Low temperature exposure influences nitrogen metabolism resulting in decreased Cry1Ac insecticidal endotoxin content in cotton seeds.

BACKGROUND: Sudden temperature drops, resulting from extreme weather events, often occur during the boll-setting period of cotton in Xinjiang, China, causing decreased expression of Bacillus thuringiensis (Bt) insecticidal proteins in cotton bolls. The precise threshold temperatures and durations that lead to significant changes in Cry1Ac endotoxin levels under low temperatures remain unclear. To address this, we investigated the effects of different temperatures and stress durations on Cry1Ac endotoxin levels in cotton bolls. In 2020-2021, two Bt transgenic cotton varieties, conventional Sikang1 and hybrid Sikang3, were selected as experimental materials. Various low temperatures (ranging from 16 to 20 °C) with different durations (12 h, 24 h and 48 h) were applied during the peak boll-setting period. RESULTS: As the temperature decreased, the Cry1Ac endotoxin content in the boll shell, fiber, and seed exhibited a declining trend. Moreover, the threshold temperature which caused a significant reduction in Cry1Ac endotoxin content increased with the prolonged duration of low-temperature stress. Among the components of cotton bolls, seeds were most affected by low-temperature stress, with the threshold temperature for a significant reduction in Cry1Ac endotoxin content ranging from 17 °C to 19 °C. Correlation analysis indicated that low temperatures led to a decrease in protein synthesis capacity and an increase in degradation ability, resulting in reduced Cry1Ac endotoxin content. Pathway analysis revealed that both free amino acid and peptidase had significant negative effects on Cry1Ac endotoxin content. CONCLUSION: In summary, when the daily average temperature was ≤ 19 °C, implementing cultural practices to reduce free amino acid content and peptidase activity could serve as effective cold defense strategies for Bt cotton production.

An Adjustable Pneumatic Planter with Reduced Source Vibration for Better Precision in Field Seeding.

The growing demand for agricultural output and limited resources encourage precision applications to generate higher-order output by utilizing minimal inputs of seed, fertilizer, land, and water. An electronically operated planter was developed, considering problems like ground-wheel skidding, field vibration, and the lack of ease in field adjustments of ground-wheel-driven seed-metering plates. The seed-metering plate of each unit of the developed planter is individually driven by a brushless direct current (BLDC) motor, and a BLDC motor-based aspirator is attached for pneumatic suction of seeds. The revolutions per minute (RPM) of the seed-metering plate are controlled by a microcontroller as per the received data relating to RPM from the ground wheel and the current RPM of the seed-metering plate. A feedback loop with proportional integral derivative (PID) control is responsible for reducing the error. Additionally, each row unit is attached to a parallelogram-based depth control system that can provide depth between 0 and 100 mm. The suction pressure in each unit is regulated as per seed type using the RPM control knob of an individual BLDC motor-based aspirator. The row-to-row spacing can be changed from 350 mm to any desired spacing. The cotton variety selected for the study was RCH 659, and the crucial parameters like orifice size, vacuum pressure, and forward speed were optimized in the laboratory with the adoption of a central composite rotatable design. An orifice diameter of 2.947 mm with vacuum pressure of 3.961 kPa and forward speed of 4.261 km/h was found optimal. A quality feed index of 93% with a precision index of 8.01% was observed from laboratory tests under optimized conditions. Quality feed index and precision index values of 88.8 and 12.75%, respectively, were obtained from field tests under optimized conditions.

Silver and gold nanoparticles: Eco-friendly synthesis, antibiofilm, antiviral, and anticancer bioactivities.

For the first time in this study, silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) were green synthesized by the cost-effective and eco-friendly procedure using Cotton seed meal and Fodder yeast extracts. The biosynthesized NPs were characterized by UV-Vis spectroscopy, dynamic light scattering analysis (DLS), transmission electron microscopy (TEM), selected area electron diffraction (SAED), and fourier-transform infrared (FTIR) spectroscopy. Furthermore, the biosynthesized NPs were tested in vitro against biofilm formation by some pathogenic negative bacteria (Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., and Pseudomonas aeruginosa) and negative bacteria (staphylococcus aureus) as well as against human denovirus serotype 5 (HAdV-5) and anticancer activity using HepG2 hepatocarcinoma cells. UV-Vis absorption spectra of reaction mixture of AgNPs and AuNPs exhibited maximum absorbance at 440 nm and 540 nm, respectively. This finding was confirmed by DLS measurements that the highest intensity of the AgNPs and AuNPs were 84 nm and 73.9 nm, respectively. FTIR measurements identified some functional groups detected in Cotton seed meal and Fodder yeast extracts that could be responsible for reduction of silver and gold ions to metallic silver and gold. The morphologies and particle size of AgNPs and AuNPs were confirmed by the TEM and SAED pattern analysis. Biosynthesized AgNPs and AuNPs showed good inhibitory effects against biofilms produced by Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., Pseudomonas aeruginosa, and Staphylococcus aureus. In addition, they showed anticancer activities against hepatocellular carcinoma (HepG-2) and antiviral activity against human adenovirus serotype 5 infection in vitro. Finally, the results of this study is expected to be extremely helpful to nano-biotechnology, pharmaceutical, and food packing applications through developing antimicrobial and/or an anticancer drugs from ecofriendly and inexpensive nanoparticles with multi-potentiality.

Methods to differentiate between cotton tract area ghee and cotton seed oil adulterated ghee.

Ghee, the clarified butter fat is one of the principal dairy products in India. In some places of India, cotton seed is fed extensively to dairy animals which changes the physico-chemical constants and fatty acid profile of the milk fat. Ghee is often adulterated with cotton seed oil and is marketed as cotton tract area ghee. Physico-chemical constants like RM value, Polenske value, BR reading, saponification value, iodine value and colorimetric tests i.e., Halphen and DPPH radical test were employed to differentiate ghee adulterated with cotton seed oil and that from cotton tract area. Chromatographic techniques like HPLC and GC-MS were also explored. Physico-chemical constants were not useful to differentiate the two types of ghee. Cyclopropenoic acids were observed only in ghee adulterated with cotton seed oil and not in cotton tract ghee. The RP-HPLC could able to distinguish the cotton tract area ghee from ghee adulterated with cotton seed oil on the basis of presence of ß-sitosterol in the latter. Halphen test was positive for cotton tract ghee, but not for the cotton seed oil adulterated ghee. Methylene blue reduction and DPPH radical test were also found to be useful to distinguish both types of ghee.

Engineering Trienoic Fatty Acids into Cottonseed Oil Improves Low-Temperature Seed Germination, Plant Photosynthesis and Cotton Fiber Quality.

Alpha-linolenic acid (ALA, 18:3Δ9,12,15) and γ-linolenic acid \ (GLA, 18:3Δ6,9,12) are important trienoic fatty acids, which are beneficial for human health in their own right, or as precursors for the biosynthesis of long-chain polyunsaturated fatty acids. ALA and GLA in seed oil are synthesized from linoleic acid (LA, 18:2Δ9,12) by the microsomal ω-3 fatty acid desaturase (FAD3) and Δ6 desaturase (D6D), respectively. Cotton (Gossypium hirsutum L.) seed oil composition was modified by transforming with an FAD3 gene from Brassica napus and a D6D gene from Echium plantagineum, resulting in approximately 30% ALA and 20% GLA, respectively. The total oil content in transgenic seeds remained unaltered relative to parental seeds. Despite the use of a seed-specific promoter for transgene expression, low levels of GLA and increased levels of ALA were found in non-seed cotton tissues. At low temperature, the germinating cottonseeds containing the linolenic acid isomers elongated faster than the untransformed controls. ALA-producing lines also showed higher photosynthetic rates at cooler temperature and better fiber quality compared to both untransformed controls and GLA-producing lines. The oxidative stability of the novel cottonseed oils was assessed, providing guidance for potential food, pharmaceutical and industrial applications of these oils.

Evaluation of feeding value of combination of alternate protein sources in White Leghorn layers.

1. A study was conducted to evaluate the effects of including graded levels of an alternate protein mixture (APM) containing cottonseed meal, distillery dried grain with solubles and mustard seed meal in a fixed ratio as a substitute for soybean meal in layer diets (hens aged 27 to 46 weeks). The trial studied the possibility of higher inclusion levels of APM with increased dietary concentrations of crude protein (CP) or critical amino acids (CAA). 2. Maize-soybean meal diet was used as the control diet. In experiment 1, APM was included at 0, 120, 160 and 200 g/kg diet (145 g/kg CP). In experiment 2, the control and three test diets (200 g/kg APM) with three levels of CP (145, 155 and 165 g/kg) were used. In experiment 3, a control and four test diets (200 g/kg APM) with four concentrations of CAA (100%, 105%, 110% and 115% relative to the control) were used. In all experiments, each diet was fed ad libitum to nine replicates of 88 birds each, housed in colony cages. 3. In experiment 1, egg production (EP) and feed intake (FI) were not affected by feeding up to 160 g/kg APM, but, at 200 g/kg, EP was significantly reduced. Feed conversion ratio (FCR) for the hens fed the highest level of APM was similar to the control. Egg weight (EW) and egg mass (EM) in the 160 and 200 g/kg AMS groups were lower than the control group. 4. Increase in dietary CP from 145 to 155 g/kg improved EP and reduced FCR to levels similar to the control group. EM in hens fed 165 g/kg APM was similar to the control in experiment 2. 5. Increasing concentrations of CAA progressively improved EP and reduced FCR in experiment 3. The EP at CAA 110% and FCR at CAA 105% were similar to the control group. 6. Based on the results, it was concluded that the APM can be included up to 160 g/kg diet without affecting layer performance, as long as the diet supplied the recommended levels of protein and amino acids. By increasing either protein from 145 to 155 g/kg or CAA concentrations by 10%, the dietary levels of the APM could be increased to 200 g/kg without affecting the layer performance.

A Novel Nanoemulsion Intermediate Gel as a Promising Approach for Delivery of Itraconazole: Design, In Vitro and Ex Vivo Appraisal.

The aim the study was to design, formulate, and evaluate self-nanoemulsifying drug delivery systems (SNEDDS) of itraconazole for improving the topical antifungal properties of the drug by adopting the nanoemulsion intermediate gel of the optimized system. Solubility study was conducted to select the most appropriate oils and surfactants for formulation. Different possible systems were created. Ternary phase diagrams were constructed to select the most promising system for further study. The nanoemulsion intermediate gel of the selected system was evaluated for stability, dilution effect, viscosity, pH, antifungal activity, droplet size, PDI, and zeta potential. In vitro release of the drug from the selected intermediate gel was investigated, and the kinetic model of drug release was determined. Ex vivo permeation of itraconazole was studied, and the amount of drug accumulated in the skin was calculated. Solubility and phase diagrams revealed that the system consisting of 60% cotton seed oil and 40% span 80 provided the nanoemulsion intermediate gel with the highest drug concentration. The selected system had a droplet size of about 236 nm and zeta potential of - 59.8. The viscosity of the corresponding intermediate gel was 1583.47 cp. The system exhibited high stability at 4°C and 25°C for 12 months and improved antifungal activity. In vitro release study showed complete release of itraconazole within 4 h, while the ex vivo permeation study revealed accumulation of the majority of the drug within the skin layers (72.5%).

Supplementary value of ensiled brewers spent grain used as replacement to cotton seed cake in the concentrate diet of lactating crossbred dairy cows.

The aim of this study was to investigate the effects of ensiled brewers spent grain (BSG) when used as replacement to cotton seed cake in the concentrate diet of lactating crossbred dairy cows. Eight early lactating F1 Boran X Friesian cows were used in a short term feeding trial to identify optimum level of ensiled BSG substitution of cotton seed cake (0, 33, 66, and 100%) in iso-nitrogenous diets. A 4 × 4 double Latin square design was used to analyze the data set generated from the feeding and digestibility trials. The results showed that as the level of ensiled BSG replacement to cotton seed cake increased, daily intakes on the natural pasture hay: 8.1 (T1) vs 7.6 (T2), 6.0 (T3), and 5.1 (T4); total feed dry matter: 14.6 (T1) vs 14.0 (T2), 12.9 (T3), and 12.2 (T4); crude protein (CP): 2.0 (T1) vs 1.9 (T2), 1.8 (T3), and 1.7 (T4); neutral detergent fiber (NDF): 8.4 (T1) vs 8.2 (T2), 7.4 (T3), and 6.8 (T4); and acid detergent fiber (ADF): 4.8 (T1) vs 4.5 (T2), 3.8 (T3), and 3.3 (T4) decreased (P < 0.05). However, estimated metabolizable energy (EME) intakes (129 vs 126) and body weight of the animals (465 vs 467) on the control diet and dietary T2 (33% BSG replacement for cottonseed cake) were comparable (P > 0.05). Thereafter, differences in daily EME intakes and body weight changes decreased with an increase in the level of ensiled BSG in the concentrate diet (P < 0.05) compared with both the control and animals on T2. Ensiled BSG, on the other hand, substantially improved (P < 0.05) total ration's apparent digestibility of dry matter (DM): 629 (T1) vs 659 (T3), 686 (T4); CP: 676(T1) vs 690(T3), 738(T4); NDF: 524 (T1) vs 544 (T3), 581 (T4); and ADF: 341 (T1) vs 350 (T2), 392 (T3), 440(T4) and daily milk yield: 14.5 (T1) vs 15.4 (T4) and milk production efficiency: 0.98 (T1) vs 1.11 (T3) and 1.26 (T4). So, ensiled BSG can be recommended to fully replace cotton seed cake from the concentrate diet of lactating dairy cows under local conditions. Additional research is needed to minimize and/or avoid body weight loss, milk fat, and total solids.

Senescence-induced expression of ZmSUT1 in cotton delays leaf senescence while the seed coat-specific expression increases yield.

KEY MESSAGE: Sink-specific expression of a sucrose transporter protein gene from the C4 plant maize can promote carbohydrate accumulation in target tissues and increase both fiber and seed yield of cotton. Sucrose is the principal form of photosynthetic products transported from source tissue to sink tissue in higher plants. Enhancing the partition of carbohydrate to the target organ is a promising way to improve crop productivity. The C4 plant Zea mays exhibits a substantially higher rate of export of photosynthates than many C3 plants, and its sucrose transporter protein ZmSut1 displays important role in sucrose allocation. To investigate how use of ZmSUT1 gene to increase the fiber and seed yield of cotton, in this study, we expressed the gene in cotton under a senescence-inducible promoter PSAG12 and a seed coat-specific promoter BAN, respectively. We show that senescence-induced expression of ZmSUT1 results in an increase of sugar accumulation in leaves. Although the leaf senescence was postponed in PSAG12::ZmSUT1 cotton, the photosynthetic rate of the leaves was decreased. In contrast, seed coat-specific expression of the gene leads to an increase of sugar accumulation in fibers and bolls, and the leaf of transgenic BAN::ZmSUT1 cotton displayed higher photosynthetic capacity than the wild type. Importantly, both fiber and seed yield of transgenic BAN::ZmSUT1 cotton are significantly enhanced. Our data indicate the potential of enhancing yield of carbohydrate crops by the regulation of sugar partitioning.

Near-Infrared Hyperspectral Imaging Combined with Deep Learning to Identify Cotton Seed Varieties.

Cotton seed purity is a critical factor influencing the cotton yield. In this study, near-infrared hyperspectral imaging was used to identify seven varieties of cotton seeds. Score images formed by pixel-wise principal component analysis (PCA) showed that there were differences among different varieties of cotton seeds. Effective wavelengths were selected according to PCA loadings. A self-design convolution neural network (CNN) and a Residual Network (ResNet) were used to establish classification models. Partial least squares discriminant analysis (PLS-DA), logistic regression (LR) and support vector machine (SVM) were used as direct classifiers based on full spectra and effective wavelengths for comparison. Furthermore, PLS-DA, LR and SVM models were used for cotton seeds classification based on deep features extracted by self-design CNN and ResNet models. LR and PLS-DA models using deep features as input performed slightly better than those using full spectra and effective wavelengths directly. Self-design CNN based models performed slightly better than ResNet based models. Classification models using full spectra performed better than those using effective wavelengths, with classification accuracy of calibration, validation and prediction sets all over 80% for most models. The overall results illustrated that near-infrared hyperspectral imaging with deep learning was feasible to identify cotton seed varieties.

Physical, chemical and biological behaviour of fumigants on cottonseed.

Fumigation is required to protect cottonseed in storage and pre-shipment from insect pests and/or microorganisms. Fumigation of cottonseed with carbon disulphide (CS2), carbonyl sulphide (COS), ethanedinitrile (C2N2), ethyl formate (EF), methyl bromide (MB) and phosphine (PH3) showed that >85% of the fumigants disappeared within 5 h of exposure. COS maintained >20 mg L-1 for 24 h. After 1 day of aeration, 75%-85% of the absorbed COS and MB and 20%-40% of the absorbed CS2, EF and PH3 were released from treated cottonseed. The fumigant residues were reduced by 80% for COS, 50% for EF or MB and 25% for CS2 after 1 day of aeration. After 13 days of aeration, fumigant residues were reduced by 95% for MB, 65% for EF, 55% for CS2 and to natural levels in the COS residue. Carbon disulphide, COS, PH3, EF and C2N2 had no effect on the germination of cottonseed, but germination was reduced to 50% by MB. COS has potential as a fumigant for control of insect pests in cottonseed because it dissipates quickly and does not negatively impact germination. On the other hand, MB appears to strongly absorb and requires an extended period for residues to dissipate, and it negatively impacts germination.

Genetic enhancement of palmitic acid accumulation in cotton seed oil through RNAi down-regulation of ghKAS2 encoding ß-ketoacyl-ACP synthase II (KASII).

Palmitic acid (C16:0) already makes up approximately 25% of the total fatty acids in the conventional cotton seed oil. However, further enhancements in palmitic acid content at the expense of the predominant unsaturated fatty acids would provide increased oxidative stability of cotton seed oil and also impart the high melting point required for making margarine, shortening and confectionary products free of trans fatty acids. Seed-specific RNAi-mediated down-regulation of ß-ketoacyl-ACP synthase II (KASII) catalysing the elongation of palmitoyl-ACP to stearoyl-ACP has succeeded in dramatically increasing the C16 fatty acid content of cotton seed oil to well beyond its natural limits, reaching up to 65% of total fatty acids. The elevated C16 levels were comprised of predominantly palmitic acid (C16:0, 51%) and to a lesser extent palmitoleic acid (C16:1, 11%) and hexadecadienoic acid (C16:2, 3%), and were stably inherited. Despite of the dramatic alteration of fatty acid composition and a slight yet significant reduction in oil content in these high-palmitic (HP) lines, seed germination remained unaffected. Regiochemical analysis of triacylglycerols (TAG) showed that the increased levels of palmitic acid mainly occurred at the outer positions, while C16:1 and C16:2 were predominantly found in the sn-2 position in both TAG and phosphatidylcholine. Crossing the HP line with previously created high-oleic (HO) and high-stearic (HS) genotypes demonstrated that HP and HO traits could be achieved simultaneously; however, elevation of stearic acid was hindered in the presence of high level of palmitic acid.

Tea waste: an effective and economic substrate for oyster mushroom cultivation.

BACKGROUND: Tea waste is the residue that remains after tea leaves have been extracted by hot water to obtain water-soluble components. The waste contains a re-usable energy substrate and nutrients which may pollute the environment if they are not dealt with appropriately. Other agricultural wastes have been widely studied as substrates for cultivating mushrooms. In the present study, we cultivated oyster mushroom using tea waste as substrate. To study the feasibility of re-using it, tea waste was added to the substrate at different ratios in different experimental groups. Three mushroom strains (39, 71 and YOU) were compared and evaluated. Mycelia growth rate, yield, biological efficiency and growth duration were measured. RESULTS: Substrates with different tea waste ratios showed different growth and yield performance. The substrate containing 40-60% of tea waste resulted in the highest yield. CONCLUSION: Tea waste could be used as an effective and economic substrate for oyster mushroom cultivation. This study also provided a useful way of dealing with massive amounts of tea waste.

Determination of strobilurin fungicides in cotton seed by combination of acetonitrile extraction and dispersive liquid-liquid microextraction coupled with gas chromatography.

The simultaneous determination of four strobilurin fungicides (picoxystrobin, kresoxim-methyl, trifloxystrobin, and azoxystrobin) in cotton seed by combining acetonitrile extraction and dispersive liquid-liquid microextraction was developed prior to GC with electron capture detection. Several factors, including the type and volume of the extraction and dispersive solvents, extraction condition and time, and salt addition, were optimized. The analytes were extracted with acetonitrile from cotton seed and the clean-up was carried out by primary secondary amine. Afterwards, 60 µL of n-hexane/toluene (1:1, v/v) with a lower density than water was mixed with 1 mL of the acetonitrile extract, then the mixture was injected into 7 mL of distilled water. A 0.1 mL pipette was used to collect a few microliters of n-hexane/toluene from the top of the aqueous solution. The enrichment factors of the analytes ranged from 36 to 67. The LODs were in the range of 0.1 × 10(-3) -2 × 10(-3) mg/kg. The relative recoveries varied from 87.7 to 95.2% with RSDs of 4.1-8.5% for the four fungicides. The good performance of the method, compared with the conventional pretreatments, has demonstrated it is suitable for determining low concentrations of strobilurin fungicide residues in cotton seed.

Effect of carbohydrate sources and levels of cotton seed meal in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in young dairy bulls.

The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and cassava chip+rice bran in the ratio of 3:1 (CR3:1), and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM) and 328 g CP/kg (HCM) at similar overall CP levels (490 g CP/kg). Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull.

Integrated microbiology and metabolomics analysis reveal responses of cotton rhizosphere microbiome and metabolite spectrum to conventional seed coating agents.

Fludioxonil (FL) and metalaxyl-M·fludioxonil·azoxystrobin (MFA) are conventional seed coating agents for controlling cotton seedling diseases. However, their effects on seed endophytic and rhizosphere microecology are still poorly understood. This study aimed to assess the effects of FL and MFA on cotton seed endophytes, rhizosphere soil enzymatic activities, microbiome and metabolites. Both seed coating agents significantly changed seed endophytic bacterial and fungal communities. Growing coated seeds in the soils originating from the Alar (AL) and Shihezi (SH) region inhibited soil catalase activity and decreased both bacterial and fungal biomass. Seed coating agents increased rhizosphere bacterial alpha diversity for the first 21 days but decreased fungal alpha diversity after day 21 in the AL soil. Seed coating reduced the abundance of a number of beneficial microorganisms but enriched some potential pollutant-degrading microorganisms. Seed coating agents may have affected the complexity of the co-occurrence network of the microbiome in the AL soil, reducing connectivity, opposite to what was observed in the SH soil. MFA had more pronounced effects on soil metabolic activities than FL. Furthermore, there were strong links between soil microbial communities, metabolites and enzymatic activities. These findings provide valuable information for future research and development on application of seed coatings for disease management.

Melatonin promotes seed germination under salt stress by regulating ABA and GA3 in cotton (Gossypium hirsutum L.).

Although previous studies have found that melatonin can promote seed germination, the phytohormone regulation mechanism by which exogenous melatonin mediates salt tolerance during cotton seed germination is still largely unknown. The effects of melatonin on germination traits and physiological parameters of GXM9 cotton seeds (Gossypium hirsutum L.) under three salt stress treatments (CK, germination of seeds pretreated with water alone; S, germination of seeds pretreated in 150 mM NaCl under salt stress; SM, germination of seeds pretreated in 20 µM melatonin under 150 mM NaCl solution) in the laboratory was investigated. The results showed that salt stress (150 mM) inhibited cotton seed germination and endogenous melatonin accumulation, and pretreatment with 20 µM exogenous melatonin enhanced the cotton germination rate and hypocotyl length as well as the content of endogenous melatonin during seed germination. This suggests that exogenous melatonin promotes seed germination from a morphological perspective. The contents of starch, α-amylase (EC3.3.1.1), ß-galactosidase (EC3.2.1.23), abscisic acid (ABA), and gibberellin (GA) were determined simultaneously. The results showed that the α-amylase and ß-galactosidase contents in the cotton seeds decreased by 56.97% and 20.18%, respectively, under salt stress compared with the control, while the starch content increased by 11.53% compared with the control at day 7. The ABA content increased by 25.18% and GA content decreased by 27.99% under salt stress compared with the control at 24 h. When exogenous melatonin was applied to the cotton seeds, the content of α-amylase and ß-galactosidase increased by 121.77% and 32.76%, respectively, whereas the starch contents decreased by 13.55% compared with the S treatment at day 7. Similarly, the ABA content increased by 12.20% and the GA content increased by 4.77% at 24 h. To elucidate the molecular mechanism by which melatonin promotes seed germination under salt stress, the effects of ABA- and GA-related genes on plant hormone signal transduction were analyzed by quantitative real-time PCR and RNA sequencing. The results indicated that melatonin regulated the expression of ABA and GA genes in the plant signal transduction pathway, induced embryo root development and seed germination, and alleviated dormancy. The expression of the ABA signaling gene GhABF2 was up-regulated and GhDPBF2 was down-regulated, and the expression of GA signaling genes (e.g., GhGID1C and GhGID1B) was up-regulated by melatonin. In conclusion, melatonin enhances salt tolerance in cotton seeds by regulating ABA and GA and by mediating the expression of hormone-related genes in plant hormone signal transduction. This should help us to explore the regulatory mechanisms of cotton resistance and provide a foundation for the cultivation of new varieties.

Aflatoxins in cotton seeds and cotton seed cake from Pakistan.

A study was conducted to evaluate the natural occurrence of aflatoxins (AFB1, AFB2, AFG1, AFG2) in cotton seeds (n = 110) and cotton seed cake (CSC; n = 110) from Pakistan. All samples were screened by Enzyme-Linked ImmunoSorbent Assay (ELISA). Positive samples were further quantified by IAC-HPLC-FLD. Total contamination frequency and aflatoxins mean levels were 80% and 69 µg/kg in cotton seeds and the corresponding values for cotton seed cake 88% and 89 µg/kg, respectively. Aflatoxin B1was found in all positive samples and co-occurred with AFB2, AFG1, and AFG2. Sixty-four cotton seeds and 71 CSC samples contained aflatoxins levels higher than the ML set for animal feed (20 µg/kg). The results of the present study will help the regulatory authorities to formulate strategies for monitoring aflatoxins in animal feeds.

Purification and Valorization of Waste Cotton Seed Oil as an Alternative Feedstock for Biodiesel Production.

This article is focused on the production of biodiesel from the waste cotton seed oil (WCSO), after purification, as an alternative to fossil fuels. Waste oil was collected from Sodecoton, a factory producing cotton seed oil in the Far North Cameroon. The WCSO was subjected to purification using activated coal, followed by transesterification under basic conditions (potassium hydroxide (KOH)), using methanol and ethanol. Some physico-chemical properties of biodiesel, such as absorbance of waste and purified oil, density, viscosity, water content, acid value, and its energy content were determined. The result of treating the WCSO with activated coal indicated that purification efficiency of activated coal increased with the contact time and the mass of the absorbent. Absorbance results directly proved that activated coal removed unwanted components. In the same way, activated coal concentration and exposure time influenced the level of free fatty acids of WCSO. The yield of methyl ester was 97%, while that of ethyl ester was 98%. The specific gravity at 25 °C was 0.945 ± 0.0601. An evaluation of the lower calorific value (PCI) was done in order to study the energy content of biodiesel. This was found to be a value of 37.02 ± 3.05 MJ/kg for methyl ester and 36.92 ± 7.20 MJ/kg for ethyl ester. WCSO constitutes feedstock for high volume, good quality, and sustainable production of biodiesel, as well as a realistic means of eliminating the pollution resulting from the indiscriminate disposal of waste oils from both household and industrial users.

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after-ripening and germination.

The dormancy of seeds of upland cotton can be broken during dry after-ripening, but the mechanism of its dormancy release remains unclear. Freshly harvested cotton seeds were subjected to after-ripening for 180 days. Cotton seeds from different days of after-ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H2 O2 and phytohormones and activities of antioxidant enzymes and glucose-6-phosphate dehydrogenase were measured during after-ripening and germination. Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T50 (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H2 O2 increased during after-ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H2 O2 and changed peroxidase content during germination. For cotton seed, the alleviation of coat-imposed dormancy required 30 days of after-ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H2 O2 acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.