Search Results (43)

Flavonoids are key bioactive compounds in plants that play important defense roles against abiotic stress and are involved in plant growth and development. Methyl jasmonate (MeJA) is a significant growth regulator that promotes the accumulation of flavonoids in a variety of plants, but the effect of MeJA in Hedera helix remains poorly understood. In the present study, the flavonoid content was significantly increased after MeJA treatment and peaked at 6 h post-treatment. A total of 31,931 genes were identified using transcriptome, and 6484 DEGs were identified at 6 h post-treatment. Through GO and KEGG enrichment analysis, it was shown that DEGs were primarily enriched in phenylpropanoid biosynthesis pathways. Based on the putative transcription factors derived from DEGs, growth-regulating factor (GRF), a transcription factor potentially linking MeJA signaling to flavonoid accumulation and participating in plant growth and stress responses, was further identified. A total of 20 Hh-GRFd genes were identified on the whole genome level and clustered into five phylogenetic groups with conserved subfamily characteristics. Abundant MeJA-responsive cis-elements were presented in the promoter regions of HhGRF1-HhGRF20. They exhibited a tissue-specific expression variation, and HhGRF10 was dominantly expressed in leaves of H. helix. Notably, HhGRF10 exhibited MeJA-induced expression that correlated temporally with flavonoid accumulation, suggesting that HhGRF10 might play a potential role in promoting flavonoid biosynthesis, and overexpression and knockout assay substantiated this conclusion. The finding provides the first transcriptome-wide resource for flavonoid biosynthesis in H. helix and identifies the candidate GRF-mediated regulator for flavonoid accumulation. Full article

The Cerasus humilis, a perennial shrub belonging to the Cerasus genus, is native to China and holds significant ecological and economic importance. Growth regulation factors (GRF) are a family of transcription factors (TF) that play a key role in plant growth and development. This research entailed an in-depth examination of the GRF family in C. humilis, exploring its significance in the evolution of C. humilis. Twelve GRF genes were identified in the C. humilis genome. Named separately as ChGRF1-Chumilis15987.1, ChGRF2-Chumilis25207.1, ChGRF3-Chumilis26233.1, ChGRF4-Chumilis08578.3, ChGRF5-Chumilis18808.1, ChGRF6-Chumilis12052.1, ChGRF7-Chumilis10417.1, ChGRF8-Chumilis01608.1, ChGRF9-Chumilis14057.1, ChGRF10-Chumilis12169.1, ChGRF11-Chumilis14952.1, and ChGRF12-Chumilis07534.1. Phylogenetic analysis divided twelve GRF genes into five subfamilies. The gene structure, pattern, and cis-regulatory components of the GRF gene family were analyzed. In addition, according to collinearity analysis, there are six collinearity with Arabidopsis, twelve collinearity with Malus pumila, eight collinearity with Vitis vinifera, and three collinearity with Oryza sativa. Intraspecific collinearity analysis revealed the presence of three pairs of tandem repeat genes in the dwarf cherry genome. Identifying cis-acting elements revealed the prominent presence of gibberellin reaction elements, which are widely distributed in the promoter region. Cluster heatmap analysis showed that ChGRF2 had the highest expression levels in fruits and stems. ChGRF3 is highly expressed in red fruits of different colors, while ChGRF6 and ChGRF12 are highly expressed in yellow fruits. This study mainly focused on dwarf cherries treated with gibberellin. As the treatment time increased, the ChGRF gene showed different expression levels. ChGRF2, ChGRF3, ChGRF6, and ChGRF12 were up-regulated under gibberellin treatment. These genes all contain hormone-responsive cis-acting elements, indicating tht the ChGRF gene family plays a vital role under gibberellin treatment in C. humilis. The results laid the foundation for further research on the biological functions of the GRF genes in C. humilis. Full article

Small RNAs (sRNAs) are pivotal in regulating gene expression and are involved in a diverse array of biological processes. Among these, microRNAs (miRNAs) and phased small interfering RNAs (phasiRNAs) have been extensively investigated over the past decades. We conducted an in-depth analysis of deep sequencing data from the gymnosperm Ginkgo biloba, encompassing sRNA, transcriptome, and degradome libraries. Our analysis identified a total of 746 miRNAs and 654 phasiRNA precursor (PHAS) loci, with 526 (80%) of the PHAS loci predicted to be triggered by 515 miRNAs (69%). Several miRNA-PHAS modules, particularly the miR159/miR319-PHAS module, were found to potentially regulate reproductive development by targeting GAMYB genes and triggering phasiRNA biogenesis. The miR390-PHAS module appears to be involved in flavonoid biosynthesis by targeting key enzyme genes such as chalcone synthase (CHS) and anthocyanin synthase (ANS). Through target gene identification and coexpression analysis, we uncovered two distinct models of complex regulatory networks: growth-related factors like ARF and GRF seem to be regulated exclusively by miRNAs (Model 1), while certain disease resistance-related genes are predicted to be regulated by both miRNAs and phasiRNAs (Model 2), indicating diverse regulatory mechanisms across different biological processes. Overall, our study provides a comprehensive annotation of miRNA and PHAS loci in G. biloba and elucidates a post-transcriptional regulatory network, offering novel insights into sRNA research in gymnosperms. Full article

Prunus avium is a woody plant of economic importance within the genus Prunus, the family Rosaceae, which is affected by various environmental factors during its long growth period. Growth-regulating factors (GRFs) and GRF-interacting factors (GIFs) are essential in regulating plant growth and development, responding to environmental stresses, and responding to exogenous hormone induction. Genome-wide analysis showed 13 GRF genes on eight chromosomes and three GIF genes on three chromosomes in P. avium, clustered into three and two branches, respectively. Cis-acting element analysis indicated that the PavGRF promoters contained regulatory elements associated with hormones, light stress, and growth development. Therefore, we evaluated the effects of gibberellin and light stress on the GRF and GIF genes in P. avium at different stages. Transcriptome data revealed that five PavGRFs exhibited elevated expression levels during the green ripening and color conversion stages in P. avium, PavGRF9 and PavGIF1 displayed higher expression during the full red stage, and gibberellin treatment led to the upregulation of these five PavGRFs and PavGIF1 during the full red stage. However, light stress did not significantly impact the expression of PavGRFs and PavGIFs. Additionally, miR396 could bind to the PavGRFs, thereby regulating the expression level of PavGIF after transcription. This study revealed the potential roles of the GRF and GIF transcription factor families in P. avium fruit growth and development, exogenous hormone treatment, and light stress, laying the foundation for further research on the roles of the GRF and GIF gene families in P. avium. Full article

Soybean is an important and versatile crop worldwide. Enhancing soybean architecture offers a potential method to increase yield. Plant-specific transcription factors play a crucial, yet often unnoticed, role in regulating plant growth and development. GRF-INTERACTING FACTOR (GIF) genes are plant-specific transcription factors; however, their functions in soybean remain poorly understood. Eight GmGIF members were identified in soybean (Glycine max L.). Phylogenetic analysis divided the eight GmGIF proteins into three groups. In this study, we focused on the role of GmGIF5 owing to its high expression level in the meristem. Subcellular localization and transcriptional activity analysis showed that GmGIF5 was localized to the nucleus and has self-transactivation ability. To elucidate the biological function of GmGIF5, we constructed transgenic Arabidopsis lines overexpressing the gene. Phenotype observations indicated that the overexpression of GmGIF5 contributed to larger leaves, higher plants, wider stems, and larger seeds. The organs of GmGIF5 overexpression lines exhibited larger sizes primarily due to an increase in cell size rather than cell number. RNA sequencing was performed to investigate the underlying mechanism for these effects, showing that differentially expressed genes in overexpression lines were mainly enriched in cell wall modification processes. Our study provides new clues for an understanding of the roles of the GmGIF family in soybean, which can promote the further application of these genes in genetic breeding. Full article

Populus euphratica Oliv. typically has four kinds of heteromorphic leaves: linear (Li), lanceolate (La), ovate (Ov) and broad ovate (Bo). Heteromorphic leaves help P. euphratica adapt to extreme desert environments and further contribute to protection against land desertification in Northwest China. In the authors’ previous research, growth-regulating factors (GRFs) were speculated to be related to the development of P. euphratica heteromorphic leaves via multi-omics integrated analysis. However, the genomic features and biological role of the P. euphratica GRF gene family in heteromorphic leaves are still unclear. In this study, 19 PeGRF genes were genome-widely identified and characterized in P. euphratica, and their physicochemical properties, gene structure and phylogenetic evolution were analyzed. An analysis of the research showed that PeGRFs were unevenly distributed on 11 chromosomes and that PeGRF proteins contained conserved motif 1 (WRC) and motif 2 (QLQ). Moreover, 19, 15, 19 and 22 GRFs were identified in Populus deltoides Marshall, Populus pruinosa Schrenk, Salix sinopurpurea C. Wang et C. Y. Yang and Salix suchowensis W. C. Cheng, respectively. A collinearity analysis showed that the PeGRF family evolved slowly within Populus species. A phylogenetic tree of the GRF family was also constructed, and GRFs were divided into four subfamilies. A large number of cis-acting elements were related to plant growth and development, plant hormone response and stress response on the promoter of PeGRFs. The expression pattern of PeGRFs showed significant up-regulation in broad leaves (Ov and Bo) compared with narrow leaves (Li and La). In combination with the predicted gene regulatory network, PeGRF9 (PeuTF06G01147.1) may have an important contribution to the leaf shape development of P. euphratica. The heterologous expression of PeGRF9 in wild-type plants (Col-0) of Arabidopsis thaliana (L.) Heynh was also studied, showing a significant increase in the leaf area of overexpressing plants compared with the wild type. Nineteen PeGRF gene members were identified and characterized in P. euphratica, and a comparison of the genomic analysis of Populus GRF members revealed their evolutionary features. The further overexpression of PeGRF9 in A. thaliana revealed its biological role in the heteromorphic leaves of P. euphratica. This study not only provides new insights into the evolution and function of PeGRFs in P. euphratica heteromorphic leaves but also helps in an understanding of the adaptive evolution of P. euphratica in drought desert environments. Full article

All multicellular organisms undergo senescence, but the continuous division of the vascular cambium in plants enables certain tree species to survive for hundreds or even thousands of years. Previous studies have focused on the development of the vascular cambium, but the mechanisms regulating age-related changes remain poorly understood. This study investigated age-related changes in the vascular cambium of P. euphratica trees aged 50 to 350 years. The number of cambium cells in the 50-year-old tree group was 10 ± 2, while the number of cambium cells in the 200-year-old and 350-year-old tree groups significantly decreased. The thickness of the cambium cells exhibited a similar trend. In addition, the net photosynthetic and transpiration rates continue to increase with age, but no notable differences were found in factors like average leaf area, palisade tissue thickness, and stomatal density. A total of 6491 differentially expressed genes (DEGs) were identified in the vascular cambium of P. euphratica at three distinct ages using RNA sequencing. The expression patterns of DEGs associated with cell division and differentiation, lignin biosynthesis, plant hormones, and transcription factors were analyzed. DEGs related to XTH, EXP, PAL, C4H, ABA, Br, GA, and others are highly expressed in older trees, whilst those encoding expansins, kinases, cyclins, 4CL, Auxin, Eth, SA, and others are more prevalent in younger trees. Gene family members, such as NAC, MYB, HD-ZIP III, WRKY, and GRF, have various regulatory functions in the vascular cambium. The findings offer insights into how ancient P. euphratica trees maintain vitality by balancing growth and aging, providing a foundation for future research on their longevity mechanisms. Full article

The key genes involved in plant regeneration play a crucial role in enhancing reproductive capabilities, plant genetic transformation, and gene editing efficiency. Camellia oleifera, a vital woody oil crop, faces challenges in genetic improvement efficiency due to its slow growth and the difficulties it experiences during the regeneration process. This study focused on the expression levels of mRNA and miRNA during the somatic embryogenesis of C. oleifera, and the core genes of plant regeneration involved in the C. oleifera somatic embryogenesis process were identified, including WUSCHEL-related homeobox 4 (WOX4), WOX13, and DNA-Binding One Zinc Finger 5.6 (DOF5.6) during the somatic embryo callus induction phase; WOX11, PLETHORA2 (PLT2), and Growth-regulating factor (GRF) during the somatic embryogenesis and bud regeneration phase; and miRNAs such as miR156, miR319, and miR394. These key regulatory factors may participate in the regulation of plant auxin and cytokinin and play a core role in the regeneration process of C. oleifera. The research data elucidate the process of somatic embryogenesis in C. oleifera at the molecular level. The key regulatory genes identified provide potential targets for improving the regeneration efficiency of C. oleifera and other woody oilseed plants. Full article

Drought stress affects crop growth and development, significantly reducing crop yield and quality. Alfalfa (Medicago sativa L.), the most widely cultivated forage crop, is particularly susceptible to drought. The general regulatory factor (GRF) protein 14-3-3, a highly conserved family in plants, specifically recognizes and binds to phosphoserine residues in target proteins, regulating both plant development and responses to environmental stressors. In this study, 66 alfalfa 14-3-3 proteins were identified, and the full-length MsGRF2 gene was cloned and functionally analyzed. The expression of MsGRF2 was highest in alfalfa inflorescences and lowest in roots. Transgenic tobacco overexpressing MsGRF2 exhibited increased tolerance to low temperature and drought stress, evidenced by physiological indicators including low levels of active oxygen species and increased activity of antioxidant enzymes and osmoregulatory substances. Under drought stress conditions, compared to wild-type plants, MsGRF2-overexpressing tobacco plants exhibited significantly increased expression of drought stress-related genes ERD10B and TIP, while the expression of BRI1, Cu/Zn-SOD, ERF2, and KC1 was significantly reduced. Together, these results provide new insights into the roles of the 14-3-3 protein MsGRF2 in plant drought response mechanisms. Full article

Growth-regulating factor (GRF) is a plant-specific family of transcription factors crucial for meristem development and plant growth. Sorghum (Sorghum bicolor L. Moench) is a cereal species widely used for food, feed and fuel. While sorghum stems are important biomass components, the regulation of stem development and the carbohydrate composition of the stem tissues remain largely unknown. Here, we identified 11 SbGRF-encoding genes and found the SbGRF expansion driven by whole-genome duplication events. By comparative analyses of GRFs between rice and sorghum, we demonstrated the divergence of whole-genome duplication (WGD)-derived OsGRFs and SbGRFs. A comparison of SbGRFs’ expression profiles supports that the WGD-duplicated OsGRFs and SbGRFs experienced distinct evolutionary trajectories, possibly leading to diverged functions. RNA-seq analysis of the internode tissues identified several SbGRFs involved in internode elongation, maturation and cell wall metabolism. We constructed co-expression networks with the RNA-seq data of sorghum internodes. Network analysis discovered that SbGRF1, 5 and 7 could be involved in the down-regulation of the biosynthesis of cell wall components, while SbGRF4, 6, 8 and 9 could be associated with the regulation of cell wall loosening, reassembly and/or starch biosynthesis. In summary, our genome-wide analysis of SbGRFs reveals the distinct evolutionary trajectories of WGD-derived SbGRF pairs. Importantly, expression analyses highlight previously unknown functions of several SbGRFs in internode elongation, maturation and the potential involvement in the metabolism of the cell wall and starch during post-anthesis stages. Full article

Growth-regulating factor (GRF) is a multi-gene family that plays an important role in plant growth and development and is widely present in plants. Currently, GRF gene members have been reported in many plants, but the GRF gene family has not been found in sweet potato. In this study, ten GRF genes were identified in sweet potato (Ipomoea batatas), twelve and twelve were identified in its two diploid relatives (Ipomoea trifida) and (Ipomoea triloba), which were unevenly distributed on nine different chromosomes. Subcellular localization analysis showed that GRF genes of sweet potato, I. trifida, and I. triloba were all located in the nucleus. The expression analysis showed that the expression of IbGRFs was diverse in different sweet potato parts, and most of the genes were upregulated and even had the highest expression in the vigorous growth buds. These findings provide molecular characterization of sweet potato and its two diploid relatives, the GRF families, further supporting functional characterization. Full article

The roots, stems, leaves, and seeds of Eucommia ulmoides contain a large amount of trans-polyisoprene (also known as Eu-rubber), which is considered to be an important laticiferous plant with valuable industrial applications. Eu-rubber used in industry is mainly extracted from leaves. Therefore, it is of great significance to identify genes related to regulating the leaf size of E. ulmoides. Plant growth-regulating factors (GRFs) play important roles in regulating leaf size, and their functions are highly conserved across different plant species. However, there have been very limited reports on EuGRFs until now. In this study, eight canonical EuGRFs with both QLQ and WRC domains and two putative eul-miR396s were identified in the chromosome-level genome of E. ulmoides. It is found that, unlike AtGRFs, all EuGRFs contain the miR396s binding site in the terminal of WRC domains. These EuGRFs were distributed on six chromosomes in the genome of E. ulmoides. Collinearity analysis of the E. ulmoides genome revealed that EuGRF1 and EuGRF3 exhibit collinear relationships with EuGRF2, suggesting that those three genes may have emerged via gene replication events. The collinear relationship between EuGRFs, AtGRFs, and OsGRFs showed that EuGRF5 and EuGRF8 had no collinear members in Arabidopsis and rice. Almost all EuGRFs show a higher expression level in growing and developing tissues, and most EuGRF promoters process phytohormone-response and stress-induced cis-elements. Moreover, we found the expression of EuGRFs was significantly induced by gibberellins (GA₃) in three hours, and the height of E. ulmoides seedlings was significantly increased one week after GA₃ treatment. The findings in this study provide potential candidate genes for further research and lay the foundation for further exploring the molecular mechanism underlying E. ulmoides development in response to GA₃. Full article

Transgenic technology is a crucial tool for gene functional analysis and targeted genetic modification in the para rubber tree (Hevea brasiliensis). However, low efficiency of plant regeneration via somatic embryogenesis remains a bottleneck of successful genetic transformation in H. brasiliensis. Enhancing expression of GROWTH-REGULATING FACTOR 4 (GRF4)-GRF-INTERACTING FACTOR 1 (GIF1) has been reported to significantly improve shoot and embryo regeneration in multiple crops. Here, we identified endogenous HbGRF4 and HbGIF1 from the rubber clone Reyan7-33-97, the expressions of which dramatically increased along with somatic embryo (SE) production. Intriguingly, overexpression of HbGRF4 or HbGRF4-HbGIF1 markedly enhanced the efficiency of embryogenesis in two H. brasiliensis callus lines with contrasting rates of SE production. Transcriptional profiling revealed that the genes involved in jasmonic acid response were up-regulated, whereas those in ethylene biosynthesis and response as well as the S-adenosylmethionine-dependent methyltransferase activity were down-regulated in HbGRF4- and HbGRF4-HbGIF1-overexpressing H. brasiliensis embryos. These findings open up a new avenue for improving SE production in rubber tree, and help to unravel the underlying mechanisms of HbGRF4-enhanced somatic embryogenesis. Full article

Flax is an important cash crop globally with a variety of commercial uses. It has been widely used for fiber, oil, nutrition, feed and in composite materials. Growth regulatory factor (GRF) is a transcription factor family unique to plants, and is involved in regulating many processes of growth and development. Bioinformatics analysis of the GRF family in flax predicted 17 LuGRF genes, which all contained the characteristic QLQ and WRC domains. Equally, 15 of 17 LuGRFs (88%) are predicted to be regulated by lus-miR396 miRNA. Phylogenetic analysis of GRFs from flax and several other well-characterized species defined five clades; LuGRF genes were found in four clades. Most LuGRF gene promoters contained cis-regulatory elements known to be responsive to hormones and stress. The chromosomal locations and collinearity of LuGRF genes were also analyzed. The three-dimensional structure of LuGRF proteins was predicted using homology modeling. The transcript expression data indicated that most LuGRF family members were highly expressed in flax fruit and embryos, whereas LuGRF3, LuGRF12 and LuGRF16 were enriched in response to salt stress. Real-time quantitative fluorescent PCR (qRT-PCR) showed that both LuGRF1 and LuGRF11 were up-regulated under ABA and MeJA stimuli, indicating that these genes were involved in defense. LuGRF1 was demonstrated to be localized to the nucleus as expected for a transcription factor. These results provide a basis for further exploration of the molecular mechanism of LuGRF gene function and obtaining improved flax breeding lines. Full article

Secondary xylem produced by stem secondary growth is the main source of tree biomass and possesses great economic and ecological value in papermaking, construction, biofuels, and the global carbon cycle. The secondary xylem formation is a complex developmental process, and the underlying regulatory networks and potential mechanisms are still under exploration. In this study, using hybrid poplar (Populus alba × Populus glandulosa clone 84K) as a model system, we first ascertained three representative stages of stem secondary growth and then investigated the regulatory network of secondary xylem formation by joint analysis of transcriptome and miRNAs. Notably, 7507 differentially expressed genes (DEGs) and 55 differentially expressed miRNAs (DEMs) were identified from stage 1 without initiating secondary growth to stage 2 with just initiating secondary growth, which was much more than those identified from stage 2 to stage 3 with obvious secondary growth. DEGs encoding transcription factors and lignin biosynthetic enzymes and those associated with plant hormones were found to participate in the secondary xylem formation. MiRNA-target analysis revealed that a total of 85 DEMs were predicted to have 2948 putative targets. Among them, PagmiR396d-PagGRFs, PagmiR395c-PagGA2ox1/PagLHW/PagSULTR2/PagPolyubiquitin 1, PagmiR482d-PagLAC4, PagmiR167e-PagbHLH62, and PagmiR167f/g/h-PagbHLH110 modules were involved in the regulating cambial activity and its differentiation into secondary xylem, cell expansion, secondary cell wall deposition, and programmed cell death. Our results give new insights into the regulatory network and mechanism of secondary xylem formation. Full article