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Background Flowering is a tightly regulated process influencing yield and promoting plant genetic diversity and conservation. Orchardgrass ( Dactylis glomerata ) exhibits excellent yield traits and stress resistance, making it ideal for animal husbandry and ecological restoration. However, the molecular regulatory factors of the flowering time of orchardgrass are still unknown, limiting its molecular breeding. Results To speed up molecular breeding to enhance flowering traits in orchardgrass, we conducted a genome-wide association study (GWAS). A diverse panel of 249 orchardgrass accessions was phenotyped for heading stage and flowering time. GWAS analysis identified 359 candidate genes that overlapped or were adjacent to effective single-nucleotide polymorphisms (SNPs), which were considered potential flowering time-related genes. Furthermore, we validated that formin-like protein 18 ( DgFH18 ) and choline monooxygenase ( DgCMO-like ) was two important flowering candidate genes by overexpressing them in Arabidopsis to unravel their potential functions. Overexpression of DgFH18 and DgCMO-like positively regulated flowering time by inducing the expression of flowering-related genes. Moreover, sucrose treatment could significantly promote the expression of flavonoid pathway genes and enhance the content of total flavonoids and anthocyanins in the DgCMO-like -overexpressing lines compared to the wild type. Conclusion These results provide valuable resources for future orchardgrass breeding programs and broaden the current comprehension of flowering time regulation in perennial grasses.

Auxin response factor (ARF), a transcription factor, is crucial in controlling growth, development, and response to environmental stress. Orchardgrass ( Dactylis glomerata ) is an economically significant, widely cultivated forage grass. However, information on the genome-wide information and functional characterization of ARFs in orchardgrass is limited. This study identified 27 ARF genes based on the orchardgrass genome database. These DgARFs were unevenly distributed across the seven orchardgrass chromosomes and clustered into four classes. Phylogenetic analysis with multispecies of ARF proteins indicated that the ARFs exhibit a relatively conserved evolutionary path. Focusing on hormone signaling responses, DgARF7 demonstrated a potential positive regulatory role in response to 3-indole acetic acid, methyl jasmonate, gibberellin, salicylic acid, and abscisic acid signals. Additionally, exposure to drought stress induced noticeable oscillatory changes in DgARF7 gene. Notably, DgARF7 enhanced drought tolerance through heterologous expression in yeast and overexpression in Arabidopsis. Overexpressed Arabidopsis lines of DgARF7 exhibited a markedly higher relative water content and superoxide dismutase activity, while the malondialdehyde content was significantly decreased compared to wild type under drought stress. DgARF7 also accelerated flowering time by inducing the flowering-related gene expression levels in Arabidopsis. This research provides important insights into the role of DgARF7 in orchardgrass and provides further understanding in molecular breeding.

Three different DNA-based techniques, Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Amplified Fragment Length Polymorphism (AFLP markers, were used for fingerprinting Dactylis glomerata genotypes and for detecting genetic variation between the three different subspecies. In this study, RAPD assays produced 97 bands, of which 40 were polymorphic (41.2%). The ISSR primers amplified 91 bands, and 54 showed polymorphism (59.3%). Finally, the AFLP showed 100 bands, of which 92 were polymorphic (92%). The fragments were scored as present (1) or absent (0),and those readings were entered in a computer file as a binary matrix (one for each marker). Three cluster analyses were performed to express–in the form of dendrograms–the relationships among the genotypes and the genetic variability detected. All DNA-based techniques used were able to amplify all of the genotypes. There were highly significant correlation coefficients between cophenetic matrices based on the genetic distance for the RAPD, ISSR, AFLP, and combined RAPD-ISSR-AFLP data (0.68, 0.78, 0.70, and 0.70, respectively). Two hypotheses were formulated to explain these results; both of them are in agreement with the results obtained using these three types of molecular markers. We conclude that when we study genotypes close related, the analysis of variability could require more than one DNA-based technique; in fact, the genetic variation present in different sources could interfere or combine with the more or less polymorphic ability, as our results showed for RAPD, ISSR and AFLP markers. Our results indicate that AFLP seemed to be the bestsuited molecular assay for fingerprinting and assessing genetic relationship among genotypes of Dactylis glomerata.

Background Orchardgrass ( Dactylis glomerata L.) is one of the four most economically important forage grasses cultivated globally and serves as an excellent perennial forage with high ecological value. Plant height is a key determinant of both biomass and grain yield. While numerous genes regulating plant height have been identified in annual crops, no such genes have been reported for orchardgrass. Results In this study, we analyzed the relationship between plant height and biomass yield in a natural population of 264 orchardgrass genotypes and found that a plant height of 90–110 cm contributed to the maximum biomass yield. Genome-wide association analysis (GWAS) identified 23 candidate loci associated with plant height, corresponding to 62 candidate genes. Among these, DgSAUR71 , a member of the small auxin-up RNA (SAUR) gene family, emerged as a novel candidate gene associated with plant height. Functional analysis revealed that DgSAUR71 slightly reduced plant height in rice ( Oryza sativa L.) and was involved in regulating plant height in orchardgrass. Conclusions This study demonstrates that plant height is an important contributor for optimizing biomass yield in orchardgrass, with an optimal range identified. DgSAUR71 was identified as a gene associated with plant height through GWAS and shown to negatively regulate plant height. These findings provide new insights into plant height regulation in orchardgrass and contribute to advancing crop height diversification research.

Summary Orchardgrass (Dactylis glomerata L.) is an important forage grass for cultivating livestock worldwide. Here, we report an ~1.84‐Gb chromosome‐scale diploid genome assembly of orchardgrass, with a contig N50 of 0.93 Mb, a scaffold N50 of 6.08 Mb and a super‐scaffold N50 of 252.52 Mb, which is the first chromosome‐scale assembled genome of a cool‐season forage grass. The genome includes 40 088 protein‐coding genes, and 69% of the assembled sequences are transposable elements, with long terminal repeats (LTRs) being the most abundant. The LTRretrotransposons may have been activated and expanded in the grass genome in response to environmental changes during the Pleistocene between 0 and 1 million years ago. Phylogenetic analysis reveals that orchardgrass diverged after rice but before three Triticeae species, and evolutionarily conserved chromosomes were detected by analysing ancient chromosome rearrangements in these grass species. We also resequenced the whole genome of 76 orchardgrass accessions and found that germplasm from Northern Europe and East Asia clustered together, likely due to the exchange of plants along the ‘Silk Road’ or other ancient trade routes connecting the East and West. Last, a combined transcriptome, quantitative genetic and bulk segregant analysis provided insights into the genetic network regulating flowering time in orchardgrass and revealed four main candidate genes controlling this trait. This chromosome‐scale genome and the online database of orchardgrass developed here will facilitate the discovery of genes controlling agronomically important traits, stimulate genetic improvement of and functional genetic research on orchardgrass and provide comparative genetic resources for other forage grasses.

Background Vernalization and the transition from vegetative to reproductive growth involve multiple pathways, vital for controlling floral organ formation and flowering time. However, little transcription information is available about the mechanisms behind environmental adaption and growth regulation. Here, we used high-throughput sequencing to analyze the comprehensive transcriptome of Dactylis glomerata L. during six different growth periods. Results During vernalization, 4689 differentially expressed genes (DEGs) significantly increased in abundance, while 3841 decreased. Furthermore, 12,967 DEGs were identified during booting stage and flowering stage, including 7750 up-regulated and 5219 down-regulated DEGs. Pathway analysis indicated that transcripts related to circadian rhythm, photoperiod, photosynthesis, flavonoid biosynthesis, starch, and sucrose metabolism changed significantly at different stages. Coexpression and weighted correlation network analysis (WGCNA) analysis linked different stages to transcriptional changes and provided evidence of inner relation modules associated with signal transduction, stress responses, cell division, and hormonal transport. Conclusions We found enrichment in transcription factors (TFs) related to WRKY, NAC, AP2/EREBP, AUX/IAA, MADS-BOX, ABI3/VP1, bHLH, and the CCAAT family during vernalization and floral bud development. TFs expression patterns revealed intricate temporal variations, suggesting relatively separate regulatory programs of TF modules. Further study will unlock insights into the ability of the circadian rhythm and photoperiod to regulate vernalization and flowering time in perennial grass.

Plant-specific TCP transcription factors regulate several plant growth and development processes. Nevertheless, little information is available about the TCP family in orchardgrass (Dactylis glomerata L.). This study identified 22 DgTCP transcription factors in orchardgrass and determined their structure, phylogeny, and expression in different tissues and developmental stages. The phylogenetic tree classified the DgTCP gene family into two main subfamilies, including class I and II supported by the exon–intron structure and conserved motifs. The DgTCP promoter regions contained various cis-elements associated with hormones, growth and development, and stress responses, including MBS (drought inducibility), circadian (circadian rhythms), and TCA-element (salicylic acid responsiveness). Moreover, DgTCP9 possibly regulates tillering and flowering time. Additionally, several stress treatments upregulated DgTCP1, DgTCP2, DgTCP6, DgTCP12, and DgTCP17, indicting their potential effects regarding regulating responses to the respective stress. This research offers a valuable basis for further studies of the TCP gene family in other Gramineae and reveals new ideas for increasing gene utilization.

BACKGROUND AND AIMS: Functional traits are indicators of plant interactions with their environment and the resource-use strategies of species can be defined through some key functional traits. The importance of genetic variability and phenotypic plasticity in trait variations in response to a common environmental change was investigated in two subalpine species. METHODS: Two species with contrasted resource-use strategies, Dactylis glomerata and Festuca paniculata, were grown along a productivity gradient in a greenhouse experiment. Functional traits of different genotypes were measured to estimate the relative roles of phenotypic plasticity and genetic variability, and to compare their levels of phenotypic plasticity. KEY RESULTS: Trait variability in the field for the two species is more likely to be the result of phenotypic plasticity rather than of genetic differentiation between populations. The exploitative species D. glomerata expressed an overall higher level of phenotypic plasticity compared with the conservative species F. paniculata. In addition to different amplitudes of phenotypic plasticity, the two species differed in their pattern of response for three functional traits relevant to resource use (specific leaf area, leaf dry matter content and leaf nitrogen content). CONCLUSIONS: Functional trait variability was mainly the result of phenotypic plasticity, with the exploitative species showing greater variability. In addition to average trait values, two species with different resource-use strategies differed in their plastic responses to productivity.

Dactylis glomerata L. (orchardgrass) is an important perennial forage species in temperate areas of the world. It is usually used for silage, grazing and hay because of its high nutritional value and reproducibility. Central Asia, Xinjiang and Tibetan Plateau in China possess various special micro-environments that harbor many valuable resources, while different degrees of degradation of the grassland ecosystem occurred due to climatic changing and human activities. Investigating the genetic diversity of wild D. glomerat could provide basis for collection, protection, and utilization of some excellent germplasm resources. Totally 210 individuals from 14 populations-five from Xinjiang, two from Kangding (Tibetan Plateau), and seven from Central Asia were identified using AFLP technology. The average values of Nei's genetic diversity (Hj) and Shannon information index (Ho) were 0.383 and 0.394 respectively. UPGMA tree, STRUCTURE analysis and principal coordinate analysis (PCoA) showed populations from same region clustered together. AMOVA revealed 35.10% of the genetic differentiation (Fst) occurred among populations. Gene flow (Nm) was limited among all populations. Genetic diversity of D. glomerata was high but limited under isolation-by-distance pattern, resulting in high genetic differentiation and low gene flow among populations. Adjacent regions also exhibited similar results because of the barriers of high mountains. The environmental factors, such as precipitation, elevation, latitude and longitude also had some impacts on genetic diversity and structure pattern of populations.

Main conclusion : According to the results presented in this paper the fungal endophyte Epichloee typhina significantly improves the growth, PSII photochemistry and C assimilation efficiency of its host Dactylis glomerata. In this paper, we present a comprehensive study of the impact of the endophytic fungi Epichloee typhina on its plant hosts' photosynthesis apparatus. Chlorophyll a fluorescence, gas exchange, immuno-blotting and spectrophotometric measurements were employed to assess photosynthetic performance, changes in pigment content and mechanisms associated with light harvesting, carbon assimilation and energy distribution in Dactylis glomerata colonized with Epichloee typhina. According to the results presented in this study, colonization of D. glomerata results in improved photosynthesis efficiency. Additionally, we propose a new mechanism allowing plants to cope with the withdrawal of a significant fraction of its energy resources by the endophytic fungi. The abundance of LHCI, LHCII proteins as well as chlorophyll b was significantly higher in E+ plants. Malate export out of the chloroplast was shown to be increased in colonized plants. To our knowledge, we are the first to report this phenomenon. Epichloee colonization improved PSII photochemistry and C assimilation efficiency. Elevated energy demands of E+ D. glomerata plants are met by increasing the rate of carbon assimilation and PSII photochemistry.