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Integration of GWAS and WGCNA reveals novel candidate genes for cottonseed oil content in Gossypium hirsutum L.

Genetic improvement of cottonseed oil content represents a crucial strategy for enhancing the comprehensive utilization of cotton. Here, genome-wide association study (GWAS) and weighted gene co-expression network analysis (WGCNA) were integrated to elucidate the genetic control underlying oil content. Phenotypic evaluation of 159 cotton accessions revealed extensive genetic variation, with kernel oil content ranging from 17.81% to 39.50%. Population structure analysis based on 20,213 single nucleotide polymorphisms (SNPs) classified the accessions into two major subpopulations. A total of 18 SNPs exhibited significant associations with oil content, two of which were stably detected across multiple environments using the FarmCPU model. Further haplotype analysis within linkage disequilibrium (LD) blocks confirmed a favorable haplotype on chromosome A05 that was strongly correlated with elevated oil content. Integration of publicly available transcriptome data from 11 ovule developmental stages with WGCNA identified modules significantly linked to oil content. Of the 74 candidate genes within LD intervals, 17 were assigned to WGCNA modules. Functional annotation and enrichment analyses highlighted four putative candidate genes (GH_A05G1503, GH_A05G1506, GH_A05G1531, and GH_A10G2150) involved in oil biosynthesis. These findings deepen our understanding of the genetic mechanisms governing cottonseed oil biosynthesis and lay a foundation for breeding high-oil cotton varieties.

Gossypium

The influence of crude cottonseed oil in the feed on the blood and egg yolk lipoproteins of laying hens.

Lipovitellin, very low density lipoproteins (VLDL), low density lipoproteins (LDL), high density lipoproteins (HDL), and proteins of d greater than 1.20 were isolated from blood plasma and egg yolks obtained from hens fed a normal diet or one containing 2.5% of crude cottonseed oil. The amounts and compositions of each fraction were determined. Hen blood plasma and egg yolk VLDL and LDL obtained from hens fed a normal diet contained similar levels of lipid, and the fatty acid compositions of those lipids were, for the most part, similar. The percentages of VLDL and LDL in total lipoproteins were similar for plasma and egg yolk obtained from hens fed the normal diet. Separation of VLDL from LDL was not clear-cut in eggs from hens fed the diet which contained crude cottonseed oil. Lipovitellin, as it is isolated from egg yolk, did not appear to be present in any appreciable amount in hen blood plasma. Hen plasma appeared to contain about 10% of a liproprotein d greater than 1.20, the lipid of which was similar in fatty acid composition to that of lipovitellin except for palmitic and oleic acids. Lipids of all the different lipoproteins isolated from plasma and egg yolks of hens fed diets which contained 2.5% of crude cottonseed oil contained more stearic acid and less palmitoleic and oleic acids than did those from normal hens. The increased content of stearic acid increased the density of the lipoproteins so that a larger proportion of the lipoproteins were in the LDL and a smaller proportion were in the VLDL than in lipoproteins from normal plasma and eggs.

Animals

Effects of dietary fats on the fatty acid composition of enamel and dentinal lipids of rabbit molars.

The effect of dietary fats such as butter, lard, cottonseed oil, and hydrogenated cottonseed oil on the fatty acid composition of enamel and dentin lipids of rabbit molars was studied. The enamel and dentin of molars of all of the rabbits contained a variety of fatty acids, palmitic, stearic, oliec, linoleic, and eicosapentaenoic being the major ones. Enamal lipids were more unsaturated than dentinal lipids in all of the groups. A significant difference was noticed in the fatty acid composition of both the enamel and dentin between these four groups of rabbits. Linoleic acid content was highest in both the enamel and dentin lipids of rabbits fed cottonseed oil. There was no marked difference in the saturated-unsaturated fatty acid ratio in both the enamel and dentin among rabbits fed cottonseed oil, lard, and butter. It is concluded that tooth lipids are responsive to diets and are in direct equilibrium with dietary components.

Animals

Feeding polyunsaturated vegetable oils to lactating cows.

Holstein cows fed concentrate:hay diets also were fed for 14 days supplements of soybean oil plus casein, soybean oil protected from ruminal hydrogenation by encapsulation in a casein-formaldehyde matrix, cottonseed oil plus casein, or cottonseed oil protected with casein formaldehyde. The supplements were fed at rates to give a linoleic acid (18:2) intake of 225 g/day. Yields of milk and milk protein were not affected by treatment. Milk 18:2 was not increased by the unprotected soybean oil or cottonseed oil but was increased by protected soybean and cottonseed oil from a control of 2.3 to 5.7% of total milk fat. Milk 18:0 and 18:1 also increased. Compensatory declines were observed in milk 16:0 and 14:0 acids. In fecal fatty acids during the treatment periods, percentage of 18:2 of the total fat decreased and 18:0 markedly increased. These results indicate hydrogenation of the dietary oils in the alimentary tract or a differential absorption. Fecal 16:0 and 14:0 decreased.

Animals

The effect of a non-absorbable fat, sucrose polyester, on the metabolism of vitamin A by the rat.

Sucrose polyester (SPE) is a fat-like material that is not absorbed. The effect of this material on vitamin A metabolism was determined by measuring the amount of the vitamin that was stored in the liver of rats following the ingestion of a known amount of vitamin A. In one study, the vitamin A was administered as an oral dose in a vehicle consisting of various proportions of cottonseed oil and SPE. Each 1% replacement of cottonseed oil by SPE resulted in a 0.26% decrease in the amount of vitamin A found in the liver. In the second study, the vitamin A was incorporated into diets in which the fat component consisted of various proportions of cottonseed oil and SPE. When these diets were consumed for 1 week, each 1% replacement of cottonseed oil by SPE resulted in a 0.84% decrease in the storage of vitamin A by the liver. It is proposed that in the lumen of the intestine vitamin A distributes between the customary micellar phase and the unhydrolyzed oil phase of SPE. The vitamin A in this latter phase is eliminated in the feces.

Absorption

Effects of nutritional factors on the development of ethanol-induced fatty liver in KK and KK-Ay mice.

KK and KK-Ay mice developed a steady and reproducible fatty liver when they were given free access to an ethanol solution as a drinking fluid for 10 to 20 days. The present studies were undertaken to elucidate effects of nutritional factors on liver fat contents of the mice given water or ethanol solution. In contrast to cornstarch, sucrose tended to increase the liver fat of control mice. A higher concentration of dietary casein lowered the liver fat of control mice, whereas the dietary concentration of cottonseed oil did not significantly affect the liver rat levels either in control or ethanol groups. Thus the standard basal diets favorable for the development of the alcoholic fatty liver have been established, for example, 10% cottonseed oil, 25 and 30% casein, 58.4 and 53.4% cornstarch for KK (12-15 weeds old) and KK-Ay (5-10 weeks old), respectively. Neither choline, myoinositol, nor any lipotropic agent tested prevented the development of the alcoholic fatty liver. Unlike in rats, orotic acid did not induce a fatty liver but rather alleviated the ethanol-induced fatty liver in these mice.

Age Factors

Clinical use of 10% soybean oil emulsion.

The pharmaceutical and therapeutic aspects of the use of 10% soybean oil emulsion in total parenteral nutrition (TPN) are reviewed. The composition of the commercial preparation (intralipid), comparison with cottonseed oil emulsion, advantages and disadvantages of use, metabolism, toxicity, therapeutic monitoring, clinical uses, and evaluative studies are discussed. Soybean oil emulsion is a useful source of calories and essential fatty acids in TPN. The product is more expensive than other solutions used in TPN. Woybean oil emulsion should be considered for use in small hospitals and in home therapy because of the advantage of peripheral administration.

Adult

Quantitative cell culture biocompatibility testing of medical devices and correlation to animal tests.

The biocompatibility of a wide variety of biomaterials was quantitatively assessed, in a physiologically normal environment" as to cytotoxicity induced in WI-38 cells by cell culture medium extracts. Materials tested included PVC plastic, rubber, silicone rubber, polyethylene, polypropylene, acetal, polyurethane, Teflon, nylon, epoxy, and polystyrene. Cell culture test results were correlated to U.S.P. animal tests. Potential test artifacts, lead, barium, cadmium, and endotoxin were tested for cytotoxicity in WI-38 cells. Cell culture methods yielded more positive tests, particularly rubber, PVC plastic and silicone rubber compounds, than observed in U.S.P. animal tests. Positivity in animal tests did not correlate quantitatively to cytotoxic titers in cell culture. Discrepancies between cell cultures tests and animal tests, specifically rubber compounds, were attributable, in some instances, to differentials in elution efficiency between saline, cottonseed oil, and complete MEM cell culture medium. In other instances, particularly PVC plastics, differences between cell culture and animal test results were due to an inherent difference in the two indicator systems to respond to specific toxic moieties.

Acetals

Molecular sizes of egg yolk very low density lipoproteins fractionated by ultracentrifugation.

Egg yolk very low density lipoproteins isolated from normal eggs were separated into six fractions by preparative ultracentrifugation. The sizes of the molecules in three of the fractions were estimated by gel chromatography and by electron microscopy of the lipoproteins using a negative staining technique. The three fractions had average diameters of 39 nm., 32 n., and 25 nm. and average molecular weights of 17 X 10(6), 10 X 10(6), and 5 X 10(6). There was considerable overlap of size of molecules in the three fractions. Size differences were not observed in the ultracentrifuge separated fractions of very low density lipoproteins isolated from eggs laid by hens receiving crude cottonseed oil in their diets. However, the fatty acids from these fractions differed in stearic and oleic acid contents while those from the normal egg lipoproteins did not. The floating very low density lipoproteins contained more oleic acid and less stearic acid then the sedimenting ones did.

Animals

Stearoyl-CoA desaturase activity in mammary adenocarcinomas carried by C3H mice.

Transplantable mammary adenocarcinomas and livers of C3H mice fed a stock diet or a linoleate rich diet (15% corn oil) contain similar amounts of oleate (ca 3 mg/gm tissue). On feeding either a high carbohydrate, fat free or a high carbohydrate, saturated fat-containing (15% hydrogenated coconut or cottonseed oil) diet for 6 weeks, oleate levels increased 2-fold in tumor and 5-fold in liver. The specific activity of stearoyl-CoA desaturase in liver microsomes was similar to that in the corresponding fractions of mammary glands of lactating mice. In liver, this activity was enhanced 2- to 3-fold by feeding a high carbohydrate, fat free or a high carbohydrate, saturated fat diet. The desaturase activity in mammary tumor microsomes, while only 10% of that in hepatic microsomes, remained unaltered regardless of the type of diet fed. These observations suggest that (a) a major portion of the oleate in the mammary tumor is not produced within the tissue, (b) dietary adaptation is not a general characteristic of stearoyl-CoA desaturase in neoplastic tissues, and (c) enhanced desaturase activity in liver is directly related to the absence of linoleate or oleate, or to a large decrease in oleate in the diet.

Adenocarcinoma

Effects of the unsaturation of dietary fat and of arachidonate supplementation on cholesterol pool expansion in the guinea pigs.

We have studied the effects of methyl arachidonate supplementation on the lipid metabolism of guinea pigs fed cholesterol. Four groups of guinea pigs were fed a purified diet containing 9.5% hydrogenated coconut oil (HCNO), a highly saturated fat with or without the addition of 1% cholesterol, for 15 weeks. One half of the animals fed the control and the cholesterol-containing diets were supplemented with 15 mg methyl arachidonate three times per week. Supplementation with methyl arachidonate did not alter the concentration of plasma total (TC) or unesterified (FC) cholesterol, erythrocyte cholesterol and plasma phospholipid or the ratio of plasma FC/TC. Accumulation of cholesterol in the major organs of the cholesterol-fed groups was also unchanged. In both control and cholesterol-fed groups, methyl arachidonate decreased the proportion of oleic (18:1) and linoleic acids (18:2) and increased arachidonic acid (20:4) content of plasma and liver phospholipid. A comparison between the results of this study and studies using cottonseed oil showed that the type of dietary fat modifies the effects of cholesterol: plasma cholesterol levels were higher and liver cholesterol storage was lower in animals fed the saturated fat than in those fed the fat rich in polyunsaturated fatty acids (PUFA). Furthermore, in spite of similar changes in erythrocyte cholesterol content and shape abnormalities, no overt hemolytic anemia was observed in the groups fed cholesterol and saturated fat, in contrast to those fed cholesterol + PUFA-containing fat. We conclude that in guinea pigs supplementary methyl arachidonate had no hypocholesterolemic effect at the levels we fed, that circulating cholesterol levels are not a measure of cholesterol accumulation by organs and that the decrease of serum cholesterol in response of PUFA is due in part to an increase of cholesterol storage in the liver.

Animals

Wheat-based diets: effect of short-term consumption on serum cholesterol and triglyceride levels in infants.

The effect of consumption of wheat-based diets on serum cholesterol and triglyceride levels was studied in eight previously malnourished children. While consuming a control diet of casein, soy-cottonseed oil blend, and a mixture of sucrose and starch, the serum cholesterol level was 169 +/- 42 mg/dl (mean +/- SD). This decreased significantly (P less than .001) to 108 +/- 30 mg/dl after nine days' consumption of an isoenergetic-isonitrogenous diet in which whole wheat or white flour provided all the protein and +/- 50% of carbohydrate, and remained at this level for the 27-day dietary period. On changing again to the casein-based diet, the serum cholesterol level rose within nine days to 154 +/- 42 mg/dl. There was no similar diet-related change in serum triglyceride values. None of the known mechanisms whereby diet affects serum cholesterol adequately explains these findings.

Body Weight

The cockerel as an animal model for atherosclerosis research.

The chicken is a good animal model for the study of atherosclerosis research because it is: 1. Omnivorous. 2. Small and suitable for prolonged laboratory investigation. 3. Able to develop spontaneous atherosclerosis. 4. Capable of producing atherosclerosis after cholesterol feeding with elevated hypercholesterolemia. A diet of 1/4% cholesterol plus 5% cottonseed oil added to starter-grower-mash resulted in aortic atherosclerosis with a slight but significant increase in plasma cholesterol. 5. Plasma levels of cholesterol and triglyceride are similar to those in humans. 6. Lipid composition of high and low density lipoproteins as well as chylomicrons resembles those of humans. 7. Has been noted that there is no essential difference between vascular lesions seen in chickens as a result of cholesterol diet and that of atherosclerosis observed in man.

Animals

Response to the hypobetalipoproteinemic agent adamantyloxyphenyl piperidine in hyperlipemic rats.

The hypobetalipoproteinemic activity of U-41,792 (1-[p-(1-adamantyloxy)-phenyl]-piperidine) is a marked and selective reduction of heparin precipitating lipoproteins (low density plus very low density lipoproteins) in cholesterol-cholic acid induced hypercholesterolemic rats. This activity consists of both a reduction in heparin precipitating lipoproteins (HPL) and an increase in high density lipoproteins that are not precipitated by heparin. The increase in high density lipoproteins is routinely noted by decreases in HPL/cholesterol ratios. The pattern of response following single 100 mg/kg doses of U-418792 was determined. After an I.V. dose was administered in a cottonseed oil emulsion, serum cholesterol levels were reduced, beginning at 8 hr after administration and persisting for 96 hr. Similar results, though delayed somewhat, were obtained after a single oral dose. Activity was accompanied by increases in weight and cholesterol content of livers. After multiple, daily, oral doses, liver weights, total lipids, and cholesterol contents were reduced. Hypobetalipopreteinmeic activity was enhanced by prolonged treatments as demonstrated by analyses of serum obtained weekly throughout 7 wk.

Adamantane

Cholesteryl ester metabolism in fat- and cholesterol/fat-fed guinea pigs.

Guinea pigs were fed a semisynthetic diet containing 10% (by weight) cottonseed oil with or without 1% cholesterol. In response to cholesterol/fat feeding there was a significant accumulation of cholesteryl ester (CE), particularly in the liver, but also in the kidney, spleen and suprarenal glands. The hepatic acyl-CoA:cholesterol acyltransferase (ACAT) increased 5-10 times when the animals were fed cholesterol fat during 11 weeks while the acid cholesterol esterase (CE-ase) was similar in the two dietary groups. Intestinal lymph showed the highest content of cholesterol (both free and esterified) in guinea pigs fed cholesterol/fat. A low activity of lecithin:cholesterol acyltransferase (LCAT) was present in the intestinal lymph, irrespective of dietary composition. Triglyceride-rich lipoproteins seem to inhibit LCAT activity in the intestinal lymph. Plasma cholesterol levels in animals fed cholesterol/fat increased markedly while LCAT remained unaffected by the diets. Activity of ACAT and CE-ase in kidney and spleen was low compared to liver tissue and the enzyme activities were not affected by the cholesterol/fat feeding.

Adrenal Glands