Plants are constantly challenged by diverse pathogens and respond through multilayered immune mechanisms described by the classical "zig-zag" model. Apoplast nanoparticles (ANs), extracellular vesicle (EV)-like entities,...Plants are constantly challenged by diverse pathogens and respond through multilayered immune mechanisms described by the classical "zig-zag" model. Apoplast nanoparticles (ANs), extracellular vesicle (EV)-like entities, are dynamically remodeled during plant-pathogen interactions. Although EVs are known to carry immune-regulatory cargos, the specificity and coordination of ANs protein responses to distinct pathogens remain largely unresolved. Here, we show that the glycosylation profiles of ANs and their N-glycoproteins undergo distinct remodeling upon infection with Pseudomonas syringae and Botrytis cinerea, as revealed by lectin microarray and mass spectrometry analyses. We identify SPILR, an ANs protein that modulates bacterial morphology and motility by reducing polygalacturonase activity and inhibiting the bacterial flagellar P-ring protein, FlgI. Additionally, the ESM1 protein enhances Arabidopsis resistance to B. cinerea through modulation of lipase activity and lipid metabolism. Disruption of N-glycosylation sites on ANs proteins compromises their antimicrobial function and alters host resistance to both bacterial and fungal pathogens. Together, our findings uncover N-glycosylation as a critical determinant of ANs-mediated extracellular immunity, highlighting glycosylation as an integrative mechanism linking vesicle biology, pathogen specificity, and immune signaling. This work establishes a framework for glycoengineering-based strategies to enhance crop resistance and advance nano-agricultural applications.
Long non-coding RNAs (lncRNAs) play an important role in regulating plant growth and development, and stress response. However, their roles in Botrytis cinerea resistance remain poorly characterized. As a devastating nec...Long non-coding RNAs (lncRNAs) play an important role in regulating plant growth and development, and stress response. However, their roles in Botrytis cinerea resistance remain poorly characterized. As a devastating necrotrophic fungus, gray mold is one of the most serious diseases that affects crop production worldwide. In this study, through screening of B. cinerea resistance in 18 strawberry germplasms, we identified Fragaria nilgerrensis as a resistant species and Fragaria nubicola as a susceptible one. We conducted lncRNA-seq on resistant and susceptible strawberries at 4 d after B. cinerea inoculation and untreated controls. lincRNA6679 and its target gene FnWRKY14 were identified in F. nilgerrensis. Coding potential assessment and RNA pull-down experiments revealed that lincRNA6679 is a long non-coding RNA that positively regulates FnWRKY14 expression by forming molecular complexes with FnWRKY50 or FnMYB59. Genetic transformation demonstrated that both lincRNA6679 and FnWRKY14 enhance resistance to B. cinerea in strawberries. Moreover, FnWRKY14 is bound to the FnPR1B promoter, activating its expression. FnWRKY14 was phosphorylated and activated by FnMAPK3 or FnMAPK6, further upregulating the expression of FnPR1B and enhancing disease resistance. This study revealed how long non-coding RNAs regulate strawberry resistance to B. cinerea, broadening the scope of research on strawberry resistance mechanisms and providing new strategies for molecular breeding.
Soil salinization is a global challenge threatening agricultural production, food security, and sustainable development. As a pioneer crop on saline-alkali land, sunflower plays a crucial role in the improvement and util...Soil salinization is a global challenge threatening agricultural production, food security, and sustainable development. As a pioneer crop on saline-alkali land, sunflower plays a crucial role in the improvement and utilization of salt-affected soils. However, the molecular mechanisms underlying sunflower salt tolerance remain poorly understood. In this study, we identified a key R2R3-MYB gene, HaMYB22, through a combination of genome and transcriptome analyses. Functional characterization demonstrates that overexpression of HaMYB22 significantly enhances salt tolerance in both Arabidopsis and sunflower, whereas its silencing decreases salt resistance. Protein interaction assays revealed that HaMYB22 interacts with HaMYB120 and HaMYB181. Glutathione S-transferase HaGST3.2 was identified as a direct target of HaMYB22, and superior haplotype HaMYB22 can strongly increase HaGST3.2 transcripts. Moreover, HaMYB120 and HaMYB181 synergistically strengthen HaMYB22-mediated HaGST3.2 activation. HaGST3.2 silencing in sunflower decreases salt tolerance. Our findings revealed the importance of the HaMYB22-HaGST3.2 module in sunflower salt tolerance.
Calcium (Ca), a dual-functional mineral that serves both as an essential structural factor and a signaling molecule, plays a critical role in regulating fundamental physiological processes in plants, including developmen...Calcium (Ca), a dual-functional mineral that serves both as an essential structural factor and a signaling molecule, plays a critical role in regulating fundamental physiological processes in plants, including development, stress response, and fruit quality traits. However, a comprehensive and systematic summary of calcium's regulatory functions in fruit quality is still lacking. This review aims to clarify the pivotal roles of calcium in regulating key fruit quality attributes, including external traits such as morphology and coloration; internal nutritional properties, such as flavor-related metabolites and bioactive compounds; and physiological disorders such as cracking, softening, browning, chilling injury, blossom-end rot, water core, and bitter pit. Considering its diverse regulatory functions, genetic manipulation of Ca signaling pathways and the application of nano-calcium formulations offer promising strategies for improving fruit yield and quality in commercial production systems. This review further outlines the underlying mechanisms through which calcium influences fruit quality and suggests future research directions to address existing knowledge gaps.
Auxin plays a pivotal role in regulating crop nitrogen (N)-use efficiency (NUE), coordinating both N-responsive root development and the expression of N metabolism genes. In our previous work, we identified DULL NITROGEN...Auxin plays a pivotal role in regulating crop nitrogen (N)-use efficiency (NUE), coordinating both N-responsive root development and the expression of N metabolism genes. In our previous work, we identified DULL NITROGEN RESPONSE1 (DNR1) as a repressor within the auxin-mediated NUE network in rice. Here, we further delineate this pathway by identifying the SUMO E3 ligase OsSIZ1 as a key upstream regulator of DNR1. Contrary to its canonical role, we discovered that OsSIZ1 exhibits ubiquitin E3 ligase activity toward DNR1, facilitating its polyubiquitination at lysine 314 and subsequent degradation. This degradation promotes auxin accumulation, thereby enhancing NUE and grain yield. Notably, the yield advantage driven by OsSIZ1 is most pronounced under low-N conditions, underscoring its potential as a breeding target for developing resilient crops that require less N fertilizer, enabling a more sustainable agriculture.
Alkaline stress is a major constraint on crop growth and development and negatively impacts soybean (Glycine max) production and yield. Despite the remarkable progress that has been made in investigating beneficial micro...Alkaline stress is a major constraint on crop growth and development and negatively impacts soybean (Glycine max) production and yield. Despite the remarkable progress that has been made in investigating beneficial microbes that facilitate plant growth and development, the role of rhizobacteria in regulating alkaline tolerance in soybean remains poorly understood. Here, we isolated Klebsiella sp. strain B7 from the Suaeda glauca roots and found that it enhances the alkaline tolerance of soybean by secreting pyruvic acid. Metabolome and RT-qPCR analysis of soybean roots indicated that high levels of pyruvic acid secreted by B7 activated the expression of genes involved in pyruvic acid metabolism and increased L-malic acid accumulation in soybean roots, thereby effectively mitigating reactive oxygen species induced by alkaline stress. Overexpression of these pyruvic acid metabolism-associated genes greatly enhanced alkaline tolerance of soybean and ATP-citrate lyase activity, further confirming the positive role of pyruvic acid in L-malic acid biosynthesis and alkaline tolerance in soybean. Notably, the B7 application to alkaline soil enhanced the soybean yield. Moreover, B7 recruited more beneficial microbes and shaped the composition of the rhizosphere bacterial community of soybean plants. These findings highlight the vital function of rhizobacteria strain B7 in enhancing alkaline tolerance in soybean, thus providing further evidence for the crucial role of plant growth-promoting rhizobacteria in the abiotic stress response of soybean.
Leaf angle is a key agronomic trait for improving planting density and yield in lettuce, particularly in controlled-environment agriculture and high-density field cultivation. Leaf angle regulation is well studied in mon...Leaf angle is a key agronomic trait for improving planting density and yield in lettuce, particularly in controlled-environment agriculture and high-density field cultivation. Leaf angle regulation is well studied in monocots; however, the genetic and molecular mechanisms in dicots remain largely unknown. Here, we genetically clone and functionally characterize LsOFP6a, an OVATE family protein gene, as a key regulator of leaf angle in lettuce. A nonsense mutation in LsOFP6a in large-leaf-angle cultivars produces a truncated protein with impaired function. CRISPR/Cas9 knockout and complementary tests confirmed that LsOFP6a negatively regulates leaf angle in lettuce. LsOFP6a physically interacts with the BELL-like homeodomain transcription factor LsBLH2. Genetic analyses revealed that LsOFP6a regulates leaf angle through an LsBLH2-dependent pathway, and LsBLH2 is recessive-epistatic to LsOFP6a. LsBLH2 directly upregulates the expression of the cytokinin oxidase gene LsCKX5a. LsOFP6a represses the transcriptional activity of LsBLH2 on LsCKX5, leading to elevated cytokinin levels and small leaf angle. Furthermore, LsOFP6a inhibits the effects of LsBLH2 on repressing abaxial gene LsYAB1, leading to enhanced abaxial cell elongation and erect leaves. Loss of function of LsOFP6a decreases the cytokinin level and represses abaxial cells, resulting in large leaf angles. In summary, the LsOFP6a-LsBLH2 module orchestrates cytokinin catabolism and leaf dorsiventrality to regulate lettuce leaf angle. Our study suggests potential novel strategies for the breeding of lettuce with compact architecture and suitable for high-density planting in the open field and plant factories.
Seed dormancy (SD) is the primary genetic determinant of pre-harvest sprouting (PHS) resistance. However, the molecular mechanisms underlying SD remain incompletely understood. Here, we identified a wheat cytochrome P450...Seed dormancy (SD) is the primary genetic determinant of pre-harvest sprouting (PHS) resistance. However, the molecular mechanisms underlying SD remain incompletely understood. Here, we identified a wheat cytochrome P450 gene, TaCYP94-A1, that is expressed at significantly higher levels in weak-dormancy varieties than in strong-dormancy varieties. TaCYP94-A1 expression increased during SD release and decreased during dormancy establishment. Knockout of TaCYP94-A1 markedly enhanced SD and PHS resistance without adversely affecting yield-related traits. Two key single-nucleotide polymorphisms (T/C at -1,895 bp and T/C at -1,225 bp) in the TaCYP94-A1 promoter were significantly associated with SD variation, with the TaCYP94-A1 and TaCYP94-A1 allele combination (haplotype Hap4) strongly associated with enhanced dormancy. Two transcription factors, TaABI4 and TaNAC-A1, bind directly to the 5'-ACCGC-3' (C, -1,895 bp) and 5'-GACTTC-3' (C, -1,225 bp) motifs in the TaCYP94-A1 promoter, respectively, and regulate its transcription through antagonistic protein-protein interactions in the nucleus. Physiological, biochemical, and gene expression analyses revealed that the TaABI4/TaNAC-A1-TaCYP94-A1 module regulates SD through crosstalk with the gibberellic acid, abscisic acid, and jasmonic acid pathways. Together, these findings uncover a previously uncharacterized regulatory module controlling SD and provide valuable genetic resources and molecular markers for developing PHS-resistant wheat cultivars through molecular design breeding.
The rice blast fungal effector AVR-PikC binds to the rice protein HIPP19, which may contribute to plant susceptibility. The compound B93 induces the interaction between the rice E3 ligase APIP6 and AVR-PikC, which result...The rice blast fungal effector AVR-PikC binds to the rice protein HIPP19, which may contribute to plant susceptibility. The compound B93 induces the interaction between the rice E3 ligase APIP6 and AVR-PikC, which results in the ubiquitination and degradation of AVR-PikC, thereby facilitating plant resistance.
The brown planthopper (Nilaparvata lugens Stål, BPH) is a major rice pest that feeds on sieve tubes, where plants respond by depositing callose to restrict phloem sap ingestion. However, the molecular basis of how rice s...The brown planthopper (Nilaparvata lugens Stål, BPH) is a major rice pest that feeds on sieve tubes, where plants respond by depositing callose to restrict phloem sap ingestion. However, the molecular basis of how rice stabilizes callose at plasmodesmata and how BPH overcomes this defense remains poorly understood. Here, we identify OsPDCB1, a plasmodesmal callose-binding protein that positively regulates BPH resistance by anchoring callose through its X8 domain. Loss- and gain-of-function analyses demonstrate that OsPDCB1 is essential for callose accumulation and effective phloem defense. We further identified NlVRSP1, a BPH salivary effector that is highly conserved across rice planthopper species. This effector directly interacts with OsPDCB1 and disrupts its callose-binding activity, revealing a previously uncharacterized effector-host interaction module at the plasmodesmal interface. Importantly, haplotype analysis uncovered a resistance-associated allele (OsPDCB1), enriched in Indica rice, which enhances resistance when introgressed into susceptible Japonica backgrounds. Collectively, these findings identify OsPDCB1 as a key mediator of callose-based defense and a promising genetic target for breeding BPH-resistant rice cultivars, while providing mechanistic insight into how insect effectors subvert plasmodesmal immunity.
O-Methyltransferases (OMTs) play crucial roles in plant defense, environmental adaptation, and quality formation by catalyzing the biosynthesis of diverse methylated metabolites. Although OMT (COMT and CCoAOMT) genes hav...O-Methyltransferases (OMTs) play crucial roles in plant defense, environmental adaptation, and quality formation by catalyzing the biosynthesis of diverse methylated metabolites. Although OMT (COMT and CCoAOMT) genes have been functionally characterized in various plant species, the evolutionary trajectory of the entire OMT gene family and the functional divergence of the CCoAOMT subfamily remain to be systematically elucidated. In this study, we performed pan-genome analysis of the OMT gene family in 61 tomato (Solanum spp.) accessions and conducted phylogenetic analysis across 20 plant species (from algae to angiosperms), identifying 2,882 OMT genes. Phylogenetic reconstruction revealed that all extant plant CCoAOMT genes evolved from a single ancestral lineage (Clade I) originating before the divergence of red and green algae. In tomato, 2,199 OMT genes were classified into 42 orthogroups: nine core, five soft-core, 22 dispensable, and six private orthogroups, with 52.4% classified as dispensable genes. OMT genes in the Solanum genus have predominantly undergone purifying selection. Among all COMT orthogroups, a single tandem duplicate cluster stands out as exclusively conserved. Members of this cluster have evolved a distinct catalytic role, as evidenced by the finding that SlCOMT2c exclusively catalyzes the formation of kaempferide via the 4'-O-methylation of kaempferol. Ion mobility spectrometry showed that SlAOMT, a member of the CCoAOMT-like subfamily, catalyzes the methylation of luteolin to produce two isomeric products identified as diosmetin and chrysoeriol while losing the canonical catalytic function of the CCoAOMT subfamily. In addition, we identified a potential gene regulatory network associated with methylated flavonoid biosynthesis. This study establishes an integrative framework for elucidating OMT evolution and provides analytical tools for identifying genes involved in isomeric methylated flavonoid biosynthesis, paving the way for studying adaptive evolution and specialized metabolic pathways in plants.
The BARELY ANY MERISTEM 1-AVRPPHB SUSCEPTIBLE 1 (PBS1) module is an early plasma membrane switch activating reactive oxygen species-dependent heat responses. PBS1 may integrate heat and immunity, offering targets for bre...The BARELY ANY MERISTEM 1-AVRPPHB SUSCEPTIBLE 1 (PBS1) module is an early plasma membrane switch activating reactive oxygen species-dependent heat responses. PBS1 may integrate heat and immunity, offering targets for breeding climate-resilient crops.
The selective degradation of aberrant mRNAs plays a vital role in ensuring cellular survival under stress conditions. Here, we investigated the role of OsFKBP20-1b, a splicing factor, in dehydration stress response in ri...The selective degradation of aberrant mRNAs plays a vital role in ensuring cellular survival under stress conditions. Here, we investigated the role of OsFKBP20-1b, a splicing factor, in dehydration stress response in rice (Oryza sativa). We show that OsFKBP20-1b associates with the core nonsense-mediated mRNA decay (NMD) components, UP-FRAMESHIFT1 (OsUPF1) and OsUPF2, enhances their stability, thereby supporting the efficient degradation of aberrant transcripts during dehydration stress. These associations were demonstrated using bimolecular fluorescence complementation (BiFC), co-immunoprecipitation (Co-IP), and in vitro binding assays. Integrative analyses combining ribosome profiling and transcriptome sequencing further revealed that OsFKBP20-1b influences both alternative splicing (AS) patterns and translational dynamics of stress-responsive transcripts. Notably, loss of OsFKBP20-1b compromises OsUPF1- and OsUPF2-mediated decay of aberrant mRNAs under dehydration conditions. Consistent with these molecular defects, osfkbp20-1b mutant plants exhibited heightened sensitivity to dehydration stress. Together, our findings identify OsFKBP20-1b as a key regulator linking pre-mRNA splicing with cytoplasmic RNA surveillance during dehydration stress, thereby providing mechanistic insight into post-transcriptional control of stress adaptation in rice. These results advance our understanding of RNA quality control pathways in plants and suggest potential molecular targets for improving drought-resilience in crops.
Cadmium (Cd) is a toxic heavy metal that poses serious risks to human health and the ecological environment. Perilla frutescens (L.) Britt. has important medicinal and culinary value, yet its seedlings are highly sensiti...Cadmium (Cd) is a toxic heavy metal that poses serious risks to human health and the ecological environment. Perilla frutescens (L.) Britt. has important medicinal and culinary value, yet its seedlings are highly sensitive to cadmium exposure. Carbohydrates, which mediate key aspects of plant-microbe interactions, play an essential role in recruiting rhizosphere microbiota. In this study, we examined how inoculation with Oceanobacillus picturae alleviates cadmium toxicity by secreting carbohydrate metabolites that reshape the rhizosphere microbial community of perilla. Inoculation markedly reduced cadmium-induced root damage, increasing fresh and dry plant weights by 2.3-fold and 1.1-fold, and enhancing root length by 14% compared with the control. In addition, root exudate profiles showed clear changes following inoculation. Metabolomic analyses revealed that stachyose was a key exudate enriched under stress conditions and acted synergistically with Azospirillum brasilense and Acinetobacter pittii to enhance perilla growth and cadmium tolerance. These findings demonstrate that perilla recruits specific plant growth-promoting rhizobacteria through stachyose-mediated chemical signaling in response to cadmium stress. This work advances our understanding of plant-microbe interactions under heavy metal stress and provides a foundation for microbiome-based phytoremediation technologies. It also offers practical value for developing sustainable agricultural practices and supporting ecological conservation.
A strategy coupling high-activity nucleases with dimeric TadA-8e optimizes plant Cas12-based adenine base editors, boosts editing efficiency, and provides precise editors for crop breeding and genomics research.A strategy coupling high-activity nucleases with dimeric TadA-8e optimizes plant Cas12-based adenine base editors, boosts editing efficiency, and provides precise editors for crop breeding and genomics research.
Du C, Yun Y, Li W
… +10 more, Wu X, Liu X, Li M, Li Y, Wang S, Li W, He Q, Gong Z, Du H, Sun Q
J Integr Plant Biol
· 2026 May · PMID 41663343
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Salt stress is a major abiotic constraint limiting global crop production. Oat (Avena sativa L.), an allohexaploid cereal renowned for robust stress tolerance, remains poorly understood in terms of the molecular mechanis...Salt stress is a major abiotic constraint limiting global crop production. Oat (Avena sativa L.), an allohexaploid cereal renowned for robust stress tolerance, remains poorly understood in terms of the molecular mechanisms underlying its response to salt stress. Here, we perform transcriptome profiling across multiple developmental stages and tissues of oat under salt stress, and construct the co-expression regulatory network to identify salt tolerance-associated gene modules. Notably, 10 salt-responsive transcription factor (SRTF) families with dynamic expression patterns are identified as core regulators, showing extensive subgenomic functional divergence, characterized by subgenome-dominant expression, as well as subgenome-specific duplication or loss events. Further integration with a genome-wide association study (GWAS) of the germination rate under salt stress in 225 oat accessions identified a 3-bp InDel variation within the duplicated gene AsWRKY49-D2, which specifically modulates its expression by facilitating binding of the TF AsZAT18, with AsWRKY49-D2 further mediating oat salt tolerance through targeted regulation of AsSOS2 and AsSOS3. Intriguingly, the salt-tolerant allele of AsWRKY49 is scarcely distributed in Chinese oat accessions, highlighting its considerable potential for breeding application. These results shed light on the regulatory mechanisms underlying oat salt tolerance, providing valuable information for exploring salt tolerance genes and breeding new salt-tolerant oat varieties.
Despite decades of research on NAC-MYB master regulators, the transcription factor (TF)-mediated network governing wood development remains fragmented, hindering a systems-level understanding of secondary xylem formation...Despite decades of research on NAC-MYB master regulators, the transcription factor (TF)-mediated network governing wood development remains fragmented, hindering a systems-level understanding of secondary xylem formation. In this study, we identified GROWTH REGULATING FACTOR 20 (PagGRF20) as a key regulator within the poplar wood formation network. Compared to wild-type (WT) poplars, the PagGRF20-overexpressing lines showed a 22.1%-23.9% increase in secondary xylem thickness, a 37.7%-43.3% increase in cell wall thickness, and higher cellulose and xylan content. These phenotypic changes coincided with upregulation of genes involved in cellulose and xylan biosynthetic pathways. Conversely, PagGRF20-RNAi lines displayed opposite phenotypic traits. Molecular analyses revealed that PagGRF20 binds to the TGT[C/T]AGA cis-regulatory elements in the promoters of wood polymer biosynthesis genes, modulating polysaccharide production by activating the glycosyltransferase IRREGULAR XYLEM 14-LIKE (PagIRX14L). Crucially, overexpression of PagIRX14L in poplar increased the xylan content and decreased lignin levels, suggesting that PagGRF20-mediated transcriptional activation of PagIRX14L significantly contributes to the observed shifts in cell wall composition. Additionally, the R2R3-MYB repressor PagMYB4 physically interacts with PagGRF20, forming a transcriptional complex that suppresses lignin biosynthesis genes, including LACCASE 11 (PagLAC11), potentially altering the balance of cell wall components and affecting cellulose and xylan accumulation. Finally, we built a three-layer, PagGRF20-centered regulatory network to uncover the mechanisms and target genes underlying wood formation. Our results suggest that the PagGRF20-PagMYB4 module coordinates secondary cell wall (SCW) biosynthetic pathways, providing novel insights into engineering wood with enhanced polysaccharide deposition and low lignin content.
Plant viruses frequently reprogram conserved growth-defense regulatory hubs to promote infection. Here, we show that the rice grassy stunt virus (RGSV) suppresses salicylic acid (SA)-mediated antiviral immunity by target...Plant viruses frequently reprogram conserved growth-defense regulatory hubs to promote infection. Here, we show that the rice grassy stunt virus (RGSV) suppresses salicylic acid (SA)-mediated antiviral immunity by targeting the miR156-SPL-ICS1 module. The viral effector P3 directly binds a conserved 12-bp cis-element in the miR156a promoter, activating its transcription and increasing miR156 accumulation. Increased miR156 represses SPL14 and SPL17 transcripts, while RGSV infection is also associated with a pronounced reduction in SPL14/17 protein abundance. P3 physically associates with SPL14 and SPL17, indicating an additional post-transcriptional layer contributing to SPL attenuation. Genetic analyses demonstrate that SPL14 and SPL17 positively regulate ICS1, a key enzyme in SA biosynthesis, and that loss of SPL14/17 function compromises SA accumulation and antiviral defense. Conversely, overexpression of SPL14 or SPL17 mitigates RGSV symptoms and restricts viral accumulation, whereas exogenous SA restores immunity and partially rescues disease-associated architectural defects. Together, our findings reveal a dual-layer virulence strategy in which RGSV P3 coordinately suppresses the miR156-SPL14/17-ICS1 pathway at transcriptional and post-transcriptional levels, uncovering a central regulatory node that links rice development and antiviral immunity and providing actionable targets for engineering RGSV-resistant rice.
THESEUS1 (THE1) is a component of the CrRLK1L-RALF signaling complex specifically responsible for establishing the polytubey block at the Arabidopsis septum. Genetic and biochemical analyses demonstrate that THE1, togeth...THESEUS1 (THE1) is a component of the CrRLK1L-RALF signaling complex specifically responsible for establishing the polytubey block at the Arabidopsis septum. Genetic and biochemical analyses demonstrate that THE1, together with FERONIA, ANJ and HERK1, forms a receptor complex that senses pollen tube-derived RALF peptides, thereby establishing a barrier to prevent the emergence of multiple pollen tubes from the septum.