Plants

Ruta chalepensis is an herb used to treat various ailments, and its potential cytotoxic effects on different tumor cell lines have been extensively studied. The present study aimed to evaluate the cytotoxic activity of R. chalepensis methanol extract (RCME), sub-partitions obtained from solvents of increasing polarity, and major compounds, as well as their hemolytic, anti-hemolytic, and antioxidant potential. The in vitro cytotoxic activity against the human hepatocarcinoma (HEP-G2) and the murine lymphoma cell line (L5178Y-R) was evaluated using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, whereas selectivity indices (SIs) were determined by comparing cytotoxicity against normal African green monkey kidney cells (VERO) and human peripheral blood mononuclear cells (PBMC). Hemolytic and anti-hemolytic activities were evaluated on human erythrocytes. The most effective cytotoxic treatment was evaluated for nitric oxide release by J774A.1 macrophages. Antioxidant activity of R. chalepensis material was also determined. Results showed that RCME produced significant (p < 0.05) cytotoxicity in HEP-G2 (IC₅₀ = 1.79 µg/mL) and L5178Y-R (IC₅₀ = 1.60 µg/mL) cells and exhibited high SIs (291.50 and 114.80, respectively). In addition, the n-hexane fraction (RCHF) showed an IC₅₀ of 18.31 µg/mL in HEP-G2 cells and an SI of 9.48 in VERO cells, whereas the chloroform fraction (RCCF) evidenced an IC₅₀ of 1.60 µg/mL in L5178Y-R cells and an SI of 34.27 in PBMC cells. Chalepensin (CHL), rutamarin (RTM), and graveolin (GRV), which are major components of R. chalepensis, showed high activity against L5178Y-R cells, with IC₅₀ of 9.15, 15.13 and SI of 45.08 µg/mL, respectively. In addition, CHL, RTM, and GRV showed SIs of 24.76, 9.98, and 3.52, respectively, when compared with PBMC cells. RCME at concentrations of 125 µg/mL and 250 µg/mL, significantly (p < 0.05) decreased nitrite production in J774A.1 cells, when exposed to lipopolysaccharide. This study demonstrated that RCME showed significant cytotoxic activity against HEP-G2 and L5178Y-R cells, without affecting normal VERO, PBMC, and J774A.1 cells. Full article

Compatibility interactions between the host and the fungal proteins are necessary to successfully establish a disease in plants by fungi or other diseases. Photochemical and antimicrobial substances are generally known to increase plant resilience, which is essential for eradicating fungus infections. Through homology modeling and in silico docking analysis, we assessed 50 phytochemicals from cucumber (Cucumis sativus), 15 antimicrobial compounds from botanical sources, and six compounds from chemical sources against two proteins of Pseudoperonospora cubensis linked to cucumber downy mildew. Alpha and beta sheets made up the 3D structures of the two protein models. According to Ramachandran plot analysis, the QNE 4 effector protein model was considered high quality because it had 86.8% of its residues in the preferred region. The results of the molecular docking analysis showed that the QNE4 and cytochrome oxidase subunit 1 proteins of P. cubensis showed good binding affinities with glucosyl flavones, terpenoids and flavonoids from phytochemicals, antimicrobial compounds from botanicals (garlic and clove), and chemically synthesized compounds, indicating the potential for antifungal activity. Full article

Plant awareness disparity (PAD, formerly plant blindness) is the human inability to notice plants in everyday life. It is suggested that the main underlying factors of PAD are: 1. the inability to recognize individual plants and 2. stronger preferences for animals, which prevents building positive attitudes toward them. The presentation of individual plants should trigger more positive responses toward them than the presentation of groups of plants. Strong preferences for animals predict that the presence of an animal on a plant might enhance positive perceptions of the plant by people. We experimentally investigated the perceived attractiveness and willingness to protect (WTP) plants presented individually and in groups and with or without various pollinators in a sample of Slovak people (N = 238). In contrast to the first prediction, only one of four plants (dog rose, but not saffron, spruce, or beech tree) received higher attractiveness scores when presented individually than in a group. None of these species received higher WTP scores when presented individually, rather than in a group. The effect of the presence of pollinators on flower attractiveness and WTP was distinguished between vertebrates and invertebrates; only flowers with birds and bats increased their attractiveness scores, while flowers with invertebrates, including a butterfly, honeybee, beetle, and the syrphid fly, received similar or lower scores than the same plant species without pollinators. WTP plants significantly increased only when the scarlet honeycreeper and the cave nectar bat were present on flowers as pollinators. People showed significantly stronger preferences for items that associate 1. plants with pollinators and 2. plants with animals that distribute animal seed than for items focused solely on plants. Connecting animals and plants should help reduce PAD. This aim cannot be achieved, however, by presenting individual plants and/or plants with randomly chosen pollinators. Full article

Solanum section Leptostemonum is an ideal lineage to test the theoretical framework regarding proposed evolutionary benefits of outcrossing sexual systems in comparison to cosexuality. Theoretically, non-cosexual taxa should support more genetic diversity within populations, experience less inbreeding, and have less genetic structure due to a restricted ability to self-fertilize. However, many confounding factors present challenges for a confident inference that inherent differences in sexual systems influence observed genetic patterns among populations. This study provides a foundational baseline of the population genetics of several species of different sexual systems with the aim of generating hypotheses of any factor—including sexual system—that influences genetic patterns. Importantly, results indicate that dioecious S. asymmetriphyllum maintains less genetic structure and greater admixture among populations than cosexual S. raphiotes at the same three locations where they co-occur. This suggests that when certain conditions are met, the evolution of dioecy may have proceeded as a means to avoid genetic consequences of self-compatibility and may support hypotheses of benefits gained through differential resource allocation partitioned across sexes. Arguably, the most significant finding of this study is that all taxa are strongly inbred, possibly reflective of a shared response to recent climate shifts, such as the increased frequency and intensity of the region’s fire regime. Full article

The yerba mate leaf metabolic composition depends mainly on genetics, sex, plant and leaf age, light intensity, harvest time, climate, and fertilization. In yerba mate, the secondary sexual dimorphism (SSD), the leaf metabolic SSD association with the frequency of leaf harvests, and the stability of the metabolites in the two genders over the years is not known. It was hypothesized that (1) the SSD in the metabolite segregation would differ among the winter and summer growth pauses, (2) females would show lower metabolite concentrations, and (3) the metabolic concentrations would show stability over the years on the same plants, not obligatorily associated with the SSD stability expression. Variations in theobromine, caffeine, chlorogenic and caffeic acids were correlated to the increasing time since the previous harvest, especially in females. However, the frequency of the metabolic SSD were associated with the studied growth pauses, rejecting the first hypothesis. No regular gender superiority was expressed in the yerba mate leaf secondary metabolites, rejecting our second hypothesis, even though more cases of superior female metabolite accumulation were identified. The stability of the leaf protein was preserved over the four years, with no SSD cases observed. The leaf methylxanthines were time stable, while the decrease in the phenolic content occurred with tree aging, which was not associated with the SSD expression, partially proving our third hypothesis. The novelty was related to the time stability of the leaf metabolic SSD observed over the winter and summer growth pauses, and over the four consecutive years without a regular expression of the male- or female-biased concentrations in the studied metabolites. To demystify the random metabolic gender responses in yerba mate, gender-orientated experiments with a high number of tree repetitions must be conducted, including clonal plants grown in various environments, such as monoculture and agroforestry, or on plantations in different climates and altitudes. Full article

Grewia lasiocarpa E. Mey. Ex Harv., Malvaceae (forest raisin) is a tropical small tree or shrub valued for its ecological importance as well as its nutritional, antioxidant, antibacterial, and anti-cancer properties as well as its ecological and ornamental importance. Glandular and non-glandular trichomes are present on the fruits, stem bark and leaves of G. lasiocarpa and these trichomes are the first line of defense. They are important structures that plants use to combat biotic and abiotic stress. The development of G. lasiocarpa trichomes and the biomechanics of the exudates present in the glandular (capitate) trichome were investigated for the first time using advanced microscopy techniques [Scanning electron microscope (SEM) and Transmission electron microscope (TEM)]. The pressurized cuticular striations may play a role in the exudates’ biomechanics, i.e., releasing secondary metabolites present in the capitate trichome, which was observed to be multidirectional. The presence of many glandular trichomes on a plant implies an increase in the amount of phytometabolites. A common precursor for the development of trichomes (non-glandular and glandular) was observed to be DNA synthesis associated with a periclinal cell division, thus the final fate of the cell is determined by cell cycle regulation, polarity, and expansion. The glandular trichomes of G. lasiocarpa are multicellular and polyglandular, while the non-glandular (glandless) trichomes are either single-celled or multicellular. Since, trichomes ‘house’ phytocompounds of medicinal, nutritional, and agronomical benefits; the molecular and genetic study of the glandular trichomes of Grewia lasiocarpa will be beneficial to humanity. Full article

Soil salinity is a major abiotic stress in global agricultural productivity with an estimated 50% of arable land predicted to become salinized by 2050. Since most domesticated crops are glycophytes, they cannot be cultivated on salt soils. The use of beneficial microorganisms inhabiting the rhizosphere (PGPR) is a promising tool to alleviate salt stress in various crops and represents a strategy to increase agricultural productivity in salt soils. Increasing evidence underlines that PGPR affect plant physiological, biochemical, and molecular responses to salt stress. The mechanisms behind these phenomena include osmotic adjustment, modulation of the plant antioxidant system, ion homeostasis, modulation of the phytohormonal balance, increase in nutrient uptake, and the formation of biofilms. This review focuses on the recent literature regarding the molecular mechanisms that PGPR use to improve plant growth under salinity. In addition, very recent -OMICs approaches were reported, dissecting the role of PGPR in modulating plant genomes and epigenomes, opening up the possibility of combining the high genetic variations of plants with the action of PGPR for the selection of useful plant traits to cope with salt stress conditions. Full article

Mangroves are ecologically significant plants in marine habitats that inhabit the coastlines of many countries. Being a highly productive and diverse ecosystem, mangroves are rich in numerous classes of phytochemicals that are of great importance in the field of pharmaceutical industries. The red mangrove (Rhizophora stylosa Griff.) is a common member of the Rhizophoraceae family and the dominant species in the mangrove ecosystem of Indonesia. R. stylosa mangrove species are rich in alkaloids, flavonoids, phenolic acids, tannins, terpenoids, saponins, and steroids, and are widely used in traditional medicine for anti-inflammatory, antibacterial, antioxidant, and antipyretic effects. This review aims to provide a comprehensive understanding of the botanical description, phytochemical profiles, pharmacological activities, and medicinal potentials of R. stylosa. Full article

Plant invasion has severely damaged ecosystem stability and species diversity worldwide. The cooperation between arbuscular mycorrhizal fungi (AMF) and plant roots is often affected by changes in the external environment. Exogenous phosphorus (P) addition can alter the root absorption of soil resources, thus regulating the root growth and development of exotic and native plants. However, it remains unclear how exogenous P addition regulates the root growth and development of exotic and native plants mediated by AMF, affecting the exotic plant invasion. In this experiment, the invasive plant Eupatorium adenophorum and native plant Eupatorium lindleyanum were selected and cultured under intraspecific (Intra-) competition and interspecific (Inter-) competition conditions, involving inoculation with (M⁺) and without AMF (M⁻) and three different levels of P addition including no addition (P₀), addition with 15 mg P kg⁻¹ soil (P₁₅), and addition with 25 mg P kg⁻¹ soil (P₂₅) for the two species. Root traits of the two species were analyzed to study the response of the two species’ roots to AMF inoculation and P addition. The results showed that AMF significantly promoted the root biomass, length, surface area, volume, tips, branching points, and carbon (C), nitrogen (N), and P accumulation of the two species. Under M⁺ treatment, the Inter- competition decreased the root growth and nutrient accumulation of invasive E. adenophorum but increased the root growth and nutrient accumulation of native E. lindleyanum relative to the Intra- competition. Meanwhile, the exotic and native plants responded differently to P addition, exhibiting root growth and nutrient accumulation of invasive E. adenophorum increased with P addition, whereas native E. lindleyanum reduced with P addition. Further, the root growth and nutrition accumulation of native E. lindleyanum were higher than invasive E. adenophorum under Inter- competition. In conclusion, exogenous P addition promoted the invasive plant but reduced the native plant in root growth and nutrient accumulation regulated by AMF, although the native plant outcompeted the invasive plant when the two species competed. The findings provide a critical perspective that the anthropogenic P fertilizer addition might potentially contribute to the successful invasion of exotic plants. Full article

Rosa roxburghii f. eseiosa Ku is a variety of Rosa roxburghii, with two known genotypes: Wuci 1 and Wuci 2. The lack of prickle on the peel of R. roxburghii f. eseiosa makes it easy to pick and process, but its fruit size is small. Therefore, we aim to induce polyploidy in order to obtain a larger fruit variety of R. roxburghii f. eseiosa. In this study, current-year stems of Wuci 1 and Wuci 2 were used as materials for polyploid induction, which was carried out through colchicine treatment coupled with tissue culture and rapid propagation technology. Impregnation and smearing methods were effectively used to produce polyploids. Using flow cytometry and a chromosome counting method, it was found that one autotetraploid of Wuci 1 (2n = 4x = 28) was obtained by the impregnation method before primary culture, with a variation rate of 1.11%. Meanwhile, seven Wuci 2 bud mutation tetraploids (2n = 4x = 28) were produced by smearing methods during the training seedling stage. When tissue-culture seedlings were treated with 20 mg/L colchicine for 15 days, the highest polyploidy rate was up to 60%. Morphological differences between different ploidys were observed. The side leaflet shape index, guard cell length, and stomatal length of the Wuci 1 tetraploid were significantly different from those of the Wuci 1 diploid. The terminal leaflet width, terminal leaflet shape index, side leaflet length, side leaflet width, guard cell length, guard cell width, stomatal length, and stomatal width of the Wuci 2 tetraploid were significantly different from those of the Wuci 2 diploid. Additionally, the leaf color of the Wuci 1 and Wuci 2 tetraploids changed from light to dark, with an initial decrease in chlorophyll content followed by an increase. In summary, this study established an effective method for inducing polyploids in R. roxburghii f. eseiosa, which could provide a foundation for the breeding and development of new genetic resources for R. roxburghii f. eseiosa and other R. roxburghii varieties in the future. Full article

We aimed to explore how the invasion of the alien plant Solanum elaeagnifolium affects soil microbial and nematode communities in Mediterranean pines (Pinus brutia) and maquis (Quercus coccifera). In each habitat, we studied soil communities from the undisturbed core of both formations and from their disturbed peripheral areas that were either invaded or not by S. elaeagnifolium. Most studied variables were affected by habitat type, while the effect of S. elaeagnifolium was different in each habitat. Compared to maquis, the soil in pines had higher silt content and lower sand content and higher water content and organic content, supporting a much larger microbial biomass (PLFA) and an abundance of microbivorous nematodes. The invasion of S. elaeagnifolium in pines had a negative effect on organic content and microbial biomass, which was reflected in most bacterivorous and fungivorous nematode genera. Herbivores were not affected. In contrast, in maquis, organic content and microbial biomass responded positively to invasion, raising the few genera of enrichment opportunists and the Enrichment Index. Most microbivores were not affected, while herbivores, mostly Paratylenchus, increased. The plants colonizing the peripheral areas in maquis probably offered a qualitative food source to microbes and root herbivores, which in pines was not sufficient to affect the much larger microbial biomass. Full article

Wheat is a staple crop; its production must achieve both high yield and good quality due to worldwide demands for food security and better quality of life. It has been found that the grain qualities vary greatly within the different layers of wheat kernels. In this paper, the spatial distributions of protein and its components, starch, dietary fiber, and microelements are summarized in detail. The underlying mechanisms regarding the formation of protein and starch, as well as spatial distribution, are discussed from the views of substrate supply and the protein and starch synthesis capacity. The regulating effects of cultivation practices on gradients in composition are identified. Finally, breakthrough solutions for exploring the underlying mechanisms of the spatial gradients of functional components are presented. This paper will provide research perspectives for producing wheat that is both high in yield and of good quality. Full article

The structure of phytobenthic diatom communities was studied to reveal differences between natural and channelized river sections in Slovenia. As part of the national monitoring of surface waters, samples of phytobenthos were collected at 85 sites throughout the country according to standard protocols. At the same time, basic environmental parameters were also assessed. Trophic (TI) and saprobic (SI) indices were calculated based on diatoms and other algae, while diversity indices and gradient analyses were performed only for the diatom community. The results showed that channelized rivers harbor significantly more diverse benthic diatom communities than natural sections, mainly due to the significantly higher number of motile diatom taxa that are able to take advantage of more nutrient-rich and less-shaded river sections because of their high adaptability. Selected environmental parameters explained 34% of the variability in diatom community structure when taxa were classified into ecological types. The removal of Achnanthidium minutissimum yielded clearer results (24.1%) than the total species matrix (22.6%). Therefore, we suggest excluding this taxon from calculations of TI, SI, or other indices when it is determined as A. minutissimum complex, because A. minutissimum complex was most abundant in both types of reaches in our study and has a wide ecological amplitude, which reduces the indicative power of the diatom community in the evaluation of environmental conditions and ecological status. Full article

The application of silicon (Si) fertilizer positively impacts crop health, yield, and seed quality worldwide. Si is a “quasi-essential” element that is crucial for plant nutrition and stress response but is less associated with growth. This study aimed to investigate the effect of Si on the yield of cultivated soybean (Glycine max L). Two locations, Gyeongsan and Gunwi, in the Republic of Korea were selected, and a land suitability analysis was performed using QGIS version 3.28.1. The experiments at both locations consisted of three treatments: the control, Si fertilizer application at 2.3 kg per plot (9 m × 9 m) (T1), and Si fertilizer application at 4.6 kg per plot (9 m × 9 m) (T2). The agronomic, root, and yield traits, as well as vegetative indices, were analyzed to evaluate the overall impact of Si. The results demonstrated that Si had consistently significant effects on most root and shoot parameters in the two experimental fields, which led to significantly increased crop yield when compared with the control, with T2 (22.8% and 25.6%, representing an output of 2.19 and 2.24 t ha⁻¹ at Gyeongsan and Gunwi, respectively) showing a higher yield than T1 (11% and 14.2%, representing 1.98 and 2.04 t ha⁻¹ at Gyeongsan and Gunwi, respectively). These results demonstrate the positive impact of exogenous Si application on the overall growth, morphological and physiological traits, and yield output of soybeans. However, the application of the optimal concentration of Si according to the crop requirement, soil status, and environmental conditions requires further studies. Full article

With increasing throughput in both the generation and phenotyping of mutant lines in plants, it is important to have an efficient and reliable genotyping method. Traditional workflows, still commonly used in many labs, have time-consuming and expensive steps, such as DNA purification, cloning and growing E. coli cultures. We propose an alternative workflow where these steps are bypassed, using Phire polymerase on fresh plant tissue, and ExoProStar treatment as preparation for sequencing. We generated CRISPR-Cas9 mutants for ZAS (ZAXINONE SYNTHASE) in rice with two guide RNAs. Using both a traditional workflow and our proposed workflow, we genotyped nine T1 plants. To interpret the sequencing output, which is often complex in CRISPR-generated mutants, we used free online automatic analysis systems and compared the results. Our proposed workflow produces results of the same quality as the old workflow, but in 1 day instead of 3 days and about 35 times cheaper. This workflow also consists of fewer steps and reduces the risk of cross contamination and mistakes. Furthermore, the automated sequence analysis packages are mostly accurate and could easily be used for bulk analysis. Based on these advantages, we encourage academic and commercial labs conducting genotyping to consider switching over to our proposed workflow. Full article

The carnivorous pitcher plants of the genus Nepenthes exhibit many ethnobotanical uses, including treatments of stomachache and fever. In this study, we prepared different extracts from the pitcher, stem, and leaf extracts of Nepenthes miranda obtained using 100% methanol and analyzed their inhibitory effects on recombinant single-stranded DNA-binding protein (SSB) from Klebsiella pneumoniae (KpSSB). SSB is essential for DNA replication and cell survival and thus an attractive target for potential antipathogen chemotherapy. Different extracts prepared from Sinningia bullata, a tuberous member of the flowering plant family Gesneriaceae, were also used to investigate anti-KpSSB properties. Among these extracts, the stem extract of N. miranda exhibited the highest anti-KpSSB activity with an IC₅₀ value of 15.0 ± 1.8 μg/mL. The cytotoxic effects of the stem extract of N. miranda on the survival and apoptosis of the cancer cell lines Ca9-22 gingival carcinoma, CAL27 oral adenosquamous carcinoma, PC-9 pulmonary adenocarcinoma, B16F10 melanoma, and 4T1 mammary carcinoma cells were also demonstrated and compared. Based on collective data, the cytotoxic activities of the stem extract at a concentration of 20 μg/mL followed the order Ca9-22 > CAL27 > PC9 > 4T1 > B16F10 cells. The stem extract of N. miranda at a concentration of 40 μg/mL completely inhibited Ca9-22 cell migration and proliferation. In addition, incubation with this extract at a concentration of 20 μg/mL boosted the distribution of the G2 phase from 7.9% to 29.2% in the Ca9-22 cells; in other words, the stem extract might suppress Ca9-22 cell proliferation by inducing G2 cell cycle arrest. Through gas chromatography–mass spectrometry, the 16 most abundant compounds in the stem extract of N. miranda were tentatively identified. The 10 most abundant compounds in the stem extract of N. miranda were used for docking analysis, and their docking scores were compared. The binding capacity of these compounds was in the order sitosterol > hexadecanoic acid > oleic acid > plumbagin > 2-ethyl-3-methylnaphtho[2,3-b]thiophene-4,9-dione > methyl α-d-galactopyranoside > 3-methoxycatechol > catechol > pyrogallol > hydroxyhydroquinone; thus, sitosterol might exhibit the greatest inhibitory capacity against KpSSB among the selected compounds. Overall, these results may indicate the pharmacological potential of N. miranda for further therapeutic applications. Full article

Pre-sowing seed priming is one of the methods used to improve the performance of tomato plants under salt stress, but its effect photosynthesis, yield, and quality have not yet been well investigated. This experiment aimed to alleviate the impact of sodium chloride stress on the photosynthesis parameters of tomato cv. Micro−Tom (a dwarf Solanum lycopersicum L.) plants exposed to salt stress conditions. Each treatment combination consisted of five different sodium chloride concentrations (0 mM, 50 mM, 100 mM, 150 mM, and 200 mM) and four priming treatments (0 MPa, −0.4 MPa, −0.8 MPa, and −1.2 MPa), with five replications. Microtome seeds were subjected to polyethylene glycol (PEG6000) treatments for 48 hours for priming, followed by germination on a moist filter paper, and then transferred to the germination bed after 24 h. Subsequently, the seedlings were transplanted into the Rockwool, and the salinity treatments were administered after a month. In our study salinity significantly affected tomato plants’ physiological and antioxidant attributes. Primed seeds produced plants that exhibited relatively better photosynthetic activity than those grown from unprimed seeds. Our findings indicated that priming doses of −0.8 MPa and −1.2 MPa were the most effective at stimulating tomato plant photosynthesis, and biochemical contents under salinity-related conditions. Moreover, primed plants demonstrated relatively superior fruit quality features such as fruit color, fruit Brix, sugars (glucose, fructose, and sucrose), organic acids, and vitamin C contents under salt stress, compared to non-primed plants. Furthermore, priming treatments significantly decreased the malondialdehyde, proline, and hydrogen peroxide content in plant leaves. Our results suggest that seed priming may be a long-term method for improving crop productivity and quality in challenging environments by enhancing the growth, physiological responses, and fruit quality attributes of Micro-Tom tomato plants under salt stress conditions. Full article

Catharanthus roseus L. (G.) Don is the most widely studied plant because of its high pharmacological value. In vitro culture uses various plant parts such as leaves, nodes, internodes and roots for inducing callus and subsequent plant regeneration in C. roseus. However, till now, little work has been conducted on anther tissue using plant tissue culture techniques. Therefore, the aim of this work is to establish a protocol for in vitro induction of callus by utilizing anthers as explants in MS (Murashige and Skoog) medium fortified with different concentrations and combinations of PGRs. The best callusing medium contains high α-naphthalene acetic acid (NAA) and low kinetin (Kn) concentrations showing a callusing frequency of 86.6%. SEM–EDX analysis was carried out to compare the elemental distribution on the surfaces of anther and anther-derived calli, and the two were noted to be nearly identical in their elemental composition. Gas chromatography–mass spectrometry (GC–MS) analysis of methanol extracts of anther and anther-derived calli was conducted, which revealed the presence of a wide range of phytocompounds. Some of them are ajmalicine, vindolinine, coronaridine, squalene, pleiocarpamine, stigmasterol, etc. More importantly, about 17 compounds are exclusively present in anther-derived callus (not in anther) of Catharanthus. The ploidy status of anther-derived callus was examined via flow cytometry (FCM), and it was estimated to be 0.76 pg, showing the haploid nature of callus. The present work therefore represents an efficient way to produce high-value medicinal compounds from anther callus in a lesser period of time on a larger scale. Full article

Lindera megaphylla, a broad-leaved evergreen that is used as a landscape ornamental plant and medicinal plant, is an ecologically important and dominant tree species. However, little is known about the molecular mechanisms of its growth, development, and metabolism. The selection of suitable reference genes is critical for molecular biological analyses. To date, no research on reference genes as a foundation for gene expression analysis has been undertaken in L. megaphylla. In this study, 14 candidate genes were selected from the transcriptome database of L. megaphylla for RT-qPCR assay under different conditions. Results showed that helicase-15 and UBC28 were most stable in different tissues of seedlings and adult trees. For different leaf developmental stages, the best combination of reference genes was ACT7 and UBC36. UBC36 and TCTP were the best under cold treatment, while PAB2 and CYP20-2 were the best under heat treatment. Finally, a RT-qPCR assay of LmNAC83 and LmERF60 genes were used to further verify the reliability of selected reference genes above. This work is the first to select and evaluate the stability of reference genes for the normalization of gene expression analysis in L. megaphylla and will provide an important foundation for future genetic studies of this species. Full article