Search Results (41)

Flowering time is an important trait affecting regional adaptation and yield stability in rapeseed (Brassica napus). To dissect the genetic basis of flowering time and vernalization-type differentiation in B. napus, this study integrated genome-wide SNPs, flowering-time phenotypes, vernalization types, and geographic origin information to establish a candidate QTL discovery framework combining genome-wide association study (GWAS) and machine learning. Based on 8,387,529 high-quality SNPs, GWAS was performed using the FarmCPU model under two covariate settings: a PC-corrected model and a PC + vernalization type-corrected model. These two models identified 32 and 27 suggestive SNPs, respectively. In parallel, ExtraTreesRegressor was used to estimate the feature contribution of SNPs to flowering-time prediction, leading to the identification of 18,854 ML_delta10SD high-contribution SNPs, which were further merged into 997 ML-derived candidate QTL intervals, hereafter referred to as ML-QTLs. By integrating GWAS-derived QTLs and ML-QTLs at the interval level, 34 GWAS- and ML-supported candidate regions were prioritized. Candidate gene annotation revealed that these regions contained genes related to flowering regulation, including FLC, FRIGIDA-like, VIN3-like, SOC1, ELF3, CONSTANS-like, APETALA1-like, and genes involved in the gibberellin pathway. Top-SNP genotypic effect analysis and multi-SNP genotype combination analysis further showed that genetic variation within representative candidate regions was significantly associated with flowering-time differences and vernalization-type differentiation. Predictive modeling indicated that candidate markers selected by GWAS and machine learning showed predictive potential within the present population. Overall, this study prioritized a set of GWAS- and ML-supported candidate regions and putative candidate genes associated with flowering time in B. napus from the complementary perspectives of statistical association and machine learning-based predictive contribution, providing candidate resources for future marker development, functional validation, and adaptive improvement in rapeseed. Full article

The miRNA172–APETALA2 (AP2) regulatory module is a conserved mechanism governing floral development in plants. Disruption of the miR172 target sites in AP2 genes has been shown to be key to the domestication of double flowers in ornamental species. Camellia japonica, a woody ornamental plant with diverse floral forms, serves as an important model for studying double-flower formation. In this study, we characterized two AP2-like transcription factors, CjAP2-1 and CjAP2-2, which possess evolutionarily conserved miR172-binding sites and exhibit broad expression across floral tissues. To investigate the role of the miR172–AP2 module in C. japonica, we identified four members of the miR172 family and demonstrated that miR172 is directly involved in the cleavage of CjAP2-1 and CjAP2-2 transcripts. Through bulked amplicon sequencing of cultivars with diverse floral forms, we uncovered natural variations at the miR172-binding sites of CjAP2-1 and CjAP2-2, which can potentially disrupt miR172-mediated mRNA cleavage. We showed that two dinucleotide mutations (CjAP2-1-mut5 and CjAP2-1-mut9) significantly reduced the miR172-mediated repression of CjAP2-1 transcripts. Functional analysis in Arabidopsis revealed that overexpression of the CjAP2-1-mut5 variant caused significant floral abnormalities, including ectopic formation of reproductive organs, loss of floral determinacy, and fusion of floral organs. Further analysis of downstream genes indicated that key regulators of floral homeotic and meristem activity were markedly altered in the transgenic plants. Our findings demonstrate that perturbations in the miR172–AP2 regulatory relationship underlie the formation of double flowers in C. japonica by altering floral meristem determinacy and organ identity. Full article

The APETALA2/Ethylene Response Factor (AP2/ERF) family of transcription factors (TF) is characterized by their participation in various biological processes related to growth, development, and response to stress. ERFs are ideal candidates for crop improvement because they regulate defense genes like JERF1, JERF3, LeERF2, NtERF5, and Tsil which confer tolerance to drought, salinity, osmotic stress, and pathogen attack, respectively. The ERF subfamily includes the TF Pti4, whose activity is regulated by different signaling pathways, thus providing tolerance response to multiple factors such as drought, salinity, cold, and pathogen attack in tomato. In this work we evaluated the effect of overexpression of TF SlPti4 from Solanum lycopersicum in transgenic tobacco plants when subjected to saline, osmotic, and drought stress. Our results from this study demonstrated that transgenic lines overexpressing Pti4 tolerate abiotic stress during germination and in plants. The transgenic lines showed improvements in photoinhibition, electron transport rate, chlorophyll content, and biomass, as well as a reduction in malondialdehyde content. Full article

The double-flower trait is highly valued in ornamental plants due to its unique aesthetic appeal, yet its genetic basis varies significantly across different species. While AGAMOUS (AG) and APETALA2 (AP2)-like genes have been demonstrated to play crucial roles in floral organ identity regulation in the model plant Arabidopsis thaliana, the underlying mechanisms governing double-flower formation in many ornamental species remain largely unexplored. In this study, we examined the inheritance pattern of this trait and identified a genetic variant associated with petal number variation. Crosses between the single-flowered cultivar ‘Baccarat White’ (BW) and the semi-double cultivar ‘Duo Lavender’ (DL) produced a 1:1 segregation of single and semi-double flowers in the F1 generation, while self-pollination of DL yielded a 1:2:1 segregation of single, semi-double, and double flowers. These results indicate that the double-flower trait follows a single-gene, semi-dominant inheritance model. Whole-genome sequencing of BW and DL followed by sequence analysis of floral organ identity genes revealed no significant differences in B-class (PhGLO1, PhGLO2, PhDEF, and PhTM6) or C-class (pMADS3 and FBP6) genes between the two cultivars. Notably, a 10 kb insertion upstream of the miR172 target site in the PhBOB gene was detected in DL. PCR genotyping of 192 F1 progenies demonstrated complete co-segregation between this insertion and the double-flower phenotype, suggesting a strong genetic association. Moreover, qRT-PCR analysis showed that PhBOB expression was significantly elevated in DL—exhibiting a 69-fold increase in petals compared to BW—implying that its overexpression disrupts the petal-to-stamen identity transition. Additionally, another AP2 family gene, PhROB3, was upregulated in semi-double flowers, with a 10-fold higher expression in the petals and stamens of DL relative to BW, suggesting its potential role in floral organ differentiation. This study elucidates the molecular regulatory mechanism underlying the double-flower trait in petunia, highlighting the role of PhBOB in floral organ identity specification and providing new insights into the potential function of PhROB3 in double-flower development. Full article

ERF56, a member of the APETALA2/ETHYLENE-RESPONSIVE FACTOR (AP2/ERF) transcription factor (TF) family, was reported to be an early nitrate-responsive TF in Arabidopsis. But the function of ERF56 in nitrate signaling remains not entirely clear. This study aimed to investigate the role of ERF56 in nitrate-dependent plant growth and nitrate signaling. We confirmed with reverse transcription quantitative PCR (RT-qPCR) that the transcription of ERF56 is quickly induced by nitrate. ERF56 overexpressors displayed decreased nitrate-dependent plant growth, while erf56 mutants exhibited increased plant growth. Confocal imaging demonstrated that ERF56 is localized into nuclei. Assays with the glucuronidase (GUS) reporter showed that ERF56 is mainly expressed at the region of maturation of roots and in anthers. The dual-luciferase assay manifested that the transcription of ERF56 is not directly regulated by NIN-LIKE PROTEIN 7 (NLP7). The transcriptome analysis identified 1038 candidate genes regulated by ERF56 directly. A gene ontology (GO) over-representation analysis showed that ERF56 is involved in the processes of water transport, inorganic molecule transmembrane transport, secondary metabolite biosynthesis, and cell wall organization. We revealed that ERF56 represses nitrate-dependent growth through regulating the processes of inorganic molecule transmembrane transport, the secondary metabolite biosynthesis, and cell wall organization. Full article

Background/Objectives: BABY BOOM (BBM), a transcription factor from the APETALA2 (AP2) protein family, plays a critical role in somatic embryo induction and apomixis. BBM has now been widely applied to induce apomixis or enhance plant transformation and regeneration efficiency through overexpression or ectopic expression. However, the structural and functional evolutionary history of BBM genes in plants is still not well understood. Methods: The protein sequences of 10 selected plant species were used to locate the branch of BBM-Like by key domain identification and phylogenetic tree construction. The identified BBML genes were used for further conserved motif identification, gene structural analysis, miRNA binding site prediction, cis-acting element prediction, collinear analysis, protein–protein interaction network construction, three-dimensional structure modeling, molecular docking, and expression pattern analysis. Results: A total of 24 BBML proteins were identified from 10 representative plant species. Phylogenetic relationship analysis displayed that BBML proteins from eudicots and monocots were divided into two clusters, with monocots exhibiting a higher number of BBMLs. Gene duplication events indicated that whole genome/segmental duplication were the primary drivers of BBML genes’ evolution in the tested species, with purifying selection playing a key role during evolution processes. Comparative analysis of motif, domains, and gene structures revealed that most BBMLs were highly evolutionarily conserved. The expression patterns of BBML genes revealed significant tissue specificity, particularly in the root and embryo. We also constructed protein–protein interaction networks and molecular docking models to identify functional pathways and key amino acid residues of BBML proteins. The functions of BBMLs may differ between monocots and eudicots, as suggested by the functional enrichment of interacting proteins. Conclusions: Our research delved into the molecular mechanism, evolutionary relationships, functional differentiation, and expression patterns of BBML genes across plants, laying the groundwork for further investigations into the molecular properties and biological roles of BBMLs. Full article

In a warm winter due to climate warming, it is necessary to suppress early flowering of autumn-sown wheat plants. Here, we propose the use of cytoplasmic genome effects for this purpose. Alloplasmic lines, or cytoplasmic substitution lines, of bread wheat (Triticum aestivum) have cytoplasm from a related wild Aegilops species through recurrent backcrossing and exhibit altered characteristics compared with the euplasmic lines from which they are derived. Thus, alloplasmic lines with Aegilops mutica cytoplasm show delayed flowering compared with lines carrying normal cytoplasm. In the wheat flowering pathway, VERNALIZATION 1 (VRN1) encodes an APETALA1/FRUITFULL-like MADS box transcription factor that plays a central role in the activation of florigen genes, which induce floral meristems in the shoot apex. Here, we compared expression of VRN1 alleles in alloplasmic and euplasmic lines after vernalization. We found that alloplasmic wheat showed a lower level of VRN1 expression after vernalization compared with euplasmic wheat. Thus, nuclear-cytoplasm interactions affect the expression levels of the nuclear VRN1 gene; these interactions might occur through the pathway termed retrograde signaling. In warm winters, autumn-sown wheat cultivars with spring habit can pass through the reproductive growth phase in very early spring, resulting in a decreased tiller/ear number and reduced yield performance. Here, we present data showing that an alloplasmic line of ‘Fukusayaka’ can avoid the decrease in tiller/ear numbers during warm winters, suggesting that this alloplasmic line may be useful for development of varieties adapted to global warming. Full article

Somatic embryogenesis is valuable for clonal propagation and genetic improvement, and it also serves as an ideal system for studying plant development mechanisms. In Larix kaempferi, microRNA171 and its target gene L. kaempferi SCARECROW-LIKE6 (LaSCL6), which has two alternative splicing variants, can regulate somatic embryogenesis; however, the underlying molecular mechanism is still unknown. In this study, we overexpressed these two LaSCL6 variants in Oryza sativa and Arabidopsis thaliana and then used the RNA-Seq method to screen genes from O. sativa and A. thaliana, whose expression patterns are related to those of LaSCL6 variants. The screened genes were then used to search L. kaempferi proteins to identify the candidate target genes of LaSCL6. After yeast one-hybrid and dual- luciferase transcriptional activity assays, cytochrome P450, family 89, subfamily A, polypeptide 5 (CYP89A5), and wall-associated receptor kinase-like 20 (WAKL20) were confirmed to be the target genes of LaSCL6-var1; in addition, WAKL20 and UDP-glycosyltransferase 85A3 (UGT85A3) were confirmed to be the target genes of LaSCL6-var2. Moreover, APETALA2-like protein 2, a transcription factor from the AP2/ERF family, was shown to interact with LaSCL6-var1 and LaSCL6-var2. Taken together, our results suggest a regulatory network of miR171-LaSCL6. The findings presented here not only provide novel insights into the regulation of the miR171-LaSCL6 module but also explain the mechanism underlying larch somatic embryogenesis and other biological processes. Full article

Grapevine (Vitis vinifera) is an important and popular perennial fruit tree cultivated worldwide. Grapevine ripening is affected by flowering time, and although members of the MADS-box protein family play vital roles in regulating flowering in plants, the functions of MADS-box proteins in grapevine remain largely unknown. AGAMOUS-LIKE 11 (VvAGL11), a MADS-box gene in grapevine, was reported to be a regulator of seed morphogenesis. In this study, heterologous overexpression of VvAGL11 was found to significantly promote flowering in Arabidopsis, suggesting that its active expression in grapevine may induce early flowering and ripening. Transcriptome analysis showed that VvAGL11 overexpression affected the expression of genes involved in stress responses, hormonal signaling responses, and flowering regulation. Notably, VvAGL11 significantly increased the expression of key flowering genes such as FLOWERING LOCUS T (FT), APETALA3 (AP3), and SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 5 (SPL5), which might have contributed to the early flowering in Arabidopsis. In summary, we characterized a novel flowering regulator, VvAGL11, which could be a potential target for early ripening breeding in grapevine. Full article

In the rosid species Arabidopsis thaliana, the AP2-type AP2 transcription factor (TF) is required for specifying the sepals and petals identities and confers a major A-function to antagonize the C-function in the outer floral whorls. In the asterid species Petunia, the AP2-type ROB TFs are required for perianth and pistil development, as well as repressing the B-function together with TOE-type TF BEN. In Long-homostyle (LH) Fagopyrum esculentum, VIGS-silencing showed that FaesAP2 is mainly involved in controlling filament and style length, but FaesTOE is mainly involved in regulating filament length and pollen grain development. Both FaesAP2 (AP2-type) and FaesTOE (TOE-type) are redundantly involved in style and/or filament length determination instead of perianth development. However, neither FaesAP2 nor FaesTOE could directly repress the B and/or C class genes in common buckwheat. Moreover, the FaesAP1_2 silenced flower showed tepal numbers, and filament length decreased obviously. Interestingly, yeast one-hybrid (Y1H) and dual-luciferase reporter (DR) further suggested that FaesTOE directly up-regulates FaesAP1_2 to be involved in filament length determination in LH common buckwheat. Moreover, the knockdown of FaesTOE expression could result in expression down-regulation of the directly target FaesAP1_2 in the FaesTOE-silenced LH plants. Our findings uncover a stamen development pathway in common buckwheat and offer deeper insight into the functional evolution of AP2 orthologs in the early-diverging core eudicots. Full article

Exogenous ethylene is commonly utilized to initiate flower induction in pineapple (Ananas comosus (L.) Merr.). However, the molecular mechanisms and metabolic changes involved are not well understood. In this study, we explored the genetic network and metabolic shifts in the ‘Comte de Paris’ pineapple variety during ethylene-induced flowering. This was achieved through an integrative analysis of metabolome and transcriptome profiles at vegetative shoot apexes (0 d after ethephon treatment named BL_0d), the stage of bract primordia (8 d after ethephon treatment named BL_8d), stage of flower primordia (18 d after ethephon treatment named BL_18d), and the stage of stopped floret differentiation (34 d after ethephon treatment named BL_34d). We isolated and identified 804 metabolites in the pineapple shoot apex and inflorescence, categorized into 24 classes. Notably, 29, 31, and 46 metabolites showed significant changes from BL_0d to BL_8d, BL_8d to BL_18d, and BL_18d to BL_34d, respectively. A marked decrease in indole was observed, suggesting its role as a characteristic metabolite during flower induction. Transcriptomic analysis revealed 956, 1768, and 4483 differentially expressed genes (DEGs) for BL_0d vs. BL_8d, BL_8d vs. BL_18d, and BL_18d vs. BL_34d, respectively. These DEGs were significantly enriched in carbohydrate metabolism and hormone signaling pathways, indicating their potential involvement in flower induction. Integrating metabolomic and transcriptomic data, we identified several candidate genes, such as Agamous-Like9 (AGL9), Ethylene Insensitive 3-like (ETIL3), Apetala2 (AP2), AP2-like ethylene-responsive transcription factor ANT (ANT), and Sucrose synthase 2 (SS2), that play potentially crucial roles in ethylene-induced flower induction in pineapple. We also established a regulatory network for pineapple flower induction, correlating metabolites and DEGs, based on the Arabidopsis thaliana pathway as a reference. Overall, our findings offer a deeper understanding of the metabolomic and molecular mechanisms driving pineapple flowering. Full article

Intramuscular fat (IMF) deposits help improve meat quality such as marbling, juicy, flavor and tenderness. Long-chain acyl-CoA dehydrogenase (ACADL) is a key enzyme for catalyzing fatty acid oxidation, and studies have shown ACADL is involved in the deposition and differentiation of intramuscular adipocytes. However, the effect of ACADL on intramuscular adipocytes differentiation in goats needs further study. In this study, to explore the mechanism of ACADL on the development of goat intramuscular adipocytes, we constructed an over-expression plasmids and a SI-RNA of ACADL to explore the function of ACADL on the development of goat IMF. It was found that overexpression of ACADL promoted the differentiation of goat intramuscular adipocytes, and promoted the expression of fat cell differentiation marker genes lipoprotein lipase (LPL), peroxisome proliferator activated receptor gamma (PPARγ), APETALA-2-like transcription factor gene (AP2), CCAT enhancer binding protein (CEBPα), preadipocyte Factor 1 (Pref-1) and CCAT enhancer binding protein (CEBPβ), and the opposite trend occurred after interference. In addition, we screened of this related tumor necrosis factor (TNF) signaling pathway by RNA-Seq. So, we validate the signaling pathway with inhibitor of TNF signaling pathway. In summary, these results indicate that ACADL promotes intramuscular adipocytes differentiation through activation TNF signaling pathway. This study provides an important basis for the mechanism of IMF development. Full article

Tree peony (Paoenia ostii) is a famous Chinese traditional flower well-known in many countries of the world. However, the short and concentrated flowering period of tree peony greatly affects the ornamental and economic value of the flowers. Exogenous brassinosteroid (BR) treatment can delay the flowering period of ostii T. Hong et J. X. Zhang var. lishizhenenii B. A. Shen for 3 days, but the underlying regulatory mechanism remains elusive. Here, full-length transcriptome and transcriptome sequencing were used to mine key genes related to BR-induced delayed flowering in tree peony. The transcriptome sequencing of the petals yielded 21.27 G clean data and 62,229 isoforms. Among them, 58,218 isoforms were annotated in NR, NT, SwissProt, KEGG, KOG, InterPro and GO databases. GO and KEGG analyses showed that 2460 DEGs were related to delayed flowering in response to BR. Additionally, a total of seven genes affecting flowering were annotated from 11 isoforms, which responded to BR through three pathways to delay the flowering of P. ostii var. lishizhenii. BR treatment increased the expression of BRASSINOSTEROID-SIGNALING KINASE3 (BSK3), potentially by promoting BRI1 ASSOCIATED KINASE RECEPTOR 1 (BAK1). Moreover, BR treatment suppressed the expression of SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE 1 (SPL1), possibly to inhibit the expressions of downstream WRKY genes and APETALA 2 (AP2). Meanwhile, BR treatment promoted the expression of DELLA, which might inhibit the expressions of CONSTANS (CO) and SPL. These results suggest a theoretical basis for further analyses of the molecular mechanism of flowering regulation in tree peony. Full article

The tropical common bean (Phaseolus vulgaris L.) is an obligatory short-day plant that requires relaxation of the photoperiod to induce flowering. Similar to other crops, photoperiod-induced floral initiation depends on the differentiation and maintenance of meristems. In this study, the global changes in transcript expression profiles were analyzed in two meristematic tissues corresponding to the vegetative and inflorescence meristems of two genotypes with different sensitivities to photoperiods. A total of 3396 differentially expressed genes (DEGs) were identified, and 1271 and 1533 were found to be up-regulated and down-regulated, respectively, whereas 592 genes showed discordant expression patterns between both genotypes. Arabidopsis homologues of DEGs were identified, and most of them were not previously involved in Arabidopsis floral transition, suggesting an evolutionary divergence of the transcriptional regulatory networks of the flowering process of both species. However, some genes belonging to the photoperiod and flower development pathways with evolutionarily conserved transcriptional profiles have been found. In addition, the flower meristem identity genes APETALA1 and LEAFY, as well as CONSTANS-LIKE 5, were identified as markers to distinguish between the vegetative and reproductive stages. Our data also indicated that the down-regulation of the photoperiodic genes seems to be directly associated with promoting floral transition under inductive short-day lengths. These findings provide valuable insight into the molecular factors that underlie meristematic development and contribute to understanding the photoperiod adaptation in the common bean. Full article

Olive (Olea europaea L.) trees can reach a very old age and still bear fruit. Although traditional groves are planted at low density and are rainfed, many newer groves are planted at higher densities and irrigated. As expected, initial yields per area are larger in high density plantations, yet some farmers claim they experience a reduction in productivity with grove age, even in well maintained trees. In order to test the accuracy of this claim and its underlying cause, we measured several productivity parameters in selected branches of trees in seven sites differing in cultivar (‘Barnea’ or ‘Souri’), location and irrigation regime (rainfed or irrigated) for two consecutive years. For each site (cultivar/location/regime), we compared neighboring groves of different ages, altogether 14 groves. There was no consistent reduction in productivity in older groves. Differences in productivity between irrigated cultivars were mostly due to variation in the percentage of inflorescences that formed fruit. Several parameters were higher in irrigated, compared to rainfed ‘Souri’. Differences in productivity between years within the same grove was mostly due to variation in the percentage of nodes forming inflorescences. We studied the expression of OeFT2 encoding a FLOWERING LOCUS T protein involved in olive flower induction in leaves of trees of different ages, including juvenile seedlings. Expression increased during winter in mature trees and correlated with the percentage of inflorescences formed. The leaves of juvenile seedlings expressed higher levels of two genes encoding APETALA2-like proteins, potential inhibitors of OeFT2 expression. The buds of juvenile seedlings expressed higher levels of OeTFL1, encoding a TERMINAL FLOWER 1 protein, a potential inhibitor of OeFT2 function in the meristem. Our results suggest that olives, once past the juvenile phase, can retain a similar level of productivity even in densely planted well maintained groves. Full article