Assessment of housekeeping genes stability for gene transcription regulation analysis of Spodoptera littoralis (Lepidoptera: Noctuidae) under Spodoptera littoralis nucleopolyhedrovirus viral infection.

BACKGROUND: Normalization with respect to stable housekeeping genes is important to facilitate gene transcription regulation research and acquire more accurate quantitative polymerase chain reaction (qPCR) data. In the current study, five candidates housekeeping genes of the cotton leafworm, Spodoptera littoralis encoding for Actin (Actin), elongation factor 1-alpha (EF1α), ribosomal protein S3 (RPS3), ribosomal protein 49 (RP49), and Ubiquitin (Ubi), were evaluated as normalization housekeeping genes under Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) viral infection. METHODS AND RESULTS: The qPCR results confirmed the expression of all five housekeeping genes in S. littoralis viral infected larvae. The expression profiles of the housekeeping genes showed that the EF1α, Actin, and RP49 had the minimum average Ct values of 18.41 ± 0.66, 18.84 ± 0.90 and 19.01 ± 0.87 in all infected samples, respectively. While RPS3 and Ubi showed the maximum average Ct of 21.61 ± 0.51 and 21.11 ± 0.82, respectively. According to the results of ΔCt and geNorm analysis, EF1α was ranked as the most stable housekeeping gene during infection time-course. While by using BestKeeper, geNorm and NormFinder, the Ubi, RP49, and RPS3 showed the most genes transcription stability. The obtained results were also validated using the Cytochrome c oxidase (COX) gene transcripts in response to SpliNPV infection. CONCLUSIONS: The results revealed that EF1α and Ubi were the most stable housekeeping genes to be used for normalizing S. littoralis gene transcription regulation under SpliNPV infection. These findings, provide a significant addition for gene transcription regulation studies of S. littoralis upon infection using SpliNPV as a bio-agent.

The ability of selected fungal strains to produce carboxylesterase enzymes for biodegradation and use of bifenthrin insecticide as carbon source: in vitro and in silico approaches.

Bifenthrin (BF) is a broad-spectrum type I pyrethroid insecticide that acts on insects by impairing the nervous system and inhibiting ATPase activity, and it has toxic effects on non-target organisms and high persistence in the environment. This study aimed to determine the potential of six different fungi, including Pseudozyma hubeiensis PA, Trichoderma reesei PF, Trichoderma koningiopsis PD, Purpureocillium lilacinum ACE3, Talaromyces pinophilus ACE4, and Aspergillus niger AJ-F3, to degrade BF. Three different concentrations of BF, including 0.1%, 0.2%, and 0.3% w/v, were used in the sensitivity testing that revealed a significant (p ≤ 0.01) impact of BF on fungal growth. Enzymatic assays demonstrated that both intracellular and extracellular carboxylesterases hydrolyzed BF with the enzymatic activity of up to 175 ± 3 U (µmol/min) and 45 ± 1 U, respectively. All tested fungi were capable of utilizing BF as a sole carbon source producing 0.06 ± 0.01 to 0.45 ± 0.01 mg dry biomass per mg BF. Moreover, the presence of PytH was determined in the fungi using bioinformatics tools and was found in A. niger, T. pinophilus, T. reesei, and P. lilacinum. 3D structures of the PytH homologs were predicted using AlphaFold2, and their intermolecular interactions with pyrethroids were determined using MOE. All the homologs interacted with different pyrethroids with a binding energy of lesser than - 10 kcal/mol. Based on the study, it was concluded that the investigated fungi have a greater potential for the biodegradation of BF.

Sublethal concentrations of conventional nematicides alter the physiological activities of Meloidogyne incognita and suppress parasitism.

Reducing nematicide dose rates could be a useful strategy for mitigating their negative effects on health and the environment. In this study, enzymatic activities and the parasitic ability of Meloidogyne incognita after exposure to sub-lethal concentrations (0.25, 1, 2, and 5 ppm) of ethoprophos, fenamiphos, and oxamyl were investigated. Although the tested concentrations did not show nematicidal properties in vitro, they reduced root galls by at least 30% at 0.25 ppm and up to 67% at 5 ppm in pots, besides disrupting nematode fertility. For all three nematicides at 2 ppm, a chemotaxis assay showed that ≤ 11% of the nematode population was successfully oriented to the host roots, compared to 44% in the control. Ethoprophos and fenamiphos at 5 ppm showed poor inhibitory effects on acetylcholinesterase (AChE) activity (5.6% and 12.5%, respectively). In contrast, the same nematicides were shown to be strong ATPase inhibitors, causing 82.4% and 82.8% inhibition, respectively. At the same concentration, oxamyl moderately inhibited AChE and ATPase-specific activities, the inhibition being 22.5% and 35.2%, respectively. This study suggests that the use of very low nematicide concentrations could be a promising strategy for nematode management. Furthermore, it has also highlighted the role of ATPases as a possible target site for suppressing nematode activity in the development of future nematicides.

Species richness, abundance, distributional pattern and trait composition of butterfly assemblage change along an altitudinal gradient in the Gulmarg region of Jammu & Kashmir, India.

Despite enormous diversity, abundance, and role in ecosystem processes, little is known about how butterflies differ across altitudinal gradients. For this, butterfly communities were investigated along an altitudinal gradient of 2700-3200 m a.s.l, along the Gulmarg region of Jammu & Kashmir, India. We aimed to determine how the altitudinal gradient and environmental factors affect the butterfly diversity and abundance. Our findings indicate that species richness and diversity are mainly affected by the synergism between climate and vegetation. Alpha diversity indices showed that butterfly communities were more diverse at lower elevations and declined significantly with increase in elevation. Overall, butterfly abundance and diversity is stronger at lower elevations and gradually keep dropping towards higher elevations because floristic diversity decreased on which butterflies rely for survival and propagation. A total of 2023 individuals of butterflies were recorded belonging to 40 species, represented by 27 genera and 05 families. Six survey sites (S I- S VI) were assessed for butterfly diversity from 2018 to 2020 in the Gulmarg region of Jammu & Kashmir. Across the survey, Nymphalidae was the most dominant family represented by 16 genera and 23 species, while Papilionidae and Hesperiidae were least dominant represented by 01 genera and 01 species each. Among the six collection sites selected, Site I was most dominant, represented by 16 genera and 21 species, while Site VI was least dominant, represented by 04 genera and 04 species.

Chrysin abrogates gibberellic acid-induced testicular oxidative stress and dysfunction via the regulation of antioxidants and steroidogenesis- and apoptosis-associated genes.

GA3 is widely used as a growth stimulant in agricultural regions. The long-term use of GA3 can cause organs damage. Chrysin is a flavonoid found in nature that is commonly used to treat organ toxicity. In this study, we examined the effect of chrysin on the testes function of GA3-affected rats. A total of 24 male Wistar rats were divided into 4 groups. Saline was given to the control group. The chrysin group was given orally 50 mg/kg/BW of chrysin in saline. The GA3 group received a daily oral gavage of GA3 (55 mg/kg/BW). The protective group (chrysin + GA3) was given chrysin and GA3 as those described in chrysin and GA3 groups. There were an increase in MDA levels in the serum and testicular tissue of GA3-treated group. Catalase, GSH, and SOD levels were all lowered in the GA3-treated rats. Chrysin dramatically reduced the harmful effects of GA3 by restoring reproductive hormone levels, altered sperm parameters, and antioxidant capabilities. Furthermore, GA3 reduced the quantitative expression of steroidogenesis genes StAR and 3-HSD, as well as Bcl2 genes, while it increased the apoptotic marker BAX; all were alleviated by the pre-administration of chrysin. The pre-administration of chrysin protected the GA3 group from spermatogenic vacuolation, interstitial edema, necrosis, and depletion. Chrysin inhibited oxidative stress and modulated antioxidant activity, as well as apoptosis-/anti-apoptosis-related mediators in the testes. Chrysin has the potential to repair GA3-induced testicular dysfunctions. This suggests that chrysin is preferable as a medication to mitigate GA3-induced oxidative damage in the testes. PRACTICAL APPLICATIONS: Chrysin has the potential to repair GA3-induced testicular dysfunctions. This suggests that chrysin is preferable as a medication to mitigate GA3-induced oxidative damage in the testes.

Chemical evaluation of partially acidulated phosphate rocks and their impact on dry matter yield and phosphorus uptake of maize.

Previous studies investigated the direct application of phosphate rock and its partially acidulated to enhance its solubility compared to soluble fertilizers. However, the interaction between the effect of particles diameter and partial acidulation of phosphate rock on phosphorus (P) availability and its effect on dry matter yield and P uptake is still elusive. This study was conducted to assess the effect of partially acidulated Egyptian phosphate rocks with different particle size diameters on P availability and its effect on dry matter yield and P uptake of maize (Zea mays L.). A pot experiment was conducted on maize plants grown on light clay soil for 42 days. Acidulation was done by mixing phosphate rock with single superphosphate or triple superphosphate at a total rate of 200 mg P kg-1 with five acidulation mix ratios (100:0, 75:25, 50:50, 25:75, and 0:100). Different particle size diameters of phosphate rocks (500, 212, 75, and <45 µm included nano-particles ranged from 69.3 to 25.7 nm) were used. We found that dry matter yield and P uptake increased significantly due to the use of partially acidulated phosphate rocks especially when triple superphosphate was used for acidulation and the mixing ratio of 50:50 was the best. We also found that maize yield and P uptake increased significantly with decreasing particle size. It is recommended to use finely grounded partially acidulated phosphate rocks with particles diameter less than 45 µm at acidulation ratio 50% and no need to increase acidulation ratio above that as a slow-release phosphate fertilizer.

Influence of Ascophyllum nodosum Extract Foliar Spray on the Physiological and Biochemical Attributes of Okra under Drought Stress.

Drought stress restricts the growth of okra (Abelmoschus esculentus L.) primarily by disrupting its physiological and biochemical functions. This study evaluated the role of Ascophyllum nodosum extract (ANE) in improving the drought tolerance of okra. Drought stress (3 days (control), 6 days (mild stress), and 9 days (severe stress)) and 4 doses of ANE (0, 0.1%, 0.2%, and 0.3%) were imposed after 30 days of cultivation. The results indicate that drought stress decreases the chlorophyll content (total chlorophyll, chlorophyll a, chlorophyll b, and carotenoid) but increases the activity of anthocyanin, proline, and antioxidant enzymes such as ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT). Physiological and biochemical plant disturbances and visible growth reduction in okra under drought stress were significantly decreased by the application of ANE foliar spray. ANE spray (0.3%) significantly increased the chlorophyll abundance and activity of anthocyanin, proline, and antioxidants (APX, POD, and CAT). ANE regulated and improved biochemical and physiological functions in okra under both drought and control conditions. The results of the current study show that ANE foliar spray may improve the growth performance of okra and result in the development of drought tolerance in okra.

Controlling of Meloidgyne incognita (Tylenchida: Heteroderidae) using nematicides, Linum usitatissimum extract and certain organic acids on four peppers cultivars under greenhouse conditions.

Organic acids and plant extracts, which have a nemacidal action and may be used instead of nematicides that pollute the environment, are one way for controlling the pepper root-knot nematode. We provide in this study for a first time a new strategy for management Meloidgyne incognita (Kofoid and White) by using organic acids and plant extract compared to nematicides on four peppers cultivars (Super amarr, Super mard, Super noura and Werta) under greenhouse conditions compared to nematicides. This study aimed to evaluate 0.1% of organic acids (humic and salicylic acid) and 0.1% of Linum usitatissimum extract on plant parameters of pepper varieties (Super amarr, Super mard, Super noura and Werta) and control of M. incognita under greenhouse conditions compared to four nematicides (Oxamyl 24% SL, Fosthiazates 75% EC, Ethoprophos N40% EC and Fenamiphos 40% EC). Our data obtained four nematicides were more effectiveness than other treatments in reduced galls and egg masses of M. incognita. Whilst, humic and salicylic acids have remarkably higher nematicidal activity than L. usitatissimum in all lines of pepper. Therefore, plant extract and organic acids may be used a best alternative of nematicides to control PPNs and caused the longitudinal growth of plant. Also, ultimately reduce environmental risk from nematicide pollution.

Utilizing biomass energy for improving summer squash greenhouse productivity during the winter season.

The objective of this present research is to use agricultural residues as a source of energy for heating greenhouses during winter seasons and sequestrating soil carbon dioxide through adding biochar to the soil media. To fulfill the objective of the research work, summer squash was transplanted in a constructed greenhouse and heated using an attached biomass-burning system. The performance of the attached biomass-burning system was experimentally studied under different agricultural residues (corn stalks, cotton stalks and okra stalks), heating fluids (water and oil) and air fan operating periods (10, 15 and 20 min/h). Results indicated that the biomass-burning system allowed increasing temperature and relative humidity inside the greenhouse up to 27.2 and 80 %, respectively. The maximum biomass-burning system efficiency of 81 % was achieved with the use of okra stalks as a source of energy and oil as a heating fluid side by side with adjusting the suction fan operating period at 15 min/h. Adding bio-charcoal to the soil media, enhanced the soil carbon, resulting in a total fresh yield of 3.7 and 2.9 kg/pot with a total number of leaves per plant of 55 and 47 leaves under conditions of with and without charcoal addition, respectively.

In vivo and in vitro management of Meloidogyne incognita (Tylenchida: Heteroderidae) using rhizosphere bacteria, Pseudomonas spp. and Serratia spp. compared with oxamyl.

Biological control using rhizosphere bacteria, Pseudomonas spp. and Serratia spp. is a prospective alternative technique to overcome plant parasitic nematodes infection. So, the current study was conducted in vitro on five egg-masses, 100 free eggs and 100 infective juveniles (IJs) of Meloidogyne incognita as well as greenhouse treatments on Luffa aegyptiaca L. to evaluate the nematicidal potential of six strains belong to Pseudomonas spp. and Serratia spp. as compared to oxamyl. Results showed that the inhibitory effect and juvenile mortality varied according to bacteria species, strains and exposure time. All the tested bacteria significantly (P ≤ 0.05) inhibited egg hatching and increased juvenile mortality in vitro. After 3 days of treatment, Pseudomonas spp. were more effective against eggs (48.31to 55.15%) and IJs (20.98 to 25.30%) than Serratia spp. (44.55 to 49.75% with eggs) and (19.06 to 21.61% with IJs), respectively. In the pot experiment, Luffa aegyptiaca L. treated with Serratia spp. and Pseudomonas spp. displayed significantly higher (P ≤ 0.05) levels of growth (as indicated by root length, fresh roots weight and fresh shoots weight) compared to control plants and significantly (P ≤ 0.05) suppressed galling (number of galls) and reproduction (as indicated by number of egg-masses on roots and number of eggs and juveniles in pot soil). Meanwhile, among the treated plants, Serratia spp. and Pseudomonas spp. gave the best results in shoot weight of pots infected by eggs of M. incognita than those infected with IJs as compared with positive control. While, oxamyl treatment gave the best results in pots infected by eggs and IJs. The lowest galling (gall index), number of eggs and juveniles in soil was observed in the treatment with mixture of Serratia spp. and Pseudomonas spp. as well as, enhanced growth of sponge gourd more than application each of them alone. Pots treated with oxamyl overwhelmed those treated with mixture of Serratia spp. and Pseudomonas spp.

The impact of insecticides and plant extracts on the suppression of insect vector (Bemisia tabaci) of Mungbean yellow mosaic virus (MYMV).

Mungbean yellow mosaic virus (MYMV) is an important constraint in successful production of mungbean (Vigna radiata L.) in many countries, including Pakistan. The MYMV spreads by insect vector whitefly (Bemisia tabaci Gennadius). The use of resistant cultivars is the most effective management tactics for MYMV. Twenty mungbean varieties/lines were screened against insect vector of MYMV under field condition in the current study. Resistance levels for varieties/lines were assessed through visual scoring of typical disease symptoms. Furthermore, the impacts of two insecticides 'Imidacloprid' and 'Thiamethoxam' and two plant extracts, i.e., neem (Azadirachta indica), and Eucalyptus (Eucalyptus camaldulensis) were tested on the suppression of whitefly. Field screening indicated that none of the tested varieties/lines proved immune/highly resistant, while significant variations were recorded among varieties/lines for resistance level. All varieties/lines were systemically infected with MYMV. The varieties 'AARI-2006' and 'Mung-14043' were considered as resistant to MYMV based on visual symptoms and the lowest vector population. These varieties were followed by 'NM-2006' and 'NL-31', which proved as moderately resistant to MYMV. All remaining varieties/lines were grouped as moderately to highly susceptible to MYMV based on visual symptoms' scoring. These results revealed that existing mungbean germplasm do not possess high resistance level MYMV. However, the lines showing higher resistance in the current study must be exploited in breeding programs for the development of resistant mungbean varieties/lines against MYMV. Imidacloprid proved as the most effective insecticide at all concentrations to manage whitefly population. Therefore, use of the varieties with higher resistance level and spraying Imidacloprid could lower the incidence of MYMV.

Biochemical changes in grape rootstocks resulted from humic acid treatments in relation to nematode infection.

OBJECTIVE: To investigate the effect of humic acid on nematode infected, resistant and susceptible grapes in relation to lipid peroxidation and antioxidant mechanisms on selected biochemical parameters known as proactive substances. METHODS: The grape rootstocks, superior, superior/freedom and freedom were reacted differently to Meloidogyne incognita and Rotylenchulus reniformis according to rootstock progenitor. Two weeks after inoculation, two commercial products of humic acid were applied at the rate of (2, 4 mL or grams/plant) as soil drench. After 4 months, nematode soil populations were extracted and counted. A subsample of roots from each plant was stained and gall numbers, embedded stages per root were calculated, final population, nematode build up (Pf/Pi), average of eggs/eggmass were estimated. Subsamples of fresh root of each treatment were chemically analyzed. RESULTS: Freedom reduced significantly the nematode criteria and build up. Humic acid granules appeared to be more suppressive to nematode build up on superior and the higher dose on superior/freedom than liquid treatments. On freedom, all treatments reduced significantly the nematode build up regardless to the material nature. The higher dose was more effective than the lower one. As a result of humic acid applications, the malondialdehyde (MDA) and H2O2 contents were significantly reduced after humic acid treatments while the antioxidant compounds glutathione (GSH), ascorbic acid (ASA) and total phenol contents were significantly increased when compared with check. Antioxidant defense enzymes ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT) and polyphenol oxidase (PPO)showed significant increase in their specific activities in treated plants compared with nematode treated check. CONCLUSIONS: Humic acid treatments improve the yield of grape by increasing the contents of antioxidant compounds and the specific activities of antioxidant enzymes.