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Insect pollination is critical for fruit production, particularly for pears, most of which have high self-incompatibility and are less preferred by pollinators than other fruit trees. Research has focused on fruit traits, and less is known about how floral traits may influence pollination and fruit production. Hypothesising that pollination-related traits differ between Asian and European pears, which exhibit large variation in pollination and distinct domestication processes, we conducted a systematic review of pear pollination studies published globally from 1922 to 2025. Research on pollination has increased rapidly over the last 25 years. Compared to European pears, we found that Asian pears are more pollinator-dependent and exhibit different floral traits: lower nectar sugar concentration and lower relative content of attractive floral scents, but higher pollen production and a higher relative content of N-containing floral scents. Although Asian pear flowers attract a similar number of insect pollinator taxa as European pears, current Asian pear production still relies mainly on artificial pollination. Moreover, various artificial pollination techniques and pollination management practices, including introducing indigenous Asian honey bees, which outperformed in collecting pear pollen, had uncertain effects on pollination and fruit production, as less attention was paid to understanding the nature of plant-pollinator interactions. Further studies are needed to investigate how the domestication process and the co-adaptations of Asian honey bees influence the pollination-related floral traits of pears. This may provide valuable insights into enhancing floral attractiveness, improving pollination efficiency, and increasing fruit production in monoculture Asian pears.

Tulips are one of the best-known geophytes, but their taxonomy remains convoluted and their evolutionary history poorly understood. Here, we used plastid genomes to identify some issues with current classification, understand the diversification history of the genus, and identify potential species linked to historical cultivation. We gathered a large number of tulip specimens from living collections, herbaria, and online sources and through extensive fieldwork to infer maximum likelihood phylogenies of the plastid genomes of Tulipa (Liliaceae), representing ~86% of accepted species alongside a range of occasionally accepted synonyms. We used BEAST and secondary calibration points to date the tree, before assessing the biogeographical history of the genus using BioGeoBEARS. Based on our results, we described a fifth subgenus, Eduardoregelia, and identified a range of species where our results do not align with current taxonomy. Our data suggests the genus diverged from a clade containing Amana and Erythronium around 32.5 million years ago (Mya), with the most recent common ancestor existing around 22.8 Mya in the broader Central Asia region, a period that saw rapid uplift of mountain ranges and associated aridification in the region. The genus primarily diversified in Central Asia with rapid radiations within the last 10 My, possibly as a response to further orogenesis. Several migrations outside of the ancestral region have occurred, primarily via the steppes of Kazakhstan and Russia and along the Caucasus and Iranian and Anatolian Mountains to the eastern Mediterranean region. We also provide evidence that the maternal lineage of the garden tulip involved several species including T. scardica and T. suaveolens. Many tulip species present limited diagnostic characters, making their taxonomy difficult, compounded by their long use in horticulture and widespread hybridization, especially in gardens. We provide an up-to-date phylogenetic framework for this genus that may help resolve many taxonomic issues in the future, while highlighting further work to be done. We reinforce the importance of Central Asia in the evolutionary history of this genus and provide an important foundation for further study of Tulipa and more effective conservation of wild tulip species. ПРЕДПОСЫЛКИ ИССЛЕДОВАНИЯ: Тюльпаны—одни из самых известных геофитов, но их таксономия остается запутанной, а их эволюционная история—малоизученной. В этой работе, мы используем изучение пластидного генома для оценки современной таксономии, понимание характера диверсификации рода, и выявление потенциальных видов, являющихся предками культиваров. МЕТОДЫ: Мы собрали большое количество образцов тюльпанов из живых коллекций, гербариев, онлайн‐источников, и в ходе обширной полевой работы, для того чтобы составить филогению максимального соответствия пластидных геномов рода Tulipa (Liliaceae), представляющих ~86% принятых видов, наряду с видами, иногда принимаемыми в качестве синонимов. Сначала мы использовали программу BEAST и вторичные калибровочные точки для датирования филогенетического дерева, а затем оценили биогеографическую историю рода с помощью программы BioGeoBEARS. ОСНОВНЫЕ РЕЗУЛЬТАТЫ: На основании полученных результатов мы описываем пятый подрод‐ Eduardoregelia и выявляем ряд видов, для которых наши данные не согласуются с современной таксономией рода Tulipa. Наши данные показывают, что род отделился от клады, содержащей Amana и Erythronium, около 32,5 млн. лет назад, при этом наиболее недавний общий предок существовал около 22,8 млн. лет назад в регионе в широком смысле Центральной Азии—в период, характеризовавшийся быстрым поднятием горных систем и связанной с этим аридизации климата в регионе. Диверсификация рода Tulipa преимущественно происходила в Центральной Азии, а быстрая эволюционная радиация имела место в течение последних десяти миллионов лет как результат дальнейшего орогенеза. Несколько миграций за пределы исходного ареала происходили в основном через степи Казахстана и России, а также вдоль гор Кавказа, Ирана и Анатолии в восточное Средиземноморье. Мы также получили данные, свидетельствующие о том, что материнская линия садового тюльпана включала несколько видов, в том числе T. scardica и T. suaveolens. ВЫВОДЫ: Многие виды тюльпанов имеют ограниченное число характерных признаков, что затрудняет их таксономию, что усугубляется их длительным использованием в садоводстве и широким распространением гибридизации, особенно в садах и огородах. Мы представляем современную филогенетическую систематику этого рода, которая поможет решить многие таксономические проблемы, при этом подчеркивая необходимость дальнейшей работы. Мы подтверждаем важность Центральной Азии в эволюционной истории этого рода и закладываем основу для дальнейшего изучения рода Tulipa и более эффективного сохранения диких видов тюльпанов.

Flowering is a crucial process in the growth and development of plants, playing an essential role in their life cycles. Thus, research into the regulation of flowering time holds significant importance. While CmBBX7 has been identified as a flowering activator regulated by photoperiod, the molecular mechanisms underlying its regulation at the protein level remain unclear. In this study, we utilized yeast two-hybrid screening to identify two proteins that interact with CmBBX7: the phosphatase CmPP6 and the E3 ubiquitin ligase CmMIEL1. Ubiquitination assays confirmed that CmMIEL1 promotes the degradation of CmBBX7, thereby affecting its protein stability and influencing flowering. Genetic evidence indicated that both CmPP6 and CmMIEL1 delay flowering. By interacting with CmBBX7, they indirectly repress the expression of CmFTL1, which is a key flowering gene in chrysanthemum. Our findings provide preliminary evidence that ubiquitin modification affects the stability of the CmBBX7 protein and thereby regulates flowering in chrysanthemum.

Soil salinity poses a major threat to global food security, impairing plant physiology through ionic toxicity, osmotic stress, and oxidative damage. However, conventional breeding and genetic engineering approaches face limitations due to the complexity of stress responses and regulatory hurdles. Nanotechnology, through the unique properties of nanomaterials (NMs), offers a promising alternative. This review synthesizes recent advances in nanoscale strategies for early salt stress sensing, the regulation of ion homeostasis, and reactive oxygen species scavenging. It further examines how NMs enhance nutrient use efficiency under saline conditions and discusses emerging applications in microbiome engineering and nontransgenic genetic improvements. The integration of machine learning to optimize NM design and application is also discussed. By synthesizing insights across these disciplines, this review provides a holistic framework for developing next-generation, nanotechnology-driven solutions to mitigate salinity stress, bridging the fundamental research with field-scale application for sustainable agriculture.

Cluster bean (Cyamopsis tetragonoloba L.) is usually grown for its high galactomannan content as an industrial crop. Young edible cluster bean pods are also eaten as a nutritious vegetable in many areas. There is little research on the effects of drought on pod yield or the nutritional content of edible cluster bean pods, even with the growing importance of producing vegetable cluster bean in dry and semiarid regions. In this study, the effect of silicon nanoparticles (Si NPs) and methyl jasmonate (MeJA) applied concurrently on the growth performance, pod productivity, and nutritional quality of vegetable cluster bean was evaluated at three irrigation rates. A three-factor experiment was done in a completely randomized block design with three replications under field conditions, with three irrigation rates (100% field capacity, 50% field capacity, and 25% field capacity), three Si NP levels (0, 50, and 100 ppm), and three MeJA levels (0, 20, and 40 μmol L-1). Drought stress (with irrigation at 25% FC) significantly reduced the growth of the plant, pod yield, crude protein, crude fat, fiber, dry weight, and mineral composition of the edible cluster bean pods. Pod weight reduction of approximately 50% occurred when they were irrigated at 25% FC, compared to pod yield from well-watered plants. The addition of Si NPs and MeJA, when applied together, was able to reduce the adverse effects of low and moderate drought on pod yield and improve the productivity and nutritional quality of the edible cluster bean pods. The combined application of 100-ppm Si-NPs and 40-μmol L-1 MeJA was the most effective treatment for improving the productivity and nutritional quality of vegetable cluster bean under deficit irrigation. These findings suggest that this combination for cluster bean cultivation under moderate drought conditions after economic feasibility and field-scale validation studies.

Bergamotene, a key sesquiterpene compound, is crucial for defining the fruity aroma. However, the precise regulatory mechanisms governing its biosynthesis have been largely elusive. In this study, we elucidated that the DOF transcription factor LcDOF5.8 plays a central role in the biosynthesis of α-bergamotene by directly enhancing the expression of the terpene synthase gene LcTPSbms in litchi fruit. Our findings revealed that the concentration of α-bergamotene varies significantly among four distinct litchi cultivars: 'Guanyinlv', 'Bingli', 'Guiwei' and 'Nuomici'. Notably, a strong correlation was observed between the expression level of LcTPSbms and the content level of α-bergamotene across these cultivars. Furthermore, both in vitro and in vivo catalytic assays confirmed that LcTPSbms is capable of catalyzing the synthesis of α-bergamotene. Importantly, electrophoretic mobility shift assays (EMSA) and Dual-LUC assays demonstrated that LcTPSbms is positively regulated by LcDOF5.8. Silencing LcDOF5.8 in litchi aril resulted in decreased LcTPSbms expression, whereas transient overexpression of LcDOF5.8 led to a significant upregulation of LcTPSbms and a concomitant increase in α-bergamotene biosynthesis. In summary, our research uncovers a regulatory module involving LcDOF5.8 and LcTPSbms that plays a critical role in the biosynthesis of α-bergamotene, offering valuable insights into the molecular mechanisms underlying the fruity aroma of litchi fruit.

The growing demand for natural, sustainable food preservation strategies has intensified research into plant-derived antimicrobial proteins with multifunctional bioactivities. This study reports the functional characterisation of TdGASA2, a cysteine-rich protein of the Snakin/GASA family isolated from Triticum turgidum ssp. durum, as a candidate bioactive ingredient for antimicrobial food packaging. TdGASA2 displayed broad-spectrum antibacterial activity against all tested strains, with the lowest minimal inhibitory concentration (MIC) values observed against Listeria monocytogenes and Pseudomonas aeruginosa (10.50 and 11.25 μg/mL, respectively), and retained a mean of 77.8% of its native antibacterial potency after autoclaving (121 °C, 20 min), reflecting the thermal resilience of its disulfide-stabilised Snakin/GASA scaffold. The protein additionally exhibited concentration-dependent α-amylase inhibition (IC₅₀ = 72.4 ± 0.31 μg/mL) and enhanced bactericidal inactivation of L. monocytogenes under oxidative stress. Incorporation of TdGASA2 into chitosan-based films improved tensile strength and reduced water vapor permeability and water solubility, consistent with protein-polysaccharide interactions inferred from established structure-property relationships in the literature; the films also maintained a high rate of soil-burial mass loss and macro-disintegration (>92% after 10 days), which reflects overall gravimetric mass reduction rather than confirmed biodegradation in the absence of an abiotic soil control. In a 10-day refrigerated chicken breast storage trial, TdGASA2-enriched films (2× MIC against L. monocytogenes) reduced total aerobic microbial growth by approximately 2.4 log CFU/g and attenuated lipid and protein oxidation relative to the unfunctionalised control, corresponding to an exploratory, Random Forest-predicted shelf-life extension of 3-4 days. Collectively, these results identify TdGASA2 as a promising, multifunctional bioactive candidate for biodegradable active food packaging; direct spectroscopic confirmation of the proposed protein-polymer interactions, migration behaviour in regulatory food simulants, toxicological and allergenicity profiling, sensory impact, and formal food-contact regulatory assessment remain to be established before its suitability for practical or commercial application can be confirmed.

The widespread use of commercial active dry yeasts (ADYs) in winemaking is well-established. Its successful implantation during industrial fermentation varies significantly according to wineries and regions. This inconsistency frequently causes undesirable quality fluctuations and deterioration. However, their interactions with native microbiota has not been fully elucidated. This study deciphers microbiota and metabolite profiles during industrial Cabernet Sauvignon wine fermentations with different implantation percentage of commercial active dry yeasts (ADYs) through multi-omics. Strain-level implantation percentages of ADYs were found to differ significantly among the studied wineries. High-throughput sequencing further showed differences in fungal and bacterial community structures between different dominance of ADYs. Metabolome analysis showed that 291 non-volatile metabolites represented the variations between high (80%) and low (50%) implantation percentage of ADYs. These metabolites were mainly involved in 20 different pathways, such as amino acids synthesis, cutin, suberine and wax biosynthesis. Also, significant differences in aroma profiles were observed between high and low dominance samples. Correlation analysis among microorganisms, non-volatile and volatile profiles revealed the effect of the implantation of ADYs on both microbial communities interactions and the resulting metabolite profiles, and highlighted the pivotal role of key microorganisms in shaping characteristic aromas. This study enhances understanding of how ADYs implantations affect microbial communities and metabolite profiles in wines, providing insights into a microbial "terroir" of relevance to wine character and wine quality.

Agrobacterium-mediated transformation remains challenging for economically important citrus crops because of low transformation and regeneration efficiencies. REGENERATION FACTOR1 (REF1), a plant elicitor peptide, is a local wound signal perceived by PROPEP RECEPTOR-LIKE KINASE 1 (PORK1) and transduced via WOUND-INDUCED DEDIFFERENTIATION 1 (WIND1). Exogenous REF1 peptide application has recently been reported to improve transformation and regeneration of herbaceous plants including tomato, which was explored on Agrobacterium-mediated citrus transformation in this study. We identified REF1, PORK1, and WIND1 orthologs in sweet orange (Citrus sinensis), which encodes two REF1 isoforms, CsREF1-1 and CsREF1-2. REF1 of tomato (SlREF1) and the two CsREF1 isoforms of different concentrations were tested for their effect on Agrobacterium-mediated transformation of citrus via exogenous application. CsREF1-1 at 10 or 100 nM and CsREF1-2 at 100 nM significantly increased GFP-positive transgenic callus formation, whereas SlREF1 had no significant effect. In addition, CsREF1-1, CsREF1-2, and SlREF1 reduced shoot regeneration. To overcome this trade-off, we exposed epicotyl explants to 100 nM CsREF1-2 only during the first month and then transferred them to REF1-free regeneration medium, yielding 4.1-fold more GFP-positive shoots than the untreated control. Such a staged application of CsREF1-2 also improved GFP-positive shoot recovery in Carrizo citrange by 3.3-fold. Reverse transcription-quantitative PCR (RT-qPCR) showed that CsREF1-2 induced CsWIND1 and CsESR1, which are critical for callus formation and shoot regeneration, but continuous CsREF1-2 exposure also suppressed shoot/meristem regulators, which was mitigated by removal of CsREF1-2 in the later stage. AlphaFold3 and PRODIGY predictions indicated the highest binding affinity for CsREF1-2 with CsPORK1, consistent with its superior activity. Overall, both CsREF1 peptides enhance citrus transformation but suppress regeneration, and staged exogenous application of CsREF1 peptides improves genetic improvement of recalcitrant perennial crops such as citrus, which decouples early dedifferentiation from later organogenesis, substantially expanding the toolbox for high-throughput functional genomics in citrus.

Improving glucosinolate (GSL) profiles in rapeseed (Brassica napus)-high in leaves for pathogen resistance but low in seeds for meal quality-is a key breeding goal, yet its genetic basis remains unclear. Here, we present a chromosome-level genome assembly for ZY821, an elite high-GSL variety, generated using long-read sequencing and Hi-C scaffolding. Comparative analysis with the low-GSL variety ZS11 identified three major homoeologous exchange (HE) events and extensive structural variation. Notably, an A09-C09 HE event replaced the low-expression BnaC09.MYB28 allele with the high-expression BnaA09.MYB28 allele, resulting in elevated MYB28s expression and thereby increased GSL accumulation in ZY821, whereas a deletion of BnaA09.MYB28 in ZS11 significantly reduced the expression of multiple putative downstream targets in the GSL biosynthesis pathway, leading to a reduction in GSL content. This mechanism was supported by population-level HE analysis and time-course transcriptomes across 116 RNA-Seq samples. Furthermore, joint differential expression and co-expression network analyses uncovered several novel candidate genes implicated in GSL metabolism. Collectively, our study provides new mechanistic insights into the genetic control of GSL accumulation, with significant implications for breeding optimized GSL profiles.

Cadmium (Cd), a highly toxic heavy metal, poses significant threats to agricultural productivity and human health by accumulating in the food chain. Selenium (Se), an essential micronutrient, has shown promise in mitigating Cd toxicity in plants. However, the underlying molecular mechanisms remain largely unknown. This study elucidates how exogenous selenite (Na2SeO3) reduces Cd accumulation in the edible parts of flowering Chinese cabbage. Our results demonstrate that Se application (2.5 µM) dramatically mitigated Cd-induced growth inhibition, recovering shoot and root biomass to 96.3% and 90.7% of those of the control levels, respectively, while significantly decreasing Cd concentration in the shoots. Mechanistically, Se orchestrates a multi-layered defense strategy in the roots under Cd stress. Ultrastructural and physiological analyses revealed that, Se reduced the average net Cd2 + influx by 24.3%, increased root cell wall thickness by 40%, and significantly boosted cell wall lignification under Cd stress. Furthermore, Se promoted Cd compartmentalization by shifting Cd from metabolically sensitive organelles to the soluble fraction under Cd stress. At the molecular level, transcriptome analysis combined with Weighted Gene Co-expression Network Analysis (WGCNA) identified a potential regulatory module. The transcription factor bHLH39 acts as a key repressor, directly binding to the promoter of the lignin biosynthesis gene CAD5. Se co-application suppresses the Cd-induced expression of bHLH39, thereby de-repressing CAD5 expression to promote lignification and physically restrict Cd translocation. This study identified the bHLH39-CAD5 regulatory module as a potential mechanism underlying Se-mediated Cd detoxification, providing a strategy for improving low-Cd leafy vegetables crops to ensure food safety.

Drought and salt stress are significant environmental limitations that severely constrain plant growth and productivity, therefore, enhancing stress tolerance is a key goal in crop improvement. The plant-specific FCS-like zinc finger (FLZ) proteins have been identified as important regulators of stress adaptation. In this study, we conducted a genome-wide characterization of the FLZ gene family in apple and functionally characterized MdFLZ2. qRT-PCR analysis revealed that MdFLZ2 was differentially expressed across various tissues and transcriptionally induced by both drought and salt stress. Subcellular localization assays demonstrated that the MdFLZ2 protein is localized to both the nucleus and the cytoplasm. The overexpression of MdFLZ2 in apple calli, Arabidopsis and tomato conferred increased resistance to drought and salt stress. In addition, yeast two-hybrid (Y2H) assays confirmed that MdFLZ2 interacted with MdSnRK1.1, and similar interactions were also detected between other MdFLZ family members and MdSnRK1.1. Collectively, our findings suggest MdFLZ2 as a positive regulator of drought and salt tolerance and highlight its potential to serve as a genetic resource for abiotic stress improvement.

This study used multi-omics technologies to analyze aroma formation during Congou black tea fermentation. Volatile compounds were analyzed by headspace solid phase microextraction coupled with gas chromatography mass spectrometry using two columns of different polarity. Fermentation increased total volatile normalized peak area fivefold, with alcohols, aldehydes, and acids increasing over sevenfold. 29 differential metabolites were screened, including amino acid derived phenylacetaldehyde, phenylethanol, and 2-methylbutanal; fatty acid derived (E,E)-2,4-heptadienal, hexanal, and 1-hexanol; and isoprenoid derived linalool, geraniol, and beta ionone. Transcriptomic, proteomic, and enzyme analyses revealed that biosynthesis contributed to early accumulation of amino acid and isoprenoid derived aromas, whereas ortho quinone mediated Strecker degradation and free radical induced fatty acid auto oxidation dominated generation of amino and fatty acid derived aromas during middle and late fermentation. In conclusion, aroma formation during fermentation results from biosynthesis and non-enzymatic oxidation, with the latter possibly dominating amino and fatty acid derived aromas.

Solanum torvum, a superior vegetable grafting rootstock and medicinal Solanaceae plant, exhibits strong seed dormancy, which limits its commercial cultivation. Among various strategies explored to improve the germination rate of S. torvum, exogenous application of gibberellin (GA) has been shown to be effective. In this study, a GA concentration of 2.5 mM was established as the optimal for breaking dormancy in S. torvum seeds. Transcriptome analysis of dry, water-soaked, GA-soaked, and GA-induced germinated seeds was conducted to investigate the molecular mechanism of GA-mediated dormancy release. During the soaking period, GA application significantly induced transcriptome changes in processes including protein processing, translation, and peptide biosynthesis. Concurrently, GA treatment promoted plant hormone signal transduction, enhanced DNA-binding transcription factor activity, and activated monocarboxylic acid biosynthetic process, all of which facilitated seed water absorption. Furthermore, the differentially expressed genes (DEGs) induced by GA during soaking primarily functioned in signal transduction or activation. While most of these DEGs returned to their pre-treatment expression levels before subsequent recovery, a subset persisted until seed germination. During radicle protrusion, the persistent DEGs were associated with energy metabolism and cell structure establishment. Notably, heat shock protein (HSP) genes showed dynamic expression across all stages (soaking, germination, and radicle penetration). Furthermore, by adjusting germination conditions, temperature was confirmed to be a necessary but not sufficient condition for GA-induced S. torvum seed germination. However, functional validation (e.g., using HSP inhibitors or genetic approaches) is still required to confirm the causal role of HSPs. Collectively, these findings not only clarify the molecular basis of GA-regulated seed dormancy breaking in S. torvum but also provide practical guidance for optimizing its commercial propagation protocols.

Although ERF transcription factors (TFs) play critical roles in abiotic stress tolerance, the molecular mechanisms underlying this role remain incompletely understood. This study identified BpERF1A in birch (Betula platyphylla) as a drought-responsive TF through co-expression regulatory network analysis. Expression of BpERF1A was stalwartly induced by drought stress, and drought treatment markedly boosted its promoter activity. Functional analyses demonstrated that overexpression of BpERF1A markedly improved drought tolerance compared with wild-type (WT) birch, whereas its repression increased drought sensitivity. Overexpression lines also exhibited higher antioxidant enzyme activities and proline content relative to WT. Chromatin immunoprecipitation-polymerase chain reaction, yeast one-hybrid and dual-luciferase (dual-LUC) assays confirmed that BpERF1A directly binds to G-box elements in the promoters of BpDHN (dehydrin) and BpAOS (allene oxide synthase), activating their transcription. Furthermore, overexpression of BpDHN/BpAOS enhanced drought tolerance and promoted reactive oxygen species (ROS) scavenging in transgenic plants. Protein-protein interaction analysis using bimolecular fluorescence complementation revealed that BpERF1A interacts with BpRAV1 to form a heterodimer, which further enhances BpERF1A binding to the BpDHN promoter and its transcriptional activation. Collectively, these findings establish a central role for the BpERF1A regulatory module in drought acclimation and highlight its synergistic interaction with BpRAV1, providing an effective strategy to enhance drought tolerance through improved ROS scavenging capacity.

Soilless cultivation has emerged as an effective approach for improving cucumber productivity under protected cultivation by enhancing root-zone conditions and minimizing soil-borne constraints. However, the performance of different growing media combinations remains insufficiently understood. Therefore, this study investigated the effects of different growing media on cucumber growth, yield, fruit quality, and root-knot nematode incidence under protected cultivation. A completely randomized design (CRD) experiment was conducted with 13 different growing media treatments, including soil, perlite, vermiculite, cocopeat, sand, and their combinations. According to the findings, the Vermiculite + Cocopeat in the ratio of 1:1 (T9) combination reduced root-knot nematode infection while dramatically increasing vine length, fruit set, fruit retention, fruit yield (28.05 kg/m2), and quality metrics. Vine length and yield were found to be strongly positively correlated by correlation analysis, while productivity was negatively correlated with root-knot infestation. Fruit set, flowering period, and fruit retention all directly increased yield, according to path analysis. Strong genetic control over these variables was shown by estimates of variance components, which showed moderate heritability for fruit set and retention and high heritability (114.18%) for days to flower initiation. However, yield-related traits exhibited low heritability viz yield per vine (14.24%) and YPP (12.89%), highlighting the significant role of environmental factors. These findings suggest that combination of Vermiculite and Cocopeat is the most suitable growing medium for cucumber cultivation in protected conditions, optimizing yield and reducing disease susceptibility.

Volatile esters are key contributors to the characteristic aroma in fruit, and their accumulation directly dictates fruit quality and consumer acceptance. Alternative oxidase pathway is known to its role in regulating fruit quality, but its molecular mechanism in ester aroma accumulation remains unclear. Here, we demonstrated that the alternative oxidase pathway acts as the dominant respiratory pathway in Nanguo pear (Pyrus ussuriensis) during ripening, and promotes ester aroma accumulation by increasing histone acetylation levels of PuAAT1 (alcohol acetyltransferase 1), the key gene governing ester synthesis. Further, we identified PuHDAC15 as a critical histone deacetylase that modulates acetylation levels and interacts with the transcription factor PuWRKY7. Mechanistically, PuWRKY7 directly binds to the W-box elements in the PuAAT1 promoter. The PuHDAC15-PuWRKY7 complex acts synergistically to repress PuAAT1 transcription, thereby decreasing its histone acetylation levels and gene expression, and consequently inhibiting ester aroma accumulation in Nanguo pear. This study reveals how the alternative oxidase pathway integrates into fruit aroma formation via epigenetic regulation and gene expression, thereby providing a scientific basis for targeted improvement of fruit quality.

Chlorophyll, the pigment in plant leaves, is crucial for capturing light during photosynthesis. Mutations that disrupt chlorophyll production or chloroplast development frequently lead to changes in leaf color. Here, we report the identification and characterization of a chlorotic mutant in cucumber (Cucumis sativus L.), named tnyl3, which exhibits chlorotic cotyledons and seedling lethality. Map-based cloning revealed that the tnyl3 mutation results from a Tnt1 retrotransposon insertion in a gene encoding nuclear factor Y subunit B3 (NF-YB3), a transcription factor subunit. Compared with the wild type, the mutant exhibited dramatic decreases in chlorophyll a and b levels and a significantly lower net photosynthetic rate. Ultrastructural analysis revealed that the chloroplasts in the tnyl3 mutants are structurally abnormal and characterized by underdeveloped thylakoid membranes. Gene expression analyses revealed that CsNF-YB3 is highly expressed in young seedling tissues and is upregulated by light. CRISPR/Cas9 analyses subsequently confirmed that the tnyl3 phenotype is caused by the loss of CsNF-YB3. Yeast two-hybrid and split-LUC assays demonstrated that CsNF-YB3 interacts directly with the cucumber NF-YC2 protein and promotes CsTIC21 transcription, suggesting that it functions as part of an NF-Y transcriptional complex. Transcriptome profiling of the mutant revealed extensive downregulation of photosynthesis-related genes, which is consistent with its impaired chloroplast function. Our findings establish CsNF-YB3 as a crucial genetic factor for chloroplast development and pigment synthesis in cucumber. This work provides new insight into the NF-Y-mediated regulatory network controlling chloroplast biogenesis and offers a potential genetic target for improving plant photosynthetic performance and vigor.

Aromatic esters are key determinants of apple fruit aroma and consumer preference. Here, we investigated the molecular mechanisms underlying ester biosynthesis by characterizing MdAAT2-like, a critical ester-synthesizing gene in apple. MdAAT2-like expressions were significantly upregulated during fruit ripening and positively correlated with ester accumulation. Functional validation in apple and tomato demonstrated that MdAAT2-like overexpression enhanced ester contents, while silencing or knockout reduced ester production. Enzymatic assays revealed that MdAAT2-like exhibits higher catalytic efficiency for medium- and short-chain acyl-CoAs compared to other AAT family members. Two transcription factors, MdMYB98-like and MdWRKY21, were identified as direct activators of MdAAT2-like. Both factors bind its promoter and synergistically enhance transcription through protein interaction. Notably, we uncovered a positive feedback model wherein MdMYB98-like and MdWRKY21 reciprocally activate each other's expression, reinforcing rapid ester synthesis during ripening. This study reveals a MYB-WRKY regulatory module controlling ester biosynthesis in apple.