Drought stress significantly alters the metabolic homeostasis of tea plants; however, few studies have examined the role of specific metabolites, particularly tea polyphenols, in drought resistance. This study reveals that the tea polyphenol content in drought-tolerant tea cultivars tends to increase under drought conditions. Notably, in environments characterized by staged and repeated drought, changes in tea polyphenol are significantly positively correlated with drought resistance. To investigate this further, we irrigated the roots with exogenous tea polyphenols before subjecting the plants to drought. Our findings indicated that the absorptive roots of the experimental group exhibited enhanced development, improved cellular integrity, and a significant increase in peroxidase activity. A comprehensive analysis of the transcriptome and metabolome revealed that tea polyphenols are closely associated with the phenylpropanoid metabolism pathway. Notably, CsMYB77 and CsPOD44 genes were identified as highly correlated with this pathway. Overexpression experiments in Arabidopsis thaliana demonstrated that CsMYB77 promotes the expression of phenylpropanoid pathway genes, thereby enhancing drought resistance. Conversely, antisense oligonucleotide silencing of CsMYB77 decreased drought resistance in tea plants. Additional experiments, including yeast one-hybrid assays, luciferase complementation imaging, dual-luciferase assays, and electrophoretic mobility shift assays, confirmed that CsMYB77 positively regulates the expression of CsPOD44. In summary, our findings indicate that the differences in drought tolerance among tea cultivars are closely linked to phenylpropanoid metabolism. Specifically, tea polyphenols may mediate the regulatory network involving CsMYB77 and CsPOD44, thereby enhancing stress resistance by promoting root development. This study offers new insights into the breeding of drought-resistant tea cultivars.

Flesh fruits are essential for human health, but pathogen infection poses a threat to fruit production and postharvest storage. The hormone salicylic acid (SA) and its metabolites, such as sugar conjugates and methyl salicylate (MeSA), play a crucial role in regulating plant immune responses. However, the UDP-glycosyltransferases (UGTs) responsible for modulating SA metabolism in fruit have yet to be identified, and further investigation is needed to elucidate its involvement in fruit immune response. Here, we identified PpUGT74F2 as an enzyme with the highest transcription level in peach fruit, responsible for catalyzing the biosynthesis of SA glucoside (SAG), but not for MeSAG formation in fruit. Furthermore, infection of peach fruit with Monilinia fructicola, which causes brown rot disease, led to reduced expression of PpUGT74F2, resulting in a significant decrease in SAG content and an increase in MeSA levels. Transgenic tomatoes expressing heterologous PpUGT74F2 increased susceptibility to gray mold. Interestingly, overexpressing PpUGT74F2 did not affect SA levels but dramatically reduced MeSA levels in response to pathogen infection, accompanied by significantly reduced expression of pathogen-related (PR) genes in transgenic tomatoes. This study highlights that PpUGT74F2 acts as a negative regulatory factor for fruit immunity through a distinct mechanism not previously reported in plants.

Tea plant (Camellia sinensis) is among the most significant beverage crops globally. The size of tea buds not only directly affects the yield and quality of fresh leaves, but also plays a key role in determining the suitability of different types of tea. Analyzing the genetic regulation mechanism of tea bud size is crucial for enhancing tea cultivars and boosting tea yield. In this study, a digital phenotyping technology was utilized to collected morphological characteristics of the apical buds of 280 tea accessions of representative germplasm at the ‘two and a bud’ stage. Genetic diversity analysis revealed that the length, width, perimeter, and area of tea buds followed a normal distribution and exhibited considerable variation across natural population of tea plants. Comparative transcriptomic analysis of phenotypic extreme materials revealed a strong negative correlation between the expression levels of four KNOX genes and tea bud size. A key candidate gene, CsKNOX6, was confirmed by further genome-wide association studies (GWAS). Its function was preliminarily characterized by heterologous transformation of Arabidopsis thaliana. Overexpression of CsKNOX6 reduced the leaf area in transgenic plants, which initially determined that it is a key gene negatively regulating bud size. These findings enhance our understanding of the role of KNOX genes in tea plants and provide some references for uncovering the genetic regulatory mechanisms behind tea bud size.

Blackberries (Rubus spp.) are globally consumed and well known for their rich anthocyanin and antioxidant content and distinct flavors. Improving blackberries has been challenging due to genetic complexity of traits and limited genomic resources. The blackberry genome has been particularly challenging to assemble due to its polyploid nature. Here, we present the first chromosome-scale and haplotype-phased assembly for the primocane-fruiting, thornless tetraploid blackberry selection BL1 (Rubus L. subgenus Rubus Watson). The genome assembly was generated using Oxford Nanopore Technology and Hi-C scaffolding, resulting in a 919 Mb genome distributed across 27 pseudochromosomes, with an N50 of 35.73 Mb. This assembly covers >92% of the genome length and contains over 98% of complete BUSCOs. Approximately, 58% of the assembly consists of repetitive sequences, with long terminal repeats being the most abundant class. A total of 87,968 protein-coding genes were predicted, of which, 82% were functionally annotated. Genome mining and RNA-Seq analyses identified possible candidate genes and transcription factors related to thornlessness and the key structural genes and transcription factors for anthocyanin biosynthesis. Activator genes including PAP1 and TTG1 and repressor genes such as ANL2 and MYBPA1 play an important role in the fine tuning of anthocyanin production during blackberry development. Resequencing of seven tetraploid blackberry cultivars/selections with different horticultural characteristics revealed candidate genes that could impact fruiting habit and disease resistance/susceptibility. This tetraploid reference genome should provide a valuable resource for accelerating genetic analysis of blackberries and facilitating the development of new improved cultivars with enhanced horticultural and nutritional traits.

Artificial tomato pollination methods rely on cellular vibrations from air displacement, electric vibration wands and trellis tapping, which have potential to spread pathogens. Bioacoustic frequencies emitted from buzzing bees to ultrasonication can vibrate plant cells without physical contact. The effects of frequency-dependent sonication on the poricidal anther cone sheath, self-pollination, seed set, and fruit size remain unclear. We engineered devices to investigate the frequency-dependent power-law behaviour of floral living cells from greenhouse-grown tomato varieties—contrasting contact-induced oscillations from a vibrating wand and mechanical shaker arm with precision noncontact sonication emitted by a subwoofer speaker. The velocity and acceleration of vibrating flowers and impact on poricidal anther cone sheath cellular structures, self-pollination, and fruit development were assessed. Sonic frequencies ranging from 50 to 10 000 Hz increased pollination, fruit size, weight, and seed set in Sweetelle, Endeavour, Paulanca, and Managua commercial varieties. Scanning electron microscopy revealed sonication separated the intertwined trichomes and unzipped their meshed network that locks the lobes of the anther cone sheath together thereby releasing pollen grains. Near ultra-sonic frequencies boosted fruit size, whereas seed set remained constant thereby challenging the floral cell power-law rheological characteristics under different frequency scales. Tomato flowers displayed a low power-law cell behaviour to frequency-dependent sonication enabling its effectiveness as a precision noncontact technology to boost pollination and tomato fruit size without a substrate-borne component.

The color diversity of non-purple carrot taproots is mainly affected by carotenoid species and content. Carrot cytochrome P450 carotene β-ring hydroxylase (DcCYP97A3) may influence carotenoid accumulation in carrots; however, the roles of DcCYP97A3 in carrot remain unclear. Compared to the orange carrot ‘Kurodagosun, KRD’, the yellow carrot ‘Yellowstone, YST’ had greater relative transcript levels of DcCYP97A3. DcCYP97A3 was shown to catalyze the β-ring hydroxylation of α-carotene to create zeaxanthin when it was expressed in Escherichia coli accumulating α- and β-carotene. Expression of the DcCYP97A3 of ‘YST’ in DcCYP97A3 functionally deficient orange carrot ‘KRD’ resulted in yellow taproots, decreased α-carotene and β-carotene content, decreased α-/β-carotene ratio, and increased lutein content. In carrots overexpressing the DcCYP97A3 gene, the transcript levels of DcLCYE and DcLCYB1 were significantly upregulated and downregulated, respectively. Gene editing of DcCYP97A3 in ‘YST’ resulted in DcCYP97A3 knockout mutants with significantly reduced levels of lutein and β-carotene and significantly upregulated transcript levels of DcCHXB2 and DcCCD4. These findings advance our knowledge of the molecular mechanisms behind carrot carotenoid metabolism.

Mgaloblishvili, a grapevine variety from Georgia (Southern Caucasus), exhibits a unique resistance mechanism against downy mildew. Mgaloblishvili resistance mechanism, involving pathogen recognition, activation of ethylene signalling pathway, and structural and chemical defences, is mediated by the resistance loci Rpv29, Rpv30, and Rpv31. Mgaloblishvili genome was sequenced using PacBio HiFi, resulting in a chromosome-scale diploid assembly of 986 Mbp, including 58 912 predicted protein-coding genes across two phased chromosome sets. Comparative analysis with the susceptible PN40024 genome allowed us to identify differences in structure, gene content, and gene expression, as well as the impact of structural variants (SVs) and single nucleotide polymorphisms (SNPs) between Mgaloblishvili and PN40024 loci. Resistance haplotypes were identified through DNA sequencing of a self-pollinated Mgaloblishvili population. Compared to orthologous regions in PN40024, the Rpv29 locus in Mgaloblishvili exhibits reduced gene content, while the Rpv31 locus has similar gene content. In both Mgaloblishvili and PN40024, most genes within these loci are associated with plant defence pathways. While genes in both genotypes perform similar functions, SVs and SNPs were identified as key determinants of the structural differences between the genomes. Defining the Rpv30 locus was challenging due to ambiguous marker localization. DNA sequencing allowed us to identify resistance haplotypes for both Rpv30 and Rpv31 on Mgaloblishvili haplotype 2, though insights into the Rpv29 locus remain limited. Our results indicate that Mgaloblishvili’s resistance is driven by numerous small SVs and SNPs, which lead to the loss of susceptibility factors and unique transcriptional regulation of defence-related genes.

Tanshinones are bioactive diterpenoid chemicals of the herb Salvia miltiorrhiza with a characteristic furan D-ring. As a newly identified downstream enzyme, SmCYP71D375, catalyzes hydroxylation by 14,16-ether (hetero)cyclization to form the furan D-ring from the precursor of the phenolic abietane-type diterpenoids that exist widely in Lamiaceae plants. However, its transcriptional regulatory network, with SmCYP71D375 as the direct target gene, remains unclear. In the present study, the promoter of SmCYP71D375 was employed as the bait to mine the upstream regulatory protein using the cDNA yeast library of S. miltiorrhiza. An R2R3-MYB transcription factor gene, SmMYB53, was identified. Overexpressing SmMYB53 in transgenic hairy roots upregulated SmCYP71D375 expression, thereby accelerating tanshinone accumulation, whereas tanshinone accumulation was inhibited in SmMYB53-RNAi transgenic hairy root lines. To dissect the regulatory network of SmMYB53, SmbZIP51 was captured using SmMYB53 as the bait to prey for its potential interacting proteins in the cDNA yeast library. Yeast two-hybrid, glutathione S-transferase pull-down, and bimolecular fluorescence complementation assays were independently used to verify the interaction between the SmMYB53 and SmbZIP51 proteins. We further verified that the upregulation of SmCYP71D375 activated by SmMYB53 would be inhibited by the interaction of SmMYB53 and SmbZIP51. The present findings uncover the molecular regulatory network underlying SmCYP71D375 as the direct target regulating tanshinone biosynthesis and offer a basis for the genetic improvement of medicinal substance biosynthesis in S. miltiorrhiza.

Ethylene (ET) influences the synthesis of anthocyanins, although its regulatory effects can differ significantly across various plant species. In apples (Malus domestica), ET promotes anthocyanin synthesis, whereas in Arabidopsis thaliana, it inhibits its accumulation. Our research showed that ethephon (Eth), an ET derivative, promotes anthocyanin synthesis in ‘Viviana’ lilies, which has great potential in the cut flower industry. The regulatory mechanism whereby ET influences anthocyanin synthesis in lilies remains unclear. In this study, we screened and characterized an ET-induced ET response factors (ERFs), LvERF113, with inhibitory function. Our analyses suggested that LvERF113 could inhibit the negative regulatory function of LvMYB1 at transcriptional and posttranslational levels, promoting anthocyanin synthesis in ‘Viviana’ lily tepals. In addition, LvERF113 is positively regulated by LvMYB5, forming the LvMYB5-LvERF113-LvMYB1 module controlling anthocyanin synthesis by ET in ‘Viviana’ lily. These findings offer new insights into the ET regulatory network of anthocyanin synthesis and provide a theoretical basis for the application of ET derivatives in the cut flower industry.

Leaf vasculature not only acts as a channel for nutrients and signaling information but also influences leaf morphology. It consists of several distinct cell types with specialized functions. Cell type-specific characterizations based on single-cell RNA sequencing technology could aid in understanding the identities of vascular tissues and their roles in leaf morphogenesis in Brassica rapa. Here, we generated a single-cell transcriptome landscape of the Chinese cabbage leaf vasculature. A total of 12 cell clusters covering seven known cell types were identified. Different vascular cell types were characterized by distinct identities. The xylem parenchyma and companion cells exhibited an active expression pattern of amino acid metabolism genes. Tracheary elements and sieve elements were enriched in many genes related to cell wall biosynthesis, and the phloem parenchyma was enriched in many sugar transporter-encoding genes. Pseudo-time analyses revealed the developmental trajectories of the xylem and phloem and the potential roles of auxin and ethylene in xylem development. Furthermore, we identified key candidate regulators along the differentiation trajectory of the sieve elements and companion cells. Most of the homoeologous genes in the syntenic triads from the three subgenomes showed asymmetric gene expression patterns in different vascular cell types. Collectively, our study revealed that Chinese cabbage leaf vasculature cells had highly heterogeneous transcriptomes, providing new insights into the complex processes of leaf vasculature development in B. rapa leafy vegetables and other Brassica crops.

The interaction between ethylene and melatonin in the regulation of polyphenol metabolism and the underlying mechanism remain largely unclear. This work demonstrated that ethylene treatment increased melatonin biosynthesis by inducing the VvASMT expression in grape seeds. Ethylene-induced VvERF5 transactivated VvASMT via binding to the ethylene response element in its promoter. VvERF5 overexpression led to an increase in melatonin biosynthesis while its suppression generated the opposite results in grape seeds, calli, and/or Arabidopsis seeds. A melatonin-responsive element (MTRE) was identified, and melatonin-induced VvERF104 was found to bind to the MTRE of the VvMYB14 promoter and activate its expression. VvMYB14 overexpression widely modified the expression of genes in the phenylpropanoid pathway and phenolic compound content in grape seeds. DNA affinity purification sequencing revealed that the MEME-1 motif was the most likely binding sites of VvMYB14. VvPAL, VvC4H, and VvCHS were verified to be the target genes of VvMYB14. Additionally, the overexpression of VvERF5 or VvERF104 increased the expression of VvPAL, VvC4H, and VvCHS, as well as the levels of the corresponding metabolites. Moreover, the roles of VvERF5, VvASMT, and VvERF104 in mediating ethylene-induced changes in the phenylpropanoid pathway were elucidated using their suppressing seeds. Collectively, ethylene increased the VvMYB14 expression via the pathway of ERF5-melatonin-ERF104 and thereby modified the phenylpropanoid pathway.

Rapid growth of Moso bamboo (Phyllostachys edulis) shoots (offspring ramet) is primarily fuelled by nitrogen (N) derived from parent ramet and absorbed by rhizome roots. However, the extent to which each N source supports the growth of offspring ramet and the underlying molecular mechanisms of N transport remain unclear. Here, clonal fragments consisting of a parent ramet, an offspring ramet, and an interconnected rhizome were established in a Moso bamboo forest. Additionally, ¹⁵N isotope tracing and transcriptome profiling were conducted concurrently to quantify the N contribution from the parent ramet and rhizome roots to the offspring ramet, and to reveal the molecular mechanisms underlying N transport during rapid growth (i.e. early, peak, branching, and leafing stages). The N acquisition strategy of offspring ramet shifted from being primarily provided by the parent ramet (72.53%) during early stage to being predominantly absorbed by rhizome roots (69.85%) during the leafing stage. Approximately equal N contributions (45.82%-54.18%) from the parent ramet and rhizome roots were observed during peak and branching stages. PeAAP29123 was identified as a key gene for N transport, being most closely correlated with ¹⁵N content. Biomolecular assays demonstrated that PeHDZ23987 could activate the expression of PeAAP29123 via two types of HD-motifs. Overexpression of PeHDZ23987 and PeAAP29123 significantly enhanced N starvation tolerance in transgenic rice with significantly improved N uptake efficiency. Our findings clarify the pattern and mechanisms of N supply for the rapid growth of Moso bamboo offspring ramet and provide transcriptomic evidence for long-distance N transport between clonal ramets.

The plant immune response plays a central role in maintaining a well-balanced and healthy microbiome for plant health. However, insights into how the fruit immune response and the fruit microbiome influence fruit health after harvest are limited. We investigated the temporal dynamics of the fruit microbiota and host defense gene expression patterns during postharvest storage of apple fruits at room temperature. Our results demonstrate a temporal dynamic shift in both bacterial and fungal community composition during postharvest storage that coincides with a steep-decline in host defense response gene expression associated with pattern-triggered immunity. We observed the gradual appearance of putative pathogenic/spoilage microbes belonging to genera Alternaria (fungi) and Gluconobacter and Acetobacter (bacteria) at the expense of Sporobolomyces and other genera, which have been suggested to be beneficial for plant hosts. Moreover, artificial induction of pattern-triggered immunity in apple fruit with the flg22 peptide delayed the onset of fruit rot caused by the fungal pathogen Penicillium expansum. Our results suggest that the fruit immune response helps to orchestrate a microbiome and that the collapse of the immunity results in the proliferation of spoilage microbes and fruit rot. These findings hold implications for the development of strategies to increase fruit quality and prolong shelf life in fruits and vegetables.

Spermidine (Spd) is one of the predominant polyamines in higher plants and plays a crucial role in combating various abiotic stresses. However, the molecular functions and underlying regulatory mechanisms associated with plant Spd synthase (SPDS) genes in cold tolerance remain poorly understood. In this study, cold treatment markedly induced Spd accumulation and enhanced SPDS activity in Ichang papeda (Citrus ichangensis), a cold-hardy plant in Citrus genus. Exogenous Spd supply led to dramatically improved cold tolerance. Two SPDS genes (CiSPDS1 and CiSPDS2) were identified in Ichang papeda, but only CiSPDS1 was substantially upregulated by cold. Overexpressing of CiSPDS1 in both tobacco (Nicotiana tabacum) and lemon (Citrus limon), a cold-sensitive Citrus species, promoted Spd synthesis and enhanced cold tolerance in the transgenic plants. In contrast, knockdown of CiSPDS1 in Ichang papeda by virus-induced gene silencing (VIGS) repressed Spd synthesis and greatly impaired the cold tolerance, which was restored by exogenous replenishment of Spd. In addition, we demonstrated that WRKY27 of Ichang papeda (CiWRKY27) directly bound to and activated the CiSPDS1 promoter through interacting with a W-box cis-acting element. Meanwhile, VIGS-mediated silencing of CiWRKY27 resulted in marked reduction of CiSPDS1 transcript levels and Spd contents and significantly impaired the cold tolerance in Ichang papeda. Taken together, this study illustrated the role of CiSPDS1 in cold tolerance and identified it as a direct target of CiWRKY27. These findings provide insight into the regulatory mechanism by which the molecular module CiWRKY27-CiSPDS1 regulates Spd accumulation for modulation of cold tolerance.

It is frequently observed that plant sexes differ in their response to environmental stress. Poplars are dioecious plants, and sex separation of poplars is triggered by the sex-limited expression of the poplar sex-determining gene FERR. In this study, we over-expressed FERR in a male poplar and knocked it out in a female poplar. The over-expression lines exhibited distinct morphological and physiological changes rendering the transformed plants more tolerant to drought stress. By contrast, no obvious change in drought tolerance was observed in the knockout lines. Transcriptome sequencing and molecular interaction analysis demonstrated that the effect of FERR on drought tolerance was conferred by competitive interaction with protein phosphatase 2C and SNF1-related protein kinase 2 (SnRK2). Under drought stress, an FERR-SnRK2s-ARR5 complex forms and activates the ABA signaling pathway. Our results provide direct evidence that the expression of the poplar sex-determining gene pleiotropically affects plant drought tolerance.

Kiwifruits, belonging to the genus Actinidia, are acknowledged as one of the most successfully domesticated fruits in the twentieth century. Despite the rich wild resources and diverse phenotypes within this genus, insights into the genomic changes are still limited. Here, we conducted whole-genome sequencing on seven representative materials from highly diversified sections of Actinidia, leading to the assembly and annotation of 14 haplotype genomes with sizes spanning from 602.0 to 699.6 Mb. By compiling these haplotype genomes, we constructed a super pan-genome for the genus. We identified numerous structural variations (SVs, including variations in gene copy number) and highly diverged regions in these genomes. Notably, significant SV variability was observed within the intronic regions of the MED25 and TTG1 genes across different materials, suggesting their potential roles in influencing fruit size and trichome formation. Intriguingly, our findings indicated a high genetic divergence between two haplotype genomes, with one individual, tentatively named Actinidia × leiocacarpae, from sect. Leiocacarpae. This likely hybrid with a heterozygous genome exhibited notable genetic adaptations related to resistance against bacterial canker, particularly through the upregulation of the RPM1 gene, which contains a specific SV, after infection by Pseudomonas syringae pv. actinidiae. In addition, we also discussed the interlineage hybridizations and taxonomic treatments of the genus Actinidia. Overall, the comprehensive pan-genome constructed here, along with our findings, lays a foundation for examining genetic compositions and markers, particularly those related to SVs, to facilitate hybrid breeding aimed at developing desired phenotypes in kiwifruits.

Salt stress is an important abiotic stress affecting the growth and fruit quality of apple fruits. Although jasmonic acid (JA) hormones and WRKY transcription factors (TFs) have both been reported to be involved in plant salt stress responses, the molecular mechanisms by which JA-mediated WRKY TFs regulate salt stress in apples remain unclear. Here, we report the identification of a WRKY family TF from apple, MdWRKY9, and its involvement in apple salt tolerance by regulating the expression of Na⁺/H⁺ antiporters, MdNHX1, and MdSOS2. Furthermore, we show that the protein repressors MdJAZ5 and MdJAZ10 in the JA signaling pathway can both interact with MdWRKY9 to form a complex and inhibit its DNA-binding and transcriptional activation activity. The JA signal triggers the degradation of MdJAZ5 and MdJAZ10 proteins by the 26S proteasome, disrupting the JAZ-WRKY protein complex and thereby releasing MdWRKY9 to activate downstream gene expression, promoting salt tolerance in apples. These findings provide important insights into the molecular mechanism of the WRKY TFs in JA-mediated salt tolerance in plants.

The current genetic model explaining berry skin color in Vitis vinifera is incomplete and fails to predict berry skin color phenotypes for one allele of VvMybA1, referred to as VvMybA1_SUB. Our study focuses on this specific allele, revealing that the haplotype containing VvMybA1_SUB (haplotype F) represents an ancient lineage of the berry color locus. Within haplotype F, we identified two functional subhaplotypes, HapF1 and HapF2, associated with black-skinned phenotype, and one non-functional subhaplotype, HapF^DEL, responsible for white-skinned phenotype. HapF1 likely originated from wild populations domesticated in the Near East and subsequently spread globally with the expansion of viticulture. In contrast, HapF2 has a more restricted distribution and may have emerged from hybridization events between cultivated grapevines and local wild populations as viticulture migrated to the Italian peninsula. Furthermore, we found that in white-skinned berry cultivar, HapF has undergone a large deletion at the berry color locus, removing the majority of the VvMybA genes. Previous works suggested a single common origin for white-skinned varieties during grapevine domestication. Our results challenge this notion, instead proposing that white-skinned grape cultivars arose at least twice during grapevine domestication history. Alongside the major haplotype A, some white-skinned cultivars, such as cv. ‘Sultanina’ harbor HapF^DEL. Since HapF^DEL is present only in table grape varieties, we suggest that it likely arose from a recent mutational event and dispersed along the ancient Silk Road into East Asia. These findings enhance our understanding of the genetic diversity and evolutionary trajectory of grapevine cultivars, offering insights into their domestication and spread across different geographical regions.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a highly conserved enzyme in the glycolytic pathway, also acts as a moonlighting protein, performing various functions beyond its classical role in glycolysis, such as regulating gene expression, participating in cell signal transduction, and responding to environmental stress. By interacting with various signaling molecules, GAPDH plays a regulatory role in hormone signaling pathways, influencing plant growth and development. Functional plasticity in GAPDH is modulated mainly through redox-driven post-translational modifications, which alter the enzyme’s catalytic activity and influence its subcellular distribution. This review explores the diverse functionalities of GAPDHs in plants, highlighting their significance in plant metabolic processes and stress adaptation.

Anthocyanins are important specialized fruit metabolites and major pigments, whose abundance depends on co-regulation of activators and repressors, primarily transcription factors (TFs) of the MYB family. Herein, a KANADI-like TF PuKAN4 was characterized in pear. This TF could be transcriptionally up-regulated by the anthocyanin-related R2R3-MYBs PuMYB10/PuMYB114 and exhibited high expression within red-skinned pears. Interestingly, PuKAN4 repressed anthocyanin biosynthesis in transiently overexpressed pear fruit, and stable transformation in pear calli and tobacco plants. The PuKAN4 had a conserved EAR repression domain in C-terminal, while the repression function of PuKAN4 could be offset by a transcription activation domain VP64. The dual luciferase analysis proved that PuMYB114/PuMYB10 up-regulated expression of PuKAN4. Furthermore, the PuKAN4 could physically interact with PuMYB10/PuMYB114 and did not affect the combination of MYB10/MYB114-bHLH3, as demonstrated by Y2H, pull-down and firefly luciferase complementation. Thus, the PuKAN4 should play the role of active repressor, the formation of PuKAN4-PuMYB10/PuMYB114-PubHLH3 complex inhibited pear anthocyanin biosynthesis. Our findings unveiled an activator-and-repressor feedback loop between PuMYB114/PuMYB10 and PuKAN4, which possibly balance biosynthesis activity and prevents over-accumulation of phenylpropanoids.

Fluoride (F) is a nonessential but potentially harmful element for plants, especially when present in excess. The tea plant is known for its ability to hyperaccumulate F from the soil and eventually accumulates in the leaves; however, how the tea plant transports F to the leaves remains unclear. Here, we found that Se can significantly decrease the transport efficiency of F from root to leaf. Therefore, RNA-Sequencing was performed on tea roots cotreated with selenite and fluoride, and then we isolated a plasma membrane-localized F transporter CsNPF2.3 from tea plant roots and examined its role in transport of F in tea plants. The results showed that CsNPF2.3 exhibited F transport activity when heterologously expressed in yeast. Expression pattern analysis revealed that CsNPF2.3 is expressed in epidermal cells, cortex cells, and xylem parenchyma cells in roots. Overexpression of CsNPF2.3 in tea roots significantly increased F content in the root, stem, and leaf, and enhanced the transport efficiency of F from root to leaf. Furthermore, in nine tea cultivars, CsNPF2.3 expression in the root was significantly positively correlated with F content in the leaf and root, and the transport efficiency of F from root to leaf. Altogether, these findings suggest that CsNPF2.3 was involved in uptake and transport of F in tea plants.

Plant resistance inducers represent an alternative strategy that mitigate stress-induced damage in plants. Previously, 2-amino-3-methylhexanoic acid (AMHA), a novel natural plant resistance inducer, was shown to significantly bolster cold tolerance, thermotolerance, and pathogen resistance in plants. However, the intricate mechanisms underlying AMHA’s response to cold stress remain elusive. Thus, we investigated the physiological and transcriptomic analyses of AMHA pretreatment on tea plant to determine its substantial role of AMHA under cold stress. The results showed that pretreatment with 100 nM AMHA effectively mitigated the detrimental effects of cold stress on photosynthesis and growth. Furthermore, differentially expressed genes were identified through RNA-seq during pretreatment, cold stress, and 2 days of recovery. These genes were mainly enriched in pathways related to flavonoid/anthocyanin, carotenoid, and ascorbic acid-glutathione (AsA-GSH) cycle, including GST (encoding glutathione S-transferase). Potential regulatory relationships between the identified genes and transcription factors were also established. Antisense oligodeoxynucleotide-silencing and overexpression experiments revealed that CsGSTU7 enhances cold resistance by maintaining redox homeostasis. In conclusion, our study suggests that antioxidant-related signaling molecules play a critical role in the signaling cascades and transcriptional regulation mediating AMHA-induced cold-stress resistance in tea plant.

Phenolic compounds are derived from the phenylpropanoid metabolic pathways of plants and include phenylpropionic acids, lignins, coumarins, and flavonoids. These compounds are among the most abundant and diverse classes of secondary metabolites found throughout the plant kingdom. Phenolic compounds play critical roles in the growth, development, and stress resistance of horticultural plants. Moreover, some phenolic compounds exhibit substantial biological activities, and they are widely utilized across various sectors, such as the pharmaceutical and food industries. The cytochrome P450 monooxygenase 98A subfamily (CYP98A) is involved mainly in the biosynthesis of phenolic compounds, mediating the meta-hydroxylation of aromatic rings in the common phenylpropane biosynthesis pathways of phenolic compounds. However, research on this family of oxidases is currently fragmented, and a systematic and comprehensive review has not yet been conducted. This review offers an exhaustive summary of the molecular features of the CYP98A family and the functions of CYP98A monooxygenases in the biosynthesis of different types of phenolic compounds. In addition, this study provides a reference for the exploration and functional study of plant CYP98A family enzymes. An enhanced understanding of CYP98A monooxygenases can help in the cultivation of high-quality horticultural plants with increased resistance to biotic and abiotic stresses and enhanced accumulation of natural bioactive compounds via metabolic engineering strategies. Moreover, the structural optimization and modification of CYP98A monooxygenases can provide additional potential targets for synthetic biology, enabling the efficient in vitro production of important phenolic compounds to address production supply conflicts.

As an important noncereal food crop grown worldwide, the genetic improvement of potato in tuber yield and quality is largely constrained due to the lacking of a high-quality reference genome and understanding of the regulatory mechanism underlying the formation of superior alleles. Here, a chromosome-scale haplotype-resolved genome assembled from an anther-cultured progeny of ‘Ningshu 15’, a tetraploid variety featured by its high starch content and drought resistance was presented. The assembled genome size was 1.653 Gb, with a contig N50 of approximately 1.4 Mb and a scaffold N50 of 61 Mb. The long terminal repeat assembly index score of the two identified haplotypes of ‘Ningshu 15’ was 11.62 and 11.94, respectively. Comparative genomic analysis revealed that positive selection occurred in gene families related to starch, sucrose, fructose and mannose metabolism, and carotenoid biosynthesis. Further genome-wide association study in 141 accessions identified a total number of 53 quantitative trait loci related to fructose, glucose, and sucrose content. Among them, a tonoplast sugar transporter encoding gene, StTST2, closely associated with glucose content was identified. Constitutive expression of StTST2 in potato and Arabidopsis increased the photosynthetic rate, chlorophyll and sugar content, biomass tuber and seed production in transgenic plants. In addition, co-immunoprecipitation assays demonstrated that StTST2 directly interacted with SUT2. Our study provides a high-quality genome assembly and new genetic locus of potato for molecular breeding.

Water spinach (Ipomoea aquatica) can accumulate cadmium (Cd) even in mildly contaminated soils, but the roles of its root tip cell types in Cd fixation and transport remain unclear. Single-cell RNA sequencing revealed nine cell types in root tips in both the QLQ cultivar (low Cd accumulation) and the T308 cultivar (high Cd accumulation). High expression of LAC2 and PER72 in the QLQ epidermis was associated with enhanced lignin deposition, which may facilitate fixation of Cd and reduce its translocation to the shoot. In T308, PER72 and hormone-related genes (PIN1, ARF8, IAA17, and EIN3) were upregulated, which was hypothesized to promote xylem and trichoblast development, potentially facilitating Cd uptake and transport. Fluorescence assays suggested that the higher pectin demethylation and lignin content in QLQ may limit Cd movement, whereas the more developed tissues in T308 may contribute to increased Cd accumulation in the shoots. These findings clarify the mechanisms by which Cd accumulates in water spinach and offer insights into mitigating Cd uptake in crops.

Plant immunity involves complex regulatory mechanisms that mediate the activation of defense responses against pathogens. Protein degradation via ubiquitination plays a crucial role in modulating these defenses, with E3 ubiquitin ligases functioning as central regulators. This study investigates the role of SlATL2, an ARABIDOPSIS TÓXICOS EN LEVADURA (ATL)-type E3 ubiquitin ligase localized in the plasma membrane, in the immune response of tomato plants against Pseudomonas syringae pv. tomato (Pst) DC3000. Our findings demonstrate that SlATL2 expression is induced upon Pst DC3000 infection and treatment with defense hormones salicylic acid and jasmonic acid. Functionally, SlATL2 negatively regulates immune responses, impairing resistance to Pst DC3000 and suppressing flg22-triggered immunity. In addition, SlATL2 limits pathogen-induced reactive oxygen species and callose accumulation by targeting the COP9 signalosome subunit 5a (SlCSN5a), a key positive regulator of tomato defense responses against Pst DC3000. This interaction, which occurs via the N-terminal residue of SlATL2, results in the ubiquitination and 26S proteasomal degradation of SlCSN5a, thereby suppressing SA-dependent expression of defense response genes associated and limiting reactive oxygen species production. This work sheds light on the molecular mechanism through which the E3 ubiquitin ligase SlATL2 attenuates tomato immune responses by targeting a COP9 signalosome subunit for degradation. These discoveries deepen our insights into the post-translational mechanisms governing plant immune responses and provide fresh opportunities to bolster crop resistance against bacterial pathogens.