Plants

Exposure to Ultraviolet-B (UV-B) light can adversely affect plant growth and cellular integrity by inducing oxidative stress and DNA damage. In this study, we investigated UV-B-induced DNA damage and repair responses in the aquatic monocotyledonous plant species Spirodela polyrhiza (duckweed). We exposed 13-day-old duckweed plantlets to broadband UV-B light for 1-10 min, followed by recovery periods of up to 24 h under normal growth conditions. We observed progressive chlorosis, wilting, and diminished plant vigor with longer durations of UV-B light exposure. Agarose gel electrophoresis demonstrated compromised genomic DNA integrity immediately after UV-B light treatment, with partial restoration of DNA quality during recovery. Immuno-slot blot assays established the accumulation of two major UV light-induced photoproducts, cyclobutane pyrimidine dimers (CPDs) and 6-4 pyrimidine-pyrimidone photoproducts [(6-4)PPs], in a dose-dependent manner following UV-B light exposure. Notably, the abundance of these DNA lesions declined substantially after recovery, indicating activation of endogenous DNA repair mechanisms. Staining with 3,3-diaminobenzidine revealed elevated accumulation of hydrogen peroxide immediately following UV-B exposure, suggesting enhanced oxidative stress. Collectively, these findings demonstrate that S. polyrhiza possesses efficient mechanisms for sensing, repairing, and mitigating DNA damage induced by oxidative stress resulting from UV-B light exposure. This study highlights the potential of duckweed as an effective model system for investigating DNA damage and repair pathways under UV-B light stress in plants.

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.

Ardisia mela is formally described from the forests of the Crystal Mts, Gabon based on a specimen collected in 1968 and informally described in 1979 by Taton as Species 2. Despite the proximity of the collection to a road, and a resurgence of botanical surveys in the Crystal Mts in recent years, the species appears not to have been refound, and as it occurs in a logging concession and areas of cleared forest can be seen in the vicinity of the collection site, the species is considered threatened and provisionally assessed as Critically Endangered using the 2012 IUCN standard.

Central to understanding biodiversity is the systematic classification of biological taxa where concepts on integrating omics data have not yet been comprehensively executed. The data-driven approach is aiming at integrating phylogenetic data from three or more scales aiming at enriching the robustness of species delimitations while also revealing evolutionary drivers of diversification and speciation. This study investigates complex-thallose liverworts as reference and combines the analysis of DNA marker sequencing, morphometrics utilizing bioimaging measurements and chemometrics using liquid chromatography high-resolution mass-spectrometry (UPLC/ESI-QTOF-MS) with data-dependent acquisition of tandem mass-spectra (DDA-MS), with character variation evaluated with dendrograms and data mining. The comparative analysis revealed similar tree topologies albeit with diverging positions of taxa that were attributed to flavonoid-glycosides, auronidins and phenanthrenes resulting from evolutionary radiation and adaptations to stressful bioclimatic conditions, whereas unique fatty acyls in Riccia link to ecological causes of speciation. Characteristic amino acid motifs confirm the intermediary position of liverworts between algae and land plants. By functionally attributing and mapping chemometric markers to the taxonomy, the data-driven approach integrating omics into integrative taxonomy is opening opportunities for plant systematics to identify evolutionary drivers and form new hypotheses on the diversification and speciation of species.

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.

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.

Utricularia gibba L. is an aquatic carnivorous plant with a diverse set of capabilities. Reddening of traps frequently occurs in old in vitro cultures. While anthocyanins are often responsible for red coloration in plants, not every plant turns red. Stress factors like high light or excess sucrose have previously been shown to induce the formation of anthocyanins. Here, we hypothesized the red trap formation to be dependent on nutrient deprivation and tested nitrogen deprivation. The results suggest, that only in combination with light, nitrogen deficiency leads to the activation of the complete anthocyanin biosynthesis pathway and visible red coloration. However, in darkness, anthocyanin biosynthesis appears generally less active compared to light conditions and expression of most anthocyanin biosynthesis genes is not significantly upregulated under nitrogen deficiency.

Three new species of sun orchid (Thelymitra) endemic to Aotearoa New Zealand are here described. These are T. palustris, T. scabrifolia and T. semaphora. The morphological distinctiveness of these three species has been acknowledged for decades; however, their taxonomic status has remained unresolved. Evidence from existing karyological data, recently generated DNA sequence data (LFY and ycf1) and morphological studies from historical and fresh collections are used here to support their formal description. Both, T. palustris and T. semaphora are restricted to wet habitats north of Auckland (North Island). Thelymitra scabrifolia inhabits mostly scrub, and it has a similar northern North Island distribution, but is has been found also in Manawat[a]whi / Three Kings Islands and historically in Otago (South Island). All three species are polyploids and are of conservation concern.

Palmer amaranth (Amaranthus palmeri S. Watson) is a highly competitive weed that has evolved resistance to several modes of action and has recently become a significant problem in Turkiye. This study was designed to determine the distribution of common target-site resistance (TSR) mechanisms to the acetolactate synthase (ALS)-inhibiting herbicide, nicosulfuron, and the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)-inhibiting herbicide, glyphosate, in A. palmeri accessions collected from two agricultural regions of Turkiye; the Cukurova Region and the Gediz Basin. A total of 96 accessions were analyzed for ALS gene mutations and relative EPSPS gene copy number variation. Whole-plant responses were also evaluated based on relative dry weight following treatment with nicosulfuron and glyphosate applied at twice the field rate. The ALS mutations Pro197Ser, Trp574Leu, and Ser653Asn were far more prevalent in plants from the Cukurova Region than those from the Gediz Basin. Elevated relative EPSPS gene copy numbers were more common and higher in the Gediz Basin. Single rate herbicide screening experiments confirmed that accessions from the Cukurova Region exhibited higher dry weight response to nicosulfuron, whereas those from the Gediz Basin had higher to glyphosate Multivariable regression analyses revealed that nicosulfuron response was primarily associated with the Pro197Ser and Trp574Leu mutations, while glyphosate response was correlated with EPSPS gene copy number. These findings reveal distinct regional patterns of TSR mechanisms in A. palmeri in Turkiye that are consistent with differences in herbicide use and/or introduction history, but the roles of local selection, demographic processes, and gene flow remain unclear.

Maize (Zea mays) is a staple food for millions in sub-Saharan Africa (SSA), contributing to both caloric intake and essential micronutrients. Recent breeding efforts have focused on enhancing its nutritional value by increasing provitamin A (PVA) carotenoid content to address vitamin A deficiency (VAD), a common problem among children under 5 years, pregnant and lactating mothers across SSA. However, high rainfall, and both warm and humid conditions in various SSA regions result in devastating foliar diseases such as maize streak virus (MSV), northern corn leaf blight (NCLB), southern corn leaf blight (SCLB), southern corn rust, grey leaf spot (GLS), and Curvularia leaf spot (CLS). Breeding for resistance can aid in mitigating yield losses caused by those diseases. Rapid, efficient screening methods can allow for examining large germplasm collections. The current study evaluated 21 maize inbred lines with contrasting PVA content, along with two commercial inbred controls, for resistance to Exserohilum turcicum, Bipolaris maydis, and Curvularia lunata, causal agents of NCLB, SCLB, and CLS, respectively, using a detached leaf assay (DLA) and under natural field infestation. Significant variation was detected among the inbreds, seven were classified as resistant to certain diseases, ten as moderately resistant, and six as susceptible to all foliar diseases. Overall, high PVA inbred lines had less susceptibility to NCLB, SCLB, and CLS. The results suggest that PVA-enriched maize inbred lines possess improved resistance to multiple foliar diseases. The field assessments of disease severity validated the effectiveness of the DLA in distinguishing resistant from susceptible inbred lines, and resistance to other diseases in field conditions was detected in parallel. The results indicate that high PVA maize has the potential to simultaneously address VAD, mycotoxin contamination in SSA with improved resistance to multiple foliar diseases.

Background: Weed control is one of the main challenges in agriculture today, particularly due to the increasing occurrence of herbicide-resistant populations. Among the most problematic species are Lolium multiflorum (L.) and Echinochloa crus-galli (L.) Beauv., for which glyphosate-resistant populations have been reported. In this context, glufosinate ammonium has emerged as an alternative for their control; however, its efficacy may vary depending on species and photosynthetic metabolism. Objective: The objective of this study was to evaluate the differential sensitivity of ryegrass (C3) and barnyardgrass (C4) to ammonium glufosinate by analyzing physiological responses associated with leaf senescence and photosystem II activity. Methods: Visual injury, chlorophyll fluorescence, and ammonium accumulation were assessed. Results: Results revealed a differential response between species. Barnyardgrass exhibited earlier symptom onset and a greater reduction in the quantum yield of photosystem II ({Phi}PSII), whereas ryegrass showed a slower senescence process. These differences indicate a higher sensitivity of barnyardgrass to glufosinate ammonium, possibly associated with its C4 photosynthetic metabolism. Conclusions: It is concluded that the effectiveness of glufosinate ammonium depends on the type of photosynthetic metabolism and on the ability of each species to cope with herbicide-induced oxidative stress. This information contributes to optimizing glufosinate ammonium use and to the development of management strategies aimed at delaying the evolution of herbicide resistance.

The rapid spread of herbicide-resistant Amaranthus palmeri S. Watson across the Mediterranean region poses a severe threat to crop production. This study characterizes the resistance profile of an A. palmeri population from Gonen, Israel, which survived multiple post-emergence herbicide applications in maize. Greenhouse dose-response and screening trials were conducted across four modes of action (MOAs). The Gonen population exhibited three-way multiple resistance to acetolactate synthase (ALS), photosystem II (PSII) and 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors, marking the first confirmed case outside the Americas. High-level resistance was demonstrated for the ALS inhibitor foramsulfuron and the PSII inhibitor metribuzin, maintaining 40-70% and 80-100% survival, respectively, at double the labelled field rate. Sequencing of the ALS gene revealed a Trp574 to Leu substitution in resistant plants. The population showed differential PSII sensitivity, as atrazine provided complete control despite high metribuzin survival. Sequencing revealed no known psbA mutations, and malathion pre-treatment caused only a slight reduction in metribuzin survival. Partial resistance was observed for the HPPD inhibitor tembotrione (30-50% survival), where malathion pre-treatment reduced survival by 20-30%. In contrast, pre-mergence applications of S-metolachlor (very long chain fatty acid inhibitor), isoxaflutole (HPPD inhibitor), and a fomesafen + terbutryn mixture [protoporphyrinogen oxidase (PPO) + PSII inhibitors] significantly reduced emergence. These findings confirm multiple-resistant A. palmeri in the region and highlight the urgent need for integrated weed management incorporating effective pre-emergence herbicides rather than relying solely on post-emergence chemical control in summer cereal rotations.

LED spectral composition is an important tool for improving the growth and nutritional quality of microgreens cultivated in controlled environments. This study evaluated the effects of three LED light treatments on growth, morphology, pigments, primary metabolites, phenolic composition, and antioxidant capacity in arugula (Eruca sativa), mustard (Brassica juncea), and radish (Raphanus sativus) microgreens. Microgreens were cultivated under controlled environmental conditions and exposed to broad-spectrum white (W), blue-enriched white (WB), and red-enriched white (R) light at a photosynthetic photon flux density of 200 micromol/m2/s. Light quality did not affect yield in any species. However, R increased cotyledon area in arugula by 50 to 60% and promoted hypocotyl elongation in both arugula and radish, whereas W resulted in the longest hypocotyls in mustard. Photosynthetic pigment composition responded differently among species. In mustard, WB increased the chlorophyll a/b ratio (1.12 to 1.18), whereas lutein concentration decreased from 7.06 to 4.20 mg 100 g/FW. Primary metabolism also responded to light treatments in a species-dependent manner. In mustard, W increased glucose (0.43 vs. 0.26 and 0.29 g 100 g/ FW) and fructose (0.33 vs. 0.20 and 0.22 g 100 g/ FW) concentrations compared with WB and R. Organic acid composition was more responsive to light treatments in radish, with higher concentrations under R. Phenolic metabolism also responded in a species-dependent manner. In mustard, W increased total phenolic content to 0.25 mg GAE g/FW compared with 0.15 mg GAE g/FW under WB and R, and ABTS antioxidant capacity to 1.17 mg TE g/FW compared with 0.74 and 0.75 mg TE g/FW under WB and R, respectively. Individual phenolic compounds were also affected by light treatments, particularly in arugula and mustard. These findings demonstrate that the effects of LED spectral composition on microgreen quality are highly species-dependent. Therefore, LED light spectra should be optimized according to the target species and the desired quality attributes rather than applying a single lighting strategy to all Brassicaceae microgreens.

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.

Ultraviolet-C (UV-C) radiation can be highly damaging to plants, yet its effects on gibberellin (GA) homeostasis are not well understood. In this study, we show that short daily UV-C pulse treatments (12 s, 1,200 J m-2) applied for seven days reduce plant height and delay flowering in Arabidopsis thaliana. Endogenous levels of the GA biosynthesis precursors GA12, GA53, GA15, and GA24, the bioactive GA4, and the GA catabolites GA34 and GA110 are all lower in UV-C treated plants than in untreated controls. These changes were accompanied by lower transcript levels of the GA biosynthesis genes KS, GA13ox1, GA20ox1, and GA3ox1, together with opposing changes in the expression of GA2ox genes. Exogenous GA4 restores growth in UV-C-treated plants, suggesting that reduced GA availability contributes to UV-C-induced growth inhibition. Consistent with this finding, the GA-signalling mutant gdella and the GA-biosynthesis mutants kao1 and kao2 show strongly reduced UV-C responses. Together, these findings highlight the importance of GA metabolism and signalling in the developmental response to repeated UV-C exposure, and suggest that exposure regimen influences the dynamics of UV-C-induced hormonal responses.

BackgroundCymbidium tortisepalum var. longibracteatum is a Class II nationally protected endangered plant in China with significant economic value. AimThis study aimed to establish an efficient in vitro flowering system and identify key hormonal factors regulating flower bud differentiation. MethodsOrthogonal experiments and hormone treatments were performed to evaluate the effects of TDZ, 6-BA, NAA, IBA, PP333, ABA, and GA3 on flower bud induction in tissue-cultured plantlets. ResultsTDZ was identified as the key initiator of flower bud differentiation, as no flower buds were induced in TDZ-free treatments. The optimal hormone combination was 1.0 mg{middle dot}L-1 6-BA + 0.4 mg{middle dot}L-1 TDZ + 0.8 mg{middle dot}L-1 NAA + 1.6 mg{middle dot}L-1 IBA, achieving a flower bud induction rate of 23.33% and a normal flower bud rate of 20.00%. PP333 inhibited flower bud differentiation but reduced malformation; at 0.2 mg{middle dot}L-1 PP333, the normal flower bud rate reached 20.00%. GA3 pretreatment resulted in a flower bud induction rate of 12.04%, whereas ABA pretreatment showed no significant promoting effect on flower bud induction. ConclusionsThis study provides an efficient in vitro flowering system and key hormonal parameters for shortening the breeding cycle and elucidating orchid flowering mechanisms.

Understanding the evolutionary origins of globally noxious weeds provides insights into the genetic background of weediness. Among species of Echinochloa, the hexaploid E. crus-galli was previously thought to have arisen through hybridization and allopolyploidization between tetraploid E. oryzicola and an undiscovered diploid species, although they exhibit marked differences in both ecological and morphological characteristics. Recently, in Japan, unknown tetraploid Echinochloa plants were discovered that exhibit distinct ecological and morphological characteristics from those of E. oryzicola, with some traits showing greater similarity to those of E. crus-galli. In this study, we investigated the relationships between well-known Echinochloa species and these unknown tetraploid plants, hereafter referred to as ''ninja''. Subgenome-based read mapping analysis and phylogenetic analysis revealed that ninja possesses a distinct genomic composition, while showing close relationships to E. oryzicola in both its chloroplast and nuclear genomes. Furthermore, the identification of a very small number of ninja x E. oryzicola hybrids among field-collected individuals suggests that these two species rarely hybridize in the wild. Although ninja and E. oryzicola showed similar genomic compositions, ninja has a broader habitat range, partially overlapping with that of E. oryzicola and extending into areas occupied by E. crus-galli. Our findings indicate the possibility that hexaploid E. crus-galli may have originated from a tetraploid Echinochloa species other than E. oryzicola.

Drought and low temperatures are major abiotic factors affecting key physiological and biochemical processes and limiting the yields of soybean (Glycine max L. Merr.). To in-crease soybean production in Europe, different agricultural strategies are applied to re-duce abiotic stress, including biostimulants. Therefore, studies on the effectiveness of local strains isolated in Europe are becoming increasingly relevant. In this study two bacterial strains Arthrobacter pascens (AP) and Bradyrhizobium japonicum (BJ) along with plant-derived protein hydrolysate (PH) were analysed with soybean plans under abiotic stress conditions in plant growth chambers. Six treatments (control; AP; BJ; PH; BJ+AP; BJ+AP+PH) were tested to evaluate biostimulation effect before stress induction (VC stage) and to determine stress reduction effect on soybeans after plants recovery period (V3 stage). Biostimulants application has positive effect on soyabean biometric parameters in early plant development stage and post stress periods. More stable long-term effect was found on structural plant development parameters, than on pigment accumulation. The best results on plant biometric parameters were found where (AP) and (BJ+AP+PH) com-bination was inoculated. (BJ+AP+PH) combination was the only effective treatment, which showed significantly different results in pigments indices, compared to the control, after stress period. Author summaryYasha Jamil: Conceptualization, Data curation, Formal analysis, Writing- original draft, Giuseppe Colla: Formal analysis, Writing- original draft, Writing- review & editing, Justina Kaziuniene: Data curation, Formal analysis, Sarune Ramoskaite :Writing- review & editing. Monika Toleikien[e]: Conceptualization, Data curation, Formal analysis, Writing- original draft, Funding acquisition, Supervision, Writing- review & editing.

Karsts of Southeast Asia cover ~400,000 km2, with tropical limestone karsts possessing highly biodiverse flora in many, varied microhabitats that are unique from lowland- and hill forests. Begonia L. is a pantropical mega-genus with ~1,550 described species to date, ideal for studying evolutionary patterns and factors driving tropical population- diversity and speciation. Begonias grow on multiple substrates including limestone and granite, sandstone, quartzite, on steep earth slopes and near streams in primary forests. Niche partitioning is commonly observed, resulting in many single-site endemic Begonia species especially on limestone karsts. Molecular phylogenetic analyses were performed on combined chloroplast ndhF-rpl32 and nuclear internal transcribed spacer ITS sequences that included 30 selected Begonia species representing all sections from Peninsular Malaysia (Pen. Msia; ten limestone-, thirteen forest- and seven granite species). Results of Bayesian inference and molecular dating showed evolution of Begonia from continental Asia into Pen. Msia during the mid-late Miocene period of ~ 10.65 mya (9.7 - 12.12 mya), with at least two independent dispersal events observed by the early splitting of the ancestral lineage into two major clades i.e. clades A- and B at 9.85 mya (HPD 9.7-10.36 mya) and 8.85 mya (HPD 7.7-11.23 mya) respectively; followed by multiple speciation events colonizing limestone- as well as non-limestone (forest, granite) habitats. Begonia speciation into limestone habitats occurred three times, with climate indicated as more important factor than substrate in diversification - clade A contained the Indian/Continental Asia (monsoon climate) species, i.e. Sect. Platycentrum and Parvibegonia, resolved slightly older than clade B where Sect. Petermannia, Jackia and Ridleyella formed a Sunda Shelf (equatorial climate) centred group. Strong population structuring by locality, common in Begonias, was also observed (e.g. B. kingiana, B. foxworthyi, B. nurii, B. sinuata), regardless of distribution. However, for widespread species (B. kingiana, B. sinuata) behavioural (physiological) inertia on niche evolution, and/or genome size variation and dynamics (genome evolution) may potentially explain species cohesion. Phylogenetics relationships are mostly concordant with current sectional classifications excepting Sect. Parvibegonia, Sect. Diploclinium and two paraphyletic species complexes. Conservation of Begonias and biodiverse limestone karsts in Malaysia remains concerning due to ongoing threats (large-scale quarrying/mining, human encroachment), and most are still without any legal protection.

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.