Search Results (83)

Background: Aluminum (Al) toxicity severely limits Medicago sativa (alfalfa) production on acidic soils, resulting in major yield losses worldwide. The highly conserved miRNA156 (miR156) functions by downregulating at least 11 SQUAMOSA promoter-binding protein-like (SPL) transcription factors in alfalfa, including SPL13, but its role in Al stress remains unclear. This study aimed to investigate the miR156/SPL regulatory network’s function in alfalfa under Al stress. Methods: Gene expression analyses, histochemical staining, nutrient profiling, phenotypic assays, transcriptome profiling, and ChIP-seq were conducted on alfalfa plants with altered miR156 and SPL13 expression to assess their roles in the Al stress response. Results: Al stress induced SPL13 expression while repressing miR156 in the roots. Elevated miR156 intensified Al accumulation, lipid peroxidation, and plasma membrane damage, accompanied by reduced leaf nitrogen, magnesium, sulfur, and phosphorus content. Phenotypically, increased SPL13 enhanced the root length and Al tolerance, whereas SPL13 silencing reduced tolerance. Transcriptome profiling of SPL13-silenced plants identified differentially expressed genes involved in the Al response, including aluminum-activated malate transporters and various transcription factors (GRAS, Myb-related, bHLH041, NAC, WRKY53, bZIP, and MADS-box). ChIP-seq revealed that SPL13 directly regulates genes encoding a protein kinase, cytochrome P450, and fasciclin-like arabinogalactan proteins. Conclusions: The MsmiR156/MsSPL13 network plays a crucial regulatory role in alfalfa’s response to Al toxicity. These findings provide novel genetic targets and foundational knowledge to advance molecular breeding for enhanced Al tolerance in alfalfa. Full article

The transition to ripening in non-climacteric species is governed by several signals, including hormones that enhance or counteract the abscisic acid (ABA)-promoting effect. The SQUAMOSA Promoter-binding protein-Like (SPL) transcription factors are involved in ripening through the modulation of anthocyanin biosynthesis. In sweet cherry fruits, several miR156-targeted PavSPLs are expressed before and during ripening. Recently, some PavSPLs were found in the transition from development to ripening in cultivars contrasting in maturity time. Additionally, several forms of miR156 were expressed in sweet cherry seeds of an early-season cultivar. In this work, we addressed the relevance of endocarp lignification and PavSPLs expression for the transition to ripening. First, we characterized early- and late-season sweet cherry cultivars, ‘Celeste’ and ‘Regina’, focusing on fruit and seed development, endocarp lignification, and PavSPL expression profile. Fruit growth dynamics revealed an earlier onset of color development and lignification in ‘Celeste’, while ‘Regina’ exhibited a prolonged lag phase and delayed embryo development. Transcript profiling at the light green stage showed a higher expression of PavSPL genes in fruits and identified cultivar-specific expressions, especially between ‘Regina’ and ‘Celeste’ seeds. Co-expression networks linked PavSPLs to genes involved in lignin and anthocyanin biosynthesis. We focused on PavSPL2 and PavSPL9, which were targeted by mtr-miR156a and gma-miR156f. Both PavSPLs and miRNAs were expressed in fruits and seeds at the yellow stage, an advanced point in the transition to ripening in sweet cherry. Exogenous application of auxin-related compounds in the mid-season cultivar ‘Lapins’ modulated endocarp lignification and pigmentation. Notably, p-IBA treatment, which enzymatically targets the lignin pathway, transiently increased anthocyanin accumulation and reduced lignin deposition, effects that correlated with the downregulation of PavSPL gene expression. These findings highlight the interplay between lignification, color evolution, and pigment biosynthesis during the transition from development to ripening in sweet cherry fruits, and suggest a role for PavSPL genes in this transition. Full article

In plants, SPL is a distinct family of transcription factors. Its protein structure possesses a highly conserved SBP domain comprising two zinc finger structures and nuclear localization regions, and microRNAs (miR156) control the transcriptional expression of the majority of SPL genes. SPLs are key TFs in regulating organ morphogenesis, phase transition/floral induction, and yield-related traits in agronomic and horticultural crops. These biomolecules have been functionally characterized for their role in augmenting plant responses to abiotic and biotic stresses. Present research gaps and viewpoints are addressed herein. Using these extensive data, researchers can more comprehensively understand how SPL genes modulate agronomic features in different ways. Full article

SQUAMOSA promoter-binding protein-like (SPL) transcription factors specific to plants are vital for regulating growth, development, secondary metabolite biosynthesis, and responses to both biotic and abiotic stresses. Despite their importance, no systematic investigations or identifications of the SPL gene family in Ginkgo biloba have been conducted. In this study, we identified 13 SPL genes within the Ginkgo biloba reference genome, spanning seven chromosomes, and categorized these genes into six groups based on their phylogenetic relationships with Arabidopsis thaliana SPL gene families. Our analysis of gene structure, conserved domains, motifs, and miR156 target predictions indicates that GbSPLs are highly conserved across evolutionary timelines. Furthermore, synteny analysis highlighted that dispersed duplication events have expanded the SPL gene family in Ginkgo biloba. Examination of the cis-regulatory elements revealed that many GbSPL genes possess motifs associated with light, hormones, and stress, implying their involvement in flavonoid biosynthesis and adaptation to environmental conditions. RNA-Seq and qRT-PCR expression profiles of GbSPL genes across various tissues and low- and high-flavonoid leaves and during both short-term and long-term water stress illustrated their roles in flavonoid biosynthesis and responses to water stress. Subcellular localization experiments showed that GbSPL2 and GbSPL11 proteins are situated within the nucleus. Our research offers the first systematic characterization of the SPL gene family in Ginkgo biloba, establishing a valuable foundation for understanding their evolutionary background and functional roles in flavonoid biosynthesis and water stress response. Full article

Chinese fir (Cunninghamia lanceolata (Lamb.) Hook) is one of the main afforestation tree species in southern China. Continuous planting for multiple generations has led to a decrease in the content of available phosphorus in the soil. To adapt to low phosphorus stress, plants develop a series of physiological, biochemical, and developmental responses through self-regulation. Recent studies have shown that miRNAs play a regulatory role in plants’ responses to low phosphorus stress. However, the regulatory mechanism of miRNAs in Chinese fir in response to low phosphorus stress is still unclear. Here, we performed small RNA sequencing on the Chinese fir roots treated with normal phosphorus and low phosphorus and identified a total of 321 miRNAs, including 139 known miRNAs and 182 new miRNAs, with 43 differentially expressed miRNAs (DEMs). Integrative analysis combined with degradome sequencing data revealed that 193 miRNAs (98 known and 95 new) targeted 469 genes, among which 23 DEMs targeted 44 genes. Gene enrichment analysis indicated that under low phosphorus stress, transcription and transcriptional regulation, as well as signal transduction, were significantly activated in Chinese fir. Modules in the miRNA–target pathways, such as miR166/HD-ZIP III, miR169/NFYA7, miR529/SPL, and miR399/UBC23, may be the key regulatory factors in the response to low phosphorus stress in Chinese fir. In addition, we found that PC-3p-1033_8666 was significantly downregulated and that PC-5p-3786_2830 was significantly upregulated, which presumably respond to low phosphorus stress by indirectly affecting phosphorus-related hormone signaling or PSR genes. The identified miRNA–target network and significantly activated pathways in this study provide insights into the post-transcriptional regulatory mechanisms of Chinese fir adapting to low phosphorus environments, which can offer theoretical references for the stress resistance and superior variety breeding of Chinese fir. Full article

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. Full article

The SPL (SQUAMOSA promoter-binding protein-like) transcription factor gene family plays a crucial role in the growth of plants, including fruit development. Although the SPL gene family has been widely studied in many plants, it remains unexplored in eggplant, an important Solanaceae vegetable crop. In this study, we performed a genome-wide analysis and identified 11 SmSPL genes with 10 motifs in the eggplant genome, which were assigned to five groups based on the amino acid sequences and the gene structure of the SBP domain. In addition, the expression patterns of 11 SmSPL genes were analyzed in different tissue types and at four different fruit development stages in eggplant. The results showed that all SmSPL genes were expressed differently in various tissues, suggesting that they may play different roles in the regulation of the development of different organs. Specifically, the expression level of SmSPL_5 was increased at the fruit expansion stage and showed the highest expression levels at 16 and 24 days after pollination, suggesting that it may be involved in regulating fruit development at both the fruit expansion and commercial fruit stages. Overall, this study provides an important basis for further exploring the function of the SmSPL gene family in eggplant. Full article

Plant-specific transcription factors known as SQUAMOSA promoter binding protein-like (SPL) genes are essential for development, growth, and abiotic stress responses. While the SPL gene family has been extensively studied in various plant species, a systematic characterization in Zanthoxylum bungeanum (Zb) is lacking. This study used transcriptomic and bioinformatics data to conduct a thorough genomic identification and expression investigation of the ZbSPL gene family. Eight subfamilies including 73 ZbSPL members were identified, most of which are predicted to be localized in the nucleus. Ka/Ks ratio analysis indicates that most ZbSPL genes have undergone purifying selection. According to evolutionary research, segmental duplication is a major factor in the amplification of the ZbSPL gene family. Gene structures, conserved motifs, and domains were found to be highly conserved among paralogs. Cis-element research revealed that ZbSPLs may be implicated in hormone and abiotic stress responses. Codon usage pattern analysis showed that the ZbSPL gene family was more inclined to A/T base endings; the higher the A/T content, the stronger the preference of the codons; and the use pattern was mainly affected by natural selection. Additionally, 36 ZbSPLs were found to be potential targets of miR156. RNA-seq demonstrated that SPL genes in Zb are differentially expressed in response to distinct abiotic stressors. Two ZbSPL genes (ZbSPL10 and ZbSPL17) were implicated in the response to salt stress, while four ZbSPL genes (ZbSPL06, ZbSPL43, ZbSPL60, and ZbSPL61) showed response to drought stress, based on a qRT-PCR investigation of the ZbSPL genes under various abiotic stress conditions. This study will help us gain a deeper understanding of the functions of ZbSPLs and lay a genetic foundation for future breeding of high-quality, highly abiotic resistant varieties of Z. bungeanum. Full article

Flower induction in pitaya (Hylocereus polyrhizus) is regulated by complex gene networks involving multiple signaling pathways that ensure flower bud (FB) formation, but its molecular determinants remain largely unknown. In this study, we aimed to identify key genes and pathways involved in pitaya flower induction by analyzing transcriptomics profiles from differentiating buds. Our results indicate that the flower induction process is driven by a combination of sugar, hormone, transcription factor (TF), and flowering-related genes. We found that during the FB induction period, the levels of sugar, starch, auxin (AUX), cytokinin (CTK) active forms dihydrozeatin riboside (dhZR), zeatin riboside (ZR), N6-isopentenyladenosine (iPA), and brassinosteroid (BR) increase in the late stage (LS), while active gibberellins (GA3, GA4) decrease, signaling a metabolic and hormonal shift essential for flowering. Differential gene expression analysis identified key genes involved in starch and sugar metabolism, AUX, CTK, BR synthesis, and (GA) degradation, with notable differential expression in photoperiod (COL, CDF, TCP), age-related (SPL), and key flowering pathways (FT, FTIP, AGL, SOC1). This study reveals a multidimensional regulatory network for FB formation in pitaya, primarily mediated by the crosstalk between sugar and hormone signaling pathways, providing new insights into the molecular mechanism of FB formation in pitaya. Full article

SQUAMOSA promoter-binding protein-like (SPL) transcription factors play a critical role in the regulation of gene expression and are indispensable in orchestrating plant growth and development while also improving resistance to environmental stressors. Although it has been identified across a wide array of plant species, there have been no comprehensive studies on the SPL gene family in centipedegrass [Eremochloa ophiuroides (Munro) Hack.], which is an important warm-season perennial C4 turfgrass. In this study, 19 potential EoSPL genes in centipedegrass were identified and assigned the names EoSPL1-EoSPL19. Gene structure and motif analysis demonstrated that there was relative consistency among the branches of the phylogenetic tree. Five pairs of segmental duplication events were detected within centipedegrass. Ten EoSPL genes were predicted to be targeted by miR156. Additionally, the EoSPL genes were found to be predominantly expressed in leaves and demonstrated diverse responses to abiotic stress (salt, drought, glufosinate ammonium, aluminum, and cold). This study offers a comprehensive insight into the SPL gene family in centipedegrass, creating a foundation for elucidating the functions of EoSPL genes and investigating their involvement in abiotic stress responses. Full article

Soil compaction poses a significant challenge in modern agriculture, as it constrains root development and hinders crop growth. The increasing evidence indicated that various phytohormones collaborate in distinct root zones to regulate root growth in compacted soils. However, the study of root development in maize under such conditions has been relatively limited. Here, we identified that the ZmSPL12 gene, belonging to the SPL transcription factor family, plays a crucial and positive role in regulating root development in the compacted soil. Specifically, the overexpression of ZmSPL12 resulted in significantly less inhibition of root growth than the wild-type plants when subjected to soil compaction. Histological analysis revealed that the capacity for root growth in compacted soil is closely associated with the development of the root cap. Further exploration demonstrated that ZmSPL12 modulates root growth through regulating ethylene signaling. Our findings underscored that ZmSPL12 expression level is induced by soil compaction and then enhances root penetration by regulating root cap and development, thereby enabling roots to thrive better in the compacted soil environment. Full article

Garden dahlias (Dahlia pinnata) are popular for their wide range of color variations, with polychromatic cultivars enhancing their ornamental value. Previous studies on the anthocyanin biosynthetic pathway (ABP) have indicated that the post-transcriptional suppression of the chalcone synthase gene (CHS) is involved in the formation of the white petals of dahlias. To further explore the complex mechanisms underlying polychromatic petal formation, we selected the bicolor cultivar ‘LiRen’ to identify candidate genetic factors. Through the detection of proanthocyanidin and anthocyanin, it was indicated that the white tips of the petals lacked anthocyanin but accumulated some proanthocyanidin, albeit at significantly lower levels than those at the red bases of the petals. This suggests that the upstream ABP, which involves CHS, is not entirely inactive. Transcription sequencing and quantitative reverse transcription PCR (qRT-PCR) analysis demonstrated that the inactive ABP in the white tips results from the downregulation of ABP structural genes. The low abundance of DpMYB1 appears to be the key factor influencing the lack of strong transcription activation of the structural genes. Additionally, highly upregulated DpSPL9 targeted by the downregulated miR156 in the white tips was identified through qRT-PCR. This suggests that DpSPL9 may act as an anthocyanin depressor to destabilize the MYB-bHLH-WDR complex through interaction with DpMYB1. The findings indicate that the DpMYB1 and miR156-DpSPL9 modules play potential regulatory roles in the formation of bicolor petals. Overall, these results provide new insights into the color patterning of dahlias and will be valuable for further studies regarding the mechanisms underlying polychromatic petal formation. Full article

Bamboo plants have erratic flowering habits with a long vegetative growth and an uncertain flowering cycle. The process of floral transition has always been one of the hot and intriguing topics in bamboo developmental biology. As master modulators of gene expression at the post-transcriptional level, miRNAs play a crucial role in regulating reproductive growth, especially in floral transition of flowering plants. Pleioblastus pygmaeus is a kind of excellent ground cover ornamental bamboo species. In this study, we performed miRNA expression profiling of the shoot buds and flower buds from the bamboo species, to investigate flowering-related miRNAs in bamboo plants. A total of 179 mature miRNAs were identified from P. pygmaeus, including 120 known miRNAs and 59 novel miRNAs, of which 96 (61 known miRNAs and 35 novel miRNAs) were differentially expressed in the shoots at different growth stages. Based on target gene (TG) prediction, a total of 2099 transcription factors (TFs) were annotated to be TGs of the 96 differentially expressed miRNAs (DEMs), corresponding to 839 recordings of DEM-TF pairs. In addition, we identified 23 known DEMs involved in flowering and six known miRNAs related to floral organ development based on previous reports. Among these, there were 11 significantly differentially expressed miRNAs, with 124 TF targets corresponding to 132 DEM-TF pairs in P. pygmaeus. In particular, we focused on the identification of miR156a-SPL (SQUAMOSA Promoter-Binding protein-Like) modules in the age pathway, which are well-known to regulate the vegetative-to-reproductive phase transition in flowering plants. A total of 36 TF targets of miR156a were identified, among which there were 11 SPLs. The Dual-Luciferase transient expression assay indicated miR156a mediated the repression of the PpSPL targets in P. pygmaeus. The integrated analysis of miRNAs and TGs at genome scale in this study provides insight into the essential roles of individual miRNAs in modulating flowering transition through regulating TF targets in bamboo plants. Full article

Clubroot disease, caused by the pathogen Plasmodiophora brassicae, is a serious disease that poses a critical threat to cabbage production. However, the molecular mechanism of the microRNAs (miRNAs) involved in the cabbage’s response to P. brassicae infection remains to be elucidated. Here, the mRNA and miRNA expression profiles of cabbage in response to a P. brassicae infection were analyzed. In the transcriptome analysis, 2217 and 5552 differentially expressed genes (DEGs) were identified at 7d and 21d after inoculation, which were enriched in MAPK signaling, plant–pathogen interaction, plant hormone signal transduction, and phenylpropanoid biosynthesis pathways. BolC02g057640.2J, BolC09g006890.2J, BolC02g013230.2J, BolC06g006490.2J, BolC03g052660.2J, BolC07g052580.2J, and BolC04g044910.2J were predicted to be significantly involved in the defense response or plant–pathogen interaction through co-expression network analysis. Small RNA data analysis identified 164 miRNAs belonging to 51 families. miR1515, miR166, miR159, and miR9563 had the greatest number of members among the miRNA families. Integrated analysis revealed 23 miRNA–mRNA interactions related to a P. brassicae infection. The target genes of differentially expressed miRNAs (DEMs) revealed the NAC, ARF, TCP, and SPL transcription factor members that probably participate in the defense response. This study provided new insights into the miRNA-involved regulatory system during the process of disease infection with P. brassicae in cabbage. Full article

Tomato fruit ripening is an elaborate genetic trait correlating with significant changes at physiological and biochemical levels. Sugar metabolism plays an important role in this highly orchestrated process and ultimately determines the quality and nutritional value of fruit. However, the mode of molecular regulation is not well understood. Galactinoal-sucrose galactosyltransferase (GSGT), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), can transfer the galactose unit from 1-α-D-galactosyl-myo-inositol to sucrose and yield raffinose, or catalyze the reverse reaction. In the present study, the expression of SlGSGT2 was decreased by Potato Virus X (PVX)-mediated gene silencing, which led to an unripe phenotype in tomato fruit. The physiological and biochemical changes induced by SlGSGT2 silencing suggested that the process of fruit ripening was delayed as well. SlGSGT2 silencing also led to significant changes in gene expression levels associated with ethylene production, pigment accumulation, and ripening-associated transcription factors (TFs). In addition, the interaction between SlGSGT2 and SlSPL-CNR indicated a possible regulatory mechanism via ripening-related TFs. These findings would contribute to illustrating the biological functions of GSGT2 in tomato fruit ripening and quality forming. Full article