Haoran Wang

The spring transition in bermudagrass (Cynodon dactylon) overseeded with perennial ryegrass (Lolium perenne) remains a major challenge in turf management due to persistent competition from the cool-season species. Conventional practices such as core cultivation can damage bermudagrass stands and delay recovery. This study evaluated a novel, non-damaging approach using a yeast-based fertilizer to enhance bermudagrass regrowth during the transition period. The fertilizer consisted of Saccharomyces cerevisiae and glucose applied as a soil drench. A greenhouse experiment was conducted over two years (2023–2024) using “Yangjiang” bermudagrass overseeded with “Wintergame” perennial ryegrass. Five treatments were compared: control (0 g·m⁻² yeast + 0 g·m⁻² glucose), yeast alone (200 g·m⁻²), and yeast combined with glucose at 100, 200, or 400 g·m⁻². Growth parameters were assessed at 7, 14, and 28 days after treatment. The application of 200 g·m⁻² yeast + 200 g·m⁻² glucose yielded the most significant improvements. At 14 days, bermudagrass shoot density and turf cover significantly (p < 0.05) increased by 45.81% and 129.51%, respectively, compared to the control. By 28 days, aboveground and belowground biomass significantly (p < 0.05) increased by 308.14% and 51.35%, respectively. Root system architecture was also significantly (p < 0.05) enhanced, with total root length, surface area, and volume rising by 62.05%, 40.59%, and 63.51%. These results demonstrate that yeast fertilizer strongly promotes bermudagrass shoot and root growth during spring transition, likely by generating CO₂ to improve soil porosity without physical turf injury. This method provides a practical and complementary strategy for managing overseeded turfgrass systems.

Garden dahlias (Dahlia pinnata) are popular for their rich flower color variations that have produced many typical bicolor cultivars. Previous studies on the anthocyanin biosynthetic pathway (ABP) observed that the miR156-SPL9 module contributes to the formation of white tips on dahlia petals by repressing the MYB-bHLH-WDR complex. In this study, we further detected the potential post-transcriptional regulation involved in the bicolor petal formation by the small RNA sequencing of red bases and white tips. Compared with red bases, 89 differentially expressed miRNAs and 6349 target genes were identified. And 78 up-regulated miRNAs with their 249 down-regulated target genes were involved in the formation process of white petal tips. The target genes of differentially expressed miRNAs significantly enriched in the ABPs and miRNAs of six conserved families (MIR 156, 164, 167, 169, 482 and 6114) targeted to four transcription factor families (ARF, HD-ZIP, SBP and NAC) were involved in the post-transcriptional gene silencing (PTGS) of the ABP. Transcription sequencing and quantitative reverse transcription PCR analysis demonstrated that the MIR167-ARF8 module and the MIR6114-ANL2 module were the candidate regulators of the inactive ABP in the white tips by depressing the transcription of multiple structure genes. The findings gave new insights into the post-transcriptional regulation of the ABP and would be valuable for further studies of the PTGS mechanisms of bicolor petal formation.

Barley (Hordeum vulgare L.) is the fourth-largest cereal crop widely grown for livestock feed, brewing malts and human food. The TIFY family is a plant-specific protein family with diverse functions in plant growth, development and stress responses. However, a genome-wide comprehensive analysis of the TIFY gene family has not yet been characterized in Hordeum vulgare. In the present study, 21 and 22 TIFY family members were identified in the genomes of Hv_Morex and Hv_Barke, respectively. The HvTIFY proteins could be divided into the TIFY, ZIM/ZML and JAZ groups, and the JAZ group could be further clustered into six subgroups. HvTIFY genes were conserved in the two genotypes, and all of the duplicated gene pairs in the barley TIFY family were dominated by intense purifying selection. Tandem duplication was the main driving force for the expansion of the HvTIFY gene family. In silico gene expression profiling revealed most members of the Hv_Morex JAZ group were predominantly expressed in reproductive organs and root tissues and were also more involved in the responses to cold treatment and spot blotch infection than other groups. Quite a few JAZ genes (Hv_MoJAZ1, Hv_MoJAZ4, Hv_MoJAZ6, Hv_MoJAZ9, Hv_MoJAZ11, Hv_MoJAZ12 and Hv_MoJAZ14) were found to be tightly associated with the growth of barley and the responses to cold and spot blotch infection stresses. The genome-wide comprehensive analysis of the structure and function of the HvTIFY genes will contribute further to our understanding of the functions of these genes in response to abiotic and biotic stresses in Hordeum vulgare.

The U-box protein family of ubiquitin ligases is important in the biological processes of plant growth, development, and biotic and abiotic stress responses. Plants in the genus Zoysia are recognized as excellent warm-season turfgrass species with drought, wear and salt tolerance. In this study, we conducted the genome-wide identification of plant U-box (PUB) genes in Zoysia japonica based on U-box domain searching. In total, 71 ZjPUB genes were identified, and a protein tree was constructed of AtPUBs, OsPUBs, and ZjPUBs, clustered into five groups. The gene structures, characteristics, cis-elements and protein interaction prediction network were analyzed. There were mainly ABRE, ERE, MYB and MYC cis-elements distributed in the promoter regions of ZjPUBs. ZjPUBs were predicted to interact with PDR1 and EXO70B1, related to the abscisic acid signaling pathway. To better understand the roles of ZjPUBs under salt stress, the expression levels of 18 ZjPUBs under salt stress were detected using transcriptome data and qRT-PCR analysis, revealing that 16 ZjPUBs were upregulated in the roots under salt treatment. This indicates that ZjPUBs might participate in the Z. japonica salt stress response. This research provides insight into the Z. japonica PUB gene family and may support the genetic improvement in the molecular breeding of salt-tolerant zoysiagrass varieties.

Centipedegrass (Eremochloa ophiuroides) is an important warm-season grass plant used as a turfgrass as well as pasture grass in tropical and subtropical regions, with wide application in land surface greening and soil conservation in South China and southern United States. In this study, the complete cp genome of E. ophiuroides was assembled using high-throughput Illumina sequencing technology. The circle pseudomolecule for E. ophiuroides cp genome is 139,107 bp in length, with a quadripartite structure consisting of a large single copyregion of 82,081 bp and a small single copy region of 12,566 bp separated by a pair of inverted repeat regions of 22,230 bp each. The overall A + T content of the whole genome is 61.60%, showing an asymmetric nucleotide composition. The genome encodes a total of 131 gene species, composed of 20 duplicated genes within the IR regions and 111 unique genes comprising 77 protein-coding genes, 30 transfer RNA genes, and 4 ribosome RNA genes. The complete cp genome sequence contains 51 long repeats and 197 simple sequence repeats, and a high degree of collinearity among E. ophiuroide and other Gramineae plants was disclosed. Phylogenetic analysis showed E. ophiuroides, together with the other two Eremochloa species, is closely related to Mnesithea helferi within the subtribe Rottboelliinae. These findings will be beneficial for the classification and identification of the Eremochloa taxa, phylogenetic resolution, novel gene discovery, and functional genomic studies for the genus Eremochloa.

The TIFY family is a group of novel plant-specific transcription factors involved in plant development, signal transduction, and responses to stress and hormones. TIFY genes have been found and functionally characterized in a number of plant species. However, there is no information about this family in warm-season grass plants. The current study identified 24 TIFY genes in Eremochloa ophiuroides, a well-known perennial warm-season grass species with a high tolerance to aluminum toxicity and good adaptability to the barren acidic soils. All of the 24 EoTIFYs were unevenly located on six out of nine chromosomes and could be classified into two subfamilies (ZIM/ZML and JAZ), consisting of 3 and 21 genes, respectively, with the JAZ subfamily being further divided into five subgroups (JAZ I to JAZ V). The amino acids of 24 EoTIFYs showed apparent differences between the two subfamilies based on the analysis of gene structures and conserved motifs. MCScanX analysis revealed the tandem duplication and segmental duplication of several EoTIFY genes occurred during E. ophiuroides genome evolution. Syntenic analyses of TIFY genes between E. ophiuroides and other five plant species (including A. thaliana, O. sativa, B. distachyon, S. biocolor, and S. italica) provided valuable clues for understanding the potential evolution of the EoTIFY family. qRT-PCR analysis revealed that EoTIFY genes exhibited different spatial expression patterns in different tissues. In addition, the expressions of EoTIFY genes were highly induced by MeJA and all of the EoTIFY family members except for EoJAZ2 displayed upregulated expression by MeJA. Ten EoTIFY genes (EoZML1, EoZML1, EoJAZ1, EoJAZ3, EoJAZ5, EoJAZ6, EoJAZ8, EoJAZ9, EoJAZ10, and EoJAZ21) were observed to be highly expressed under both exogenous MeJA treatment and aluminum stress, respectively. These results suggest that EoTIFY genes play a role in the JA-regulated pathway of plant growth and aluminum resistance as well. The results of this study laid a foundation for further understanding the function of TIFY genes in E. ophiuroides, and provided useful information for future aluminum tolerance related breeding and gene function research in warm-season grass plants.

Centipedegrass (Eremochloa ophiuroides (Munro.) Hack.) is a species originating in China and is an excellent warm-season turfgrass. As a native species in southern China, it is naturally distributed in the phosphorus-deficient and aluminum-toxic acid soil areas. It is important to research the molecular mechanism of centipedegrass responses to phosphorus-deficiency and/or aluminum-toxicity stress. Quantitative Real-Time PCR (qRT-PCR) is a common method for gene expression analysis, and the accuracy of qRT-PCR results depends heavily on the stability of internal reference genes. However, there are still no reported stable and effective reference genes for qRT-PCR analysis of target genes under the acid-soil-related stresses in different organs of centipedegrass. For scientific rigor, the gene used as a reference for any plant species and/or any stress conditions should be first systematically screened and evaluated. This study is the first to provide a group of reliable reference genes to quantify the expression levels of functional genes of Eremochloa ophiuroides under multiple stresses of P deficiency and/or aluminum toxicity. In this study, centipedegrass seedlings of the acid-soil-resistant strain ‘E041’ and acid-soil-sensitive strain ‘E089’ were used for qRT-PCR analysis. A total of 11 candidate reference genes (ACT, TUB, GAPDH, TIP41, CACS, HNR, EP, EF1α, EIF4α, PP2A and actin) were detected by qRT-PCR technology, and the stability of candidate genes was evaluated with the combination of four internal stability analysis software programs. The candidate reference genes exhibited differential stability of expression in roots, stems and leaves under phosphorus-deficiency and/or aluminum-toxicity stress. On the whole, the results showed that GAPDH, TIP41 and HNR were the most stable in the total of samples. In addition, for different tissues under various stresses, the selected reference genes were also different. CACS and PP2A were identified as two stable reference genes in roots through all three stress treatments (phosphate deficiency, aluminum toxicity, and the multiple stress treatment of aluminum toxicity and phosphate deficiency). Moreover, CACS was also stable as a reference gene in roots under each treatment (phosphate deficiency, aluminum toxicity, or multiple stresses of aluminum toxicity and phosphate deficiency). In stems under all three stress treatments, GAPDH and EIF4α were the most stable reference genes; for leaves, PP2A and TIP41 showed the two highest rankings in all three stress treatments. Finally, qRT-PCR analysis of the expression patterns of the target gene ALMT1 was performed to verify the selected reference genes. The application of the reference genes identified as internal controls for qRT-PCR analysis will enable accurate analysis of the target gene expression levels and expression patterns in centipedegrass under acid-soil-related stresses.

The R2R3-MYB genes in plants play an essential role in the drought-responsive signaling pathway. Plenty of R2R3-MYB S21 and S22 subgroup genes in Arabidopsis have been implicated in dehydration conditions, yet few have been covered in terms of the role of the S21 and S22 subgroup genes in poplar under drought. PdMYB2R089 and PdMYB2R151 genes, respectively belonging to the S21 and S22 subgroups of NL895 (Populus deltoides × P. euramericana cv. ‘Nanlin895′), were selected based on the previous expression analysis of poplar R2R3-MYB genes that are responsive to dehydration. The regulatory functions of two target genes in plant responses to drought stress were studied and speculated through the genetic transformation of Arabidopsis thaliana. PdMYB2R089 and PdMYB2R151 could promote the closure of stomata in leaves, lessen the production of malondialdehyde (MDA), enhance the activity of the peroxidase (POD) enzyme, and shorten the life cycle of transgenic plants, in part owing to their similar conserved domains. Moreover, PdMYB2R089 could strengthen root length and lateral root growth. These results suggest that PdMYB2R089 and PdMYB2R151 genes might have the potential to improve drought adaptability in plants. In addition, PdMYB2R151 could significantly improve the seed germination rate of transgenic Arabidopsis, but PdMYB2R089 could not. This finding provides a clue for the subsequent functional dissection of S21 and S22 subgroup genes in poplar that is responsive to drought.

R2R3-type MYB transcription factors are implicated in drought stress, which is a primary factor limiting the growth and development of woody plants. The identification of R2R3-MYB genes in the Populus trichocarpa genome has been previously reported. Nevertheless, the diversity and complexity of the conserved domain of the MYB gene caused inconsistencies in these identification results. There is still a lack of drought-responsive expression patterns and functional studies of R2R3-MYB transcription factors in Populus species. In this study, we identified a total of 210 R2R3-MYB genes in the P. trichocarpa genome, of which 207 genes were unevenly distributed across all 19 chromosomes. These poplar R2R3-MYB genes were phylogenetically divided into 23 subgroups. Collinear analysis demonstrated that the poplar R2R3-MYB genes underwent rapid expansion and that whole-genome duplication events were a dominant factor in the process of rapid gene expansion. Subcellular localization assays indicated that poplar R2R3-MYB TFs mainly played a transcriptional regulatory role in the nucleus. Ten R2R3-MYB genes were cloned from P. deltoides × P. euramericana cv. Nanlin895, and their expression patterns were tissue-specific. A majority of the genes showed similar drought-responsive expression patterns in two out of three tissues. This study provides a valid cue for further functional characterization of drought-responsive R2R3-MYB genes in poplar and provides support for the development of new poplar genotypes with elevated drought tolerance.