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Literature Sharing | SPL1 positively regulates cuticular ridge wax biosynthesis in Arabidopsis

Release time:

2025-06-06

This study investigates the molecular mechanisms underlying the formation and maintenance of cuticular ridge structures, which are characteristic of the sepals and petals in various plant species. The authors reveal that the production of cuticular ridge wax in Arabidopsis is regulated by SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors. Double mutants of SPL1 and SPL12 (spl1spl12) show severe defects in cuticular ridge formation, with significantly reduced seed-setting rates and increased organ fusion compared to wild-type plants. Further analysis demonstrates that SPL1 positively regulates the expression of key enzymes involved in wax biosynthesis, including the alkane-forming enzymes ECERIFERUM1 (CER1), CER1-like (CER1L), and the primary (1-) alcohol-forming enzyme CER4. These enzymes are essential for the synthesis of alkanes and 1-alcohols, which are crucial for the accumulation of cuticular ridge wax. Additionally, the study identifies HSP70–16 as an interacting partner of SPL1, enhancing its transcriptional activity. Together, these results uncover a regulatory network critical for cuticular ridge formation and provide new insights into plant developmental processes.

In the spl1spl12 double mutants, vegetative growth appeared normal, but there was a significant reduction in seed set during the post-reproductive phase due to flowers failing to produce siliques. Stereomicroscopy of stage 13 flowers revealed that the sepals of spl1spl12 mutants did not open properly compared to wild-type plants. Internal analysis of floral organs showed that stamens, gynoecia, and petals in the fused flowers did not extend properly, often growing twisted or curved. The restricted space in unopened buds prevented mature pollen from contacting the stigma. Additionally, papilla cells on the stigma were abnormal, leading to reduced pollination. However, pollen grains were viable at anthesis, and artificial self-pollination restored normal seed production, indicating that the male and female gametophytes were not defective. These results suggest that the reduced fertility in spl1spl12 is mainly due to the failure of sepals to open.

Fig. 1. Loss of sepal cuticular ridges wax in spl1spl12 reduces seed setting rate.

To investigate the underlying mechanisms, the researchers compared the transcriptomes of Col-0 and spl1spl12 flowers under normal conditions. KEGG annotation of differentially expressed genes (DEGs) revealed significant enrichment in pathways related to cutin, suberine, and wax biosynthesis, suggesting that the loss of SPL1 and SPL12 disrupts wax biosynthesis, leading to reduced wax accumulation in the sepal epidermis. Among the downregulated DEGs, key wax biosynthesis genes—CER1-Like (CER1L), CER1, and CER4—were notably reduced in spl1spl12 mutants. These CER genes showed higher expression levels in floral organs than in vegetative tissues, peaking at flower development stage 12. CER1L and CER1 are involved in the alkane-forming pathway of floral wax biosynthesis, while CER4, an acyl-CoA reductase, is crucial for primary alcohol formation. Mutations in CER4 lead to a significant reduction in primary alcohols and wax esters, further implicating it in the wax biosynthesis process.

Fig. 2. SPL1 activates wax biosynthetic genes.

To identify potential interacting partners of SPL1, a yeast two-hybrid screening was performed, which identified 21 candidate proteins. Notably, three independent clones encoded AT1G11660, corresponding to the molecular chaperone HSP70–16. This is significant because previous studies have shown that loss-of-function mutants of HSP70–16 exhibit impaired reproductive development, including reduced seed set due to postgenital sepal fusion defects. Subcellular localization analysis revealed that SPL1 and HSP70–16 co-localize in N. benthamiana leaf epidermal cells. The physical interaction between SPL1 and HSP70–16 was confirmed using yeast two-hybrid assays, luciferase complementation, and bimolecular fluorescence complementation. A triple mutant (spl1spl12hsp70–16) was created, and phenotypic analysis showed that it exhibited more severe developmental defects than the spl1spl12 double mutant, including reduced seed set and abnormal cuticular ridge formation at the sepal tips. These results suggest a functional synergy between SPL1 and HSP70–16 in regulating reproductive development.

Fig. 3. SPL1 interacts with HSP70–16.

Comparative transcriptomic profiling of the spl1spl12hsp70–16 triple mutants versus wild-type plants, followed by KEGG pathway enrichment analysis, revealed significant differential expression of genes linked to lipid metabolism, including fatty acid biosynthesis, elongation, and pathways related to cuticular wax and suberin biosynthesis. This transcriptional reprogramming suggests a synergistic regulatory interaction between SPL1 and HSP70–16 in modulating these metabolic processes.

Fig. 4. HSP70–16 enhances SPL1 transcriptional activity through protein interaction.

In summary, the study reveals the critical role of SPL proteins in cuticular ridge formation. The spl1spl12 double mutants show reduced wax in sepal cuticular ridges, leading to sepal fusion, and failure in flowering and seed setting. These findings highlight the importance of an intact cuticle for plant growth and development. Further analysis shows that HSP70–16 interacts with SPL1, enhancing its transcriptional activation of wax biosynthesis genes. The authors propose a novel regulatory mechanism involving the SPL1-HSP70–16 module, which is crucial for sepal cuticular ridge formation and preventing organ fusion. This work provides insights into cuticle integrity and broader developmental processes in plants.

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Literature Sharing | Interaction of PsMYB4 with PsEGL3 inhibits anthocyanin biosynthesis in tree peony yellow flowers

This study explores the molecular mechanism behind yellow flower formation in tree peony, a highly valued ornamental plant in China. Researchers identified two transcription factors, PsMYB4 and PsEGL3, that are highly expressed in a yellow-flowered cultivar.

2025-07-10

Literature Sharing | Putative Upstream Regulators DoNF-YB3 and DoIDD12 Correlate with DoGSTF11 Expression and Anthocyanin Accumulation in Dendrobium officinale

This study explores the role of the DoGSTF11 gene in Dendrobium officinale, a traditional medicinal herb. While previous research has focused on polysaccharides and alkaloids, this work addresses the lesser-known anthocyanin pathway. The researchers found that DoGSTF11 is highly expressed in the purplish variety of D. officinale and is localized in the nucleus and cell membrane, though it lacks transcriptional activation ability. Overexpressing DoGSTF11 in tomato led to increased anthocyanin accumulation, suggesting it plays a role in anthocyanin transport or sequestration. Protein interaction studies revealed that DoGSTF11 interacts with DoGST31, and that DoIDD12 and DoNF-YB3 may regulate its expression. Overall, the findings highlight DoGSTF11 as a positive regulator of anthocyanin accumulation and offer new insights for flavonoid metabolic engineering in D. officinale.

2025-07-08

Literature Sharing | Chinese cabbage orphan gene BR3 confers bolting resistance to Arabidopsis through the gibberellin pathway

Premature bolting affects yield and quality in Chinese cabbage, highlighting the importance of identifying bolting resistance genes. This study identifies an orphan gene, BR3 (BOLTING RESISTANCE 3), in Arabidopsis thaliana as a positive regulator of bolting resistance. BR3 is expressed during the seedling and flowering stages and localizes to the plasma membrane and nucleus. Overexpression of BR3 (BR3OE) delays bolting and flowering under both long-day and short-day conditions, with increased rosette leaf number and reduced plant height. Key flowering genes are downregulated in BR3OE plants. GA₃ treatment induces early flowering in both BR3OE and wild type (WT) plants, but BR3OE still flowers later than WT. In Chinese cabbage, BR3 shows co-expression with DELLA genes BrRGA1 and BrRGL3, suggesting a regulatory role through the GA pathway. This study offers new insights into bolting resistance mechanisms and provides valuable targets for breeding bolting-resistant Chinese cabbage varieties.

2025-07-03

Literature Sharing | Regulation of co-translational mRNA decay by PAP and DXO1 in Arabidopsis

This study investigates the regulation of the co-translational mRNA decay (CTRD) pathway in Arabidopsis, a critical mechanism for maintaining mRNA homeostasis. The researchers found that 3ʹ-phosphoadenosine 5ʹ-phosphate (PAP), an inhibitor of exoribonucleases, affects CTRD activity. Specifically, they showed that loss of FRY1 impairs XRN4-dependent CTRD and that exogenous PAP treatment stabilizes CTRD target mRNAs. Additionally, they discovered that another PAP-sensitive exoribonuclease, DXO1, also contributes to CTRD, likely by acting on NAD⁺-capped mRNAs involved in stress responses. These findings reveal new layers of regulation and additional players in the CTRD pathway in plants.

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Literature Sharing | Two pathogen-inducible UDP-glycosyltransferases, UGT73C3 and UGT73C4, catalyze the glycosylation of pinoresinol to promote plant immunity in Arabidopsis

This study uncovers a novel immune regulatory pathway in Arabidopsis thaliana involving two UDP-glycosyltransferases, UGT73C3 and UGT73C4, which are highly induced by Pseudomonas syringae infection. Overexpression of these genes enhances disease resistance, while their double mutation compromises immunity. Metabolomic analysis and biochemical assays reveal that UGT73C3/C4 glycosylate pinoresinol into its mono- and diglucoside forms, which promote immune responses by boosting ROS production and callose deposition. Additionally, the transcription factor HB34 directly activates UGT73C3/C4 expression, linking transcriptional regulation to glycosylation-mediated immunity. This work highlights the physiological significance of UGTs in plant defense through pinoresinol glycosylation.

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Literature Sharing | PbARP1 enhances salt tolerance of ‘Duli’ pear (Pyrus betulifolia Bunge) through abscisic acid signalling pathway

This study investigates salt stress tolerance in pears and identifies 35 salt-tolerant genes using a yeast expression library. Among them, PbARP1 was found to be significantly upregulated under salt stress in 'Duli' pear (Pyrus betulaefolia). Functional analyses showed that overexpression of PbARP1 in transgenic pear calli enhanced salt tolerance, while its suppression increased sensitivity to salt stress. Silencing PbARP1 also altered the expression of key ABA signaling genes, including PbPYL4, PbPYL9, PbPYL8, PbSRK2I, and PbABI5, suggesting that PbARP1 modulates salt stress responses through the ABA signaling pathway. Furthermore, PbPYL8, an ABA receptor, was identified as a protein interacting with PbARP1, highlighting its pivotal role in ABA-mediated salt stress regulation in pear.

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Literature Sharing | Phosphorylation of the strawberry MADS-box CMB1 regulates ripening via the catabolism of abscisic acid

This study uncovers a regulatory mechanism linking transcriptional control and posttranslational modification in strawberry fruit ripening. The MADS-box transcription factor FaCMB1 acts as a negative regulator of ripening, with both its transcript and protein levels decreasing during fruit development, a process enhanced by ABA. Functional manipulation of FaCMB1 significantly affected ripening and ABA content.

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Literature Sharing | Overexpression of soybean flavonoid 3′-hydroxylase enhances plant salt tolerance by promoting ascorbic acid biosynthesis

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