Plant Cuticle Research Articles

Fabrication and characterization of biomimetic plant cuticles from pullulan - graphene oxide (PU-GO) and beeswax - stearic acid (BW-SA) for Citrus Limon Rosso preservation

Inspired by the natural plant cuticles, a novel strategy was proposed for the fabrication of biomimetic plant cuticles from pullulan-graphene oxide (PU-GO) and beeswax-stearic acid (BW-SA), which could serve as hydrophilic polysaccharides and hydrophobic waxes, respectively. PU-GO and PU-GO/BW-SA in different GO concentrations (0, 10, 30 and 50 μg/mL) were prepared, and their structural characteristics and basic properties were investigated. Results showed that PU-GO/BW-SA possessed a hydrophilic layer and a hydrophobic structure similar to the structure of natural plant cuticles. The incorporation of GO enhanced the barrier properties of the films and PU-GO/BW-SA showed a higher contact angle, lower tensile strength and higher barrier properties compared with PU-GO. In addition, PU-GO/BW-SA in 10 μg/mL GO concentration (PU-GO10/BW-SA) possessed the lowest WVP (7.2 × 10−7 g/(m h Pa)) and a contact angle (93.78°) similar to natural plant cuticles. Applications in Citrus Limon Rosso further proved the potential of PU-GO10/BW-SA as a biomimetic plant cuticle in fruit preservation.

NAC transcription factor SlNOR-like1 plays a dual regulatory role in tomato fruit cuticle formation.

The plant cuticle is an important protective barrier on the plant surface, constructed mainly by polymerized cutin matrix and a complex wax mixture. Although the pathway of plant cuticle biosynthesis has been clarified, knowledge of the transcriptional regulation network underlying fruit cuticle formation remains limited. In the present work, we discovered that tomato fruits of the NAC transcription factor SlNOR-like1 knockout mutants (nor-like1) produced by CRISPR/Cas9 [clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9] displayed reduced cutin deposition and cuticle thickness, with a microcracking phenotype, while wax accumulation was promoted. Further research revealed that SlNOR-like1 promotes cutin deposition by binding to the promoters of glycerol-3-phosphate acyltransferase6 (SlGPAT6; a key gene for cutin monomer formation) and CUTIN DEFICIENT2 (SlCD2; a positive regulator of cutin production) to activate their expression. Meanwhile, SlNOR-like1 inhibits wax accumulation, acting as a transcriptional repressor by targeting wax biosynthesis, and transport-related genes 3-ketoacyl-CoA synthase1 (SlKCS1), ECERIFERUM 1-2 (SlCER1-2), SlWAX2, and glycosylphosphatidylinositol-anchored lipid transfer protein 1-like (SlLTPG1-like). In conclusion, SlNOR-like1 executes a dual regulatory effect on tomato fruit cuticle development. Our results provide a new model for the transcriptional regulation of fruit cuticle formation.

Mapping of the Waxy Gene in Brassica napus L. via Bulked Segregant Analysis (BSA) and Whole-Genome Resequencing

Plant cuticular wax is the covering of the outer layer of the plant. It forms a protective barrier on the epidermis of plants and plays a vital role like a safeguard from abiotic and biotic stresses. In the present study, Brassica napus L. materials with and without wax powder were observed. Genetic analysis showed that the separation ratio of waxy plants to waxless plants was 15:1 in the F2 population, which indicated that the wax powder formation was controlled by two pairs of genes. In order to identify the candidate genes associated with the wax powder trait of B. napus L., bulked segregant analysis (BSA) was performed. The homozygous waxy plants, the homozygous waxless plants, and plants from three parents were selected for establishing five DNA pools for genome-wide resequencing. The results of the resequencing showed that the site associated with wax powder trait was located in the region of 590,663–1,657,546 bp on chromosome A08. And 48 single nucleotide polymorphisms (SNPs) were found between the DNA sequences of waxy plants and waxless plants in this region. These SNPs were distributed across 16 gene loci. qRT-PCR analysis was conducted for the 16 candidate genes and three genes (BnaA08g01070D, BnaA08g02130D, and BnaA08g00890D) showed significantly differential expression between waxy and waxless parents. BnaA08g01070D and BnaA08g02130D were significantly down-regulated in the waxless parent, while BnaA08g00890D was significantly up-regulated in the waxless parent. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the BnaA08g02130D gene was enriched in lipid biosynthetic or metabolic processes. All the results in our study would provide valuable clues for exploring the genes involved in wax powder development.

Effect of Epidermis of Plant Leaves on Their Interaction Efficiency with Low-Intensity Laser Light

Effect of Epidermis of Plant Leaves on Their Interaction Efficiency with Low-Intensity Laser Light

AaRgs1 and AaRgs2 differential regulate fungal development, stress response and appressorium-like formation in Alternaria alternata

Alternaria alternata can recognize and respond to physichemical cues from the host plant epidermis through a complex signaling network to initiate infectious structures formation. The primarily functions of RGS as a negative regulator of G protein signaling has been characterized in several model fungi, but its biological function in A. alternata remains unclear. In present study three RGS proteins were identified in A. alternata by Blast alignment and designed as AaRgs1, AaRgs2 and AaRgs3. Targeted genes deletion demonstrated that AaRgs1 and AaRgs2 play the opposite function in vegetative growth, melanin production and secretion, stress response and appressorium-like formation rate in A. alternata, while AaRgs3 had no certain regulatory effect. Differently, AaRgs1 negatively regulated tolerance to high-osmosis stress response and cell wall synthesis inhibitors by down regulating the expression level of AaHog1 and AaSlt2. AaRgs2 acts as a negative role in regulating appressorium-like formation rates through decreasing intracellular cyclic adenosine monophosphate (cAMP) accumulation. Deletion of AaRgs1 and AaRgs2 also resulted in significantly reducing penetrating ability and pathogenicity. The findings indicated that RGS proteins play multiple roles in fungal development, stress response and appressorium-like formation of A. alternata.

Analysis of fossil plant cuticles using vibrational spectroscopy: A new preparation protocol

Analyses for organic “fingerprints” on fossilized plant cuticles and pollen hold valuable chemotaxonomic and palaeoclimatic information, and are thus becoming more utilized by palaeobotanists. Plant cuticle and pollen composition are generally analyzed after standard treatments with several chemical reagents for mineral and mesophyll removal. However, the potential alterations on the fossil composition caused by the different cleaning reagents used are still poorly understood. We tested the effects of commonly used palaeobotanical processing methods on the spectra of fossilized cuticles from successions of Late Triassic to Early Jurassic age, including the gymnosperms Lepidopteris, Ginkgoites, Podozamites, Ptilozamites and Pterophyllum astartense. Our study shows that standard chemical processing caused chemical alterations that might lead to erroneous interpretation of the infrared (IR) spectra. The difference in pH caused by HCl induces changes in the proportion between the two bands at ∼1720 and 1600 cm−1 (carboxylate and C-C stretch of aromatic compounds) indicating that the band at ∼1610 cm−1 at least partially corresponds to carboxylate instead of C-C stretch of aromatic compounds.Interestingly, despite being used in high concentration, HF did not cause changes in the chemical composition of the cuticles. The most alarming changes were caused by the use of Schulze's solution, which resulted in the addition of both NO2 and (O)NO2 compounds in the cuticle. Consequently, a new protocol using H2CO3, HF, and H2O2 for preparing fossil plant cuticles aimed for chemical analyses is proposed, which provides an effective substitute to the conventional methods. In particular, a less aggressive and more sustainable alternative to Schulze's solution is shown to be hydrogen peroxide, which causes only minor alteration of the fossil cuticle's chemical composition. Future work should carefully follow protocols, having in mind the impacts of different solutions used to treat leaves and other palaeobotanical material such as palynomorphs with aims to enable the direct comparison of spectra obtained in different studies.

Adaptation of yeast Saccharomyces cerevisiae to grape-skin environment

Saccharomyces cerevisiae, an essential player in alcoholic fermentation during winemaking, is rarely found in intact grapes. Although grape-skin environment is unsuitable for S. cerevisiae’s stable residence, Saccharomycetaceae-family fermentative yeasts can increase population on grape berries after colonization during raisin production. Here, we addressed adaptation of S. cerevisiae to grape-skin ecosystem. The yeast-like fungus Aureobasidium pullulans, a major grape-skin resident, exhibited broad spectrum assimilation of plant-derived carbon sources, including ω-hydroxy fatty acid, arising from degradation of plant cuticles. In fact, A. pullulans encoded and secreted possible cutinase-like esterase for cuticle degradation. When intact grape berries were used as a sole carbon source, such grape-skin associated fungi increased the accessibility to fermentable sugars by degrading and assimilating the plant cell wall and cuticle compounds. Their ability seems also helpful for S. cerevisiae to obtain energy through alcoholic fermentation. Thus, degradation and utilization of grape-skin materials by resident microbiota may account for their residence on grape-skin and S. cerevisiae’s possible commensal behaviors. Conclusively, this study focused on the symbiosis between grape-skin microbiota and S. cerevisiae from the perspective of winemaking origin. Such plant–microbe symbiotic interaction may be a prerequisite for triggering spontaneous food fermentation.

An optodigital system for visualizing the leaf epidermal surface using embedded speckle SIM: A 3D non-destructive approach

Intricate characterisation of the leaf epidermal surface is of utmost importance in agricultural crop management. The epidermal surface serves as a plant's outermost interface with the environment and is responsible for several physiological functions such as, balancing plant metabolism, regulating photosynthesis rate, and controlling plant-pathogen interactions. Existing microscopic techniques for large-scale, high-resolution characterisation of epidermal surfaces are in general destructive and time consuming, allowing only a single time point to be measured per sample. Here, the first time use of embedded speckle structured illumination microscope (ES-SIM) for the non-destructive characterisation of plant epidermal surface is reported. The developed optodigital system and analysis algorithm can enable automated characterisation of epidermal surface at multiple time points. The effectiveness of the proposed method for accurate detection of necrosis through 3D surface profiling of the leaf epidermis of a traditional medicinal plant, Piper sarmentosum, is demonstrated. ES-SIM provided 2.2 times better sensitivity for necrosis detection in comparison to conventional microscopy.

Palynology of Permian red-bed successions of Colorado and Wyoming and its influence on Laramide strata

ABSTRACTThe first reported Permian (Kungurian to Roadian) palynomorphs are described from Colorado, recovered from bedded gypsum and rare organic-rich shale intercalated in the red siltstone-dominated Lykins and State Bridge Formations. Surprisingly, these units generally lack the taeniate, saccate pollen that typifies most Permian continental rocks elsewhere, yet they contain abundant terrestrially derived palynomacerals, a low-diversity suite of sphaeromorph and acanthomorph acritarchs, and extremely rare non-taeniate, bissacate pollen grains. Acritarchs, known from one stratigraphic interval, are well-preserved and interpreted to represent autochthonous deposition during a marine incursion into the depocenter. This interpretation is consistent with their occurrence in a gray mudstone that is mantled by a mollusk-dominated coquina that bears conodonts, palaeoniscoid scales, and actinopterygian teeth. In contrast, most studied samples are dominated by wood fragments, charcoal, cuticles, and unidentified phytoclasts—all interpreted to represent dispersed plant cuticle and wood of continental origin. Fossils occur in black paper shale, gray fetid calcareous siltstone, and rhythmically bedded gypsum that is closely associated with thin limestone. Palynofacies analyses suggest that non-acritarch palynomacerals were deposited in dysoxic to anoxic waters that received minimal suspended terrigenous input. When combined with sedimentologic information, these non-acritarch fossils are hypothesized to have been deposited in shallow epicontinental lake-like settings that were periodically alkaline, hypersaline, and/or emergent.

Sooty blotch and flyspeck on mango (Mangifera indica L.) caused by Peltaster-like species in Malaysia

Fungi in the sooty blotch and flyspeck (SBFS) disease complex appear as pigmented blemishes on plant cuticles and are commonly observed on several fruits, including mango. In Malaysia, SBFS on mango has not been reported previously. However, SBFS symptoms were observed in commercial mango orchards in Perak, resulting in economic losses for sale of fresh fruit. Therefore, morphological, molecular, and pathogenicity studies were conducted to identify SBFS taxa on mango in Perak. A total of 27 mango fruit showing SBFS blemishes were collected from several commercial orchards in Perak. Colony morphologies were described on fruit and in purified culture, and DNA was extracted from the cultures. The internal transcribed spacer (ITS) and large subunit (LSU) regions of ribosomal DNA of two representative isolates were amplified, sequenced, and deposited in GenBank. Maximum parsimony (MP) and maximum likelihood (ML) methods were used to construct phylogenetic trees. A modified version of Koch's postulates was performed for the two isolates using unripe, asymptomatic fruit with two inoculation methods: in situ in an orchard and on excised fruit. In each case, two fruit per isolate were inoculated and two additional fruit were sprayed with distilled water as a control. For the in situ inoculation, an additional two non-inoculated fruit were enclosed in bags until harvest. The colonies resulting from the inoculation of mango fruit matched the original isolates in mycelial type and ITS sequence. Two previously unknown Peltaster-like species were identified as casual fungi of SBFS on mango in Malaysia. To increase management effectiveness, more extensive research on SBFS phylogenetic diversity in Malaysia should be conducted.

Detection and quantification of cotton trichomes by deep learning algorithm

Cotton is one of the most economically important agricultural crops with the harvest of cotton fibers. Trichomes are initiated from the plant epidermis, which is responsible for plant resistance to many abiotic and biotic stresses. Cotton fibers share similar underlying developmental molecular mechanisms with trichomes on the stem and leaf margins of cotton. Compared with fibers, it is easier to observe the trichome phenotype of cotton stem and leaf margins. However, because of the variability in coloration and the high occlusion of the trichomes, counting the density and observing the distribution of trichomes is still highly laborious. To help resolve this problem, the present research provides a solution based on deep learning in the screening of cotton trichomes, which makes it possible to monitor the phenotype of the cotton efficiently. With a fluorescent microscopy imaging procedure, we clearly observed that there are two types of trichome phenotype inGossypium hirsutum(‘TM-1′), which are singled and clustered trichomes. The clustered trichome originates from two or more adjacent epidermal cells with no branches. The TM1 trichome was used to create a collection of singled and clustered trichome pictures. Following data production, we evaluated the effectiveness of several you only look once (YOLO) model iterations in the context of identifying singled and clustered trichomes, respectively. Results show that the YOLOv5s model outperforms YOLOv3 and YOLOv4 in average accuracy, recall, accuracy, and F1 score. We utilized the clustered trichome data set to assess the performance of YOLOv5s and Mask R-CNN in order to more precisely detect the real trichome distribution with simultaneous detection of single and clustered trichomes. The obtained findings show that both models may be utilized to concurrently identify singled and clustered trichomes since the discrepancy between the detected values of the two models and the real values is quite minimal. Mask R-CNN has a high IOU value, whereas YOLOv5s has a low false-positive detection rate and fast detection speed. Our models and datasets can be used as a starting point to measure the trichome trait in other cotton species and establish a cotton trichome detection and counting system based on deep learning technology.

Identification and improvement of isothiocyanate-based inhibitors on stomatal opening to act as drought tolerance-conferring agrochemicals

Stomatal pores in the plant epidermis open and close to regulate gas exchange between leaves and the atmosphere. Upon light stimulation, the plasma membrane (PM) H+-ATPase is phosphorylated and activated via an intracellular signal transduction pathway in stomatal guard cells, providing a primary driving force for the opening movement. To uncover and manipulate this stomatal opening pathway, we screened a chemical library and identified benzyl isothiocyanate (BITC), a Brassicales-specific metabolite, as a potent stomatal-opening inhibitor that suppresses PM H+-ATPase phosphorylation. We further developed BITC derivatives with multiple isothiocyanate groups (multi-ITCs), which demonstrate inhibitory activity on stomatal opening up to 66 times stronger, as well as a longer duration of the effect and negligible toxicity. The multi-ITC treatment inhibits plant leaf wilting in both short (1.5 h) and long-term (24 h) periods. Our research elucidates the biological function of BITC and its use as an agrochemical that confers drought tolerance on plants by suppressing stomatal opening.

Cuticle architecture and mechanical properties: a functional relationship delineated through correlated multimodal imaging.

Cuticles are multifunctional hydrophobic biocomposites that protect the aerial organs of plants. During plant development, plant cuticles must accommodate different mechanical constraints combining extensibility and stiffness, and the corresponding relationships with their architecture are unknown. Recent data showed a fine-tuning of cuticle architecture during fruit development, with several chemical clusters which raise the question of how they impact the mechanical properties of cuticles. We investigated the in-depth nanomechanical properties of tomato (Solanum lycopersicum) fruit cuticle from early development to ripening, in relation to chemical and structural heterogeneities by developing a correlative multimodal imaging approach. Unprecedented sharps heterogeneities were evidenced including an in-depth mechanical gradient and a 'soft' central furrow that were maintained throughout the plant development despite the overall increase in elastic modulus. In addition, we demonstrated that these local mechanical areas are correlated to chemical and structural gradients. This study shed light on fine-tuning of mechanical properties of cuticles through the modulation of their architecture, providing new insight for our understanding of structure-function relationships of plant cuticles and for the design of bioinspired material.

Regulation of hair cell and stomatal size by a hair cell-specific peroxidase in the grass Brachypodium distachyon

The leaf epidermis is the outermost cell layer forming the interface between plants and the atmosphere that must both provide a robust barrier against (a)biotic stressors and facilitate carbon dioxide uptake and leaf transpiration.1 To achieve these opposing requirements, the plant epidermis developed a wide range of specialized cell types such as stomata and hair cells. Although factors forming these individual cell types are known,2,3,4,5 it is poorly understood how their number and size are coordinated. Here, we identified a role for BdPRX76/BdPOX, a class III peroxidase, in regulating hair cell and stomatal size in the model grass Brachypodium distachyon. In bdpox mutants, prickle hair cells were smaller and stomata were longer. Because stomatal density remained unchanged, the negative correlation between stomatal size and density was disrupted in bdpox and resulted in higher stomatal conductance and lower intrinsic water-use efficiency. BdPOX was exclusively expressed in hair cells, suggesting that BdPOX cell-autonomously promotes hair cell size and indirectly restricts stomatal length. Cell-wall autofluorescence and lignin stainings indicated a role for BdPOX in the lignification or crosslinking of related phenolic compounds at the hair cell base. Ectopic expression of BdPOX in the stomatal lineage increased phenolic autofluorescence in guard cell (GC) walls and restricted stomatal elongation in bdpox. Together, we highlight a developmental interplay between hair cells and stomata that optimizes epidermal functionality. We propose that cell-type-specific changes disrupt this interplay and lead to compensatory developmental defects in other epidermal cell types.

Leaf cuticular waxes of wild-type Welsh onion (Allium fistulosum L.) and a wax-deficient mutant: Compounds with terminal and mid-chain functionalities

Plant cuticles cover aerial organs to limit non-stomatal water loss and protect against insects and pathogens. Cuticles contain complex mixtures of fatty acid-derived waxes, with various chain lengths and diverse functional groups. To further our understanding of the chemical diversity and biosynthesis of these compounds, this study investigated leaf cuticular waxes of Welsh onion (Allium fistulosum L.) wild type and a wax-deficient mutant. Leaf waxes were extracted with chloroform, separated using thin layer chromatography (TLC), and analyzed using gas chromatography-mass spectrometry (GC-MS). The extracts contained typical wax compound classes found in nearly all plant lineages but also two uncommon compound classes. Analyses of characteristic MS fragmentation patterns followed by comparisons with synthetic standards identified the latter as very-long-chain ketones and primary ketols. The ketols were minor compounds, with chain lengths ranging from C28 to C32 and carbonyls mainly on C-18 and C-20 in wild type wax, and a C28 chain with C-16 carbonyl in the mutant. The ketones made up 70% of total wax in the wild type, consisting mainly of C31 isomers with carbonyl group on C-14 or C-16. In contrast, the mutant wax comprised only 4% ketones, with chain lengths C27 and C29 and carbonyls predominantly on C-12 and C-14, respectively. A two-carbon homolog shift between wild type and mutant was also observed in the primary alcohols (a major wax compound class), whilst alkanes exhibited a four-carbon shift. Overall, the compositional data shed light on possible biosynthetic pathways to wax ketones that can be tested in future studies.

WAX INDUCER 1 Regulates β-Diketone Biosynthesis by Mediating Expression of the Cer-cqu Gene Cluster in Barley.

Plant surface properties are crucial determinants of resilience to abiotic and biotic stresses. The outer layer of the plant cuticle consists of chemically diverse epicuticular waxes. The WAX INDUCER1/SHINE subfamily of APETALA2/ETHYLENE RESPONSIVE FACTORS regulates cuticle properties in plants. In this study, four barley genes homologous to the Arabidopsis thaliana AtWIN1 gene were mutated using RNA-guided Cas9 endonuclease. Mutations in one of them, the HvWIN1 gene, caused a recessive glossy sheath phenotype associated with β-diketone deficiency. A complementation test for win1 knockout (KO) and cer-x mutants showed that Cer-X and WIN1 are allelic variants of the same genomic locus. A comparison of the transcriptome from leaf sheaths of win1 KO and wild-type plants revealed a specific and strong downregulation of a large gene cluster residing at the previously known Cer-cqu locus. Our findings allowed us to postulate that the WIN1 transcription factor in barley is a master mediator of the β-diketone biosynthesis pathway acting through developmental stage- and organ-specific transactivation of the Cer-cqu gene cluster.

Analysis of plant cuticles and their interactions with agrochemical surfactants using a 3D printed diffusion chamber

BackgroundDecades of research is available on their effects of single component surfactant on active ingredient diffusion across plant cuticular membranes, but ingredient diffusion is rarely analysed in the presence of commercial surfactants. Also, diffusion studies require expensive or specialized apparatus the fabrication of which often requires skilled labour and specialized facilities. In this research we have addressed both problems where the effects of four commercially available surfactants on a known tracer molecule were investigated using a 3D printed customized diffusion chamber.ResultsAs a proof-of-concept a customized 3D printed diffusion chamber was devised using two different thermoplastics and was successfully used in a range of diffusion tests . The effect of various solvents and surfactants on S. lycopersicum cuticular membrane indicated an increased rate of flux of tracer molecules across the membranes. This research has validated the application of 3D printing in diffusion sciences and demonstrated the flexibility and potential of this technique.ConclusionsUsing a 3D printed diffusion apparatus, the effect of commercial surfactants on molecular diffusion through isolated plant membranes was studied. Further, we have included here the steps involved in material selection, design, fabrication, and post processing procedures for successful recreation of the chamber. The customizability and rapid production process of the 3D printing demonstrates the power of additive manufacturing in the design and use of customizable labware.

Site-specific analysis reveals candidate cross-kingdom small RNAs, tRNA and rRNA fragments, and signs of fungal RNA phasing in the barley-powdery mildew interaction.

The establishment of host-microbe interactions requires molecular communication between both partners, which may involve the mutual transfer of noncoding small RNAs. Previous evidence suggests that this is also true for powdery mildew disease in barley, which is caused by the fungal pathogen Blumeria hordei. However, previous studies lacked spatial resolution regarding the accumulation of small RNAs upon host infection by B. hordei. Here, we analysed site-specific small RNA repertoires in the context of the barley-B. hordei interaction. To this end, we dissected infected leaves into separate fractions representing different sites that are key to the pathogenic process: epiphytic fungal mycelium, infected plant epidermis, isolated haustoria, a vesicle-enriched fraction from infected epidermis, and extracellular vesicles. Unexpectedly, we discovered enrichment of specific 31-33-base 5'-terminal fragments of barley 5.8S ribosomal RNA in extracellular vesicles and infected epidermis, as well as particular B. hordei transfer RNA fragments in haustoria. We describe canonical small RNAs from both the plant host and the fungal pathogen that may confer cross-kingdom RNA interference activity. Interestingly, we found first evidence of phased small interfering RNAs in B. hordei, a feature usually attributed to plants, which may be associated with the posttranscriptional control of fungal coding genes, pseudogenes, and transposable elements. Our data suggest a key and possibly site-specific role for cross-kingdom RNA interference and noncoding RNA fragments in the host-pathogen communication between B. hordei and its host barley.

Endoparasitic Gall Mites: Two New Novophytoptus Species (Eriophyoidea, Phytoptidae) from Southern African Sedges (Cyperaceae, Carex) and New Hypotheses on the Phylogeny of Novophytoptines

Eriophyoid mites (Acariformes, Eriophyoidea) are microscopic chelicerates morphologically greatly preadapted to endoparasitism on plants. Members of at least six phylogenetically distant eriophyoid genera from two families homoplastically transitioned to endoparasitism and acquired the ability to penetrate under the plant epidermis and feed on parenchymatous cells, usually causing necrosis. Theoretically, endoparasites are expected to show patterns of codivergence with hosts more than ectoparasites. Novophytoptus Roivainen 1947 is the only eriophyoid genus comprising exclusively endoparasitic species living in subepidermal tissues of herbaceous monocots of three families of the order Poales: Cyperaceae, Juncaceae, and Poaceae. Here, we described two new endoparasitic species, N. limpopoensis n. sp. and N. zuluensis n. sp., from southern African sedges Carex spicatopaniculata Boeckeler ex C.B.Clarke and C. zuluensis C.B.Clarke, respectively, and investigated the Cox1 phylogeny of Novophytoptus. Contrary to expectations, molecular phylogenetics did not recover host-specific mite clades associated with Cyperaceae and Juncaceae, but revealed geographical groups of Novophytoptus species from Africa and Eurasia. Our results provide a substantial basis for future coevolutionary studies on novophytoptines, which will be possible when more species and sequences of Novophytoptus from geographically remote regions and from diverse hosts representing all major clades of Poales become available for analyses.

The plant cuticle

The plant cuticle is one of the key innovations that allowed plants to colonize terrestrial ecosystems. By limiting molecular diffusion, the cuticle provides an interface that ensures controlled interactions between plant surfaces and their environments. It confers diverse and sometimes astonishing properties upon plant surfaces at both the molecular level (from water and nutrient exchange capacities to almost complete impermeability), to the macroscopic level (from water repellence to iridescence). It takes the form of a continuous modification of the outer cell wall of the plant epidermis from early in plant development (surrounding the epidermis of the developing plant embryo) and is actively maintained and modified throughout the growth and development of most plant aerial organs - including non-woody stems, flowers, leaves, and even the root cap of emerging primary and lateral roots. The cuticle was first identified as a distinct structure in the early 19th century, and has since been the focus of intense research that, while revealing the fundamental role of the cuticle in the life of terrestrial plants, has also highlighted many unresolved mysteries regarding cuticle biogenesis and structure.