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Iron (Fe) deficiency in calcareous soils is a significant agricultural challenge, affecting crop productivity and nutritional quality. This study aimed to isolate, characterize, and evaluate Fe solubilizing rhizobacterial isolates from maize rhizosphere in calcareous soils as potential biofertilizers. Forty bacterial isolates coded as SG1, SG2, …, SG40 were isolated and screened for siderophore production, with ten showing significant Fe solubilizing capabilities. These isolates were further assessed for phosphate solubilization and exopolysaccharides production. The selected bacterial isolates were also screened under axenic conditions for their ability to improve maize growth. The isolates SG8, SG13, SG24, SG30 and SG33 significantly enhanced growth parameters of maize. Notably, SG30 showed highest increment in shoot length (58%), root length (54%), root fresh and dry biomass (67% and 76%), SPAD value (67%), relative water contents (69%), root surface area (61%), and Fe concentration in shoots (79%) as compared to control. The biochemical characterization of these strains showed that all these strains have capability to solubilize insoluble phosphorus, produce indole-3-acetic acid (IAA), and ammonia with catalase, urease and protease activity. Molecular identification through 16s rRNA gene sequencing confirmed high similarity (99.7-100%) of the selected isolates to various Bacillus species, including B. pyramidoids, B. firmicutes, and B. cereus. The study provides a strong base for developing eco-friendly, cost-effective biofertilizers to address Fe deficiency in crops and promote sustainable agriculture., (© 2024. The Author(s).)

Appropriate abiotic stress response is pivotal for plant survival and makes use of multiple signaling molecules and phytohormones to achieve specific and fast molecular adjustments. A multitude of studies has highlighted the role of alternative splicing in response to abiotic stress, including temperature, emphasizing the role of transcriptional regulation for stress response. Here we investigated the role of the core-splicing factor PORCUPINE (PCP) on temperature-dependent root development. We used marker lines and transcriptomic analyses to study the expression profiles of meristematic regulators and mitotic markers, and chemical treatments, as well as root hormone profiling to assess the effect of auxin signaling. The loss of PCP significantly alters RAM architecture in a temperature-dependent manner. Our results indicate that PCP modulates the expression of central meristematic regulators and is required to maintain appropriate levels of auxin in the RAM. We conclude that alternative pre-mRNA splicing is sensitive to moderate temperature fluctuations and contributes to root meristem maintenance, possibly through the regulation of phytohormone homeostasis and meristematic activity., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Plant root system is significantly influenced by high soil levels of ammonium nitrogen, leading to reduced root elongation and enhanced lateral root branching. In Arabidopsis, these processes have been reported to be mediated by phytohormones and their downstream signaling pathways, while the controlling mechanisms remain elusive in crops. Through a transcriptome analysis of roots subjected to high/low ammonium treatments, we identified a cytokinin oxidase/dehydrogenase encoding gene, CKX3, whose expression is induced by high ammonium. Knocking out CKX3 and its homologue CKX8 results in shorter seminal roots, fewer lateral roots, and reduced sensitivity to high ammonium. Endogenous cytokinin levels are elevated by high ammonium or in ckx3 mutants. Cytokinin application results in shorter seminal roots and fewer lateral roots in wild-type, mimicking the root responses of ckx3 mutants to high ammonium. Furthermore, CKX3 is transcriptionally activated by type-B RR25 and RR26, and ckx3 mutants have reduced auxin content and signaling in roots under low ammonium. This study identified RR25/26-CKX3-cytokinin as a signal module that mediates root responses to external ammonium by modulating of auxin signaling in the root meristem and lateral root primordium. This highlights the critical role of cytokinin metabolism in regulating rice root development in response to ammonium., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Extracellular auxin maxima and minima are important to control plant developmental programs. Auxin gradients are provided by the concerted action of proteins from the three major plasma membrane (PM) auxin transporter classes AUX1/LAX, PIN and ATP-BINDING CASSETTE subfamily B (ABCB) transporters. But neither genetic nor biochemical nor modeling approaches have been able to reliably assign the individual roles and interplay of these transporter types. Based on the thermodynamic properties of the transporters, we show here by mathematical modeling and computational simulations that the concerted action of different auxin transporter types allows the adjustment of specific transmembrane auxin gradients. The dynamic flexibility of the 'auxin homeostat' comes at the cost of an energy-consuming 'auxin cycling' across the membrane. An unexpected finding was that potential functional ABCB-PIN synchronization appears to allow an optimization of the trade-off between the speed of PM auxin gradient adjustment on the one hand and ATP consumption and disturbance of general anion homeostasis on the other. In conclusion, our analyses provide fundamental insights into the thermodynamic constraints and flexibility of transmembrane auxin transport in plants., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Hydathodes are small organs found on the leaf margins of vascular plants which release excess xylem sap through a process called guttation. While previous studies have hinted at additional functions of hydathode in metabolite transport or auxin metabolism, experimental support is limited. We conducted comprehensive transcriptomic, metabolomic and physiological analyses of mature Arabidopsis hydathodes. This study identified 1460 genes differentially expressed in hydathodes compared to leaf blades, indicating higher expression of most genes associated with auxin metabolism, metabolite transport, stress response, DNA, RNA or microRNA processes, plant cell wall dynamics and wax metabolism. Notably, we observed differential expression of genes encoding auxin-related transcriptional regulators, biosynthetic processes, transport and vacuolar storage supported by the measured accumulation of free and conjugated auxin in hydathodes. We also showed that 78% of the total content of 52 xylem metabolites was removed from guttation fluid at hydathodes. We demonstrate that NRT2.1 and PHT1;4 transporters capture nitrate and inorganic phosphate in guttation fluid, respectively, thus limiting the loss of nutrients during this process. Our transcriptomic and metabolomic analyses unveil an organ with its specific physiological and biological identity., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Lignin, a complex heterogenous polymer present in virtually all plant cell walls, plays a critical role in protecting plants from various stresses. However, little is known about how lignin modifications in sorghum will impact plant defense against sugarcane aphids (SCA), a key pest of sorghum. We utilized the sorghum brown midrib (bmr) mutants, which are impaired in monolignol synthesis, to understand sorghum defense mechanisms against SCA. We found that loss of Bmr12 function and overexpression (OE) of Bmr12 provided enhanced resistance and susceptibility to SCA, respectively, as compared with wild-type (WT; RTx430) plants. Monitoring of the aphid feeding behavior indicated that SCA spent more time in reaching the first sieve element phase on bmr12 plants compared with RTx430 and Bmr12-OE plants. A combination of transcriptomic and metabolomic analyses revealed that bmr12 plants displayed altered auxin metabolism upon SCA infestation and specifically, elevated levels of auxin conjugate indole-3-acetic acid-aspartic acid (IAA-Asp) were observed in bmr12 plants compared with RTx430 and Bmr12-OE plants. Furthermore, exogenous application of IAA-Asp restored resistance in Bmr12-OE plants, and artificial diet aphid feeding trial bioassays revealed that IAA-Asp is associated with enhanced resistance to SCA. Our findings highlight the molecular underpinnings that contribute to sorghum bmr12-mediated resistance to SCA., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

The mechanisms by which low light accelerates starch macromolecules degradation by auxin and gibberellin (GA) in geophytes during sprouting remain largely unknown. This study investigated these mechanisms in saffron, grown under low light (50 μmol m -2  s -1 ) and optimal light (200 μmol m -2  s -1 ) during the sprouting phase. Low light reduced starch concentration in corms by 34.0 % and increased significantly sucrose levels in corms, leaves, and leaf sheaths by 19.2 %, 9.8 %, and 134.5 %, respectively. This was associated with a 33.3 % increase in GA 3 level and enhanced auxin signaling. Leaves synthesized IAA under low light, which was transported to the corms to promote GA synthesis, facilitating starch degradation through a 228.7 % increase in amylase activity. Exogenous applications of GA and IAA, as well as the use of their synthesis or transport inhibitors, confirmed the synergistic role of these phytohormones in starch metabolism. The unigenes associated with GA biosynthesis and auxin signaling were upregulated under low light, highlighting the IAA-GA module role in starch degradation. Moreover, increased respiration rate and invertase activity, crucial for ATP biosynthesis and the tricarboxylic acid cycle, were consistent with the upregulation of related unigenes, suggesting that auxin signaling accelerates starch degradation by promoting energy metabolism. Upregulated of auxin signaling (CsSAUR32) and starch metabolism (CsSnRK1) genes under low light suggests that auxin directly regulate starch degradation in saffron corms. This study elucidates that low light modulates auxin and GA interactions to accelerate starch degradation in saffron corms during sprouting, offering insights for optimizing agricultural practices under suboptimal light conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Phloretin has different glycosylation modes in plants. Phlorizin (phloretin 2'-O-glucoside) is one of the glycosylation products of phloretin, and accumulates abundantly in apple plants. However, it is still unclear whether phlorizin is more beneficial for apple plants compared with other glycosylation products of phloretin. We created transgenic apple plants with different glycosylation modes of phloretin. In transgenic plants, the accumulation of phlorizin was partly replaced by that of trilobatin (phloretin 4'-O-glucoside) or phloretin 3',5'-di-C-glycoside. Compared with wild type, transgenic plants with less phlorizin showed dwarf phenotype, larger stomatal size, higher stomatal density and less tolerance to drought stress. Transcriptome and phytohormones assay indicate that phlorizin might regulate stomatal development and behaviour via controlling auxin and abscisic acid signalling pathways as well as carbonic anhydrase expressions. Transgenic apple plants with less phlorizin also showed less resistance to spider mites. Apple plants may hydrolyse phlorizin to produce phloretin, but cannot hydrolyse trilobatin or phloretin 3',5'-di-C-glycoside. Compared with its glycosylation products, phloretin is more toxic to spider mites. These results suggest that the glycosylation of phloretin to produce phlorizin is the optimal glycosylation mode in apple plants, and plays an important role in apple resistance to stresses., (© 2024 John Wiley & Sons Ltd.)

Most agricultural products are presently cultivated on marginal lands with poor soil properties and unfavorable environmental conditions (diseases and abiotic stresses), which can threaten plant growth and yield. Plant growth-promoting bacteria (PGPB) are beneficial bacteria that promote plant growth and biomass and act as biocontrols against diseases and stress. However, most isolated PGPBs have a single function and low survival rates owing to their limited growth behaviors. In this study, we isolated multifunctional PGPB from oil palm rhizosphere, quantitatively measured their activities, and evaluated their effectiveness in Brassica rapa (Komatsuna) cultivation. This is the first study to report the isolation of three multifunctional PGPB strains with ammonium production, phosphate-potassium-silicate solubilization, and indole-3-acetic acid (IAA) production from the oil palm rhizosphere, namely Kosakonia oryzendophytica AJLB38, Enterobacter quasimori AJTS77, and Lelliottia jeotgali AJTS83. Additionally, these strains showed antifungal activity against the oil palm pathogen Ganoderma boninense. These strains grow under high temperature, acidic and alkaline pH, and high salt concentration, which would result in their proliferation in various environmental conditions. The cultivation experiments revealed these strains improved the growth and biomass with half the dosage of chemical fertilizer application, which was not significantly different to the full dosage. Furthermore, the overall plant growth-promoting activities in quantitative assays and overall B. rapa growth in cultivation experiments were statistically correlated, which could contribute to the prediction of plant growth promotion without plant cultivation experiments. Thus, the selected PGPB could be valuable as a biofertilizer to improve soil health and quality and promote agricultural sustainability., (Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Perennial trees have a recurring annual cycle of wood formation in response to environmental fluctuations. However, the precise molecular mechanisms that regulate the seasonal formation of wood remain poorly understood. Our prior study indicates that VCM1 and VCM2 play a vital role in regulating the activity of the vascular cambium by controlling the auxin homoeostasis of the cambium zone in Populus. This study indicates that abscisic acid (ABA) affects the expression of VCM1 and VCM2, which display seasonal fluctuations in relation to photoperiod changes. ABA-responsive transcription factors AREB4 and AREB13, which are predominantly expressed in stem secondary vascular tissue, bind to VCM1 and VCM2 promoters to induce their expression. Seasonal changes in the photoperiod affect the ABA amount, which is linked to auxin-regulated cambium activity via the functions of VCM1 and VCM2. Thus, the study reveals that AREB4/AREB13-VCM1/VCM2-PIN5b acts as a molecular module connecting ABA and auxin signals to control vascular cambium activity in seasonal wood formation., (© 2024 John Wiley & Sons Ltd.)

Background: Soybean seeds are rich in protein and oil. The selection of varieties that produce high-quality seeds has been one of the priorities of soybean breeding programs. However, the influence of improved seed quality on the rhizosphere microbiota and whether the microbiota is involved in determining seed quality are still unclear. Here, we analyzed the structures of the rhizospheric bacterial communities of 100 soybean varieties, including 53 landraces and 47 modern cultivars, and evaluated the interactions between seed quality traits and rhizospheric bacteria., Results: We found that rhizospheric bacterial structures differed between landraces and cultivars and that this difference was directly related to their oil content. Seven bacterial families (Sphingomonadaceae, Gemmatimonadaceae, Nocardioidaceae, Xanthobacteraceae, Chitinophagaceae, Oxalobacteraceae, and Streptomycetaceae) were obviously enriched in the rhizospheres of the high-oil cultivars. Among them, Oxalobacteraceae (Massilia) was assembled specifically by the root exudates of high-oil cultivars and was associated with the phenolic acids and flavonoids in plant phenylpropanoid biosynthetic pathways. Furthermore, we showed that Massilia affected auxin signaling or interfered with active oxygen-related metabolism. In addition, Massilia activated glycolysis pathway, thereby promoting seed oil accumulation., Conclusions: These results provide a solid theoretical basis for the breeding of revolutionary soybean cultivars with desired seed quality and optimal microbiomes and the development of new cultivation strategies for increasing the oil content of seeds. Video Abstract., (© 2024. The Author(s).)

Background: Babesia duncani is a pathogen within the phylum Apicomplexa that causes human babesiosis. It poses a significant threat to public health, as it can be transmitted not only through tick bites but also via blood transfusion. Consequently, an understanding of the gene functions of this pathogen is necessary for the development of drugs and vaccines. However, the absence of conditional gene knockdown tools has hindered the research on this pathogen. The auxin-inducible degron (AID) system is a rapid, reversible conditional knockdown system widely used in gene function studies. Thus, there is an urgent need to establish the AID system in B. duncani to study essential gene functions., Methods: The endogenous genes of the Skp1-Cullin-F-box (SCF) complex in B. duncani were identified and confirmed through multiple sequence alignment and conserved domain analysis. The expression of the F-box protein TIR1 from Oryza sativa (OsTIR1) was achieved by constructing a transgenic parasite strain using a homologous recombination strategy. Polymerase chain reaction (PCR), western blot, and indirect immunofluorescence assay (IFA) were used to confirm the correct monoclonal parasite strain. The degradation of enhanced green fluorescent protein (eGFP) tagged with an AID degron was detected through western blot and live-cell fluorescence microscopy after treatment of indole-3-acetic acid (IAA)., Results: In this study, Skp1, Cul1, and Rbx1 of the SCF complex in B. duncani were identified through sequence alignment and domain analysis. A pure BdTIR1 strain with expression of the OsTIR1 gene was constructed through homologous recombination and confirmed. This strain showed no significant differences from the wild type (WT) in terms of growth rate and proportions of different parasite forms. The eGFP tagged with an AID degron was successfully induced for degradation using 500 μM IAA. Grayscale analysis of western blot indicated a 61.3% reduction in eGFP expression levels, while fluorescence intensity analysis showed a 77.5% decrease in fluorescence intensity. Increasing the IAA concentration to 2 mM accelerated eGFP degradation and enhanced the extent of degradation., Conclusions: This study demonstrated the functionality of the AID system in regulating protein levels by inducing rapid degradation of eGFP using IAA, providing an important research tool for studying essential gene functions related to invasion, egress, and virulence of B. duncani. Moreover, it also offers a construction strategy for apicomplexan parasites that have not developed an AID system., (© 2024. The Author(s).)

Plant growth-promoting rhizobacteria (PGPR) can promote lateral root formation, while the underlying mechanisms are not fully understood. Here, we found that Pseudomonas chlororaphis subsp. aurantiaca inoculation enhanced auxin accumulation in lateral root primordia (LRP). Upon reaching LRPs, auxin activated the AUXIN RESPONSE FACTOR 7 and 19 (ARF7/19) and promoted lateral root formation in Arabidopsis . Moreover, we found that reactive oxygen species (ROS) is required for auxin-dependent lateral root emergence, and P. chlororaphis upregulated the expression of RESPIRATORY BURST OXIDASE D and F ( RBOHD / F ), leading to the accumulation of ROS in LRP. Although scavenging ROS or rbohd / f mutants exhibited decreased lateral roots after P. chlororaphis inoculation, the bacteria-triggered auxin signals were not altered. Conversely, the application of auxin or mutants defective in auxin signaling disturbed P. chlororaphis -derived ROS accumulation in lateral roots. Collectively, these results suggest that ARF7/19-dependent auxin signaling activates RBOHD / F to produce ROS, coordinately facilitating lateral root development after P. chlororaphis treatment.

Plants perceive and orchestrate defense responses when herbivorous insects are ovipositing. Fruits, as a crucial reproductive organ in plants, have rarely been researched on the responses to insect eggs. Here, we found that oviposition by the specialist insect Bactrocera minax in navel oranges activated the lignin synthesis pathway and cell division, causing mechanical pressure that crushed the eggs. Transcriptome and metabolome analyses revealed an enrichment of oviposition-induced genes and metabolites within the lignin synthesis pathway, which was confirmed by histochemical staining. Furthermore, hydrogen peroxide (H 2 O 2 ) accumulation was observed at the oviposition sites. Plant defense-related hormones jasmonic acid (JA) and salicylic acid (SA) exhibited rapid induction after oviposition, while indole-3-acetic acid (IAA) activation occurred in the later stages of oviposition. Additionally, secondary metabolites induced by prior egg deposition were found to influence larval performance. Our studies provide molecular evidence that host fruits have evolved defense mechanisms against insect eggs and pave the way for future development of insect-resistant citrus varieties.

Topping, an important tree shaping and pruning technique, can promote the outgrowth of citrus axillary buds. However, the underlying molecular mechanism is still unclear. In this study, spring shoots of Citrus reticulata 'Huagan No.2' were topped and transcriptome was compared between axillary buds of topped and untopped shoots at 6 and 11 days after topping (DAT). 1944 and 2394 differentially expressed genes (DEGs) were found at 6 and 11 DAT, respectively. KEGG analysis revealed that many DEGs were related to starch and sucrose metabolism, signal transduction of auxin, cytokinin and abscisic acid. Specially, transcript levels of auxin synthesis, transport, and signaling-related genes (SAURs and ARF5), cytokinin signal transduction related genes (CRE1, AHP and Type-A ARRs), ABA signal responsive genes (PYL and ABF) were up-regulated by topping; while transcript levels of auxin receptor TIR1, auxin responsive genes AUX/IAAs, ABA signal transduction related gene PP2Cs and synthesis related genes NCED3 were down-regulated. On the other hand, the contents of sucrose and fructose in axillary buds of topped shoots were significantly higher than those in untopped shoots; transcript levels of 16 genes related to sucrose synthase, hexokinase, sucrose phosphate synthase, endoglucanase and glucosidase, were up-regulated in axillary buds after topping. In addition, transcript levels of genes related to trehalose 6-phosphate metabolism and glycolysis/tricarboxylic acid (TCA) cycle, as well to some transcription factors including Pkinase, Pkinase_Tyr, Kinesin, AP2/ERF, P450, MYB, NAC and Cyclin_c, significantly responded to topping. Taken together, the present results suggested that topping promoted citrus axillary bud outgrowth through comprehensively regulating plant hormone and carbohydrate metabolism, as well as signal transduction. These results deepened our understanding of citrus axillary bud outgrowth by topping and laid a foundation for further research on the molecular mechanisms of citrus axillary bud outgrowth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sugarcane (Saccharum spp.) growth is regulated by intricate gene networks and hormone secretions, positively correlating with sugarcane yield. There is a rising interest in exploring how the candidate genes found in sugarcane respond to plant growth. In this study, we simulated a typical growth environment to obtain accurate phenotypic data and screened for potential genes associated with plant growth through transcriptomics. Compared to Saccharum GuiTang 42, the other variety Saccharum GuiTang 44 exhibited earlier germination, a higher emergence rate, thicker pseudostems, taller plants, and a more extensive root system. The middle buds formed the greatest number of roots, followed by the lower and upper buds. Indole-3-acetic acid (IAA) and jasmonic acid effectively promoted bud development, while abscisic acid and trans-zeatin exhibited negative correlations with sugarcane bud growth. Transcriptome data from the upper, middle, and lower buds revealed 24,158 differentially expressed genes in all three comparisons, with MAPK signaling emerging as a critical pathway. The photosynthesis-antenna protein pathway is vital for middle and lower bud development during root germination. Lastly, key gene modules related to differences in hormone content between the two varieties were defined through weighted correlation network analysis and identified. The module significantly associated with IAA was enriched in pathways such as Proteasome and Protein processing in the endoplasmic reticulum, and the upregulation of key genes involved in this gene module had a highly significant positive correlation with bud outgrowth combined with IAA secretion. In conclusion, we have elucidated the pathways of hormones during sugarcane growth and the interactions between IAA and critical genes. These in-depth findings may guide modern sugarcane breeding., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Main Conclusion: TaZAT8-5B, a C2H2 zinc finger protein transcription factor, positively regulates drought tolerance in transgenic Arabidopsis. It promotes root growth under drought stress via the Aux/IAA-ARF module in the auxin signaling pathway. C2H2 zinc finger proteins (C2H2-ZFPs) represent the largest but relatively unexplored family of transcription factors in plants. This is particularly evident in wheat, where the functions of only a few C2H2-ZFP genes have been confirmed. In this study, we identified a novel C2H2-ZFP gene, TaZAT8-5B. This gene shows high expression in roots and flowers and is significantly induced by heat, drought, and salt stress. Under drought stress, overexpressing TaZAT8-5B in Arabidopsis resulted in increased proline content and superoxide dismutase (SOD) activity in leaves. It also led to reduced stomatal aperture and water loss, while inducing the expression of P5CS1, RD29A, and DREB1A. Consequently, it alleviated drought stress-induced malondialdehyde (MDA) accumulation and improved drought tolerance. Additionally, TaZAT8-5B promoted lateral root initiation under mannitol stress and enhanced both lateral and primary root growth under long-term drought stress. Moreover, TaZAT8-5B was induced by indole-3-acetic acid (IAA). Overexpressing TaZAT8-5B under drought stress significantly inhibited the expression of auxin signaling negative regulatory genes IAA12 and IAA14. Conversely, downstream genes (ARF7, LBD16, LBD18, and CDKA1) of IAA14 and IAA12 were upregulated in TaZAT8-5B overexpressing plants compared to wild-type (WT) plants. These findings suggest that TaZAT8-5B regulates root growth and development under drought stress via the Aux/IAA-ARF module in the auxin signaling pathway. In summary, this study elucidates the role of TaZAT8-5B in enhancing drought tolerance and its involvement in root growth and development through the auxin signaling pathway. These findings offer new insights into the functional analysis of homologous genes of TaZAT8-5B, particularly in Gramineae species., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

To explore the role and metabolic differences of plant hormones in regulating the flower bud size of daylily, we collected the flower buds from two daylily varieties 'Datong Huanghua' and 'Dongbei Huanghua' at the young bud, green, yellowing, and yellow stages for transcriptome sequencing. The differentially expressed genes (DEGs) were screened, and the exogenous plant hormone spraying experiments were conducted. A total of 199 DEGs related to the biosynthesis and metabolism of plant hormones was screened out at different flower development stages of 'Datong Huanghua' and 'Dongbei Huanghua'. These genes regulated the biosynthesis and metabolism of six plant hormones: abscisic acid, gibberellin, auxin, jasmonate, cytokinin, and ethylene. The DEGs associated with auxin were the most, which suggested that auxin played a role in regulating flower bud development. The auxin response factor ( ARF ) presented up-regulated expression at all the four stages of flower bud development, indicating that ARF played a positive regulatory role throughout the flower bud development of daylily. The experiments with exogenous spraying of six hormones further verified that indole-3-acetic acid (IAA) significantly promoted the growth and increased the nutrient content in the flower buds of 'Datong Huanghua', suggesting that IAA played a role in regulating flower bud development. Our results laid a theoretical foundation for probing into the regulatory mechanism of flower bud development of 'Datong Huanghua' and 'Dongbei Huanghua'.

To explore the rice seed microbiome, our objective was to isolate novel strains of Xanthomonas, a plant-associated bacterium with diverse lifestyles. Four isolates, anticipated to be Xanthomonas based on morphological features of yellow colonies, were obtained from healthy rice seeds. Phylo-taxono-genomic analysis revealed that these isolates formed monophyletic lineages belonging to a novel species within the genus Luteibacter. Pairwise ortho Average Nucleotide Identity and digital DNA-DNA hybridization confirmed their distinct species status. We propose Luteibacter sahnii sp. nov. as a novel species, with PPL193 T  = MTCC 13290 T  = ICMP 24807 T  = CFBP 9144 T as the type strain and PPL201, PPL552, and PPL554 as other constituent members. The fatty acid profile of the type strain is dominated by branched fatty acids like Iso-C 15:0 , consistent with other members of the genus. The novel species displays non-pathogenic attributes and exhibits plant probiotic properties, protecting rice plants from the leaf blight pathogen X. oryzae pv. oryzae. Production of Indole-3-Acetic Acid (IAA) and genomic regions encoding anti-microbial peptides emphasize its potential contributions to plant hosts. This study underscores the importance of employing a combination of phenotypic and genotypic methods in culturomics to enhance our understanding of rice seed microbiome diversity., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

PGPR has a higher potential impact on agricultural crops. It enhances plant growth and development in a variety of adverse environmental conditions, including biotic and abiotic stresses. The PGPR is commercially vital since it is more efficient, safe for the environment, and beneficial to the economy. Nowadays, salt stress has an impact on the agricultural ecosystem. Salt-tolerant PGPR can directly stimulate plant growth and development by producing a variety of metabolites and phytohormones. The current study looked at the isolation of salt-tolerant bacterial species and their ability to stimulate plant development. Four bacterial species were chosen for their better salt stress tolerance (0-5%). They were identified by 16S rRNA sequencing: Solibacillus silvestris BR1, Peribacillus frigoritolerans BR2, Paenibacillus taichungensis CR1, and Solibacillus isronensis CR2. These strains were positive production of indole acetic acid with varying incubation periods (19.66 ± 1.528 to 646.111 ± 8.058 µg/mL), salt stress (ranging from 29.556 ± 1.171 to 147.8111 ± 2.086 µg/mL), phosphate solubilization (0.145 ± 0.011 to 0.921 ± 0.007 µg/mL), ammonium production (0.299 ± 0.047 to 1.202 ± 0.142 µg/mL), HCN production (0.308 ± 0.051 to 4.269 ± 0.069 µg/mL), and siderophore production (0.190 ± 0.064 to 1.543 ± 0.108 µg/mL) for control strains were used without salt stress. The production level was expressed using a standard curve containing various standards., (© 2024. The Author(s).)

Plant-microbe communication involves a rich language of chemical signals. Among these signals are plant hormones such as auxins, which are primarily recognized for their roles in plant development. However, they also function in modulating plant-microbe interactions. Interestingly, many bacteria are capable of producing auxins too. Yet, the mechanisms by which auxins affect bacteria and the regulatory processes controlling their production are largely unknown. Rico-Jiménez and colleagues present new insights into the effects of the auxin indole-3-acetic acid on the physiology of the rhizobacterium Serratia plymuthica (M. Rico-Jiménez, Z. Udaondo, T. Krell, and M. A. Matilla, mSystems 9:e00165-24, 2024, https://doi.org/10.1128/msystems.00165-24). Their work provides a deeper mechanistic understanding of bacterial transcriptional responses to plant hormones and the impact on bacterial fitness in the context of the rhizosphere environment., Competing Interests: The author declares no conflict of interest.

Key Message: SiDVLs inhibit auxin signaling to regulate root growth by enhancing the expression of Aux/IAAs and reducing the protein accumulation of PINs. The DEVIL/ ROTUNDIFOLIA (DVL/RTFL), a small polypeptide family, is conserved in seed plants and important in regulating plant growth and development. However, the molecular mechanisms remain largely unknown. Here, 27 SiDVLs were identified in foxtail millet genome. Overexpression of three SiDVLs in Arabidopsis (Arabidopsis thaliana) strongly repressed the plant growth, especially the root growth. We demonstrate that overexpression of SiDVLs enhances Auxin/Indole-3-Acetic Acids (Aux/IAAs) transcription, thereby weakening auxin signaling in the roots. Furthermore, SiDVLs reduced the protein levels of the auxin transporters PIN-formed 1 (PIN1), PIN2, and PIN7 in the roots. The impaired auxin signaling reduces the cell division and elongation. In conclusion, SiDVLs suppress cell division and elongation in root by inhibiting auxin signaling and transport, which lead to the reduced root growth., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Developmental reprogramming allows for flexibility in growth and adaptation to changing environmental conditions. In plants, wounding events can result in new stem cell niches and lateral organs. Adventitious roots develop from aerial parts of the plant and are regulated by multiple stimuli, including wounding. Here, we find that Arabidopsis thaliana seedlings wounded at the hypocotyl-root junction reprogram certain pericycle cells to produce adventitious roots proximal to the wound site. We have determined that competence for this reprogramming is controlled; basal cells close to the wound site can produce adventitious roots, whereas cells distal from the wound site mostly cannot. We found that altering cytokinin response or indole-3-butyric acid (IBA)-to-(indole-3-acetic acid) IAA conversion resulted in an expanded adventitious root competence zone and delineated the connection between these pathways. Our work highlights the importance of endogenous IBA-derived auxin and its interaction with cytokinin in adventitious root formation and the regenerative properties of plants., Competing Interests: Declaration of interests L.C.S. is on the scientific advisory board of Prose Foods., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Microbial herbicides play a vital role in agricultural preservation, amid growing concerns over the ecological impact from extensive development and use of chemical herbicides. Utilizing beneficial microbial metabolites to combat weeds has become a significant focus of research. This study focused on isolating herbicidal active compounds from Bacillus altitudinis D30202 through activity-guided methods. First, the n-butanol extract (n-BE) of B. altitudinis D30202 underwent fractionation using macroporous adsorption resin D101 and Sephadex LH-20, identifying Fr. F as the most potent segment against wild oats (Avena fatua L.). Ultra-performance liquid chromatography - quadrupole time-of-flight mass spectrometry (UPLC - QTOF-MS) identified nine compounds in the active fraction Fr. F. Subsequently, three subfractions (Fr.F-1 to Fr.F-3) were derived from Fr.F via semi-preparative liquid chromatography, resulting in methyl indole-3-acetate (MeIAA) purification. MeIAA, functioning as an auxin analog, exhibited effects of indole-3-acetic acid (IAA) on wild oats' growth, with a root length median inhibitory concentration of 81.06 µg/ml. Furthermore, we assessed MeIAA's herbicidal impact on five weed species across diverse families and genera, providing a first-time analysis of MeIAA's mechanism on wild oats. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed structural damage to leaves and roots post-MeIAA treatment. MeIAA treatment increased superoxide anion and hydrogen peroxide levels in wild oat roots, alongside with elevated peroxidase (POD) and superoxide dismutase (SOD) activity, chlorophyll-degrading enzymes (Chlase, MDACase), malondialdehyde (MDA) content, and relative conductivity in leaves. Conversely, it decreased catalase (CAT) activity and chlorophyll content. Therefore, this study provides a new material source and theoretical foundation for ecologically sustainable agricultural weed control., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Reciprocal exchanges of metabolites between phytoplankton and bacteria influence the fitness of these microorganisms which ultimately shapes the productivity of marine ecosystems. Recent evidence suggests that plant growth-promoting hormones may be key metabolites within mutualistic phytoplankton-bacteria partnerships, but very little is known about the diversity of plant growth-promoting hormones produced by marine bacteria and their specific effects on phytoplankton growth. Here, we aimed to investigate the capacity of marine bacteria to produce 7 plant growth-promoting hormones and the effects of these hormones on Actinocyclus sp. growth., Results: We examined the plant growth-promoting hormone synthesis capabilities of 14 bacterial strains that enhance the growth of the common diatom Actinocyclus. Plant growth-promoting hormone biosynthesis was ubiquitous among the bacteria tested. Indeed all 14 strains displayed the genomic potential to synthesise multiple hormones, and mass-spectrometry confirmed that each strain produced at least 6 out of the 7 tested plant growth-promoting hormones. Some of the plant growth-promoting hormones identified here, such as brassinolide and trans-zeatin, have never been reported in marine microorganisms. Importantly, all strains produced the hormone indole-3 acetic acid (IAA) in high concentrations and released it into their surroundings. Furthermore, indole-3 acetic acid extracellular concentrations were positively correlated with the ability of each strain to promote Actinocyclus growth. When inoculated with axenic Actinocyclus cultures, only indole-3 acetic acid and gibberellic acid enhanced the growth of the diatom, with cultures exposed to indole-3 acetic acid exhibiting a two-fold increase in cell numbers., Conclusion: Our results reveal that marine bacteria produce a much broader range of plant growth-promoting hormones than previously suspected and that some of these compounds enhance the growth of a marine diatom. These findings suggest plant growth-promoting hormones play a large role in microbial communication and broaden our knowledge of their fuctions in the marine environment. Video Abstract., (© 2024. The Author(s).)

Key Message: Twenty-five VvIAA genes and eighteen VaIAA genes were identified from Pinot Noir and Shanputao, respectively. The overexpression of VaIAA3 in transgenic Arabidopsis increased cold tolerance by regulating auxin, ABA and ethylene signaling. Aux/IAA genes are key genes involved in regulating auxin signal transduction in plants. Although IAA genes have been characterized in various plant species, the role of IAA genes in grape cold resistance is unclear. To further explore the members of the Aux/IAA gene family in grape and their functions, in this study, using genomic data for Pinot Noir (Vitis vinifera cv. 'Pinot Noir') and Shanputao (Vitis amurensis), 25 VvIAA genes and 18 VaIAA genes were identified. The VaIAA genes presented different expression patterns at five different temperatures (28 ± 1 °C, 5 ± 1 °C, 0 ± 1 °C, -5 ± 1 °C, and -10 ± 1 °C) according to qRT‑PCR results. VaIAA3 was selected as a candidate gene for further functional analysis because of its high expression level under low-temperature stress. Subcellular localization experiments revealed that VaIAA3 was localized in the nucleus. Additionally, under 4 °C treatment for 24 h, relative expression level of VaIAA3, antioxidant enzyme activity, survival rate, and cold-responsive gene expression in three transgenic lines (OE-1, OE-2, OE-3) were greater, whereas relative electrolytic conductivity (REC), malondialdehyde (MDA) content and hydrogen peroxide (H 2 O 2 ) content were lower than those of the wild type (WT). Transcriptome sequencing analysis revealed that VaIAA3 regulated cold stress resistance in Arabidopsis thaliana (Arabidopsis) through pathways involving auxin, ABA, JA, or ethylene. Importantly, heterologous overexpression of VaIAA3 increased the resistance of Arabidopsis to cold stress, which provides a theoretical basis for the further use of VaIAA3 to improve cold resistance in grape., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: The rhizosphere microbiome is critical for promoting plant growth and mitigating soil-borne pathogens. However, its role in fighting soil-borne virus-induced diseases, such as wheat yellow mosaic virus (WYMV) transmitted by Polymyxa graminis zoospores, remains largely underexplored. In this study, we hypothesized that during viral infections, plant microbiomes engage in critical interactions with plants, with key microbes playing vital roles in maintaining plant health. Our research aimed to identify microbial taxa that not only suppress the disease but also boost wheat yield by using a blend of techniques, including field surveys, yield assessments, high-throughput sequencing of plant and soil microbiomes, microbial isolation, hydroponic experiments, and transcriptome sequencing., Results: We found that, compared with roots and leaves, the rhizosphere microbiome showed a better performance in predicting wheat yield and the prevalence of P. graminis and WYMV across the three WYMV-impacted regions in China. Using machine learning, we found that healthy rhizospheres were marked with potentially beneficial microorganisms, such as Sphingomonas and Allorhizobium-Neorhizobium-Parararhizobium-Rhizobium, whereas diseased rhizospheres were associated with a higher prevalence of potential pathogens, such as Bipolaris and Fusicolla. Structural equation modeling showed that these biomarkers both directly and indirectly impacted wheat yield by modulating the rhizosphere microbiome and P. graminis abundance. Upon re-introduction of two key healthy rhizosphere biomarkers, Sphingomonas azotifigens and Rhizobium deserti, into the rhizosphere, wheat growth and health were enhanced. This was attributed to the up-regulation of auxin and cytokinin signaling pathways and the regulation of jasmonic acid and salicylic acid pathways during infections., Conclusions: Overall, our study revealed the critical role of the rhizosphere microbiome in combating soil-borne viral diseases, with specific rhizosphere microbes playing key roles in this process. Video Abstract., (© 2024. The Author(s).)

Canopy shade enhances the activity of PHYTOCHROME INTERACTING FACTORs (PIFs) to boost auxin synthesis in the cotyledons. Auxin, together with local PIFs and their positive regulator CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1), promotes hypocotyl growth to facilitate access to light. Whether shade alters the cellular redox status thereby affecting growth responses, remains unexplored. Here, we show that, under shade, high auxin levels increased reactive oxygen species and nitric oxide accumulation in the hypocotyl of Arabidopsis. This nitroxidative environment favored the promotion of hypocotyl growth by COP1 under shade. We demonstrate that COP1 is S-nitrosylated, particularly under shade. Impairing this redox regulation enhanced COP1 degradation by the proteasome and diminished the capacity of COP1 to interact with target proteins and to promote hypocotyl growth. Disabling this regulation also generated transversal asymmetries in hypocotyl growth, indicating poor coordination among different cells, which resulted in random hypocotyl bending and predictably low ability to compete with neighbors. These findings highlight the significance of redox signaling in the control of diffuse growth during shade avoidance., Competing Interests: Competing interests statement:The authors declare no competing interest.

The problem of potentially toxic metal pollution is increasingly acute with the development of human society. In this study, we investigated the remediation of nickel (Ni) and cadmium (Cd) co-contamination through inoculating rice with three new-isolated Ni- and Cd-resistant plant growth-promoting rhizobacteria (PGPR) Y3, Y4, and Y5. These three strains possessed growth-promoting properties, including 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, the ability of indoleacetic acid (IAA) production, phosphate solubilization, siderophores production, and exopolysaccharide (EPS) development. According to 16S rDNA sequence homology, strains Y3, Y4, and Y5 were identified as Pseudomonas sp., Chryseobacterium sp., and Enterobacter sp., respectively. Based on the results of rice germination experiments conducted under combined toxicity, we set the contamination concentrations for Ni 2+ at 20 μg mL -1 and Cd 2+ at 40 μg mL -1 . Then we conducted potting experiments at these concentration levels to study the effects of strains Y3, Y4, and Y5 on rice growth under synergistic Ni and Cd stress. The results indicated that the inoculated strains Y3, Y4, and Y5 were effective in promoting the growth of rice seedlings under the combined stress of Ni and Cd, and conferring tolerance to Ni and Cd by increasing the antioxidant enzyme activities of the seedlings. Among them, strain Y3 exhibited stronger ACC deaminase activity, IAA production capacity, and EPS production capacity, showing the most pronounced growth-promoting effect on rice. It was demonstrated that after inoculation with strain Y3, the germination rate of rice seeds increased by 43 %, the fresh weight of stems improved by 35 %, and the chlorophyll content enhanced by 70 % and other growth-promoting phenomena. Additionally, under Ni and Cd stress, strain Y5 performed better than strain Y4 in terms of IAA production capacity and its influence on rice root growth, suggesting that IAA production might play a specifically essential role in root growth under Ni and Cd stress., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The auxin signaling molecule regulates a range of plant growth and developmental processes. The core transcriptional machinery responsible for auxin-mediated responses is conserved across all land plants. Genetic, physiological and molecular exploration in bryophyte and angiosperm model species have shown both qualitative and quantitative differences in auxin responses. Given the highly divergent ontogeny of the dominant gametophyte (bryophytes) and sporophyte (angiosperms) generations, however, it is unclear whether such differences derive from distinct phylogeny or ontogeny. Here, we address this question by comparing a range of physiological, developmental and molecular responses to auxin in both generations of the model fern Ceratopteris richardii. We find that auxin response in Ceratopteris gametophytes closely resembles that of a thalloid bryophyte, whereas the sporophyte mimics auxin response in flowering plants. This resemblance manifests both at the phenotypic and transcriptional levels. Furthermore, we show that disrupting auxin transport can lead to ectopic sporophyte induction on the gametophyte, suggesting a role for auxin in the alternation of generations. Our study thus identifies developmental phase, rather than phylogeny, as a major determinant of auxin response properties in land plants., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Background: Root plays an important role in the growth and development of fruit trees; however, the molecular mechanisms behind the differences among rootstock varie-ties remain unclear., Methods: This study examined the effects of different rootstocks on root structure and the endogenous hormone content of 1-year old apple seedlings in combinations of Tianhong 2 (T2)/ Malus robusta (HT), T2/G935, and T2/Jizhen 2 (J2)., Results: The results showed that the T2/HT treatment had greater root length, surface area, volume, average diameter, tips and forks, followed by G935 and J2. In T2/HT leaves and roots, the indole-3-acetic acid (IAA) and gibberellins (GA 3 ) levels were highest, and the abscisic acid (ABA) levels were the lowest. A root transcriptome analysis detected 10,064, 10,511, and 8,719 differentially expressed genes in T2/HT vs. T2/G935, T2/HT vs. T2/J2, and T2/J2 vs. T2/G935, respectively. The analysis of Gene Ontology (GO) terms indicated a significant enrichment in biological processes, cellular components, and molecular functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that plant hormone signaling, MAPK signaling pathway-plant, and plant-pathogen interaction played important roles in differences in the rooting ability of different rootstocks. In addition, some key differential genes were associated with root growth and development and were involved in these metabolic pathways. This study is important for enriching theoretical studies of fruit tree roots., Competing Interests: The authors declare there are no competing interests., (©2024 Li et al.)

Rice tiller angle is a key agronomic trait that has significant effects on the establishment of a high-yield rice population. However, the molecular mechanism underlying the control of rice tiller angle remains to be clarified. Here, we characterized the novel tiller-angle gene LAZY4 (LA4) in rice through map-based cloning. LA4 encodes a C3H2C3-type RING zinc-finger E3 ligase localized in the nucleus, and an in vitro ubiquitination assay revealed that the conserved RING finger domain is essential for its E3 ligase activity. We found that expression of LA4 can be induced by gravistimulation and that loss of LA4 function leads to defective shoot gravitropism caused by impaired asymmetric auxin redistribution upon gravistimulation. Genetic analysis demonstrated that LA4 acts in a distinct pathway from the starch biosynthesis regulators LA2 and LA3, which function in the starch-statolith-dependent pathway. Further genetic analysis showed that LA4 regulates shoot gravitropism and tiller angle by acting upstream of LA1 to mediate lateral auxin transport upon gravistimulation. Our studies reveal that LA4 regulates shoot gravitropism and tiller angle upstream of LA1 through a novel pathway independent of the LA2-LA3-mediated gravity-sensing mechanism, providing new insights into the rice tiller-angle regulatory network., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The economic part of the Hibiscus sabdariffa L. (Malvaceae) plant is the sepal. One of the main challenges in harvesting this product is separating its seeds, which are surrounded by sepals. If the parthenocarpy process occurs without seeds, the labor costs are reduced and the profits from the production of this plant are increased. In current study, the effect of gibberellic acid (GA 3 ), melatonin (M), and indole acetic acid (IAA) on the induction of parthenocarpy in H. sabdariffa plants was investigated. The study was conducted as a factorial experiment in the form of randomized complete block design with three replications. Different concentrations of GA 3 (0 (control), 700, 800, and 900 ppm), M (0 (control), 100, 200, and 400 ppm), and IAA (0 (control), 1000, 1200, and 1400 ppm) were foliar sprayed after the emergence of the flower bud on the 50th, 60th, and 70th days of planting, and the control plants were also sprayed with distilled water. Various measurements were taken including the number of seeds per boll, parthenocarpy percentage, capsule volume, number of bolls per plant, number of mature seeds, and phytochemical parameters such as anthocyanin, chlorophyll and carotenoid content, antioxidant value, total phenol and flavonoid content, and soluble solids. The application of GA 3 at 800 and 900 ppm along with IAA at 1000 ppm and M at 100 and 200 ppm led to the production of parthenocarpy fruits and showed the best results in the induction of parthenocarpy. The control group indicated the highest number of mature seeds per boll. The highest amount of phenol and flavonoid contents were obtained in plants treated with GA 3 at 800 ppm. For anthocyanin, the control group showed the highest value (1.63 mg g -1 ), and in the case of the antioxidant trait, plants exposed to 100 ppm M showed the highest IC50 (40.68%). Hence, the application of plant growth regulators with appropriate concentrations can be effective in inducing parthenocarpy in H. sabdariffa plants. Additionally, parthenocarpy-induced by GA 3 , IAA, and M had different impacts on fruit quality and quantity, suggesting that the effect depends on the type of employed hormones used and their concentrations., (© 2024. The Author(s).)

Soil salinization is currently one of the main abiotic stresses that restrict plant growth. Plant endophytic bacteria can alleviate abiotic stress. The aim of the current study was to isolate, characterize, and assess the plant growth-promoting and saline and alkaline stress-alleviating traits of Peribacillus simplex M1 ( P. simplex M1 ) isolates from maize. One endophytic bacterial isolate, named P. simplex M1 , was selected from the roots of maize grown in saline-alkali soil. The P. simplex M1 genome sequence analysis of the bacteria with a length of 5.8 Mbp includes about 700 genes that promote growth and 16 antioxidant activity genes that alleviate saline and alkaline stress. P. simplex M1 can grow below 400 mM NaHCO 3 on the LB culture medium; The isolate displayed multiple plant growth-stimulating features, such as nitrogen fixation, produced indole-3-acetic acid (IAA), and siderophore production. This isolate had a positive effect on the resistance to salt of maize in addition to the growth. P. simplex M1 significantly promoted seed germination by enhancing seed vigor in maize whether under normal growth or NaHCO 3 stress conditions. The seeds with NaHCO 3 treatment exhibited higher reactive oxygen species (ROS) levels than the maize in P. simplex M1 inoculant on maize. P. simplex M1 can colonize the roots of maize. The P. simplex M1 inoculant plant increased chlorophyll in leaves, stimulated root and leaf growth, increased the number of lateral roots and root dry weight, increased the length and width of the blades, and dry weight of the blades. The application of inoculants can significantly reduce the content of malondialdehyde (MDA) and increase the activity of plant antioxidant enzymes (Catalase (CAT), Superoxide Dismutase (SOD), and Peroxidase (POD)), which may thereby improve maize resistance to saline and alkaline stress. Conclusion: P. simplex M1 isolate belongs to plant growth-promoting bacteria by having high nitrogen concentration, indoleacetic acid (IAA), and siderophore, and reducing the content of ROS through the antioxidant system to alleviate salt alkali stress. This study presents the potential application of P. simplex M1 as a biological inoculant to promote plant growth and mitigate the saline and alkaline effects of maize and other crops.

Key Message: GhMAC3e expression was induced by various stresses and hormones. GhMAC3e may regulate plant growth by influencing auxin distribution, and play important roles in Verticillium wilt resistance via mediating SA signaling. The MOS4-Associated Complex (MAC) is a highly conserved protein complex involved in pre-mRNA splicing and spliceosome assembly, which plays a vital role in plant immunity. It comprises key components such as MOS4, CDC5, and PRL1. MAC3A/B, as U-box E3 ubiquitin ligases, are crucial for various plant processes including development, stress responses, and disease resistance. However, their roles in cotton remain largely unknown. In this study, we first cloned the GhMAC3e gene from cotton and explored its biological functions by using virus-induced gene silencing (VIGS) in cotton and transgenic overexpression in Arabidopsis. The results showed that GhMAC3e is ubiquitously expressed in cotton tissues and could be induced by salt stress, Verticillium dahliae (VD) infection, PEG, ABA, ETH, GA3, MeJA, and SA. Silencing GhMAC3e retarded primary stem growth and reduced biomass of cotton coupled with the reduced auxin content in the petioles and veins. Silencing GhMAC3e up-regulated expression of cell growth-related genes GhXTH16 and Gh3.6, while down-regulated GhSAUR12 expression. Ectopic expression of GhMAC3e in Arabidopsis significantly enhanced its resistance to Verticillium wilt (VW) in terms of decreased pathogen biomass and lowered plant mortality. Overexpression of GhMAC3e dramatically upregulated AtPR1 by around 15 fold and more than 262 fold under basal and VD inoculation condition, respectively. This change was not associated with the expression of GhNPR1. In conclusion, GhMAC3e may not only regulate plant growth by influencing auxin distribution and growth-related gene expression, but also play important roles in VW resistance via mediating SA signaling independent of NPR1 transcription level., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Plants are frequently exposed to herbivory and mechanical damage that result in wounding. Two fundamental strategies, regeneration and healing, are employed by plants upon wounding. How plants make different decisions and how wound healing is sustained until the damaged tissues recover are not fully understood. In this study, we found that local auxin accumulation patterns, determined by wounding modes, may activate different recovery programs in wounded tissues. Wounding triggers transient jasmonic acid (JA) signaling that promotes lignin deposition in the first few hours after wounding occurs. This early response is subsequently relayed to ABA signaling via MYC2. The induced JA signaling promotes ABA biosynthesis to maintain the expression of RAP2.6, a key factor for sustained lignin biosynthesis and the later wound-healing process. Our findings provide mechanistic insights into how plants heal from wounding and clarify the molecular mechanisms that underlie the prolonged healing process following wounding., (Copyright © 2024 The Author. Published by Elsevier Inc. All rights reserved.)

Endophytes have drawn attentions due to their effectiveness in providing benefits to host and non-host plants. In this study endophytic bacteria were isolated from stem and leaf samples of medicinally important plants Rhododendron griffithianum Wight and Rhododendron arboreum Smith subsp. cinnamomeum (Wall. ex G. Don) grown at higher altitudes of Darjeeling, India. Two endophytic bacteria, Pseudomonas lurida RGDS03 and Bacillus velezensis RCDL12 were identified based on 16S rRNA gene sequencing analysis. The endophytes exhibited indole acetic acid (IAA), gibberellic acid (GA), siderophore production, phosphate solubilization, nitrogen-fixing abilities, though B. velezensis RCDL12 showed superior production of IAA (126.04 ± 0.40 µg/mL), GA (241.00 ± 0.44 µg/mL), and phosphate (74.4 ± 0.41 µg/mL) solubilization as compared to P. lurida RGDS03. Purity of extracted IAA from these two endophytes was confirmed by HPLC and LC-MS analysis. In this study, P. lurida RGDS03 inhibited mycelial growth of two tested phytopathogens Phytophthora sp. and Pestalotiopsis sp. of broad host range. However, only against Pestalotiopsis sp. did B. velezensis RCDL12 exhibit antifungal activity. Study was conducted on growth promotion capabilities of isolates on rice and mung bean seedlings. P. lurida RGDS03, B. velezensis RCDL12 and consortium of both the strains reported with promising growth promotion on both rice (85-97%) and mung bean (86-99%) in terms of their seed germination, vegetative growth (root and shoot length, fresh and dry weight), and chlorophyll content as compared to the control plants (untreated). This study has emphasized growth-promoting and biocontrol activities of endophytic bacteria from rhododendrons, and application to enhance crop development for sustainable agriculture., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Plants generally enhance their root growth in the form of greater biomass and/or root length to boost nutrient uptake in response to short-term low nitrogen (LN). However, the underlying mechanisms of short-term LN-mediated root growth remain largely elusive. Our genome-wide association study, haplotype analysis, and phenotyping of transgenic plants showed that the crucial nitrate signaling component NIN-LIKE PROTEIN3.2 (ZmNLP3.2), a positive regulator of root biomass, is associated with natural variations in root biomass of maize (Zea mays L.) seedlings under LN. The monocot-specific gene AUXIN/INDOLE-3-ACETIC ACID14 (ZmAux/IAA14) exhibited opposite expression patterns to ZmNLP3.2 in ZmNLP3.2 knockout and overexpression lines, suggesting that ZmNLP3.2 hampers ZmAux/IAA14 transcription. Importantly, ZmAux/IAA14 knockout seedlings showed a greater root dry weight (RDW), whereas ZmAux/IAA14 overexpression reduced RDW under LN compared with wild-type plants, indicating that ZmAux/IAA14 negatively regulates the RDW of LN-grown seedlings. Moreover, in vitro and vivo assays indicated that AUXIN RESPONSE FACTOR19 (ZmARF19) binds to and transcriptionally activates ZmAux/IAA14, which was weakened by the ZmNLP3.2-ZmARF19 interaction. The zmnlp3.2 ZmAux/IAA14-OE seedlings exhibited further reduced RDW compared with ZmAux/IAA14 overexpression lines when subjected to LN treatment, corroborating the ZmNLP3.2-ZmAux/IAA14 interaction. Thus, our study reveals a ZmNLP3.2-ZmARF19-ZmAux/IAA14 module regulating root biomass in response to nitrogen limitation in maize., Competing Interests: Conflict of interest statement. The authors declare no conflict of interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Root parasitic plants in the Orobanchaceae, such as Striga and Orobanche, cause significant damage to crop production. The germination step of these root parasitic plants is induced by host-root-derived strigolactones. After germination, the radicles elongate toward the host and invade the host root. We have previously discovered that a simple amino acid, tryptophan (Trp), as well as its metabolite, the plant hormone indole-3-acetic acid (IAA), can inhibit radicle elongation of Orobanche minor. These results suggest that auxin plays a crucial role in the radicle elongation step in root parasitic plants. In this report, we used various auxin chemical probes to dissect the auxin function in the radicle growth of O. minor and Striga hermonthica. We found that synthetic auxins inhibited radicle elongation. In addition, auxin receptor antagonist, auxinole, rescued the inhibition of radicle growth by exogenous IAA. Moreover, a polar transport inhibitor of auxin, N-1-naphthylphthalamic acid, affected radicle bending. We also proved that exogenously applied Trp is converted into IAA in O. minor seeds, and auxinole partly rescued this radicle elongation. Taken together, our data demonstrate a pivotal role for auxin in radicle growth. Thus, manipulation of auxin function in root parasitic plants should offer a useful approach to combat these parasites., (© The Author(s) 2024. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

Drought-induced stress represents one of the most economically detrimental natural phenomena impacting grapevine (Vitis vinifera ) development, yield, and fruit characteristics. Also, auxin is one of the most important plant growth regulators that can reduce damage caused by stress in plants. In this study, the impact of exogenously sprayed auxin (0, 50, and 200mgL-1 ) on growth, biochemical, and anatomical parameters was investigated in two grapevine varieties (cvs. 'Rashe' and 'Fakhri') under water deficit. According to our findings, water deficit led to a notable decrease in growth, protein content, and anatomical parameters; but significantly enhanced electrolyte leakage. Grapevines exposed to water deficit exhibited substantial increases in total phenolic compounds and antioxidant activity. Applying 50mgL-1 napthalene acetic acid (NAA) reduced the effects of water deficit in both grapevine cultivars by decreasing electrolyte leakage (15% in 'Rashe' and 20% in 'Fakhri'), and accumulating protein content (22% 'Rashe' and 32% 'Fakhri'), total phenolic compounds (33%'Rashe' and 40% 'Fakhri'), and antioxidant capacity (11% 'Rashe' and 39% 'Fakhri'); anantomical parameters were also improved. However, application of 200mgL-1 NAA had adverse effects on growth and biochemical traits of grapevines, with a more pronounced impact on root growth and anatomical parameters compared to other NAA concentrations. In conclusion, the application of 50mgL-1 NAA enhanced grapevine growth, enabling them to better thrive under water deficit.

Nitrogen deficiency in low organic matter soils significantly reduces crop yield and plant health. The effects of foliar applications of indole acetic acid (IAA), trehalose (TA), and nanoparticles-coated urea (NPCU) on the growth and physiological attributes of tomatoes in nitrogen-deficient soil are not well documented in the literature. This study aims to explore the influence of IAA, TA, and NPCU on tomato plants in nitrogen-deficient soil. Treatments included control, 2mM IAA, 0.1% TA, and 2mM IAA + 0.1% TA, applied with and without NPCU. Results showed that 2mM IAA + 0.1% TA with NPCU significantly improved shoot length (~ 30%), root length (~ 63%), plant fresh (~ 48%) and dry weight (~ 48%), number of leaves (~ 38%), and leaf area (~ 58%) compared to control (NPCU only). Additionally, significant improvements in chlorophyll content, total protein, and total soluble sugar, along with a decrease in antioxidant activity (POD, SOD, CAT, and APX), validated the effectiveness of 2mM IAA + 0.1% TA with NPCU. The combined application of 2mM IAA + 0.1% TA with NPCU can be recommended as an effective strategy to enhance tomato growth and yield in nitrogen-deficient soils. This approach can be integrated into current agricultural practices to improve crop resilience and productivity, especially in regions with poor soil fertility. To confirm the efficacy of 2mM IAA + 0.1% TA with NPCU in various crops and climatic conditions, additional field studies are required., (© 2024. The Author(s).)

Perennial grains, which remain productive for multiple years, rather than growing for only one season before harvest, have deep, dense root systems that can support a richness of beneficial microorganisms, which are mostly underexplored. In this work we isolated forty-three bacterial strains associated with the rhizosphere of the OK72 perennial wheat line, developed from a cross between winter common wheat and Thinopyrum ponticum. Identified using 16S rDNA sequencing, these bacteria were assessed for plant growth-promoting traits such as indole-3-acetic acid, siderophores and ACC-deaminase acid production, biofilm formation, and the ability to solubilize phosphate and proteins. Twenty-five strains exhibiting in vitro significant plant growth promoting traits, belong to wheat keystone genera Pseudomonas, Microbacterium, Variovorax, Pedobacter, Dyadobacter, Plantibacter, and Flavobacterium. Seven strains, including Aeromicrobium and Okibacterium genera, were able to promote root growth in a commercial annual wheat cultivar while strains from Pseudomonas genus inhibited the growth of Aspergillus flavus and Fusarium species, using direct antagonism assays. The same strains produced a high amount of 1-undecanol a volatile organic compound, which may aid in suppressing fungal growth. The study highlights the potential of these bacteria to form new commercial consortia, enhancing the health and productivity of annual wheat crops within sustainable agricultural practices., (© 2024. The Author(s).)

Indole-3-acetic acid (IAA) can regulate plant growth and thus modulate the accumulation of polycyclic aromatic hydrocarbons (PAHs). However, the effect of endogenous IAA on PAHs accumulation and its influencing factors remains unclear. To unravel this, two different IAA expression genotypes of Arabidopsis thaliana, i.e., IAA-underproducing yucca1D [YUC1] mutant and wild type [WT]) were selected and treated with different fluoranthene (Flu) concentrations (0 mg/L [CK], 5 mg/L [Flu5], and 20 mg/L [Flu20]) to reveal the impact mechanism of endogenous IAA on Flu uptake by plants. The results indicated that under Flu5 treatment, the bioconcentration factors (BCF) and translocation factors (TF) of Flu in WT were 41.4 % and 14.3 % higher than those in YUC1. Similarly, under Flu20 treatment, the BCF and TF of Flu in WT were also 42.2 % and 8.2 % higher than those in YUC1. In addition, the BCF and TF were 72.5 % and 35.8 % higher under Flu5 treatment compared to Flu20 treatment for WT, and 73.4 % and 28.6 % higher respectively for YUC1. Moreover, WT exhibited higher plant growth (biomass, root morphology indicators [root length, root area and number of tips]) and IAA content compared to YUC1 under identical Flu treatments. Plant growth and IAA content declined with the increase of Flu concentration in both YUC1 and WT leaves compared with CK treatment. Conversely, in WT roots, root biomass and morphology indicators promoted followed by a decrease as the concentration of Flu increased. Additionally, the antioxidant enzyme activities (SOD, POD, and CAT) of WT were 11.1 %, 16.7 %, and 28.9 % higher than those of YUC1 under Flu5 treatment, and 13.6 %, 12.9 %, and 26.5 % higher under Flu20 treatment. Compared with CK treatment, SOD and POD activities promoted with increasing Flu concentration, whereas CAT activities decreased. Variability separation analysis revealed that level of IAA primarily influenced Flu accumulation in WT or under Flu5 treatments, whereas antioxidant enzyme activity primarily affected Flu accumulation in YUC1 or under Flu20 treatments. Exploring the relationship between the IAA synthesis gene YUCCA and IAA levels, alongside Flu accumulation, could yield novel insights into the regulation of PAH accumulation in plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Plant hormones are chemical signals governing almost every aspect of a plant's life cycle and responses to environmental cues. They are enmeshed within complex signaling networks that can only be deciphered by using broad-scale analytical methods to capture information about several plant hormone classes simultaneously. Methods used for this purpose are all based on reversed-phase (RP) liquid chromatography and mass spectrometric detection. Hydrophilic interaction chromatography (HILIC) is an alternative chromatographic method that performs well in analyses of biological samples. We therefore developed and validated a HILIC method for broad-scale plant hormone analysis including a rapid sample preparation procedure; moreover, derivatization or fractionation is not required. The method enables plant hormone screening focused on polar and moderately polar analytes including cytokinins, auxins, jasmonates, abscisic acid and its metabolites, salicylates, indoleamines (melatonin), and 1-aminocyclopropane-1-carboxylic acid (ACC), for a total of 45 analytes. Importantly, the major pitfalls of ACC analysis have been addressed. Furthermore, HILIC provides orthogonal selectivity to conventional RP methods and displays greater sensitivity, resulting in lower limits of quantification. However, it is less robust, so procedures to increase its reproducibility were established. The method's potential is demonstrated in a case study by employing an approach combining hormonal analysis with phenomics to examine responses of three Arabidopsis ecotypes toward three abiotic stress treatments: salinity, low nutrient availability, and their combination. The case study showcases the value of the simultaneous determination of several plant hormone classes coupled with phenomics data when unraveling processes involving complex cross-talk under diverse plant-environment interactions., (© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

The LED Blue Light (LBL) (450 nm) effect on hormones levels and on jasmonates (JAs) metabolism in oranges was investigated. The quantum flux (2 days, 60 μmol m -2 . s -1 ) was chosen for its efficacy in reducing postharvest rot caused by this crop's main postharvest phytopathogenic fungus (Penicillium digitatum). The analysis of abscisic (ABA), salicylic (SA) and indole-3-acetic (IAA) acids, and of JAs-related metabolites, revealed that LBL modifies all studied metabolites and had major effects on JAs levels, mainly on jasmonic acid (JA) and its precursor cis-(+)-12-oxo-phytodienoic acid (OPDA). This agrees with the up-regulation of the genes participating in their synthesis. Results highlight the relevance of CsLOX1 and CsLOX5, and the contribution of CsAOC3, in the LBL-induced OPDA biosynthesis, whereas CsOPR2, CsACX1 and CsACX3 would play a part in the synthesis of JA from OPDA. Data also suggest that the applied LBL quantum flux favors fruit JA perception by increasing the expression of the coronatine insensitive 1 (COI1) receptor; and signaling by down-regulating abundant CsJAZ negative regulators. Differences in OPDA and JA between the LBL-treated oranges and their control fruit left in the dark disappeared after shifting the LBL-treated oranges to darkness for 3 more days. However, the LBL and darkness combination slightly increased IAA and SA contents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Low temperature (LT) is an important environmental factor affecting the growth and yield of plants. Melatonin (MT) can effectively enhance the LT tolerance of cucumber. This study found that LT stress induced the expression of Comt1 (caffeic acid O-methyltransferase 1), with the highest expression being about 2-times that of the control. Meanwhile, the content of MT was found to be roughly 63.16% of that in the control samples. Compared with LT treatment alone, exogenous MT pretreatment upregulated the expression levels of TOR (Target of rapamycin), PIN1 (Pin-formed 1), and YUC4 (YUCCA 4), with maximum upregulations reaching approximately 66.67%, 79.32%, and 42.86%, respectively. These results suggest that MT may modulate the tolerance of cucumber seedlings to LT stress by regulating the expression of TOR, PIN1, and YUC4. In addition, co-treatment with AZD-8055 (a TOR inhibitor) or NPA (N-1-naphthylphthalamic acid, an auxin polar transport inhibitor) and MT attenuated MT-induced resistance to LT stress, leading to higher levels of reactive oxygen species (ROS), reduced antioxidant defense capacity, and increased damage to the membrane system in cucumber seedlings. Concurrently, the content of osmoregulatory substances and the photosynthesis decreased. These results demonstrate that both TOR and auxin were required for MT to alleviate LT-induced damage in cucumber. In summary, the present study demonstrates that TOR and auxin signaling synergistically contribute to alleviating LT damage in cucumber seedlings by exogenous MT. These findings help us understand the function of MT and provide insights into the regulatory network of MT that regulates the LT tolerance of plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The strong ability of plants to regenerate wounds is exemplified by grafting when two plants are cut and joined together to grow as one. During graft healing, tissues attach, cells proliferate, and the vasculatures connect to form a graft union. The plant hormone auxin plays a central role, and auxin-related mutants perturb grafting success. Here, we investigated the role of individual cell types and their response to auxin during Arabidopsis (Arabidopsis thaliana) graft formation. By employing a cell-specific inducible misexpression system, we blocked auxin response in individual cell types using the bodenlos mutation. We found that auxin signaling in procambial tissues was critical for successful tissue attachment and vascular differentiation. In addition, we found that auxin signaling was required for cell divisions of the procambial cells during graft formation. Loss of function mutants in cambial pathways also perturbed attachment and phloem reconnection. We propose that cambial and procambial tissues drive tissue attachment and vascular differentiation during successful grafting. Our study thus refines our knowledge of graft development and furthers our understanding of the regenerative role of the cambium., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Formation of the apical hook in etiolated dicot seedlings results from differential growth in the hypocotyl apex and is tightly controlled by environmental cues and hormones, among which auxin and gibberellins (GAs) play an important role. Cell expansion is tightly regulated by the cell wall, but whether and how feedback from this structure contributes to hook development are still unclear. Here, we show that etiolated seedlings of the Arabidopsis (Arabidopsis thaliana) quasimodo2-1 (qua2) mutant, defective in pectin biosynthesis, display severe defects in apical hook formation and maintenance, accompanied by loss of asymmetric auxin maxima and differential cell expansion. Moreover, qua2 seedlings show reduced expression of HOOKLESS1 (HLS1) and PHYTOCHROME INTERACTING FACTOR4 (PIF4), which are positive regulators of hook formation. Treatment of wild-type seedlings with the cellulose inhibitor isoxaben (isx) also prevents hook development and represses HLS1 and PIF4 expression. Exogenous GAs, loss of DELLA proteins, or HLS1 overexpression partially restore hook development in qua2 and isx-treated seedlings. Interestingly, increased agar concentration in the medium restores, both in qua2 and isx-treated seedlings, hook formation, asymmetric auxin maxima, and PIF4 and HLS1 expression. Analyses of plants expressing a Förster resonance energy transfer-based GA sensor indicate that isx reduces accumulation of GAs in the apical hook region in a turgor-dependent manner. Lack of the cell wall integrity sensor THESEUS 1, which modulates turgor loss point, restores hook formation in qua2 and isx-treated seedlings. We propose that turgor-dependent signals link changes in cell wall integrity to the PIF4-HLS1 signaling module to control differential cell elongation during hook formation., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Verticillium dahliae is a destructive, soil-borne pathogen that causes significant losses on numerous important dicots. Recently, beneficial microbes inhabiting the rhizosphere have been exploited and used to control plant diseases. In the present study, Burkholderia gladioli KRS027 demonstrated excellent inhibitory effects against Verticillium wilt in cotton seedlings. Plant growth and development was promoted by affecting the biosynthesis and signaling pathways of brassinosteroids (BRs), gibberellins (GAs), and auxins, consequently promoting stem elongation, shoot apical meristem, and root apical tissue division in cotton. Furthermore, based on the host transcriptional response to V. dahliae infection, it was found that KRS027 modulates the plants to maintain cell homeostasis and respond to other pathogen stress. Moreover, KRS027 induced disruption of V. dahliae cellular structures, as evidenced by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses. Based on the comparative transcriptomic analysis between KRS027 treated and control group of V. dahliae, KRS027 induced substantial alterations in the transcriptome, particularly affecting genes encoding secreted proteins, small cysteine-rich proteins (SCRPs), and protein kinases. In addition, KRS027 suppressed the growth of different clonal lineages of V. dahliae strains through metabolites, and volatile organic compounds (VOCs) released by KRS027 inhibited melanin biosynthesis and microsclerotia development. These findings provide valuable insights into an alternative biocontrol strategy for Verticillium wilt, demonstrating that the antagonistic bacterium KRS027 holds promise as a biocontrol agent for promoting plant growth and managing disease occurrence., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Castor (Ricinus communis L.) is an importance crop cultivated for its oil and economic value. Seed size is a crucial factor that determines crop yield. Gaining insight into the molecular regulatory processes of seed development is essential for the genetic enhancement and molecular breeding of castor. Here, we successfully fine-mapped a major QTL related to seed size, qSS3, to a 180 kb interval on chromosome 03 using F2 populations (DL01×WH11). A 17.6-kb structural variation (SV) was detected through genomic comparison between DL01 and WH11. Analysis of haplotypes showed that the existence of the complete 17.6 kb structural variant may lead to the small seed characteristic in castor. In addition, we found that qSS3 contains the microRNA396b (miR396b) sequence, which is situated within the 17.6 kb SV. The results of our experiment offer additional evidence that miR396-Growth Regulating Factor 4 (GRF4) controls seed size by impacting the growth and multiplication of seed coat and endosperm cells. Furthermore, we found that RcGRF4 activates the expression of YUCCA6 (YUC6), facilitating the production of IAA in seeds and thereby impacting the growth of castor seeds. Our research has discovered a crucial functional module that controls seed size, offering a fresh understanding of the mechanism underlying seed size regulation in castor., Competing Interests: Conflict of interest statement. The authors declare that there is no conflict of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)