Rapid Identification and Quantification of Adulteration in Methyl Eugenol using Raman Spectroscopy Coupled with Multivariate Data Analysis.

Identification of adulterants in commercial samples of methyl eugenol is necessary because it is a botanical insecticide, a tephritid male attractant lure that is used to attract and kill invasive pests such as oriental fruit flies and melon flies on crops. In this study, Raman spectroscopy was used to qualitatively and quantitatively assess commercial methyl eugenol along with adulterants. For this purpose, commercial methyl eugenol was adulterated with different concentrations of xylene. The Raman spectral features of methyl eugenol and xylene in liquid formulations were examined, and Raman peaks were identified as associated with the methyl eugenol and adulterant. Principal component analysis (PCA) and partial least-squares regression analysis (PLSR) have been used to qualitatively and quantitatively analyze the Raman spectral features. PCA was applied to differentiate Raman spectral data for various concentrations of methyl eugenol and xylene. Additionally, PLSR has been used to develop a predictive model to observe a quantitative relationship between various concentrations of adulterated methyl eugenol and their Raman spectral data sets. The root-mean-square errors of calibration and prediction were calculated using this model, and the results were found to be 1.90 and 3.86, respectively. The goodness of fit of the PLSR model is found to be 0.99. The proposed approach showed excellent potential for the rapid, quantitative detection of adulterants in methyl eugenol, and it may be applied to the analysis of a range of pesticide products.

Systemic resistance induced in tomato plants by Beauveria bassiana-derived proteins against tomato yellow leaf curl virus and aphid Myzus persicae.

BACKGROUND: Tomato (Solanum lycopersicum L.) is an economically important vegetable crop around the globe. Tomato yellow leaf curling (TYLC) is the most devastating viral disease posing a serious threat to tomato production throughout the tropical and subtropical world. Induction of microbe-mediated systemic resistance in plants has been of great interest in recent years as a novel microbiological tool in disease and insect pest management. This in-vitro study aimed to determine the effectiveness of different strains (BB252, BB72 and ARSEF-2860) of a hypocreal fungus Beauveria bassiana against TYLCV disease and aphid Myzus persicae. Potted tomato plants exogenously treated with conidial and filtrate suspensions of B. bassiana strains and of their partially purified or purified proteins were exposed to TYLCV inoculum and aphid M. persicae. RESULTS: Results showed a significant suppression of TYLCV disease severity index by the exogenous application of conidial, filtrate and protein treatments of all B. bassiana strains and this response was directly proportional to the treatment concentration. Similarly, mean fecundity rate of M. persicae was also significantly reduced by the highest concentration of ARSEF-2860-derived elicitor protein PeBb1, followed by the highest concentrations of BB252- and BB72-derived partially purified proteins. Moreover, these B. bassiana-derived proteins also caused a significant upregulation of most of the plant immune marker genes associated with plant defense. CONCLUSION: Overall, the study findings suggest that these B. bassiana strains and their partially purified or purified elicitor proteins could be effective biological tools for the management of TYLCV and aphid infestation on tomato plants. © 2023 Society of Chemical Industry.

Sublethal and transgenerational effects of synthetic insecticides on the biological parameters and functional response of Coccinella septempunctata (Coleoptera: Coccinellidae) under laboratory conditions.

Synthetic insecticides have been an inevitable part of plant protection throughout the world. Sublethal effects of these chemicals on beneficial insect species are one of the contemporary issues these days. Using the age-stage, two-sex life table model, this study evaluated the sublethal and transgenerational effects of six synthetic insecticides (imidacloprid, thiamethoxam, lambda-cyhalothrin, cypermethrin, chlorpyrifos and profenofos) commonly applied to winter vegetables, on the fitness and predation of the seven-spotted ladybeetle, Coccinella septempunctata, which is an efficient predator of aphids worldwide. According to results, all insecticides at their sublethal doses (LC30) significantly suppressed the emergence of adults, adult weight, fertility and fecundity of the parental generation compared to control treatment. The larval stage was prolonged and oviposition, fecundity and total longevity of the adult beetles were decreased in unexposed progeny whose parents were exposed to sublethal doses of all insecticides. Moreover, the biological parameters of adults, including the intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R 0) were significantly reduced when exposed to sublethal doses of insecticides. The predation rate of the F1 generation adults was also decreased after exposure to the sublethal doses of insecticides. However, chlorpyrifos, profenofos, lambda-cyhalothrin and cypermethrin exhibited more deleterious effects on the fitness and population parameters of beetles than imidacloprid and thiamethoxam.

Bacterial Symbionts Confer Thermal Tolerance to Cereal Aphids Rhopalosiphum padi and Sitobion avenae.

High-temperature events are evidenced to exert significant influence on the population performance and thermal biology of insects, such as aphids. However, it is not yet clear whether the bacterial symbionts of insects mediate the thermal tolerance traits of their hosts. This study is intended to assess the putative association among the chronic and acute thermal tolerance of two cereal aphid species, Rhopalosiphum padi (L.) and Sitobion avenae (F.), and the abundance of their bacterial symbionts. The clones of aphids were collected randomly from different fields of wheat crops and were maintained under laboratory conditions. Basal and acclimated CTmax and chronic thermal tolerance indices were measured for 5-day-old apterous aphid individuals and the abundance (gene copy numbers) of aphid-specific and total (16S rRNA) bacterial symbionts were determined using real-time RT-qPCR. The results reveal that R. padi individuals were more temperature tolerant under chronic exposure to 31 °C and also exhibited about 1.0 °C higher acclimated and basal CTmax values than those of S. avenae. Moreover, a significantly higher bacterial symbionts' gene abundance was recorded in temperature-tolerant aphid individuals than the susceptible ones for both aphid species. Although total bacterial (16S rRNA) abundance per aphid was higher in S. avenae than R. padi, the gene abundance of aphid-specific bacterial symbionts was nearly alike for both of the aphid species. Nevertheless, basal and acclimated CTmax values were positively and significantly associated with the gene abundance of total symbiont density, Buchnera aphidicola, Serratia symbiotica, Hamilton defensa, Regiella insecticola and Spiroplasma spp. for R. padi, and with the total symbiont density, total bacteria (16S rRNA) and with all aphid-specific bacterial symbionts (except Spiroplasma spp.) for S. avenae. The overall study results corroborate the potential role of the bacterial symbionts of aphids in conferring thermal tolerance to their hosts.

Putative Role of a Yet Uncharacterized Protein Elicitor PeBb1 Derived from Beauveria bassiana ARSEF 2860 Strain against Myzus persicae (Homoptera: Aphididae) in Brassica rapa ssp. pekinensis.

This study reports the characterization of protein elicitor PeBb1 derived from entomopathogenic fungus Beauveria bassiana ARSEF-2860 strain and its putative role in induced systemic resistance in Brassica rapa ssp. pekinensis against green peach aphid Myzus persicae. The sequence of purified elicitor protein was matched with the genomic sequence of a hypothetical protein BBA_10269 from B. bassiana ARSEF-2860 (GenBank Accession No. XP_008603588.1). The protein-encoding gene PeBb1 contained 534 bp cDNA encoding a polypeptide of 177 amino acids with a molecular mass of 19 kDa. The recombinant elicitor protein was expressed in Escherichia coli using pET-28a (+) expression vector and induced necrosis in the leaves of tobacco. The effects of elicitor protein on aphid M. persicae was determined by applying three different concentrations of PeBb1 (i.e., 26, 35, 53 µM) on B. rapa plants at 4-leaf stage and the treated plants were exposed to newly emerged (0-6 h old) apterous adult aphids. Bioassay results showed significant (p < 0.05) sub-lethal effects of the exogenous application of PeBb1 elicitor on M. persicae. Moreover, the RT-qPCR gene expression analyses showed a significant up-regulation of most of the key genes linked to ethylene (ET)- and jasmonic acid (JA)-associated plant defense pathways in elicitor-treated plants. These results not only recommend the putative utilization of PeBb1 elicitor protein in future biological pest control strategies against phloem-feeding insect pests such as M. persicae, but also help in better comprehension of the mechanisms through which beneficial fungi trigger the induced plant resistance.

Anti-insect activity of a partially purified protein derived from the entomopathogenic fungus Lecanicillium lecanii (Zimmermann) and its putative role in a tomato defense mechanism against green peach aphid.

Many biotrophic and necrotrophic fungi synthesize proteins that may elicit induced plant resistance against different herbivore pests. This in-vitro study elucidates the sub-lethal effect of a partially-purified protein derived from the entomopathogenic fungus Lecanicillium lecanii (Zimmerman) (Hypocreales: Clavicipitaceae) against green peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae), an economically important pest of many solanaceous crops including tomato. Bioassays were conducted to determine the impact of different concentrations of protein (i.e. 0.018, 0.036 and 0.054 µM) on the survival and fecundity of M. persicae on tomato (Lycopersicon esculentum) plants. Moreover, the potential role of this exogenous protein in the plant defense mechanism was assessed by expression analyses of key genes associated with salicylic acid (SA) and jasmonic acid (JA) pathways using RT-qPCR. The results indicated a significant negative effect of all protein concentrations on the survivorship and fecundity of M. persicae. The highest concentration (0.054 µM) resulted in lowest survival (46%) of aphids at 7th day post-treatment, while two other concentrations (0.036 and 0.018 µM) resulted in 61 and 71% survival rate, respectively. Similarly, lowest and highest mean fecundity rates were recorded for the highest protein concentration and the control (1.5 and 2.4 nymphs day-1 female-1), respectively. Moreover, L. lecanii-derived protein strongly upregulated the SA associated genes PR1, BGL2 and PAL, and moderately upregulated the JA associated genes LOX, AOS and AOC in protein-treated tomato plants compared to the control plants. These findings demonstrate the systemic resistance induced in tomato plants against M. persicae by the exogenous application of partially-purified protein extracted from L. lecanii, suggesting its further purification and characterization as a novel biological pest management tool against aphids and other phloem-feeding insect pests.

Insights into the Maternal Ancestry of Côte d'Ivoire Honeybees Using the Intergenic Region COI-COII.

Honeybee populations in Côte d'Ivoire have been previously identified as belonging to one subspecies, Apis mellifera scutellata, but other studies have since reported a mixed population consisting of A. m. adansonii and A. m. jemenitica. The population structure and the geographic distribution of honeybees in Côte d'Ivoire remain unclear. This study aimed to profile the population structure of honeybees and their biogeography in Côte d'Ivoire. A total of 33 honeybee colonies were sampled from 15 localities to investigate the maternal ancestry of indigenous honeybee populations using the DraI COI-COII mtDNA test. The results revealed that the honeybee population in Côte d'Ivoire is composed of African haplotypes, all belonging to the AI sublineage. Haplotypes A1 and A4 were recorded with five new sequence variants, including three types of haplotype A1 and two types of haplotype A4. The A1e variant was the most frequent in the A. m. adansonii distributional area. The distribution of the haplotype variants was correlated with the climate pattern in Côte d'Ivoire. This is the first study in Côte d'Ivoire that gives insights into the biogeography and mitotype structure of the local honeybee populations.

Molecular and Functional Characterization of Elicitor PeBC1 Extracted from Botrytis cinerea Involved in the Induction of Resistance against Green Peach Aphid (Myzus persicae) in Common Beans (Phaseolus vulgaris L.).

Elicitors are biofactors that induce resistance in plants against different insect pests. This in vitro study evaluated the impact of a novel elicitor protein PeBC1, extracted from a necrotrophic fungus Botrytis cinerea, on the development and fecundity parameters of green peach aphid (Myzus persicae) on common beans (Phaseolus vulgaris L.). Three different concentrations of PeBC1 elicitor (i.e., 33.56, 25.43, 19.33 µg mL-1) were applied at three different temperature regimes (i.e., 18, 21, and 25 °C). Elicitor treatments were applied topically on the bean plants at 3-leaf stage and newly emerged (0⁻6 h old) apterous adult aphids were exposed to these treated leaves. In addition to the biological parameters of aphids, the relative expression levels of key genes associated with jasmonic acid (JA) and salicylic acid (SA) plant defense pathways were also determined through RT-qPCR. Results of bioassays revealed that the application of PeBC1 elicitor protein exhibited pronounced and significant (p < 0.05) sub-lethal effects on green peach aphids. The fecundity was reduced and the nymphal development time was prolonged by different concentrations of PeBC1 elicitor and temperature regimes. Gene expression studies showed that the exogenous application of PeBC1 induced a significant upregulation of the expression levels of JA and SA pathway-associated genes in bean plants. As compared to control, elicitor-treated plants exhibited an induced resistance against aphids. Our findings suggest the potential use of PeBC1 elicitor protein in future bio-intensive management strategies against sap-sucking insect pests such as green peach aphids.

Farmer field school's training on knowledge level of citrus growers regarding improved production practices / Impacto do treinamento da escola de campo para agricultores quanto ao nível de conhecimento dos produtores de citros em relação às melhores práticas de produção

ABSTRACT: The major objective of this paper was to assess the impact of the Farmer field school (FFS) training program on the knowledge level of farmers (citrus growers) regarding improved citrus production practices. Distract Sargodha was the targeted research area for this study being the leading producer of citrus in Pakistan. Quantitative data were collected through interview schedule and analyzed through Statistical Package for Social Sciences (SPSS). It was conclude that young people were more tend towards getting training under FFS program than old age farmers. Educated farmers were in majority in FFS farmers while; conversely illiterate farmers were in majority in the category of non-FFs farmers. In category of FFS farmers livestock farming was their primary income as reported by majority (31.3%) of FFS farmers and in the case of non-FFS farmer's crop sale was their primary income source as reported by 28.4% of non-FFs farmers. In addition to that both FFS and non-FFS farmers also earn income from nonfarm sources like business, job, foreign remittances and labour. FFS training program had positive and significant impact on the knowledge level of farmers regarding citrus production and protection practices. T-test statistics showed highly significant (P<0.05) difference in knowledge level of FFS farmers and non-FFS farmers.

RESUMO: O objetivo principal deste trabalho foi avaliar o impacto do programa de treinamento da escola de campo para agricultores (FFS) quanto ao nível de conhecimento dos agricultores (produtores de citros) em relação às melhores práticas de produção de citros. O Distrito de Sargodha foi a área de pesquisa escolhida para este estudo por ser o principal produtor de citros no Paquistão. Os dados quantitativos foram coletados por meio de entrevistas e analisados por meio do Statistical Package for Social Sciences (SPSS). Conclui-se que os jovens eram mais tendenciosos à obtenção de treinamento através do programa FFS do que os agricultores mais velhos. Os agricultores escolarizados estavam, em sua maioria, na categoria de agricultores FFS, enquanto que os agricultores analfabetos estavam, na categoria de não FF. Na categoria de FFS, a pecuária se mostrou o principal rendimento, conforme relatado pela maioria dos agricultores (31,3%), enquanto que, em 28% dos agricultores não-FFs, a principal fonte de renda era a venda de suas colheitas. Além disso, tanto os agricultores FFS como não FFS também ganham renda com fontes não agrícolas. O programa de treinamento FFS teve impacto positivo e significativo sobre o nível de conhecimento dos agricultores em relação às práticas de produção e proteção de citrus. O teste-T mostrou diferença altamente significativa (P<0,05) no nível de conhecimento dos agricultores FFS dos não-FFS.

Nitrous Oxide (N2O) Emissions by Termites: Does the Feeding Guild Matter?

In the tropics, termites are major players in the mineralization of organic matter leading to the production of greenhouse gases including nitrous oxide (N2O). Termites have a wide trophic diversity and their N-metabolism depends on the feeding guild. This study assessed the extent to which N2O emission levels were determined by termite feeding guild and tested the hypothesis that termite species feeding on a diet rich in N emit higher levels of N2O than those feeding on a diet low in N. An in-vitro incubation approach was used to determine the levels of N2O production in 14 termite species belonging to different feeding guilds, collected from a wide range of biomes. Fungus-growing and soil-feeding termites emit N2O. The N2O production levels varied considerably, ranging from 13.14 to 117.62 ng N2O-N d(-1) (g dry wt.)(-1) for soil-feeding species, with Cubitermes spp. having the highest production levels, and from 39.61 to 65.61 ng N2O-N d(-1) (g dry wt.)(-1) for fungus-growing species. Wood-feeding termites were net N2O consumers rather than N2O producers with a consumption ranging from 16.09 to 45.22 ng N2O-N d(-1) (g dry wt.)(-1). Incubating live termites together with their mound increased the levels of N2O production by between 6 and 13 fold for soil-feeders, with the highest increase in Capritermes capricornis, and between 14 and 34 fold for fungus-growers, with the highest increase in Macrotermes muelleri. Ammonia-oxidizing (amoA-AOB and amoA-AOA) and denitrifying (nirK, nirS, nosZ) gene markers were detected in the guts of all termite species studied. No correlation was found between the abundance of these marker genes and the levels of N2O production from different feeding guilds. Overall, these results support the hypothesis that N2O production rates were higher in termites feeding on substrates with higher N content, such as soil and fungi, compared to those feeding on N-poor wood.

Characterization of N2O emission and associated bacterial communities from the gut of wood-feeding termite Nasutitermes voeltzkowi.

Xylophagous termites rely on nitrogen deficient foodstuff with a low C/N ratio. Most research work has focused on nitrogen fixation in termites highlighting important inflow and assimilation of atmospheric nitrogen into their bodies fundamentally geared up by their intestinal microbial symbionts. Most of termite body nitrogen is of atmospheric origin, and microbially aided nitrification is the principal source of this nitrogen acquisition, but contrarily, the information regarding potent denitrification process is very scarce and poorly known, although the termite gut is considered to carry all favorable criteria necessary for microbial denitrification. Therefore, in this study, it is hypothesized that whether nitrification and denitrification processes coexist in intestinal milieu of xylophagous termites or not, and if yes, then is there any link between the denitrification product, i.e., N2O and nitrogen content of the food substrate, and moreover where these bacterial communities are found along the length of termite gut. To answer these questions, we measured in vivo N2O emission by Nasutitermes voeltzkowi (Nasutitermitinae) maintained on different substrates with varying C/N ratio, and also, molecular techniques were applied to study the diversity (DGGE) and density (qPCR) of bacterial communities in anterior and posterior gut portions. Rersults revealed that xylophagous termites emit feeble amount of N2O and molecular studies confirmed this finding by illustrating the presence of an ample density of N2O-reductase (nosZ) gene in the intestinal tract of these termites. Furthermore, intestinal bacterial communities of these termites were found more dense and diverse in posterior than anterior portion of the gut.