INTRODUCTION: The superhydrophobic structures on plant leaves are crucial for self-cleaning and anti-microbial functions, acting as both a core functional mechanism and a biomimetic blueprint for advanced materials design. Ferns, as the earliest vascular plants that transitioned from aquatic to terrestrial habitats, have evolved a remarkable diversity of these water-repellent structures, offering unique insights into their evolutionary adaptations. However, systematic studies on the hydrophobic mechanisms and adaptive significance of ferns remain relatively scarce.

 

RATIONALE: To elucidate the structural basis, ecological relevance, and evolutionary trajectory of superhydrophobicity in ferns, we combined literature review and preliminary observations to select 25 fern species for detailed analysis of leaf epidermal micromorphology. Among these, 20 species exhibiting notable hydrophobic properties were further investigated to quantify their surface wettability and correlate microstructure with function. We integrated habitat preference data and phylogenetig reconstructions based on chloroplast genomes to examine the potential influence of environmental factors and evolutionary history on the development of hydrophobic traits.

 

RESULTS: Our investigations revealed that leaf hydrophobicity in ferns is closely associated with surface microstructure. Most hydrophobic species displayed hierarchical structures comprising polygonal protrusions and epicuticular wax crystals, with a contact angle exceeding 130° serving as a reliable indicator of significant hydrophobicity. Habitat correlation analysis indicated that species endemic to humid or water-rich environments exhibited more pronounced hydrophobic properties. Furthermore, phylogenetic reconstruction demonstrated that superhydrophobicity evolved independently multiple times during fern diversification, suggesting convergent adaptation to specific ecological conditions.

 

CONCLUSION: Leaf hydrophobicity in ferns correlates directly with surface microstructure complexity, exhibiting unique multi-scale hierarchical organizations. Notably, a contact angle exceeding 130° already induces distinct hydrophobicity in ferns—lower than the conventional 140° threshold for superhydrophobicity. Species from humid habitats show enhanced water repellency, supporting superhydrophobicity as an evolutionary adaptation to environmental constraints. These findings advance the theoretical framework for plant surface hydrophobicity and provide biomimetic design principles for functional materials. Future studies should examine wax chemistry, ecological factors, and soral structures to further elucidate hydrophobic mechanisms.

 

An overview of hydrophobicity in ferns: SEM observation and contact angle measurement were conducted on the leaf epidermis of ferns, revealing that superhydrophobic characteristics have independently evolved multiple times in this plant group.

 

CRISPR/Cas9 technology, as an efficient genome editing tool, has gained increasingly widespread application in the life sciences. Incorporating this technology into university experimental teaching enables students to deepen their understanding of cutting-edge research advances while enhancing their innovative research capabilities and practical skills. In this paper, we have designed an innovative teaching experiment, “Using CRISPR/Cas9 technology to study the effect of knocking out rice SSA1(seedling stage albino1)gene on phenotype”. This experiment helps students to deepen their understanding of the principles and experimental procedures of CRISPR/Cas9 technology and to recognize the important role played by theSSA1 gene in rice growth and development. At the same time, the experiment can improve students' ability of experimental operation and rigorous scientific research thinking, and cultivate students' exploratory skills and innovative spirit.

Research on the proteomics of bryophytes is of great significance for revealing the molecular mechanisms of plant adaptation to terrestrial environments. As one of the earliest plants to colonize land, bryophytes provide a unique model system for studying how plants adapt to changes in terrestrial environments. Through proteomic approaches, we can gain in-depth insights into the molecular mechanisms underlying stress responses, growth and development, and evolutionary processes in bryophytes. This is of considerable value for elucidating the evolutionary history and mechanisms of plant adaptability. Furthermore, research on bryophyte proteomics offers new perspectives and tools for plant science, aiding our understanding of plant responses and adaptive strategies to environmental changes. This review summarizes recent advances in bryophyte proteomics, focusing on abiotic stress responses, growth and developmental regulation, protein modifications, organelle functions, metabolic processes, and evolutionary adaptations. It further highlights the significance of these findings in understanding plant adaptation mechanisms.

INTRODUCTION: Ilex cornuta ‘Fortunei’ is widely used in landscape gardening and is a common sculptured tree species. An efficient and simple protoplast preparation and separation system is of great value for breeding and gene research of woody plants. However, there are no relevant reports on the protoplast isolation and transformation system of I. cornuta ‘Fortunei’ so far, making it necessary to carry out this study. 

RATIONALE: Woody plants face challenges in protoplast isolation and transformation due to characteristics such as perennial stem lignification and leaf leatheriness. Differences in cell wall composition and tissue structure among species require optimization of protoplast systems for specific species. To establish an efficient protoplast preparation and transient transformation system for I. cornuta ‘Fortunei’, orthogonal experiments and gradient experiments were used to explore factors affecting protoplast separation, and PEG-mediated method was applied for transient transformation. 

RESULTS: Compared with other enzyme combinations, the orthogonal test showed that the optimal enzyme concentration combination was 2.5% (W/V) cellulase, 1.0% (W/V) macerozyme, 0.5% (W/V) pectinase, and 0.4% (W/V) snailase. After 3.5 h of enzymolysis, the protoplast yield was the highest (1.18×107∙g–1 FW) with a viability of 68.30%. The suitable mannitol concentration for osmotic regulation was 0.30 mol·L–1, and dark pretreatment of leaves for 12 h improved the effect. For transient transformation, 40% (W/V) PEG-4000 mediated the transformation of pORE_R4-IcFPS2-GFP plasmid into protoplasts, and green fluorescent signals in the nucleus overlapping with DAPI blue fluorescent signals were observed under a laser confocal microscope, indicating successful transformation. 

CONCLUSION: In conclusion, this study successfully established an efficient protoplast isolation system for I. cornuta ‘Fortunei’ with optimal conditions including specific enzyme combination, enzymolysis time, osmotic regulator, and dark pretreatment. The PEG-mediated transient transformation system was also successfully constructed. These results provide a foundation for future protoplast fusion breeding and gene function research of I. cornuta.

Clathrin-mediated endocytosis (CME) is one of the primary pathways for the internalization of plasma membrane proteins in animal and plant cells. Clathrin and its adaptor protein complexes play crucial roles in plant morphogenesis, root gravitropic growth, halotropism response, immune signaling and nutrient elements. This review begins with an overview of clathrin and its mediated endocytosis in animal cells, briefly summarizing the composition, localization, and biological functions of plant clathrin and its adaptor protein complexes. It focuses on the regulatory mechanisms and biological roles of the CME pathway in plants, while also summarizing and prospecting the critical contributions of CME to plant growth, development, and responses to biotic and abiotic stresses. Furthermore, future research directions are discussed, providing valuable references for ‌in-depth exploration‌ of the ‌molecular mechanisms‌ underlying the plant CME pathway.

INTRODUCTION: The global area of saline-alkali land is approximately 932 million hectares. The problem of soil salinization caused by seawater intrusion in china's coastal areas is intensifying. Salt stress is an important abiotic stress that affects plant growth and development. Throughout the long-term evolutionary process, plants have developed complex adaptation mechanisms to cope with stress, with gene expression regulation playing a central role. Among the numerous salt stress-responsive genes, the DEAD-box RNA helicase gene family has garnered significant attention due to its crucial roles in plant growth, development, and abiotic stress responses. Studies have confirmed that members of this family are involved in the regulation of salt stress in a variety of plants. Therefore, the DEAD-box RNA helicase gene family is an important object for studying the molecular mechanism of plant salt stress response.

RATIONALE: Scaevola taccada is a semi-mangrove plant widely distributed in tropical and subtropical coastal areas. It exhibits excellent salt tolerance, drought resistance, and resilience to wind and waves. This plant holds significant ecological value for coastal sand stabilization and ecological restoration. Although the DEAD-box RNA helicase gene has been extensively studied in model plants, the genome-wide identification, chromosomal distribution, and salt stress response mechanism of this gene family remain unclear in S. taccada. Therefore, based on the genome data of S. taccada, the members of the DEAD-box gene family (StRHs) were systematically identified, and the tissue expression specificity of the StRH10 gene and its response pattern under salt stress were analyzed, aiming to provide a preliminary scientific basis for the molecular breeding of high-salt tolerant S. taccada seedlings. 

RESULTS: Fifty StRH genes were identified from the genome of S. taccada. Most StRH proteins contain 10 conserved motifs, while a few contain 8 or 9. The StRH proteins comprise 18 conserved domains, primarily SrmB and DEAD. Seven StRH genes lack introns. The promoter region of the StRH genes is rich in cis-acting elements associated with the stress response. S. taccada and Arabidopsis thaliana have 20 pairs of collinear genes, and S. taccada and Helianthus annuus have 39 pairs of collinear genes. StRH10 protein is most closely related to the chloroplast-localized DEAD-box RNA helicase AtRH03 protein in A. thaliana. The subcellular localization prediction indicated that the StRH10 protein is localized to the chloroplasts. StRH10 gene was the most expressed in leaves. Under salt stress treatment, the expression level of the StRH10 gene did not show a significant difference with increasing NaCl concentration but was significantly up-regulated when the NaCl concentration reached 400 mmol·L-1. 

CONCLUSION: The 50 members of the DEAD-box gene family identified in the genome of S. taccada in this study were unevenly distributed across the chromosomes. Most StRH proteins contain 10 conserved motifs and are clustered within the same evolutionary branch. The promoter region of StRH genes is rich in stress-responsive cis-acting elements such as ABRE, TC-rich repeats, MBS, LTR, WUN-motif and ARE. Collinearity analysis showed that there were 39 pairs of collinearity genes in S. taccada and H. annuus. The StRH10 gene, which encodes a protein predicted to be located in the chloroplast, exhibits high expression in leaves.Under salt stress, the expression of the StRH10 gene was significantly up-regulated only when the NaCl concentration reached 400 mmol·L-1, indicating that this gene responds to salt stress with a certain concentration threshold.

(A) DEAD-box gene family Cis-acting elements in the promoter region; (B) Phylogenetic tree of DEAD-box gene family members in S. taccada and A. thaliana (The red pentagram indicates the RH gene of S. taccada); (C) StRH10 expression levels in different organizations; (D) StRH10 expression levels under salt stress

INTRODUCTION: Tomato wilt disease, caused by Fusarium oxysporum f. sp. lycopersici (Fol), is a soil borne fungal disease that causes significant economic losses to global tomato agricultural production. The molecular mechanism of pathogen sRNA cross-border regulation of host DNA methylation mediated tomato resistance to wilt disease is still unclear. Here, we illustrate that SlyFRG5 methylation level activated by Fol-milR1 mediates immune response against tomato wilt disease. 

RATIONALE: The latest research shows that sRNA derived from pathogens can serve as novel effectors for cross-border participation in host-pathogen interactions. Our previous research documented that Fol-milR1 was transferred horizontally into host cells as an effector. Fol-milR1 repressed the expression of a resistant gene SlyFRG4 in tomato, as well as binded to SlyAGO4a leading to interfere with the host immune system to achieve successful infection. Therefore, this research further determines whether transmission of?Fol-milR1 into tomato causes DNA methylation in the tomato genome and elucidate other aspects of the molecular mechanism involving the action of Fol-milR1.

RESULTS: The results indicate that SlyAGO4a negatively participates in the resistance to tomato wilt disease. The methylation level of Solyc02g081370 (SlyFRG5) was identified to be directly associated with Fol-milR1 using whole genome methylation sequencing and molecular biology methods. SlyFRG5 loss-of-function alleles created using CRISPR/Cas9 in resistant cultivar tomato Motelle exhibited enhanced disease susceptibility to Fol, further supporting the idea that SlyFRG4 is essential for tomato wilt disease resistance. SlyFRG5 contributed wilt disease resistance by regulating the accumulation of host ROS in tomato. 

CONCLUSION: Combining previous research, we concluded that,during the infection Fol, Fol-milR1 was transferred into host cells to hijack SlyAGO4a which leads to methylation a Fusarium disease resistant gene SlyFRG5. This study analyzed how the Fol-milR1-SlyAGO4a-SlyFRG5 functional module mediated tomato resistance to wilt disease, which will provide a new ideas for exploring the cultivation and quality improvement of tomato resistance to Fusarium disease.

The Fol-milR1-SlyAGO4a-SlyFRG5 functional module mediated tomato resistance to wilt disease. (A) Distribution of methylation type in the CDS (Partial region) region of SlyFRG5; (B) The transcription level of SlyFRG5 at different time points after pathogen infection in MM; (C) Leaf DAB (H₂O₂) and NBT (•O₂⁻) staining (bar=1 cm).

INTRODUCTION: The distribution and concentration of nutrient elements within fruits directly influence their yield and quality. Conventional analytical methods primarily rely on digestion to quantify nutrient content in different fruit tissues. However, as fruits comprise multiple tissue structures, basic quantification techniques fail to precisely resolve nutrient distribution across these microstructural compartments. Consequently, establishing an in-situ method enabling visualization of spatial nutrient concentration profiles in fruit tissues is imperative.

RATIONALE: Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS), as an advanced micro-analytical technique, enables effective visualization of in situ elemental distribution within plant tissues. LA-ICP-MS uses a femtosecond laser (wave-length 343nm) to scan the fruit sample and the standard sample according to the line of the laser beam during the measurement of the femtosecond laser (wave-length 343nm). The burned sample material is sent to the ICP with the carrier gas Ar, and the ionic strength of P³¹, K³⁹, Ca⁴⁴, Mg²⁴, Fe⁵⁷, Mn⁵⁵, Cu⁶³, Zn⁶⁶, S³⁴, Al²⁷, Cr⁵², Mo⁹⁵, Pb²⁰⁸, Sr⁸⁸ and Rb⁸⁵ in different parts of the fruit is detected. Visualize the nutrient distribution between different tissues.

RESULTS: Through LA-ICP-MS analysis, we performed in situ, spatially-resolved and semi-quantitative analysis of nutritional element distributions across distinct tissue structures in pomelo fruit during maturation. Significant tissue-specific heterogeneity was observed in elemental distributions among fruit compartments. Three structures demonstrated consistent nutrient enrichment: cuticle of flavedo, vascular bundle-like structures in albedo, epidermal cells of juice sacs. The concentrations of potassium, nitrogen, calcium, magnesium and phosphorus were higher in the fruit of Wendan pomelo, which were mainly concentrated in the cuticle of flavedo, vascular bundle tissue of albedo and epidermal tissue of juice sac. Among them, the relative average concentration of calcium in the yellow cortex was as high as 516.21 ± 387.40 mg ·L^-1, and the relative average concentration of zinc in the cuticle of the yellow cortex was 410.22 ± 279.77 mg ·L^-1. The relative concentration of the two in the epidermal cells of the juice sac was the highest, suggesting that calcium and zinc play an important nutritional role in the fruit of Wendan pomelo.

CONCLUSION: This study established a novel LA-ICP-MS methodology for in-situ, visual and semi-quantitative analysis for nutrient distribution in different tissue structures of fruits, was used to establish the spatial relationship of nutrient distribution, which filled the gap in the visualization of fruit nutrient distribution. The relative concentration of different elements in each tissue was analyzed, which provided scientific theory for the subsequent guidance of Wendan pomelo farming measures.

LA-ICP-MS in situ visualization and semi-quantitative calcium concentration in various parts of fruit. The in situ distribution of calcium element in the flavedo, albedo and juice sac and the concentration of tissue structure were analyzed.

Pre-harvest sprouting (PHS) refers to the phenomenon in which cereal crop seeds germinate on the mother plant under humidity conditions prior to harvest, representing a major constraint on crop yield and seed production. In recent years, with the increasing frequency of extreme climate, PHS have become more common, resulting in substantial economic losses in agriculture. The PHS process primarily involves two key physiological stages: the release of seed dormancy and the initiation of germination. This paper provides a systematic review of research advances in the mechanisms underlying rice seed dormancy and germination over the past two decades. Furthermore, a chromosomal distribution map of cloned rice genes associated with seed dormancy and germination has been constructed, offering precise genetic information for PHS resistance breeding in rice. Additionally, this paper summarizes the regulatory roles of epigenetic factors and non-coding RNAs in rice seed dormancy and germination. It further discusses the molecular mechanisms through which environmental factors such as light, temperature, and water influence these processes, and proposes potential research directions concerning the interaction between environmental cues and plant hormones in the regulation of PHS. Drawing on recent application cases of PHS-resistant genes, this study also outlines future prospects for rice PHS resistance breeding strategies.

ATG8h and ATG8i are an unique sub-group of the nine ATG8 proteins encoded in Arabidopsis genome that do not need ATG4 protease for cleavage to expose their C-terminal Gly residue for lipidation and subsequent recruitment to the autophagosomal membranes. We previously showed that clathrin light chain 2 (CLC2) participates autophagy through interacting ATG8h and ATG8i and knocking out ATG8h and ATG8i simultaneously compromised autophagy, and as a result, enhanced the resistance to a biotrophic fungal pathogen. In this study, we further investigate the roles of ATG8i in disease resistance by over-expressing ATG8i. Our results showed that over-expression of ATG8i enhanced the resistance to biotrophic bacterial and fungal pathogens but compromised the resistance to a toxin secreted from a necrotropic fungal pathogen. The enhanced resistance to the biotrophic pathogens were correlated with the elevated levels of both callos deposition and H2O2 induced by Golovinomyces cichoracearum infection.

INTRODUCTION: Waxes are protective substances found on the surface of plants, especially on fruits and leaves, which help prevent non-stomatal water loss from plant tissues. Lycium barbarum is a characteristic economic forest species of Ningxia. Its fruit is a berry with a high water content, and the surface of the fruit skin is covered with wax, which hinders the normal loss of moisture during the drying process. Alkanes are the main components of the wax on L. barbarum, and the CER3 gene is a key gene involved in alkane biosynthesis. Therefore, an in-depth study of the function of CER3 in L. barbarum provides a theoretical basis for revealing the metabolic synthesis of wax in goji fruits and the molecular mechanism of LbCER3. 

RATIONALE: Research on the waxy coating of L. barbarum fruit skin has mainly focused on the structure and composition of the epicuticular wax, with relatively little attention given to its molecular mechanisms. This studyusing the main cultivated varieties of L. barbarum, Ningqi No.1 and Ningqi No.5, as test materials, this study employed methods such as GC-MS, transcriptomics, and molecular biology to investigate the changes in the content and composition of cuticular waxes on the outer and inner layers of the fruit at different developmental stages. It also involved the screening, cloning, and functional validation of key genes related to fruit wax. 

 RESULTS: During the development and maturation of the fruit, the content of inner and outer cuticular waxes in Ningqi No.1 and Ningqi No.5 gradually decreased, with the total wax content in Ningqi No.1 being significantly higher than that in Ningqi No.5. The outer and inner wax components of both varieties were the same, primarily consisting of alkanes, esters, and acids. Transcriptomic analysis and weighted gene co-expression network analysis (WGCNA) identified LbCER3 as a key gene. The full-length CDS of the LbCER3 gene is 1 884 bp, encoding 628 amino acids, and contains a C-terminal WAX2 domain (Wax2_C) and a fatty acid hydroxylase domain (FA_hydroxylase). An LbCER3 overexpression vector was constructed and successfully transformed into tomato, resulting in a significant reduction in chlorophyll leaching rate in the T1 generation transgenic lines, along with a notable increase in leaf wax content and significant increases in the wax components of alkanes and acids. 

CONCLUSION: These findings indicate that LbCER3 is a key gene in the synthesis of wax in L. barbarum.

INTRODUCTION: Ajania alabasica is a wild species of the genus Ajania in the Asteraceae family, characterized by its remarkable drought resistance and ecological adaptability, making it a valuable genetic resource for breeding. Nevertheless, the absence of an effective in vitro regeneration system has impeded the advancement of rapid propagation and genetic transformation studies on this species. 

RATIONALE: The utilisation of appropriate explants constitutes a critical strategy for plant regeneration. The core of this methodology lies in optimizing the combinations of plant growth regulators (PGRs) and additives to balance differentiation and physiological disorders such as hyperhydricity. This study was conceived with the objective of systematically determining the effects of different explant types (leaves vs. stem segments) and PGRs, with a view to establishing a robust regeneration protocol for A. alabasica. 

RESULTS: This study found that leaf explants exhibited significantly higher regeneration potential than stem segments. The results demonstrated that the optimal medium for callus induction and adventitious bud differentiation from leaves was MS basal medium supplemented with 1.0 mg∙L^–1 6-BA and 1.0 mg∙L^–1 NAA. Following the culture period, this resulted in a callus induction rate of 100% (at 15 days) and an adventitious bud differentiation rate of 92.22% with a multiplication coefficient of 6.93 (at 30 days). To address hyperhydricity, the optimal proliferation medium was identified as containing 0.3 mg∙L^–1 6-BA and 0.05 mg∙L^–1 NAA, with 5 µmol∙L^–1 AgNO3 proving effective for alleviation. For the rooting phase, the optimal medium was identified as 1/2MS medium supplemented with 0.2 mg∙L^–1 NAA, which resulted in a rooting rate of 100% and a transplant survival rate of 88%. 

CONCLUSION: This study successfully established for the first time a highly efficient and stable regeneration system for A. alabasica using leaf explants. The developed system, characterized by its high differentiation and survival rates, provides crucial technical support for subsequent genetic transformation and gene function analysis.