Cross-Serological Reaction of Glandless Cottonseed Proteins to Peanut and Tree Nut Allergic IgE.

Food allergy is a potentially life-threatening health concern caused by immunoglobulin E (IgE) antibodies that mistakenly recognize normally harmless food proteins as threats. Peanuts and tree nuts contain several seed storage proteins that commonly act as allergens. Glandless cottonseed, lacking the toxic compound gossypol, is a new food source. However, the seed storage proteins in cottonseed may act as allergens. To assess this risk, glandless cottonseed protein extracts were evaluated for IgE binding by peanut and tree nut allergic volunteers. ELISA demonstrated that 25% of 32 samples had significant binding to cottonseed extracts. Immunoblot analysis with pooled sera indicated that IgE recognized a pair of bands migrating at approximately 50 kDa. Excision of these bands and subsequent mass-spectrometric analysis demonstrated peptide matches to cotton C72 and GC72 vicilin and legumin A and B proteins. Further, in silico analysis indicated similarity of the cotton vicilin and legumin proteins to peanut vicilin (Ara h 1) and cashew nut legumin (Ana o 2) IgE-binding epitopes among others. The observations suggest both the cotton vicilin and legumin proteins were recognized by the nut allergic IgE, and they should be considered for future allergen risk assessments evaluating glandless cottonseed protein products.

Chemical Composition and Thermogravimetric Behaviors of Glanded and Glandless Cottonseed Kernels.

Common "glanded" (Gd) cottonseeds contain the toxic compound gossypol that restricts human consumption of the derived products. The "glandless" (Gl) cottonseeds of a new cotton variety, in contrast, show a trace gossypol content, indicating the great potential of cottonseed for agro-food applications. This work comparatively evaluated the chemical composition and thermogravimetric behaviors of the two types of cottonseed kernels. In contrast to the high gossypol content (3.75 g kg-1) observed in Gd kernels, the gossypol level detected in Gl kernels was only 0.06 g kg-1, meeting the FDA's criteria as human food. While the gossypol gland dots in Gd kernels were visually observed, scanning electron microcopy was not able to distinguish the microstructural difference between ground Gd and Gl samples. Chemical analysis and Fourier transform infrared (FTIR) spectroscopy showed that Gl kernels and Gd kernels had similar chemical components and mineral contents, but the former was slightly higher in protein, starch, and phosphorus contents. Thermogravimetric (TG) processes of both kernels and their residues after hexane and ethanol extraction were based on three stages of drying, de-volatilization, and char formation. TG-FTIR analysis revealed apparent spectral differences between Gd and Gl samples, as well as between raw and extracted cottonseed kernel samples, indicating that some components in Gd kernels were more susceptible to thermal decomposition than Gl kernels. The TG and TG-FTIR observations suggested that the Gl kernels could be heat treated (e.g., frying and roasting) at an optimal temperature of 140-150 °C for food applications. On the other hand, optimal pyrolysis temperatures would be much higher (350-500 °C) for Gd cottonseed and its defatted residues for non-food bio-oil and biochar production. The findings from this research enhance the potential utilization of Gd and Gl cottonseed kernels for food applications.

Constitutive and induced insect resistance in RNAi-mediated ultra-low gossypol cottonseed cotton.

BACKGROUND: Besides fibers, cotton plants also produce a large amount of seeds with a high oil and protein content. The use of these seeds is restricted by their high contents of the terpenoid gossypol, which is harmful to humans and livestock. Using a genetic engineering approach, "Ultra-low gossypol cottonseed" (ULGCS) plants were produced by knocking down an enzyme that catalyzes the formation of a precursor of gossypol. This was accomplished via RNAi-mediated silencing of the target gene using a seed-specific α-globulin promotor. Since gossypol is also a crucial defense mechanism against leaf-feeding herbivores, ULGCS plants might possess lower herbivore resistance than non-engineered plants. Therefore, we tested the constitutive and inducible direct insect resistance of two ULGCS cotton lines against the African cotton leafworm, Spodoptera littoralis. RESULT: The herbivore was equally affected by both ULGCS lines and the control (Coker 312) line when feeding on fully expanded true leaves from undamaged plants and plants induced by jasmonic acid. When plants were induced by caterpillar-damage, however, S. littoralis larvae performed better on the ULGCS plants. Terpenoid analyses revealed that the ULGCS lines were equally inducible as the control plants. Levels of terpenoids were always lower in one of the two lines. In the case of cotyledons, caterpillars performed better on ULGCS cotton than on conventional cotton. This was likely caused by reduced levels of gossypol in ULGCS cotyledons. CONCLUSION: Despite those effects, the insect resistance of ULGSC cotton can be considered as largely intact and the plants may, therefore, be an interesting alternative to conventional cotton varieties.

Initial Formulation of Novel Peanut Butter-like Products from Glandless Cottonseed.

Glandless (Gl) cottonseed is a unique cotton variety with only a trace content of toxic gossypol present. This new cottonseed raises the potential of its enhanced utilization as an agro-food for human consumption. In this work, Gl cottonseed kernels were used with additional cottonseed oil to produce novel peanut butter-like products. Kernels roasted at two temperatures (140 or 150 °C) for a given time (15 or 30 min) were first ground with different ratios of cottonseed oil and two other ingredients (i.e., salt and sugar) with a food blender, and then passed through a meat grinder with a 4-mm-hole grinding plate. Per the preliminary result, the butter-like products with Gl kernels roasted at 150 °C were subject to further structural and textural evaluation. The color of the two butter-like products was comparable to a commercial peanut butter, but the formers' textural properties were significantly different (p ≤ 0.05) from the latter. Morphologic examination by Scanning Electron Microscopy (SEM) and cryo-SEM revealed that the butter product with a longer (30 min) roasting time possessed a smoother surface than the products with a shorter (15 min) roasting time. Oil stability test showed no substantial oil separation (<3%) from the butter products over 7 weeks at ambient temperature (22 °C). This work provides the basic information and parameters for lab cottonseed butter making so that optimization and characterization of cottonseed butter formation can be designed and performed in future research.

The Effects of Oil Content on the Structural and Textural Properties of Cottonseed Butter/Spread Products.

Plant-based butters from nuts and seeds have steadily increased in consumer popularity due to their unique flavors and healthy nutritional properties. Oil content is a critical parameter to measure the proper consistency and stability of plant butter and spread products. Previous work has shown that glandless cottonseed can be used to formulate cottonseed butter products to increase the values of cottonseed. As part of the efforts made in the valorization of cottonseed, this work evaluated the effects of oil content on the microstructural and textural properties of cottonseed butter/spread products. While the oil content in the raw cottonseed kernels was 35% of the kernel biomass, additional cottonseed oil was added to make cottonseed butter products with six oil content levels (i.e., 36, 43, 47, 50, 53, and 57%). The values of three textural parameters, firmness, spreadability, and adhesiveness, decreased rapidly in an exponential mode with the increasing oil content. The particle size population in these butter samples was characterized by similar trimodal distribution, with the majority in the middle mode region with particle sizes around 4.5-10 µm. Higher oil content decreased the butter particle size slightly but increased oil separation during storage. The oxidation stability with a rapid oxygen measurement was gradually reduced from 250 min with 36% oil to 65 min with 57% oil. The results of this work provide information for the further optimization of formulation parameters of cottonseed butter products.

Molecular level characterization of the effect of roasting on the extractable components of glandless cottonseed by Fourier transform ion cyclotron resonance mass spectrometry.

Roasting is a technological process in some food applications of agricultural products. To investigate the composition changes of the extractable functional/bioactive components of cottonseed, in this work, glandless cottonseed kernels were roasted at 110, 120, 140 and 150 °C for 15 min, respectively. The UV/vis data of the 80 % ethanol extracts found that roasting increased the level of phenolic compounds. Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry of the extracts identified about 44 % to 55 % of total formulas as potential phenolic compounds. Roasting (up to 140 °C) mainly increased carbohydrate-, lignin-, and tannin-like compounds while lipid-like compounds decreased. The compositional changes at 150 °C were less than those at 140 °C, attributed to devolatilization at the higher temperature. The information of chemical profiling of cottonseed and the roasting impact would be greatly useful in enhanced utilization of cottonseed as nutrient and functional foods or food supplements.

Development of the engineered "glanded plant and glandless seed" cotton.

After fiber, cottonseed is the second most important by-product of cotton production. However, high concentrations of toxic free gossypol deposited in the glands of the cottonseed greatly hamper its effective usage as food or feed. Here, we developed a cotton line with edible cottonseed by specifically silencing the endogenous expression of GoPGF in the seeds, which led to a glandless phenotype with an ultra-low gossypol content in the seeds and nearly normal gossypol in other parts of the plants. This engineered cotton maintains normal resistance to insect pests, but the gossypol content in the seeds dropped by 98%, and thus, it can be consumed directly as food. The trait of a low gossypol content in the cottonseeds was stable and heritable, while the protein, oil content, and fiber yield or quality were nearly unchanged compared to the transgenic receptor W0. In addition, comparative transcriptome analysis showed that down-regulated genes in the ovules of the glandless cotton were enriched in terpenoid biosynthesis, indicating the underlying relationship between gland formation and gossypol biosynthesis. These results pave the way for the comprehensive utilization of cotton as a fiber, oil, and feed crop in the future.

Genetics and Breeding for Glandless Upland Cotton With Improved Yield Potential and Disease Resistance: A Review.

Glandless cotton (devoid of toxic gossypol) can be grown as a triple-purpose crop for fiber, feeds, and food (as an oil and protein source). However, its sensitivity to insect pests and its low yield due to the lack of breeding activities has prevented the realization of its potential in commercial seed production and utilization. Since the mid-1990s, the commercialization of bollworm and budworm resistant Bt cotton and the eradication of boll weevils and pink bollworms have provided an opportunity to revitalize glandless cotton production in the United States. The objectives of this study were to review the current status of genetics and breeding for glandless cotton, with a focus on the progress in breeding for glandless Upland cotton in New Mexico, United States. Because there existed a 10-20% yield gap between the best existing glandless germplasm and commercial Upland cultivars, the breeding of glandless Upland cultivars with improved yield and disease resistance was initiated at the New Mexico State University more than a decade ago. As a result, three glandless Upland cultivars, i.e., long-staple Acala 1517-18 GLS, medium staple NuMex COT 15 GLS, and NuMex COT 17 GLS with Fusarium wilt race 4 resistance were released. However, to compete with the current commercial glanded cotton, more breeding efforts are urgently needed to introduce different glandless traits (natural mutations, transgenic or genome-editing) into elite cotton backgrounds with high yields and desirable fiber quality.

Antioxidant activities of the water-soluble fractions of glandless and glanded cottonseed protein.

To promote application of cottonseed protein products in food industry, this work measured antioxidant activities of two water soluble protein samples (Gl-L and Gd-L) isolated in a lab scale from glandless and common glanded cottonseed meal, respectively, and one soluble protein samples (Gd-P) in a pilot scale from glanded cottonseed meal. SDS-gel electrophoresis showed that the distribution patterns of the peptide fragments in Gl-L and Gd-L were similar, but more fragments and higher molecular mass bands were observed in the Gd-P gel image. While Gd-P showed the highest activities, Gl-L and Gd-L exhibited comparable 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical-scavenging activities in most cases. In contrast, Gd-P showed lower capability of inhibition of linoleic acid autoxidation than Gl-L and Gd-L. It would be of great interest in further research on these protein fractions in food products and processes (such as roasting) involved in the protective effects of food spoilage.

Simulated digestion of dried leaves of Artemisia annua consumed as a treatment (pACT) for malaria.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisinin (AN) is produced by Artemisia annua, a medicinal herb long used as a tea infusion in traditional Chinese medicine to treat fever; it is also the key ingredient in current artemisinin-based combination therapies (ACTs) effective in treating malaria. Recently we showed that dried leaves from the whole plant Artemisia annua that produces artemisinin and contains artemisinin-synergistic flavonoids seem to be more effective and less costly than ACT oral malaria therapy; however little is known about how digestion affects release of artemisinin and flavonoids from dried leaves. MATERIAL AND METHODS: In the current study we used a simulated digestion system to determine how artemisinin and flavonoids are released prior to absorption into the bloodstream. Various delivery methods and staple foods were combined with dried leaves for digestion in order to investigate their impact on the bioavailability of artemisinin and flavonoids. Digestate was recovered at the end of the oral, gastric, and intestinal stages, separated into solid and liquid fractions, and extracted for measurement of artemisinin and total flavonoids. RESULTS: Compared to unencapsulated digested dried leaves, addition of sucrose, various cooking oils, and rice did not reduce the amount of artemisinin released in the intestinal liquid fraction, but the amount of released flavonoids nearly doubled. When dried leaves were encapsulated into either hydroxymethylcellulose or gelatin capsules, there was >50% decrease in released artemisinin but no change in released flavonoids. In the presence of millet or corn meal, the amount of released artemisinin declined, but there was no change in released flavonoids. Use of a mutant Artemisia annua lacking artemisinin showed that the plant matrix is critical in determining how artemisinin is affected during the digestion process. CONCLUSIONS: This study provides evidence showing how both artemisinin and flavonoids are affected by digestion and dietary components for an orally consumed plant delivered therapeutic and that artemisinin delivered via dried leaves would likely be more bioavailable if provided as a tablet instead of a capsule.