Proteome-wide analyses reveal diverse functions of protein acetylation and succinylation modifications in fast growing stolons of bermudagrass (Cynodon dactylon L.).

BACKGROUND: Bermudagrass (Cynodon dactylon L.) is an important warm-season turfgrass species with well-developed stolons, which lay the foundation for the fast propagation of bermudagrass plants through asexual clonal growth. However, the growth and development of bermudagrass stolons are still poorly understood at the molecular level. RESULTS: In this study, we comprehensively analyzed the acetylation and succinylation modifications of proteins in fast-growing stolons of the bermudagrass cultivar Yangjiang. A total of 4657 lysine acetylation sites on 1914 proteins and 226 lysine succinylation sites on 128 proteins were successfully identified using liquid chromatography coupled to tandem mass spectrometry, respectively. Furthermore, 78 proteins and 81 lysine sites were found to be both acetylated and succinylated. Functional enrichment analysis revealed that acetylated proteins regulate diverse reactions of carbohydrate metabolism and protein turnover, whereas succinylated proteins mainly regulate the citrate cycle. These results partly explained the different growth disturbances of bermudagrass stolons under treatment with sodium butyrate and sodium malonate, which interfere with protein acetylation and succinylation, respectively. Moreover, 140 acetylated proteins and 42 succinylated proteins were further characterized having similarly modified orthologs in other grass species. Site-specific mutations combined with enzymatic activity assays indicated that the conserved acetylation of catalase and succinylation of malate dehydrogenase both inhibited their activities, further implying important regulatory roles of the two modifications. CONCLUSION: In summary, our study implied that lysine acetylation and succinylation of proteins possibly play important regulatory roles in the fast growth of bermudagrass stolons. The results not only provide new insights into clonal growth of bermudagrass but also offer a rich resource for functional analyses of protein lysine acetylation and succinylation in plants.

Effects of low nitrogen supply on nitrogen uptake, assimilation and remobilization in wild bermudagrass.

The natural mechanism of underlying the low nitrogen (N) tolerance of wild bermudagrass (Cynodon dactylon (L.) Pers.) germplasm was important for reducing N fertilizer input to turf while also maintaining acceptable turf quality. The growth, N uptake, assimilation and remobilization of two wild bermudagrass accessions (C291, low N tolerant and C716, low N sensitive) were determined under low N (0.5 mM) and control N (5 mM) levels. C291 exhibited lower reduction in shoot and plant dry weight than C716. Furthermore, C291 presented a lower decrease in 15NO3- influx compared with C716, maintained its root dry weight and root surface and showed obviously enhanced CyNRT2.2 and CyNRT2.3 expression resulting in higher shoot NO3--N content than the control. Moreover, in C291, nitrate reductase (NR) activity had no significant difference with control, and cytosolic glutamine synthetase (GS1) protein content, glutamate synthetase (GOGAT) activity and glutamate dehydrogenase (GDH) activity higher than control, result in the soluble protein and free amino acid contents in the shoots did not differ compared with that in the control under low N conditions. Overall, the low N tolerant wild bermudagrass accessions adopted a low N supply based on improved root N uptake ability to achieve more nitrate to kept shoot N assimilation, and meanwhile increased N remobilization in the shoots, thereby maintaining a better N status in bermudagrass. The findings may help elucidate the low N tolerance mechanisms in bermudagrass and therefore facilitate genetic improvement of N use efficiency aiming to promote low-input turfgrass management.

Physical Exercise in the Context of Air Pollution: An Emerging Research Topic.

Physical exercise (PE) brings physiological benefits to human health; paradoxically, exposure to air pollution (AP) is harmful. Hence, the combined effects of AP and PE are interesting issues worth exploring. The objective of this study is to review literature involved in AP-PE fields to perform a knowledge-map analysis and explore the collaborations, current hotspots, physiological applications, and future perspectives. Herein, cluster, co-citation, and co-occurrence analysis were applied using CiteSpace and VOSviewer software. The results demonstrated that AP-PE domains have been springing up and in rapid growth since the 21st century. Subsequently, active countries and institutions were identified, and the productive institutions were mainly located in USA, China, UK, Spain, and Canada. Developed countries seemed to be the major promoters. Additionally, subject analysis found that environmental science, public health, and sports medicine were the core subjects, and multidimensional communications were forming. Thereafter, a holistic presentation of reference co-citation clusters was conducted to discover the research topics and trace the development focuses. Youth, elite athletes, and rural population were regarded as the noteworthy subjects. Commuter exposure and moderate aerobic exercise represented the common research context and exercise strategy, respectively. Simultaneously, the research hotspots and application fields were elaborated by keyword co-occurrence distribution. It was noted that physiological adaptations including respiratory, cardiovascular, metabolic, and mental health were the major themes; oxidative stress and inflammatory response were the mostly referred mechanisms. Finally, several challenges were proposed, which are beneficial to promote the development of the research field. Molecular mechanisms and specific pathways are still unknown and the equilibrium points and dose-effect relationships remain to be further explored. We are highly confident that this study provides a unique perspective to systematically and comprehensively review the pieces of AP-PE research and its related physiological mechanisms for future investigations.

Integration of the metabolome and transcriptome reveals the mechanism of resistance to low nitrogen supply in wild bermudagrass (Cynodon dactylon (L.) Pers.) roots.

BACKGROUND: Nitrogen (N) is an essential macronutrient that significantly affects turf quality. Commercial cultivars of bermudagrass (Cynodon dactylon (L.) Pers.) require large amounts of nitrogenous fertilizer. Wild bermudagrass germplasm from natural habitats with poor nutrition and diverse N distributions is an important source for low-N-tolerant cultivated bermudagrass breeding. However, the mechanisms underlying the differences in N utilization among wild germplasm resources of bermudagrass are not clear. RESULTS: To clarify the low N tolerance mechanism in wild bermudagrass germplasm, the growth, physiology, metabolome and transcriptome of two wild accessions, C291 (low-N-tolerant) and C716 (low-N-sensitive), were investigated. The results showed that root growth was less inhibited in low-N-tolerant C291 than in low-N-sensitive C716 under low N conditions; the root dry weight, soluble protein content and free amino acid content of C291 did not differ from those of the control, while those of C716 were significantly decreased. Down-regulation of N acquisition, primary N assimilation and amino acid biosynthesis was less pronounced in C291 than in C716 under low N conditions; glycolysis and the tricarboxylic acid (TCA) cycle pathway were also down-regulated, accompanied by a decrease in the biosynthesis of amino acids; strikingly, processes such as translation, biosynthesis of the structural constituent of ribosome, and the expression of individual aminoacyl-tRNA synthetase genes, most of genes associated with ribosomes related to protein synthesis were all up-regulated in C291, but down-regulated in C716. CONCLUSIONS: Overall, low-N-tolerant wild bermudagrass tolerated low N nutrition by reducing N primary assimilation and amino acid biosynthesis, while promoting the root protein synthesis process and thereby maintaining root N status and normal growth.

Proteome analysis of bermudagrass stolons and rhizomes provides new insights into the adaptation of plant stems to aboveground and underground growth.

As an important perennial warm-season turfgrass species, bermudagrass (Cynodon dactylon L.) forms underground-growing rhizomes and aboveground-growing stolons simultaneously, making it a fast propagating clonal plant with strong regeneration ability. In the current study, we compared the internode proteomes of rhizomes and stolons at the same developmental stage in the bermudagrass cultivar Yangjiang using iTRAQ. The results indicated that 228 protein species were differentially accumulated in the two specialized stems. In agreement with the different contents of starch, chlorophyll, anthocyanin and H2O2 in the two types of stems, photosynthesis and flavonoid biosynthesis were enriched with differentially accumulated protein species (DAPs) in stolons, whereas starch and sucrose metabolism, glycolysis, and H2O2 metabolism were enriched with DAPs in rhizomes. Burying stolons in the soil resulted in the gradual degradation of chlorophyll and anthocyanin, accumulation of starch, and increment of H2O2, which is similar to the physiological characteristics of rhizomes. These results collectively revealed that stolons and rhizomes of bermudagrass have significant differences at the proteome level and light might play important regulatory roles in the discrepancy of the proteome profiles and specialization of the two stems, providing new insights into the adaptation of plant stems to aboveground and underground growth. BIOLOGICAL SIGNIFICANCE: As two types of specialized stems that grow underground and aboveground respectively, rhizomes and stolons play important roles in overwintering and ecological invasion of many perennial and clonal plants. However, because rhizomes and stolons rarely coexist in single plant species, the differences between the two stems remain unclear at the molecular level. In this study, through an iTRAQ comparative proteomic analysis, we reported the identification of 228 differentially accumulated protein species (DAPs) in rhizomes and stolons of bermudagrass for the first time. We found that the 228 DAPs were interconnected to form protein networks in regulating diverse cellular activities and biochemical reactions. We also observed that stolons growing underground showed similar physiological activities and DAP expression as those of underground-growing rhizomes, suggesting that light might play important regulatory roles in the specialization of stolons and rhizomes. These results expanded our understanding of the mysterious adaption of plant stems to different growth conditions.

Effects of Tai Chi exercise on improving walking function and posture control in elderly patients with knee osteoarthritis: A systematic review and meta-analysis.

OBJECTIVE: It remains unclear whether Tai Chi is effective for walking function and posture control improvements in aged populations with knee osteoarthritis. The aim of this study was to systematically evaluate the effects of Tai Chi on improving walking function and posture control in elderly patients with knee osteoarthritis by updating the latest trial evidence. METHODS: Web of Science, PubMed/Medline, Embase, Scopus, PEDro, and Cochrane library were searched up to October 1, 2020 to identify RCTs evaluating Tai Chi for improving walking function and posture control in older adults with knee osteoarthritis. The primary outcomes were walking function and posture control. Meta-analysis was performed with RevMan Version 5.3 software. RESULTS: A total of 603 participants with knee osteoarthritis in the 11 trials were included. The results of meta-analysis showed that: The Tai Chi group was associated with better performance in 6-minute walk test (6 MWT), time up and go test (TUG) and "Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index" Physical Function Score than the control group ([MD: 46.67, 95% CI 36.91-56.43, P < .001]), ([MD: -0.89, 95% CI -1.16 to -0.61, P < .001]), ([MD: -11.28, 95% CI -13.33 to -9.24, P < .001]). CONCLUSION: This meta-analysis provided evidence from 11 RCTs that Tai Chi could be an excellent physical training strategy for improving walking function and posture control in older adults with knee osteoarthritis. Assuming that Tai Chi is at least effective and safe in most areas, it can be used as an adjuvant and reliable physical training strategy for walking function upgrading and balance control improvements for older patients with knee osteoarthritis.

Comparative proteomic analysis provides new insights into the specialization of shoots and stolons in bermudagrass (Cynodon dactylon L.).

BACKGROUND: Bermudagrass (Cynodon dactylon L.) is an important turfgrass species with two types of stems, shoots and stolons. Despite their importance in determining the morphological variance and plasticity of bermudagrass, the intrinsic differences between stolons and shoots are poorly understood. RESULTS: In this study, we compared the proteomes of internode sections of shoots and stolons in the bermudagrass cultivar Yangjiang. The results indicated that 376 protein species were differentially accumulated in the two types of stems. Pathway enrichment analysis revealed that five and nine biochemical pathways were significantly enriched in stolons and shoots, respectively. Specifically, enzymes participating in starch synthesis all preferentially accumulated in stolons, whereas proteins involved in glycolysis and diverse transport processes showed relatively higher abundance in shoots. ADP-glucose pyrophosphorylase (AGPase) and pyruvate kinase (PK), which catalyze rate-limiting steps of starch synthesis and glycolysis, showed high expression levels and enzyme activity in stolons and shoots, respectively, in accordance with the different starch and soluble sugar contents of the two types of stems. CONCLUSIONS: Our study revealed the differences between the shoots and stolons of bermudagrass at the proteome level. The results not only expand our understanding of the specialization of stolons and shoots but also provide clues for the breeding of bermudagrass and other turfgrasses with different plant architectures.

Unbiased phosphoproteome profiling uncovers novel phosphoproteins and phosphorylation motifs in bermudagrass stolons.

As a widely used turfgrass species, bermudagrass (Cynodon dactylon L.) can be easily propagated through colonial growth of stolons. Previous studies collectively revealed that exotic environmental factors and intrinsic hormones and genes are all involved in the differentiation, development, and diageotropical growth of stolons. However, the detailed molecular mechanism how environmental and hormone signals regulate the gene expression and biochemical activities in bermudagrass stolons remains unclear. In this study, we observed that reversible phosphorylation modification plays important roles in normal growth and physiological functions of bermudagrass stolons. LC-MS/MS analyses of the total protein extracts of bermudagrass stolons without preliminary phosphopeptide-enrichment successfully identified 646 nonredundant phosphorylation sites and 485 phosphoproteins. The phosphoproteins were significantly enriched in protein phosphorylation regulation and starch metabolism processes. Motif-X analyses further revealed that phosphoproteins containing novel phosphorylation motifs might be involved in transcription regulation of bermudagrass stolons. These results greatly expanded our understanding of the growth and development of bermudagrass stolons at the post-translational level.

Impact of Beef and Beef Product Intake on Cognition in Children and Young Adults: A Systematic Review.

(1) Background: Undernutrition and micronutrient deficiency have been consistently linked to cognitive impairment among children and young adults. As a primary source of dietary animal protein, beef consumption holds the potential to improve diet quality and positively influence cognitive function. This study systematically reviewed evidence linking beef intake to cognition among children and young adults. (2) Methods: A literature search was conducted in seven electronic bibliographic databases for studies assessing the impact of beef consumption on cognition. (3) Results: We identified eight studies reporting results from five unique interventions. Two interventions were conducted in Kenya, two in the U.S. and one in four countries including Guatemala, Pakistan, Democratic Republic of the Congo and Zambia. Only one intervention employed a non-feeding control arm and found beef consumption to improve cognitive abilities compared to the control. However, the other interventions comparing beef consumption to other food types found no consistent result. (4) Conclusions: Evidence pertaining to the impact of beef consumption on cognition remains limited due to the small and heterogeneous set of studies. Future research should adopt a population representative sample and longer follow-up period, employ a non-feeding control arm and comprehensively measure nutrient intakes among study participants.

Full-length RNA sequencing reveals unique transcriptome composition in bermudagrass.

Bermudagrass [Cynodon dactylon (L.) Pers.] is an important perennial warm-season turfgrass species with great economic value. However, the reference genome and transcriptome information are still deficient in bermudagrass, which severely impedes functional and molecular breeding studies. In this study, through analyzing a mixture sample of leaves, stolons, shoots, roots and flowers with single-molecule long-read sequencing technology from Pacific Biosciences (PacBio), we reported the first full-length transcriptome dataset of bermudagrass (C. dactylon cultivar Yangjiang) comprising 78,192 unigenes. Among the unigenes, 66,409 were functionally annotated, whereas 27,946 were found to have two or more isoforms. The annotated full-length unigenes provided many new insights into gene sequence characteristics and systematic phylogeny of bermudagrass. By comparison with transcriptome dataset in nine grass species, KEGG pathway analyses further revealed that C4 photosynthesis-related genes, notably the phosphoenolpyruvate carboxylase and pyruvate, phosphate dikinase genes, are specifically enriched in bermudagrass. These results not only explained the possible reason why bermudagrass flourishes in warm areas but also provided a solid basis for future studies in this important turfgrass species.

Molecular cloning and sequence variance analysis of the TEOSINTE BRANCHED1 (TB1) gene in bermudagrass [Cynodon dactylon (L.) Pers].

TEOSINTE BRANCHED1 (TB1) encodes a TEOSINTE BRANCHED1, CYCLOIDEA, and PROLIFERATING CELL FACTOR (TCP) transcription factor that represses axillary bud outgrowth and lateral branch formation in plants. Previous studies have elucidated the essential tillering regulatory roles of TB1 in many grasses, including maize and rice; however, the functions of TB1 in turf grasses remain unclear. In this study, we cloned the CdTB1 gene from bermudagrass, an important turfgrass species, and characterized the transactivation function of the CdTB1 protein. Sequencing the CdTB1 gene locus in a mini-core germplasm collection of Chinese bermudagrasses led to the successful identification of 66 SNP and 2 indel mutations in the protein-coding region as well as 28 SNP and 11 indel mutations in the promoter region. Interestingly, mutations in the C-terminal transactivation domain of the CdTB1 protein had no significant influence on the transactivation activity, whereas a novel 335-bp insertion mutation located in the promoter region could significantly increase the expression of the CdTB1 gene. Furthermore, wild accessions of bermudagrass harboring the novel insertion mutation were found to have significantly reduced tillers compared with other accessions, suggesting a negative correlation between the mutation and tillering. The results of this study not only expanded our knowledge of TB1 gene expression regulation but also provided possible molecular markers to breed cultivars of turf and forage grasses with specific architectural features.

Comparative transcriptome analysis provides new insights into erect and prostrate growth in bermudagrass (Cynodon dactylon L.).

Bermudagrass (Cynodon dactylon L.) is a prominent warm-season turf and forage grass species with multiple applications. In most C. dactylon cultivars and accessions, erect-growing stems (shoot) and prostrate-growing stems (stolon) often coexist. These two types of stems are both formed through tillering but grow in two directions with different tiller angles. Elucidating the mechanism of tiller angle regulation in bermudagrass could provide important clues to breed cultivars with different plant architectural features for diverse usage. In this study, we compared the stem internode transcriptome of two bermudagrass wild accessions with extremely different tiller angles and stem growth directions. A total of 2088 and 12,141 unigenes were preferentially expressed in prostrate-growing wild accession C792 and erect-growing wild accession C793, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology-based Annotation System (KOBAS) analyses further indicated that light- and gravity-responsive genes were enriched in accession C792, whereas lignin synthesis-related genes were enriched in accession C793, which well explains the difference in lignification of vascular bundles and mechanical tissues in the two accessions. These results not only expand our understanding of the genetic control of tiller angle and stem growth direction in bermudagrass but also provide insight for future molecular breeding of C. dactylon and other turfgrass species with different plant architectures.

Genetic diversity and population structure of Chinese natural bermudagrass [Cynodon dactylon (L.) Pers.] germplasm based on SRAP markers.

Bermudagrass [Cynodon dactylon (L.) Pers.], an important turfgrass used in public parks, home lawns, golf courses and sports fields, is widely distributed in China. In the present study, sequence-related amplified polymorphism (SRAP) markers were used to assess genetic diversity and population structure among 157 indigenous bermudagrass genotypes from 20 provinces in China. The application of 26 SRAP primer pairs produced 340 bands, of which 328 (96.58%) were polymorphic. The polymorphic information content (PIC) ranged from 0.36 to 0.49 with a mean of 0.44. Genetic distance coefficients among accessions ranged from 0.04 to 0.61, with an average of 0.32. The results of STRUCTURE analysis suggested that 157 bermudagrass accessions can be grouped into three subpopulations. Moreover, according to clustering based on the unweighted pair-group method of arithmetic averages (UPGMA), accessions were divided into three major clusters. The UPGMA dendrogram revealed that accessions from identical or adjacent areas were generally, but not entirely, clustered into the same cluster. Comparison of the UPGMA dendrogram and the Bayesian STRUCTURE analysis showed general agreement between the population subdivisions and the genetic relationships among accessions. Principal coordinate analysis (PCoA) with SRAP markers revealed a similar grouping of accessions to the UPGMA dendrogram and STRUCTUE analysis. Analysis of molecular variance (AMOVA) indicated that 18% of total molecular variance was attributed to diversity among subpopulations, while 82% of variance was associated with differences within subpopulations. Our study represents the most comprehensive investigation of the genetic diversity and population structure of bermudagrass in China to date, and provides valuable information for the germplasm collection, genetic improvement, and systematic utilization of bermudagrass.