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Preprints posted in the last 30 days, ranked by how well they match Plants's content profile, based on 43 papers previously published here. The average preprint has a 0.05% match score for this journal, so anything above that is already an above-average fit.

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Denisea fonensis (Thismiaceae), a new genus and species of mycoheterotroph from Guinea.

Cheek, M.; Molmou, D. N.; Delhaye, G.

2026-10-01 plant biology 10.64898/2026.09.28.755082 medRxiv
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Distinct from plants which directly parasitise other plants (parasites), non-photosynthetic mycoheterophic plants (MHPs, previously known as achlorophyllous mycotrophs or saprophytes) depend upon fungal hyphae for their carbon and have been viewed as cheaters or parasites in their fungal partnerships rather than mutualistic as in most other plant-mycorrhizal interactions. Over 400 such species in 87 genera and 11 families are recorded which maintain this lifeform through their life-cycle, while an estimate 30,000 further species, mainly orchids, depend on this mode of life to establish themselves from seeds. In most cases the fungal partners and the ultimate carbon sources, usually autotrophic plants, are unknown. High fungal specificity is the norm in those MHPs researched. We describe a new MHP genus and species, Denisea fonensis that we discovered in the Simandou range of the Republic of Guinea, West Africa in October 2025 on the footprint of a mining project. While clearly placed in Dioscoreales (which has three MHP families), the features of Denisea are so unusual that it was not initially clear in which family to place it. Finally, with molecular data we placed Denisea in Thismiaceae, corroborated by morphology. The new discovery is all the more unexpected as this is the first record of Thismiaceae from West Africa. Until now the family has only been recorded in Africa from three species of the very different genus Oxygyne, of Cameroon and Central African Republic. We provisionally assess Denisea fonensis as Critically Endangered since less than ten individuals are recorded in only a few square metres at one site, where there are imminent threats from animals. We propose urgent measures to conserve this species to reduce the risk of global extinction that has eliminated other African MHP species such as Oxygyne triandra, and further research to determine the fungal and autotrophic plant partners of this remarkable plant genus.

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Acrean fish diversity back in time: Paleoichthyology of the Rio Acre fossil fauna in Brazil

Ballen, G. A.; Alencar, K.; Muniz, F.; Guilherme, E.; Aldrin, L.; Lomba, S.; Dias-Costa, L. R.; Hsiou, A. S.

2026-10-02 paleontology 10.64898/2026.09.27.754785 medRxiv
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Freshwater fishes are an important component of the Neogene fossil record of South America. They have been recorded in the Acre fossil fauna, with considerable taxonomic and ecological diversity. Novel findings from the Patos locality in Acre State have produced several specimens representing eleven taxa, including the first occurrences in the fossil record of the genera Agamyxis, Glanidium, and Trachycorystes. These taxa comprise two orders, Characiformes and Siluriformes, with four and seven taxa respectively. The taxa found in the Patos locality are representative of typical modern Amazonian assemblages found in the drainages of the region, thus suggesting substantial ecological conservatism. An updated list of the Rio Acre fossil fish fauna is consistent with previous findings in the literature regarding clustering structure in fossil assemblages during the Neogene of South America.

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Night-break photoperiod timing shapes growth trajectories, functional phenotype, and biomass partitioning in Cannabis sativa L. in a genotype-dependent manner

Rodriguez-Roncancio, M. P.; Castillo-Hernandez, L. C.; Love, N.; Melgarejo, L. M.; Sarmiento, F.

2026-09-30 plant biology 10.64898/2026.09.29.755452 medRxiv
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Cannabis sativa L. cultivation in tropical regions or under indoor conditions requires supplemental lighting to maintain plant yields. Night-break (NB) photoperiods offer a cost-effective alternative to conventional day extension, although genotypic responses have not been clearly described. This study evaluated the effects of continuous and pulsed NB treatments on the growth, phenology, physiology, productivity, and cannabinoid accumulation of three Cannabis sativa genotypes representing two chemotypes. Plants were exposed to a commercial 18-h photoperiod (T1), a continuous 1-h NB (T2), or four 15-min NB pulses (T3) with equal total duration. Continuous NB produced responses comparable to the commercial photoperiod. In contrast, pulsed NB accelerated phenological progression by two to three weeks and reduced final plant height by 15-28%, depending on genotype, without impairing photosynthetic performance or decreasing floral dry biomass. Harvest index increased under both NB treatments, indicating enhanced biomass allocation to reproductive structures, while cannabinoid accumulation remained primarily genotype-dependent with only minor treatment-specific modulation. These findings demonstrate that NB efficacy depends not only on the duration of nocturnal light but also on its temporal distribution, highlighting temporal organization of nocturnal light as a key regulator of photoperiodic responses and a promising strategy for optimizing cannabis cultivation.

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Seeding paradise: germination ecophysiology of Xyris paradisiaca Wand. (Xyridaceae), an endangered endemic species from Central Brazil

Ordonez-Parra, C. A.; Bispo, M. R. B.; Amaral-Garcia, L.; Figueiredo, R.; Almeida, T. E. B.; Cortes, C. A.; Garcia, Q. S.

2026-09-28 plant biology 10.64898/2026.09.24.754203 medRxiv
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Seed ecophysiology is essential for understanding plant regeneration and developing effective conservation and ecological restoration strategies, yet knowledge remains scarce for most threatened Cerrado species. We characterised the germination niche, desiccation tolerance and storage behaviour of Xyris paradisiaca (Xyridaceae), an Endangered species endemic to Central Brazil, to assess whether its restricted distribution is associated with narrow germination requirements and to inform seed-based conservation. Germination was tested across light regimes, constant temperatures, decreasing water potentials and short-duration heat shocks, while thermal- and hydro-time models were used to quantify thermal and hydric thresholds. Seeds exhibited an absolute light requirement but broad abiotic tolerances. Germination remained >87% between 15 and 40 {degrees}C, with estimated cardinal temperatures of Tb = 8.94 {degrees}C, To = 33.13 {degrees}C and Tc = 45.53 {degrees}C. Germination remained similar to the control down to -0.6 MPa and exceeded 40% at -1.0 MPa, with a median base water potential of {Psi}b = -1.03 MPa. Germination also remained high after 1-min heat shocks up to 200 {degrees}C. Seeds were desiccation tolerant, with 100% germination after drying and a viability loss index of -0.04, and germination remained >90% after 24 months of ambient storage. Thus, the highly restricted distribution of X. paradisiaca is not driven by a narrow physiological germination niche. Its broad environmental tolerances, desiccation tolerance and favourable storage behaviour also highlight its unexplored potential for seed-based restoration, propagation and ex situ conservation.

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Molecular characterization of a cytochrome P450 involved in the skeletal diversification of tricyclic coumarins in Angelica keiskei.

Shinya, N.; Han, J.; Kamata, Y.; Taniguchi, M.; Miura, K.; Yazaki, K.; Sugiyama, A.; Munakata, R.

2026-10-06 plant biology 10.64898/2026.10.02.756137 medRxiv
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Abstract Tricyclic coumarins (TCs) exhibit defensive properties against herbivorous insects and possess valuable pharmacological properties for humans. They are mainly distributed in the Apiaceae, Rutaceae, Moraceae, and Fabaceae families. The chemical structural diversity of TCs in these families primarily arises from differences in skeletal variety and attachment position of the third ring, as observed in unsaturated furanocoumarins (FCs) and saturated dihydrofuranocoumarins (DHFCs). In TC biosynthesis, the FC and DHFC pathways diverge through distinct oxidation patterns. The product specificity of the reported cytochrome P450 monooxygenase at the branching point suggests that these TC groups are produced in nearly identical ratios; however, some species of the genus Angelica (Apiaceae) accumulate DHFCs at higher levels than FCs. In this study, we investigated the enzymatic mechanism responsible for forming these two TC groups using A. keiskei, a Japanese native species used as a medicinal and edible herb. Organ-specific transcriptome analysis highlighted a P450 member CYP71AJ62 that exhibited a close correlation with TC accumulation across organs. Enzymatic characterization demonstrated that CYP71AJ62 predominantly forms a hydroxylated DHFC, vaginidiol, which may promote DHFC over FC skeletal formation in TC biosynthesis. Phylogenetic analysis suggested that CYP71AJ62 has been derived from a gene responsible for FC formation by functional shift without gene duplication. Subsequent three-dimensional structural modeling and site-directed mutagenesis revealed that two amino acid residues on substrate recognition sites cooperatively determine the product specificity of this P450 enzyme. This study provides new insights into the molecular evolutionary process underlying ring-structure diversification and, more broadly, the adaptation of plants to their environments.

6
Studying the role of climate and sex in flowering dynamics of black poplar across time and space

Cazenave, X.; Marc, V.; Julien, D.; Veronique, J.; Durufle, H.

2026-10-06 plant biology 10.64898/2026.10.05.756621 medRxiv
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Global change has a major impact on rivers and their associated terrestrial compartment, the riparian forest. The black poplar (Populus nigra L.), the dominant species of the European riparian forest, is considered an indicator of the biodiversity dynamics of these ecosystems, but is threatened with extinction in several parts of its range. The species is dioecious and shows wide variability in many traits, enabling it to adapt to a broad range of environmental conditions. A detailed characterization of this natural variability is therefore a key issue both for the conservation of natural populations and for biological engineering aimed at restoring riverbanks. Phenology is both an indicator of climate change and an adaptation lever for populations, particularly in perennial species such as trees. In this study, we characterized the spatial and temporal variability in flowering phenology by studying 13 natural black poplar populations along the Loire River, France. Flowering date and duration showed substantial variability both within and between populations, and males generally flowered earlier than females. Despite this variability, the ranking of individual trees' flowering dates was highly consistent across years. Most male-female pairs shared several days of overlapping flowering each year, and this overlap was moderately stable across years. Flowering dates advanced by about five to six days per degree Celsius during early spring, with no detectable difference in sensitivity between sexes. This work contributes to the characterization of flowering variability in an economically important tree species and may inform both the conservation of genetic resources and breeding.

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CRISPR/Cas9 knockout of PtaAGL16 improves post-drought recovery and shoot vitality in hybrid poplar

Fendel, A.; Wodtke, S.; Fladung, M.; Bruegmann, T.

2026-10-06 plant biology 10.64898/2026.10.04.756531 medRxiv
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Drought stress severely limits tree growth and survival, a problem intensifying under climate change due to the rising frequency of extreme weather events. Identifying genetic variants that modulate drought stress responses and enhance resilience is therefore of great interest for developing climate-adapted trees. While the MADS-box transcription factor AGL16 has been implicated in drought responses and stomatal regulation in herbaceous species, the function of its homolog in woody perennials remains largely unexplored. Here, we investigate the role of PtaAGL16 in the poplar hybrid Populus tremula x P. alba. We generated CRISPR/Cas9 knockout lines of PtaAGL16 in the poplar clone INRA 717-1B4. Plants were subjected to a severe drought stress regime followed by a re-watering recovery phase. Morphological and physiological traits, including growth, stomatal conductance, shoot vitality, and stomatal density, together with stress-associated metabolites and drought-responsive gene expression, were analyzed. All agl16 knockout lines exhibited significantly reduced drought-induced damage compared to the wild-type. While the wild-type suffered extensive defoliation and persistent stem necrosis, the knockout lines retained higher shoot vitality and resumed growth after re-watering, including the formation of new leaves. Consequently, vitality in the knockout lines reached up to 86%, compared to only 34% in the wild-type. This improved resilience was accompanied by reduced persistent oxidative membrane damage, as indicated by lower MDA levels after recovery. These findings indicate that the disruption of PtaAGL16 is associated with enhanced drought resilience in a woody perennial species and highlight its potential as a genetic target for improving climate adaptation in fast-growing forest trees.

8
Integrated anthocyanin profiling and transcriptome analysis identify candidate genes associated with petal color variation in Cyclamen persicum

Xia, D.; Yao, G.; Li, X.; Zhang, Y.; Xing, R.; Ma, X.; Han, X.; Hu, Q.; Fu, Z.; Cao, X.; Chen, P.; Xu, S.; Gui, G.

2026-09-24 plant biology 10.64898/2026.09.23.753367 medRxiv
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Petal coloration is a key ornamental trait in Cyclamen persicum Mill. and is closely associated with anthocyanin composition and relative accumulation, yet the major anthocyanins and candidate molecular factors associated with color variation remain unclear. In this study, anthocyanin profiling and transcriptome analysis were integrated to investigate three Cyclamen materials with distinct petal colors, BXK, FXK, and SHXK. Anthocyanin profiling showed clear separation among the three materials, indicating that petal color divergence was associated with differences in anthocyanin composition and relative abundance. Petunidin-, peonidin-, delphinidin-, malvidin-, and cyanidin-related derivatives exhibited distinct accumulation patterns. Transcriptome analysis revealed marked transcriptional divergence, with phenylpropanoid and flavonoid biosynthesis repeatedly enriched across comparisons, whereas isoflavonoid and flavone and flavonol biosynthesis were enriched in selected comparisons. Mapping differentially expressed genes onto the anthocyanin biosynthetic pathway revealed branch-specific expression patterns of structural genes, including CHS, F3H/F3'H, F3'5'H, DFR, and ANS. Notably, Cluster-26256.0, annotated as a putative anthocyanidin synthase gene, showed a progressive increase in expression from BXK to FXK and SHXK, consistent with increasing petal pigmentation. Several DFR-related transcripts also showed relatively high expression in SHXK. Co-expression analysis linked key anthocyanins, structural genes, and transcription factors, while qRT-PCR generally confirmed RNA-seq trends. These results suggest that late anthocyanin biosynthesis, particularly ANS-related transcriptional variation, may contribute to dark red petal coloration. This study provides a metabolite-transcript framework for Cyclamen petal coloration and identifies ANS-related genes as priority candidates for future functional studies and flower-color breeding.

9
Distinct infection and colonization strategies in cowpea leaves by three Colletotrichum species recently reported in Nigeria

Dada, A. O.; Dania, V. O.; Oyatomi, O. A.; Abberton, M. T.; Ortega-Beltran, A.

2026-10-02 plant biology 10.64898/2026.10.01.755927 medRxiv
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Cowpea, a crop of great importance in several regions in West Africa, faces many biotic and abiotic challenges. Several species of the genus Colletotrichum cause cowpea anthracnose disease (CAD), a major constraint to cowpea production severely affecting foliage and yield. Three Colletotrichum species: C. cliviicola, C. truncatum, and C. chrysophilum recently reported as causal agents of CAD in Nigeria were investigated in their infective processes and pathogenicity on cowpea leaves using histopathological and microscopic examination of stained leaf tissues. Inoculated leaves were examined at intervals to monitor fungal establishment, colonization, and reproductive development within host tissues. The three species completed the infection cycle, progressing from conidial germination and host penetration to tissue colonization, and sporulation. However, differences in both infective ability and pathogenicity were observed in representative isolates of each species. C. truncatum isolate was the most virulent, had extensive branching hyphae, produced severe tissue necrosis, abundant acervuli, and prominent seta formation. C. cliviicola also had substantial colonization and sporulation ability with dense conidial masses and mature acervuli. In contrast, C. chrysophilum had moderate infectivity with less extensive reproductive structures and tissue degradation. Here we provide the first histopathological report of infection and pathogenic development of these three species recently reported as causal agents of CAD

10
Biosynthesis of prenylated flavonoids by two membrane-bound prenyltransferases in glandular trichomes of Macaranga tanarius.

Munakata, R.; Maeda, Y.; Shimizu, R.; Sugiyama, A.; Kumazawa, S.; Fukumoto, S.; Yazaki, K.

2026-09-24 biochemistry 10.64898/2026.09.23.753730 medRxiv
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Macaranga tanarius (Euphorbiaceae), a myrmecophilic plant grown in subtropical countries, was identified as a source of Okinawan propolis, a honeybee product, which is glandular trichomes developed on the surface of the fruits. This tissue contains a high amount of characteristic geranylated flavonoids known as nymphaeols (nymphaeol A - C and isonymphaeol B), which exhibit strong antioxidant activity. In this study, we characterized the structure of glandular trichomes of M. tanarius, showing that they are sac-like structures containing drupelet-like aggregates of secretary cells, and a subcuticular cavity was also observed. From a cDNA library, we identified two prenyltransferase (PT) genes involved in biosynthesis of nymphaeols, and a PT gene grouped in a primary metabolic PT. Our biochemical experiments demonstrated that one of the former secondary metabolic PTs (MtPT1) is a B-ring-specific geranyltransferase for eriodictyol to yield nymphaeols B and isonymphaeol B. MtPT1 was unusual for a plant PT, an enzyme that yielded multiple reaction products. The other PT (MtPT3) transferred a geranyl moiety to the A-ring of eriodictyol to form nymphaeol A. Although dimethylallyl diphosphate (DMAPP) was not recognized as its prenyl donor substrate of eriodictyol, giving a B-ring geranylated eriodyctiol as the prenyl acceptor substrate, this enzyme used DMAPP to attach the prenyl moiety to the A-ring and produced nymphaeol C. These data suggest that M. tanarius has evolved its PT function to produce a diverse range of prenylated flavonoids using a limited set of genes.

11
Towards an understanding of the taxonomy of the Eurasian montane riparian genus Myricaria (Tamaricaceae) through Angiosperms353 herbariomics

Weitkaemper, K.; Lyakh, E.; Morales-Briones, D. F.; Weitkaemper, F.; Werth, S.

2026-10-06 plant biology 10.64898/2026.10.03.751413 medRxiv
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Myricaria Desv. (Tamaricaceae) is a genus of shrubs that play a keystone role in montane riparian ecosystems across Eurasia, shaping riverine habitats and supporting ecological function. The genus has a centre of origin in the Qinghai-Tibetan Plateau and exhibits a broad distribution, but many species boundaries are unresolved. Here, we present the first phylogenomic analysis of Myricaria using the Angiosperms353 target enrichment bait set, with sampling across its full geographic range. Off-target plastid loci were extracted from next-generation sequencing data using the CAPTUS bioinformatics pipeline. We infer species relationships using ASTRAL-IV and network analysis with SplitsTree, in conjunction with morphology. We also provide a taxonomic treatment of the genus, integrating our morphological observations and phylogenomic analyses for species delimitations. Based on our analyses, recognise two infrageneric sections: sect. Parallelantherae (containing only M. elegans) and sect. Myricaria (the remainder of the genus, with type species M. germanica). We also propose synonymising M. davurica, M. bracteata and M. squamosa with M. germanica. Although species boundaries in the Asian highlands remain unresolved, we conclude that the species M. elegans, M. germanica, M. laxiflora, M. longifolia, M. platyphylla, and M. pulcherrima are well-supported both phylogenomically and morphologically.

12
Hidden Medicines in Herbarium Collections: Medicinal Plant Diversity and Ethnopharmacological Knowledge from GSUH, Nigeria

Zhigila, D. A.; Abdul, S. D.; Inuwa, I.; Yuguda, U. A.; Sawa, F. B.

2026-10-09 plant biology 10.64898/2026.10.06.756981 medRxiv
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Herbarium collections represent an important but frequently underutilized source of information for documenting medicinal plant diversity and preserving ethnomedicinal knowledge. We assessed the medicinal plant diversity represented in herbarium records from the Gombe State University Herbarium (GSUH), Nigeria, and evaluated patterns in taxonomic composition, therapeutic applications, and plant parts reported for medicinal use. We also tested the hypothesis that the plant parts used medicinally are not randomly distributed across therapeutic or ailment categories, but that particular plant parts are disproportionately associated with specific therapeutic applications. The resulting dataset comprised 1,576 specimen records, including 768 recorded as medicinal plant species across 94 families and 1,426 medicinal-use records (Table S1). Fabaceae was the most species-rich family (15.8%), followed by Asteraceae (9.3%), Malvaceae (4.8%), Lamiaceae (4.4%), Rubiaceae (3.2%), and Apocynaceae (3.2%). The most frequently represented therapeutic themes were infections and sexually transmitted diseases (50.7%), gastrointestinal disorders (41.9%), pain, inflammation, and rheumatism (41.9%), fever and malaria (40.2%), and skin diseases and wounds (40.0%). Individual species had a mean of 2.98 therapeutic themes, with medicinal breadth ranging from 1 to 7 themes. A species-aware permutation test confirmed that the observed plant-part and therapeutic-theme association was significantly stronger than expected under random association (9,999 permutations, P < 0.0001). These findings demonstrate the value of herbarium collections as taxonomically anchored resources for recovering information on medicinal plant diversity. Integrating herbarium records with published ethnomedicinal knowledge provides a reproducible framework for linking plant diversity, taxonomy and traditional medicinal applications and highlights the potential of underutilised African herbaria for ethnopharmacological research.

13
Loss of starch synthase IIa alleviates the negative impact of high temperature on rice starch during grain filling

Hashimoto, S.; Matsushima, R.; Oitome, N. F.; Hosaka, Y.; Miura, S.; Crofts, N.; Yamamoto, T.; Fujita, N.

2026-09-29 plant biology 10.64898/2026.09.28.754565 medRxiv
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High temperatures during grain filling stage are becoming increasingly frequent, compromising both grain and eating quality and thereby driving demand for heat-resilient cultivars. Such conditions are known to reduce the expression of granule-bound starch synthase I (GBSSI) and starch branching enzyme IIb (BEIIb), which are involved in starch biosynthesis, resulting in a decrease in amylose content and an increase in long-chain amylopectin. Thus, the present study introduced functional mutation in starch synthase IIa (SSIIa) that increases the proportion of short amylopectin chains to genetically compensate for the high-temperature-induced increase in amylopectin long chain. Rice lines carrying the ss2a mutation were grown at two locations with cooler (Akita) and warmer (Okayama) temperatures. Their grain traits, starch structure, and eating quality were compared. The ss2a mutant lines showed an increased proportion of short amylopectin chains as well as an increased apparent amylose content. Furthermore, these alterations in starch structure varied with the grain-filling temperature of the cultivation sites, ultimately affected eating quality. These results suggest that enriching short amylopectin chain via the ss2a mutation can counteract the increase in long amylopectin chain caused by high temperatures during grain filling, thereby maintaining a desirable starch structure and eating quality.

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PDF2.2 links constitutive defensin expression to disease resistance in Arabidopsis

Singh, S.

2026-10-09 plant biology 10.64898/2026.10.04.756585 medRxiv
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Background: Plant defensins are small cysteine-rich antimicrobial peptides. In Arabidopsis thaliana, the inducible Class I gene AtPDF1.2a is widely used as a transcriptional marker of jasmonate/ethylene immunity, whereas the constitutively expressed Class II members have received less functional attention. AtPDF2.2 was compared with AtPDF1.2a during infection of Arabidopsis by the vascular fungal pathogen Verticillium longisporum, the necrotrophic fungus Sclerotinia sclerotiorum and the bacterium Pseudomonas syringae pv. tomato DC3000. Results: Profiling of the AtPDF family showed that AtPDF2.2, together with the Class II genes AtPDF2.3 and AtPDF2.5, was significantly reduced during V. longisporum infection, whereas AtPDF1.2a was induced. A promoter-reporter line showed a corresponding decrease in AtPDF2.2 promoter activity during V. longisporum infection, strongest in infected roots. KD-pdf2.2 plants contained more V. longisporum DNA, whereas OE-PDF2.2 plants contained less. The same reciprocal disease pattern was observed during S. sclerotiorum and Pst DC3000 infection. Histochemical Prom::AtPDF2.2::GUS reporter signal decreased during S. sclerotiorum infection but increased following Pst DC3000 infection. Basal AtPDF2.2 expression was lower in JA and ET-signaling mutants and higher in the SA-deficient sid2 background, while the analyzed promoter lacked the GCC-box that mediates inducible AtPDF1.2a regulation. AtPDF1.2a overexpression reduced symptoms across the three pathosystems, but loss of AtPDF1.2a increased disease only during V. longisporum infection under the conditions tested and did not alter Pst DC3000 DNA abundance. AtPDF2.2 overexpression was not associated with reduced vegetative growth under the conditions tested, whereas AtPDF1.2a overexpression reduced rosette size and leaf number. Conclusions: AtPDF2.2 is a constitutively expressed Class II defensin whose expression level is associated with disease outcome across two fungal pathogens and a bacterium. Its promoter shows tissue-dependent regulation during V. longisporum infection, while histochemical reporter signal decreases during S. sclerotiorum infection and increases during Pst DC3000 infection. This regulatory behavior differs from that of the inducible reference gene AtPDF1.2a. The reciprocal disease phenotypes of the AtPDF2.2 transgenic backgrounds support a protective association that may involve direct antimicrobial activity, modulation of host responses or both. The absence of reduced vegetative growth in OE-PDF2.2 under the conditions tested supports further evaluation of PDF2-type genes in Brassica.

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A Synergistic Terpenoid-Phenolic Defense Network Confers Resistance to Dendrolimus superans in Hybrid Larch

Panke, Y.; Maolan, L.; Linhuai, X.; Jing, L.; Chunming, L.; Hui, B.

2026-10-09 plant biology 10.64898/2026.10.07.757267 medRxiv
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To elucidate the resistance of hybrid larch (Larix spp.) clones to Dendrolimus superans and their chemical defense mechanisms, 46 clones were subjected to field feeding-infestation experiments. Insect resistance was comprehensively evaluated by the membership function method from three perspectives: insect weight change, frass weight and defoliation rate (damage consequences); pre- and post-infestation physiological indices (constitutive and induced resistance). High- and low-resistance materials screened from each perspective were further analyzed by widely targeted LC-MS metabolomics. The three damage indices differed highly significantly among clones (P<0.01) and were positively correlated, indicating abundant genetic variation in resistance. Clones such as -30-C, -35-A and -21-A showed strong overall resistance, whereas -28-E and -20-A showed high constitutive and induced resistance. Defence enzyme activities (POD, SOD, CAT, PPO and PAL) correlated closely with secondary metabolites (total phenolics, tannins and total flavonoids) and nutrients, and rose synergistically after feeding, reflecting synergy between constitutive and induced resistance. In total, 3,577 metabolites were identified, predominantly terpenoids (>90%). The three groups yielded 1,010, 1,086 and 922 differentially accumulated metabolites, and PCA separated high- from low-resistance materials in all groups. These metabolites were significantly enriched in metabolic pathways, biosynthesis of secondary metabolites, monoterpenoid and diterpenoid biosynthesis, and phenylpropanoid biosynthesis. In conclusion, resistance of hybrid larch to D. superans is jointly conferred by the defence enzyme system, secondary metabolite accumulation and a 'terpenoid-phenolic' metabolic network: terpenoids and phenolics act synergistically via the terpenoid and phenylpropanoid pathways, together forming the core of chemical defense and supporting the breeding of insect-resistant clones.

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Floristic composition, phenology, and conservation value of four peat bogs in Bucovina, with the presence of Betula nana

Trisciuc, M.

2026-09-28 ecology 10.64898/2026.09.22.752958 medRxiv
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This paper presents a comparative analysis of the floristic composition and site characteristics of four peat bogs in Bucovina, Romania: Poiana Stampei, Romanesti, Saru Dornei, and Gaina-Lucina. The research was based on phytosociological releves on 25 msq plots and direct field phenological observations, on six field visits from May to August 2026. Vegetation was characterised using the Braun Blanquet method, and floristic similarity between sites was assessed with the Sorensen and Bray Curtis indices. All four plots shared a common core of taxa characteristic of peatland vegetation: Sphagnum spp., Carex rostrata, Drosera rotundifolia, Eriophorum vaginatum, and Vaccinium species. Species richness was 13 taxa at Poiana Stampei, Romanesti, and Saru Dornei, and 12 at Gaina-Lucina. Romanesti and Saru Dornei showed the highest floristic similarity (descriptive values, not statistically tested, given a single releve per site), while Gaina-Lucina differed most markedly, not through species richness, which was similar across sites, but through species identity and through the presence of Betula nana, a glacial relict absent from the other sites. The results provide a descriptive basis for future research on the floristic composition and conservation of these habitats.

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Phylogenomics reveals distinct evolutionary lineages of Japanese Cinnamomum species in the Nansei Islands

Iizasa, S.; Anjiki, N.; Iizasa, E.

2026-09-24 plant biology 10.64898/2026.09.23.753985 medRxiv
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Although Cinnamomum includes globally important spice and medicinal plants, genome-scale knowledge of Cinnamomum species native to Japan remains limited. The Nansei Islands are an approximately 1,200-km-long archipelago located between the Pacific Ocean and the East China Sea and represent a key biogeographic region shaped by deep-sea barriers such as the Tokara and Kerama Gaps. Here, we integrated chloroplast and nuclear genome data generated in this study with public datasets to examine the phylogenetic relationships of four Cinnamomum species native to the Nansei Islands and ten related species. Chloroplast genomes showed no major structural differences among the four island species, whereas nuclear genome data resolved two distinct evolutionary lineages in the Nansei Islands: C. yabunikkei, C. x durifruticeticola, and C. daphnoides formed a distinct Nansei Islands lineage separate from Sri Lankan, Indonesian, and Taiwanese lineages, whereas C. sieboldii was placed in a Taiwanese lineage. Ancestry estimation indicated that C. x durifruticeticola and C. daphnoides share a common ancestral component, while C. sieboldii exhibited a single homogeneous ancestral component, suggesting reduced ancestral diversity after divergence from a Taiwanese lineage. These findings reveal previously unrecognized genomic differentiation among Japanese Cinnamomum species and suggest that deep-sea barriers, dispersal, and long-term isolation shaped their diversification.

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The conserved function of bHLH121 and overexpression of N-terminal bHLH121 fragment suppresses iron deficiency response in Arabidopsis

Zhou, P.; Liu, C.; Pan, Y.; Fei, Y.; Shen, R.; Wang, R.; Lan, P.

2026-09-28 plant biology 10.64898/2026.09.27.754851 medRxiv
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Iron (Fe) homeostasis is fundamental to plant growth and development and is centrally regulated by a well-conserved network of basic helix-loop-helix (bHLH) transcription factors. In Arabidopsis thaliana, bHLH121 regulates the expression of FER-like iron deficiency-induced transcription factor (FIT) and clade Ib bHLH38/39/100/101 genes, among others. However, its functional conservation remains unexplored, and some contradictory results have been reported regarding the effects of bHLH121 overexpression. Here, we investigated the evolutionary conservation of bHLH121 through complementary tests in the Arabidopsis bhlh121 mutant created by CRISPR/Cas9 gene editing. We also investigated the effects of ectopically expressing full-length, N-terminal, and C-terminal fragments of bHLH121 on plant responses to Fe deficiency in Arabidopsis. Transgenic plants overexpressing the full-length bHLH121 driven by the cauliflower mosaic virus 35S promoter (bHLH121 ox) resulted in shorter roots and more severe leaf chlorosis compared to wild-type plants under Fe limiting conditions. Moreover, overexpression of the N-terminal fragment (N121 ox) rendered plants highly sensitive to Fe deficiency, whereas overexpression of the C terminal fragment (C121 ox) did not. Consistent with the phenotypic observations, N121 ox plants accumulated lower Fe content, and the ferric chelate reductase activity was significantly reduced. In N121 ox plants, the induction of key Fe regulatory genes, especially FIT, was significantly inhibited under Fe deficiency. Confocal imaging showed that both N121-GFP and C121-GFP fusion proteins could localize to the nucleus. Electrophoretic mobility shift assay showed that both full-length bHLH121 and N121 could effectively bind the probes derived from the promoter of FIT. The present work reveals a conserved function of bHLH121 in Fe homeostasis across several dicot species and demonstrates that the N121 may possess a dominant nature in Fe deficiency responses.

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Agronomic characterisation of a common buckwheat germplasm collection unlocks resources for genomics-assisted ideotype breeding

Hess, F.; Kronenberg, L.; Rimat, G.; Heuberger, H.; Herrmann, M. H.; Benyr, V.; Yates, S.; Studer, B.

2026-10-12 plant biology 10.64898/2026.10.09.757874 medRxiv
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The mechanisation of agriculture has achieved major increases in labour productivity but has also led to an erosion of agrobiodiversity. Neglected and underutilized crops such as common buckwheat (Fagopyrum esculentum Moench) offer the potential to diversify food production systems but require major genetic improvement to compete with established crops. Although target traits for breeding such as more homogeneous ripening, reduced height, larger seeds and higher and more stable yields are well known, large-scale and field-based phenotyping data of diverse common buckwheat accessions across multiple environments are still scarce. Here, we tested the agronomic performance of a common buckwheat germplasm collection consisting of 170 accessions of diverse origin. The collection was tested at seven year-sites, and 19 agronomic traits related to phenology, growth habit and yield were assessed. Most traits showed high repeatability (0.31 < R2 < 0.98) and heritability (0.45 < H2 < 0.97), indicating genetic control of these traits. Coefficients of variation ranged between 6.19% and 46.15%, showing the high phenotypic diversity within the collection. By combining multiple traits, distinct ideotypes for different roles in the crop rotation were investigated, and accessions with promising agronomic profiles identified. Genome-wide association studies revealed marker-trait associations for 18 traits, allowing the identification of putative candidate genes influencing these traits such as a putative GRF-INTERACTING FACTOR associated with flowering time and biomass production. Our results show that the collection offers vast phenotypic diversity that can be exploited for the development of improved varieties tailored to meet specific agronomic purposes. The efficient utilization of these genetic resources should be supported by the application of genomics-based breeding approaches. The development of genetic markers and search for putative candidate genes mark important steps towards the establishment of efficient breeding schemes. Thus, these genomic resources provide a foundation for accelerating the genetic improvement of common buckwheat and developing agronomically competitive varieties.

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The bacterial pathogen causing cucurbit yellow vine disease associates with the specialized extrafascicular phloem unique to cucurbits

Mphande, K.; LaSarre, B.; Gleason, M. L.; Beattie, G. A.

2026-09-28 plant biology 10.64898/2026.09.24.754277 medRxiv
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Cucurbit yellow vine disease (CYVD) is a phloem disease caused by a group of Serratia ureilytica bacteria within the S. marcescens complex. The CYVD pathogen is distinct from other phloem pathogens in two fundamental ways: i) unlike other bacterial phloem pathogens, the CYVD pathogen readily grows on standard laboratory media; and ii) whereas most bacterial phloem diseases are transmitted solely by hemipteran insects, CYVD can be transmitted by both hemipteran (squash bug) and non-hemipteran (cucumber beetle) vectors. The phloem of cucurbits is also distinct, with cucurbits having a unique secondary phloem system, the extrafascicular phloem (EFP), for which a role in CYVD has not been investigated. The distinctive features of this pathosystem prompted a re-evaluation of pathogen localization in planta using a green fluorescent protein-tagged CYVD strain during infection of squash (Cucurbita pepo). In epifluorescence and confocal microscopy images of transverse and longitudinal sections of stem and petiole tissues, the fluorescence patterns of the CYVD pathogen were consistent with pathogen colonization of all four known EFP tissues, namely the peripheral, entocyclic, ectocyclic, and commissural sieve tubes. The CYVD pathogen may enter the EFP during herbivory by pathogen-infected vectors that cause mechanical damage, as the EFP exhibits much slower blockage following injury than the conventional fascicular (bundle) phloem. Moreover, although EFP has been proposed to have an antimicrobial function, EFP exudate collected from the stem and petioles of C. pepo supported rapid pathogen growth. These results provide evidence for a unique tissue tropism of the newly emerged plant pathogen S. ureilytica.