidase. Generally, phenolics isolated from the black currant and sour cherry were more effective in reducing activity of the aphid peroxidase, whereas phenolics from walnut reduced the polyphenol oxidase activity. Possible application of the phenolics isolated from the woody plants as modern biopesticides towards the grain aphid is discussed.
Solanaceous plants have a great economic impact in Egypt. These groups of plants include potatoes, tomatoes and eggplants. The new invasive pest of tomatoes, Tuta absoluta (Meyrick) causes the greatest crop losses which can range from 60 to 100%. After its detection in Egypt during the last half of 2009, it spread quickly to all provinces in the country. We aiming to propose a sustainable control program for this devastating pest. In this research we tested three groups of control agents. The first was microbial and natural, the second – plant extracts and the third – chemical insecticides. Our results showed that the impact of T. absoluta can be greatly reduced by the use of sustainable control measures represented by different insecticide groups. Bioassay experiments showed that this devastating pest can be controlled with some compounds that give high mortality rates. Of these compounds, spinosad and Beauveria bassiana, microbial control agents, followed by azadirachtin, gave the best results in controlling T. absoluta. Of the chemical insecticides, lambda-cyhalotrin was the most effective, followed by lufenuron and profenofos. In conclusion we encourage farmers to use microbial and natural control measures in combating the tomato leafminer, T. absoluta, in Integrated Pest Mangement (IPM) programs.
Effect of plant phenolics isolated from leaves of blackcurrant (Ribes nigrum L.) and sour cherry (Prunus cerasus L.) as potential biopesticides against the grain aphid was examined. The extracts from the sour cherry contained a twice higher level of salicylic acid than extract from leaves of the blackcurrant. Moreover, concentration of chlorogenic acid was higher in sour cherry and the content of tannic acid within both plant tissues was at the same level. Both examined extracts had a negative effect on development of the grain aphid when the aphid fed on wheat plants sprayed with 1.0%, 2.5% and 5.0% solutions. The length of prereproductive period (time from birth to maturity) of the aphids was extended from 5.9 days on control plants (not sprayed) to 8.1 days, when the aphids were fed on the treated plants. Apterous females of Sitobion avenae fed on sprayed plants showed lower fecundity and lower values of intrinsic rate of natural increase.
Our research provides novel information concerning the insecticidal activity of Brassica alba mustard oil applied to the intestinal tract via insects’ diet against pests from the order Lepidoptera: Cydia pomonella, Dendrolimus pini, and Spodoptera exigua. The LC50 value of the oil against C. pomonella was 0.422 mg ⋅ ml–1. The LC50 of the plant oil against D. pini was 11.74 mg ⋅ ml–1. The LC50 of the botanical product against S. exigua was 11.66 mg ⋅ ml–1. The plant substance was the most active against C. pomonella in comparison with D. pini and S. exigua. The LC50 values of the oil against D. pini and S. exigua were similar. The plant oil exhibited high insecticidal activity against pests from the order Lepidoptera and may prove to be an effective biopesticide.
Recent progress in plant transformation for insect resistance has increased the interest in the potential toxicity of lectins to insect pests. The aim of the present study was to examine the effects of phytohemagglutinin (PHA) on survival, development and fecundity of the bird cherry-oat aphid, when tested on artificial diet. The laboratory tests did not reveal any phagostimulating properties of lectin PHA to R. padi, although the presence of the PHA in the diet decreased fecundity of the adult aphids compared to aphids fed on control diets. Tested lectin had a negative effect on the insects’ weight and induced their mortality. The data presented here suggest that PHA possesses a high insecticidal activity towards the bird cherry-oat aphid and might be considered as protein biopesticide against that aphid pest.
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