Genome-wide identification and expression analysis of the KNOX family and its diverse roles in response to growth and abiotic tolerance in sweet potato and its two diploid relatives.

KNOXs, a type of homeobox genes that encode atypical homeobox proteins, play an essential role in the regulation of growth and development, hormonal response, and abiotic stress in plants. However, the KNOX gene family has not been explored in sweet potato. In this study, through sequence alignment, genomic structure analysis, and phylogenetic characterization, 17, 12 and 11 KNOXs in sweet potato (I. batatas, 2n = 6x = 90) and its two diploid relatives I. trifida (2n = 2x = 30) and I. triloba (2n = 2x = 30) were identified. The protein physicochemical properties, chromosome localization, phylogenetic relationships, gene structure, protein interaction network, cis-elements of promoters, tissue-specific expression and expression patterns under hormone treatment and abiotic stresses of these 40 KNOX genes were systematically studied. IbKNOX4, -5, and - 6 were highly expressed in the leaves of the high-yield varieties Longshu9 and Xushu18. IbKNOX3 and IbKNOX8 in Class I were upregulated in initial storage roots compared to fibrous roots. IbKNOXs in Class M were specifically expressed in the stem tip and hardly expressed in other tissues. Moreover, IbKNOX2 and - 6, and their homologous genes were induced by PEG/mannitol and NaCl treatments. The results showed that KNOXs were involved in regulating growth and development, hormone crosstalk and abiotic stress responses between sweet potato and its two diploid relatives. This study provides a comparison of these KNOX genes in sweet potato and its two diploid relatives and a theoretical basis for functional studies.

Effect of grape pomace supplement on growth performance, gastrointestinal microbiota, and methane production in Tan lambs.

Grape pomace (GP), a by-product in wine production, is nutritious and can be used as a feed ingredient for ruminants; however, its role in shaping sheep gastrointestinal tract (GIT) microbiota is unclear. We conducted a controlled trial using a randomized block design with 10 Tan lambs fed a control diet (CD) and 10 Tan lambs fed a pelleted diet containing 8% GP (dry matter basis) for 46 days. Rumen, jejunum, cecum, and colon bacterial and archaeal composition were identified by 16S rRNA gene sequencing. Dry matter intake (DMI) was greater (p < 0.05) in the GP than CD group; however, there was no difference in average daily gain (ADG, p < 0.05) and feed conversion ratio (FCR, p < 0.05) between the two groups. The GP group had a greater abundance of Prevotella 1 and Prevotella 7 in the rumen; of Sharpe, Ruminococcaceae 2, and [Ruminococcus] gauvreauii group in the jejunum; of Ruminococcaceae UCG-014 and Romboutsia in the cecum, and Prevotella UCG-001 in the colon; but lesser Rikenellaceae RC9 gut group in the rumen and cecum, and Ruminococcaceae UCG-005 and Ruminococcaceae UCG-010 in the colon than the CD group. The pathways of carbohydrate metabolism, such as L-rhamnose degradation in the rumen, starch and glycogen degradation in the jejunum, galactose degradation in the cecum, and mixed acid fermentation and mannan degradation in the colon were up-graded; whereas, the pathways of tricarboxylic acid (TCA) cycle VIII, and pyruvate fermentation to acetone in the rumen and colon were down-graded with GP. The archaeal incomplete reductive TCA cycle was enriched in the rumen, jejunum, and colon; whereas, the methanogenesis from H2 and CO2, the cofactors of methanogenesis, including coenzyme M, coenzyme B, and factor 420 biosynthesis were decreased in the colon. The study concluded that a diet including GP at 8% DM did not affect ADG or FCR in Tan lambs. However, there were some potential benefits, such as enhancing propionate production by microbiota and pathways in the GIT, promoting B-vitamin production in the rumen, facilitating starch degradation and amino acid biosynthesis in the jejunum, and reducing methanogenesis in the colon.

Comparative Analysis of the PYL Gene Family in Three Ipomoea Species and the Expression Profiling of IbPYL Genes during Abiotic Stress Response in Sweetpotato.

Abscisic acid (ABA), a critical phytohormone that regulates plant development and stress response, is sensed by the ABA receptors PYR/PYL/RCAR (PYLs). The PYL genes have been widely studied in multiple plant species, while a systematic analysis of PYL genes in the genus Ipomoea remains unperformed. Here, a total of 13, 14, and 14 PYLs were identified in Ipomoea batatas, Ipomoea trifida, and Ipomoea triloba, respectively. Fragment duplication was speculated to play prominent roles in Ipomoea PYL gene expansions. These Ipomoea PYLs were classified into three subfamilies via phylogenetic analysis, which was supported by exon-intron structures and conserved motif analyses. Additionally, the interspecies collinearity analysis further depicted a potential evolutionary relationship between them. Moreover, qRT-PCR analysis showed that multiple IbPYLs are highly and differentially responsive to abiotic stress treatments, suggesting their potential roles in sweetpotato stress responses. Taken together, these data provide valuable insights into the PYLs in the genus Ipomoea, which may be useful for their further functional analysis of their defense against environmental changes.

Production and characterization of a novel interspecific somatic hybrid combining drought tolerance and high quality of sweet potato and Ipomoea triloba L.

KEY MESSAGE: A novel interspecific somatic hybrid combining drought tolerance and high quality of sweet potato and Ipomoea triloba L. was obtained and its genetic and epigenetic variations were studied. Somatic hybridization can be used to overcome the cross-incompatibility between sweet potato (Ipomoea batatas (L.) Lam.) and its wild relatives and transfer useful and desirable genes from wild relatives to cultivated plants. However, most of the interspecific somatic hybrids obtained to date cannot produce storage roots and do not exhibit agronomic characters. In the present study, a novel interspecific somatic hybrid, named XT1, was obtained through protoplast fusion between sweet potato cv. Xushu 18 and its wild relative I. triloba. This somatic hybrid produced storage roots and exhibited significantly higher drought tolerance and quality compared with its cultivated parent Xushu 18. Transcriptome and real-time quantitative PCR (qRT-PCR) analyses revealed that the well-known drought stress-responsive genes in XT1 and I. triloba were significantly up-regulated under drought stress. The genomic structural reconstructions between the two genomes of the fusion parents in XT1 were confirmed using genomic in situ hybridization (GISH) and specific nuclear and cytoplasmic DNA markers. The DNA methylation variations were characterized by methylation-sensitive amplified polymorphism (MSAP). This study not only reveals the significance of somatic hybridization in the genetic improvement of sweet potato but also provides valuable materials and knowledge for further investigating the mechanism of storage root formation in sweet potato.

Effects of soil ingestion on nutrient digestibility and rumen bacterial diversity of Tibetan sheep.

Tibetan sheep (Ovis aries) are the most numerous livestock in Tibet Plateau pasture ecosystem and have strong ecological adaptability. In the natural grazing system, soil as a natural nutrient carrier and involuntarily or intentionally ingested by Tibetan sheep contribute as an important feed approach. However, quantifying the dosages of soil ingestion for the Tibetan sheep still needs to be clarified. This study aims to characterize nutrient digestibility and rumen bacterial communities by Tibetan sheep in response to different levels of soil ingestion. Thirty sheep were selected and divided into five treatments with soil ingestion (0%, 5%, 10%, 15%, and 20%). The conclusion demonstrated that soil ingestion improved the dry matter digestibility (59.3-62.97%), ether extract (59.79-67.87%) and crude protein (59.81-66.47%) digestibility, particularly 10% soil ingestion has highest nutrient digestibility. The rumen fermentation environment adjusted after soil ingestion by improvement of pH, ammonia nitrogen and volatile fatty acids. Appropriate soil ingestion reduced the bacterial diversity ranged from 946 to 1000 OUTs as compared control (1012), and the rumen bacterial community dominant by typical fiber digestion associated Firmicutes (47.48-53.56%), Bacteroidetes (34.93-40.02%) and Fibrobacteres (4.36-9.27%). Especially, the highest digestible feed capacity and stronger environment adaptability present in 10% soil ingestion Tibetan sheep. Overall, soil ingestion stimulates rumen metabolism by creating a favorable environment for microbial fermentation, improved bacterial community abundance associated with cellulose and saccharide degradation, contribute nutrient digestibility and growth performance of Tibetan sheep.

Dietary supplementation of hemp oil in teddy dogs: Effect on apparent nutrient digestibility, blood biochemistry and metabolomics.

Present study aimed to evaluate the influence of distinct concentration of dietary supplements hemp oil on apparent nutrient digestibility, blood biochemical parameters and metabolomics of teddy dogs. A total of 25 healthy teddy dogs were selected and divided into five treatments according to diet supplements hemp oil at a rate of 0% (A), 0.5% (B), 1% (C), 2% (D), and 4% (E). Appropriate added hemp oil improved apparent nutrient digestibility of dry matter, crude protein and crude fat (86.32-88.08%, 86.87-88.87% and 96.76-97.43%). The hemp oil significantly increased blood biochemical of utilization related total protein, albumin and globulin (61.33-69.54, 35.08-40.38 and 26.53-31.63 g/L), immunity capacity related immunoglobulin E and γ-interferon (203-347kU/L and 23.04-25.78ng/L), energy-related thyroxine and triiodothyronine (27.11-36.75 and 0.94-1.67 nmol/L). In addition, hemp oil improved superoxide dismutation (26.47-33.02 U/ml) and reduced malondialdehyde (5.30-3.28 nmol/ml). The differential metabolites mainly included nucleotides and metabolites of oxidized lipids, bile and other fatty acids, coenzymes and vitamins. The main metabolic pathways included purine and arachidonic acid metabolism, bile and unsaturated fatty acid biosynthesis, cell oxidative phosphorylation and rheumatoid arthritis. Overall, appropriate dietary supplements hemp oil positively to nutrient digestibility and blood metabolism, immunity and antioxidant capacity, 1% to 2% hemp oil supplements was recommended for teddy dog diet.

Research progress on inosine monophosphate deposition mechanism in chicken muscle.

With the continuous improvements in human diet, there is an ever-increasing demand for high-quality chicken, so it is particularly important for poultry breeders to carry out the breeding of high-quality broilers in a timely fashion. Inosine monophosphate (IMP) is a flavor-enhancing substance, which plays a critical role in the umami taste of the muscle, making the content of IMP an important umami taste indicator. Currently, research on the deposition mechanism of IMP in chicken is not only necessary for chicken breeders to promote the production of high-quality meat and poultry but also to meet the human demand for chicken meat. In this paper, the research history of IMP, its structure and taste mechanisms, the pathway and influencing factors of de novo IMP synthesis, and the key genes regulating IMP synthesis and metabolism are briefly summarized. Our aim was to lay a theoretical foundation and provide scientific background and research directions for further research on high-quality broiler breeding.

Modulation of broiler plasma metabolic spectrum by the addition of lysine residue to the diet.

Flavour is an important factor in evaluating meat quality, and amino acids and fats are important components affecting meat flavour. In this study, we evaluated the relationship between the variation of lysine residue addition and the slaughter performance and meat quality of broilers, which decreased with the addition of lysine residues but improved the meat quality of the broilers. 10% lysine residue addition was the most beneficial for reducing feed cost and improving meat quality. Meanwhile, the plasma metabolites of broilers fed increasing concentrations of lysine residue supplemented feeds were analysed using liquid chromatography-mass spectrometry (LC-MS). Principal component analysis (PCA) and partial least square discriminant analysis (OPLS-DA) were used screen, the differential metabolites induced by lysine residue. In the broilers 29, 37, 63, 87, 80 and 111 differential metabolites were detected (p < 0.05). Amongst them, 3-iodotyrosine, N-methyl-L-glutamic acid, coumaraldehyde, 2-dimethylphenol, N-methylnicotinamide and L-erythrone were the common differential metabolites between group A and groups B, C, D, E, F and G. The addition of lysine residue was positively correlated with alanine aminotransferase (p < 0.05, r = 0.942) and high-density lipoprotein cholesterol (p < 0.05, r = 0.798) and negatively correlated with aspartate aminotransferase (p < 0.05, r = 0.822). According to the classification of differential metabolites and their enriched pathway analysis, differential metabolites mainly caused changes in amino acid and lipid metabolism. Our study shows that a certain proportion of lysine residue in diet affects the specific metabolic pathway of broilers, which may affect amino acid and fat metabolism by regulating alanine aminotransferase, aspartate aminotransferase and high-density lipoprotein cholesterol, ultimately affecting the flavour.

Underlying mechanisms of phosphodiesterase 10A and glutamate-ammonia ligase genes that regulate inosine monophosphate deposition and thereby affect muscle tenderness in Jingyuan chickens.

OBJECTIVE: Inosine monophosphate (IMP) is a key factor that imparts of meat flavor. Differences in the IMP content in the muscles were evaluated to improve chicken meat quality. METHODS: For this study, the IMP content was detected by high performance liquid chromatography. The gene expression profiles of Jingyuan chickens with different feeding patterns and different sexes were analyzed by RNA-sequencing (RNA-seq). RESULTS: Breast muscle IMP content in free-range chickens was extremely significantly higher than that of caged chickens (p<0.01). Breast muscle IMP content in hens was also higher than that of cocks, but the difference was not significant. Correlation analysis showed that the breast muscle IMP content in caged hens and cocks was negatively correlated with the shear force, and the breast muscle IMP content in free-range hens was significantly negatively correlated with the shear force (p<0.05). The two key genes associated with IMP synthesis in chickens with different feeding patterns were glutamate-ammonia ligase (GLUL) and phosphodiesterase 10A (PDE10A). Bioinformatics analysis revealed that the GLUL and PDE10A genes are involved in glutamine biosynthesis and purine salvage pathways respectively. In addition, GLUL expression was positively correlated with the IMP content in caged and free-range chickens, and PDE10A expression was significantly positively correlated with the IMP content in caged and free-range chickens (p<0.05). CONCLUSION: These findings will facilitate the comprehension of the deposition of IMP in the muscles and thereby aid the process of selection and breeding of good quality local chickens.

Cross-talk Between Histone and DNA Methylation Mediates Bone Loss in Hind Limb Unloading.

Bone loss induced by mechanical unloading is a common skeletal disease, but the precise mechanism remains unclear. The current study investigated the role of histone methylation, a key epigenetic marker, and its cross-talk with DNA methylation in bone loss induced by mechanical unloading. The expression of G9a, ubiquitin-like with PHD and ring finger domains 1 (UHRF1), and DNA methylation transferase 1 (DNMT1) were increased in hind limb unloading (HLU) rats. This was accompanied by an increased level of histone H3 lysine 9 (H3K9) di-/tri-methylation at lncH19 promoter. Then, alteration of G9a, DNMT1, or UHRF1 expression significantly affected lncH19 level and osteogenic activity in UMR106 cells. Osteogenic gene expression and matrix mineralization were robustly promoted after simultaneous knockdown of G9a, DNMT1, and UHRF1. Furthermore, physical interactions of lncH19 promoter with G9a and DNMT1, as well as direct interactions among DNMT1, G9a, and UHRF1 were detected. Importantly, overexpression of DNMT1, G9a, or UHRF1, respectively, resulted in enrichment of H3K9me2/me3 and 5-methylcytosine at lncH19 promoter. Finally, in vivo rescue experiments indicated that knockdown of DNMT1, G9a, or UHRF1 significantly relieved bone loss in HLU rats. In conclusion, our research demonstrated the critical role of H3K9 methylation and its cross-talk with DNA methylation in regulating lncH19 expression and bone loss in HLU rats. Combined targeting of DNMT1, G9a, and UHRF1 could be a promising strategy for the treatment of bone loss induced by mechanical unloading. © 2021 American Society for Bone and Mineral Research (ASBMR).

Correlation Analysis between AK1 mRNA Expression and Inosine Monophosphate Deposition in Jingyuan Chickens.

In this study, we examined correlations between the deposition of inosine monophosphate (IMP) and mRNA expression of the adenylate kinase 1 (AK1) gene in Jingyuan chicken. The IMP content was determined by high-performance liquid chromatography. Transcriptome sequencing was used to screen the differentially expressed gene AK1 and real-time quantitative polymerase chain reaction (PCR) to determine the expression level of AK1 mRNA associated with IMP synthesis. IMP and inosine content in the breast muscles of both Jingyuan cocks and hens was found to be significantly higher than that in the leg muscles. Similarly, the expression of AK1 mRNA in the breast muscles of cocks and hens was significantly higher than that in the leg muscles. Moreover, AK1 mRNA expression in cock breast muscles was negatively correlated with IMP content, whereas its expression in cock leg muscles was positively correlated with IMP content. In contrast, the expression of AK1 mRNA in hen breast and leg muscles was significantly positively correlated with IMP content. These findings provide a scientific basis for enhancing the meat flavor of Jingyuan chicken and promoting the development and utilization of local variety resources, as well as constituting a basis for screening IMP-regulated genes. Our study will advance our current understanding of AK1 function.

Cloning expression and immunogenicity analysis of inhibin gene in Ye Mule Aries sheep.

BACKGROUND: Ye Mule Aries sheep is one of the most important sheep breeds in Xinjiang, China. This breed is well adapted to harsh environmental conditions and displays strong disease resistance, fast growth, and high cold tolerance. To analyze the clonal expression and immunogenicity of the Ye Mule Aries sheep inhibin gene, total RNA was extracted from sheep ovarian tissue and used as a template to generate a eukaryotic expression vector and study inhibin immunogenicity. METHODS: Primers were designed to amplify the inhibin A gene via polymerase chain reaction and the amplified product was cloned between the ScalI and EcoRI restriction sites of the expression vector pEGFP-N1 to construct a recombinant plasmid, pEGFP-INHα. Following the validation of successful cloning, the pEGFP-INHα plasmid was transfected into BHK cells to verify expression in eukaryotes and subsequently utilized as an antigen in rabbits. Rabbits were tested for anti-inhibin antibodies and serum follicle-stimulating hormone (FSH) concentrations. RESULTS: The analysis of the INHα gene sequence revealed that INHα is 1109 bp long and is translated to an approximately 40 KDa protein. Bioinformatics approach indicated that the INHα gene is highly conserved between organisms. Immunization with the eukaryotic expression vector, pEGFP-INHα, which expresses the INHα gene elicited immune response and generatigeneration on of anti-INHα antibody. The antibody had a significant regulatory effect on the serum concentration of FSH in rabbits and led to higher levels of FSH, indicating increased ovary function. CONCLUSIONS: The present work resulted in a successful construction of eukaryotic expression plasmid pEGFP-INHα and verified the immunogenicity of this highly conserved protein. Further, the expression of pEGFP-INHα was shown to have a significant impact on the secretion of FSH, indicating a potential regulatory role in ovarian function. In conclusion, our current findings can serve as a working model for studying the effect of INHα on the breeding performance of Ye Mule Aries sheep, providing a novel strategy to improve their reproduction rates.

[Determination of microelements from different types of grass in the habitat of Przewalski's gazelle by sealed microwave digestion ICP-AES].

For the mixture forge from different types of grassland, trace elements including copper, manganese, iron, zinc and molybdenum were separately determined by ICP-AES using high pressure system-sealed microwave digestion in the habitat of the Przewalski's gazelle in Qinghai Hudong in summer (mid-June), autumn (mid-September) and winter (mid-December). The samples of mixture forage were digested with HNO3-H2O2 acids system. The detection limits of the method for the elements varied from 0.002 to 0.008 microg x g(-1), with relative standard deviations between 0.13% and 4.29%, and the spike recovery ratios for them were in the range from 94.0% to 101.30%. This method was simple, sensitive and precise compared with conventional method, which will provide scientific basis for the research on gazelles habitat condition.

[Determination of inorganic elements in chicken muscle by sealed microwave digestion ICP-AES].

The contents of inorganic elements including K, Ca, Na, Mg, P, S, Fe, Cu, Mn, Zn, Mo, and Co from the chicken muscle were determined by ICP-AES using sealed microwave digestion. The sample of the chicken muscle was digested with HNO3-H2O2 system. The relative standard deviation was less than 5% for all the elements, and the recovery was 92.5%-110% by adding standard recovery experiment. This method was simple, sensitive and precise and can perform simultaneous multi-elements determination compared with conventional method of the chicken muscle determination, which could satisfy the sample examination request and provide scientific rationale for determining inorganic elements of chicken meat.

[Determination of inorganic elements in the soil-grass-animal system by sealed microwave digestion ICP-AES].

The contents of inorganic elements including K, Ca, Na, Mg, P, S, Fe, Cu, Mn, Zn, Mo, and Co in the soil-grass-animal mineral system from Qinghai Tibetan Plateau were determined by ICP-AES using high pressure system-sealed microwave digestion. The sample of soil was digested with HNO3-HF-H2O2 acids system, but others including pasture, animal fur, liver, and serum were digested with HNO3-H2O2 acids system. The operation would be simplified and the blank value would be decreased with the above acids systems. The results were proved to be reliable with the relative standard deviation being 0.271%-2.633% for Ca, 2.971%-4.854% for Co, 0.372%-2.874% for Cu, 0.600%-3.683% for Fe, 0.347%-2.829% for K, 0.626%-2.593% for Mg, 0.705%-4.828% for Mn, 2.946%-4.622% for Mo, 0.689%-3.621% for Na, 0.422%-3.890% for P, and 0.143%-4.622% for S, 0.166%-2.399% for Zn, and all of them were less than 5% for all the elements, and the recovery being 97.1%-101.4% for Ca, 93.5%-112.5% for Co, 95.2%-104.0% for Cu, 96.9%-104.2% for Fe, 96.1%-105.6% for K, 96.2%-102.8% for Mg, 91.5%-108.9% for Mn, 95.0%-113.5% for Mo, 95.2%-101.8% for Na, 94.7%-100.7% for P, 98.3%-108.4% for S, and 97.5%-102.0% for Zn by adding standard recovery experiment. The results of determination were proved that the method of sealed microwave digestion ICP-AES was sensitive, precise, easy to operate and rapid for the determination of inorganic elements in the soil-grass-animal mineral system, and could satisfy the sample examination request. The methods and results were meaningful to research on the soil-pasture-animal mineral system including the contents of mineral elements, the circulation of mineral elements, the interaction, and the application of mineral additive.