Epicuticular Wax Deposition Research Articles

Amphistomy: stomata patterning inferred from 13C content and leaf-side-specific deposition of epicuticular wax.

The benefits and costs of amphistomy (AS) vs. hypostomy (HS) are not fully understood. Here, we quantify benefits of access of CO2 through stomata on the upper (adaxial) leaf surface, using 13C abundance in the adaxial and abaxial epicuticular wax. Additionally, a relationship between the distribution of stomata and epicuticular wax on the opposite leaf sides is studied. We suggest that the 13C content of long-chain aliphatic compounds of cuticular wax records the leaf internal CO2 concentration in chloroplasts adjacent to the adaxial and abaxial epidermes. This unique property stems from: (1) wax synthesis being located exclusively in epidermal cells; and (2) ongoing wax renewal over the whole leaf lifespan. Compound-specific and bulk wax 13C abundance (δ) was related to amphistomy level (ASL; as a fraction of adaxial in all stomata) of four AS and five HS species grown under various levels of irradiance. The isotopic polarity of epicuticular wax, i.e. the difference in abaxial and adaxial δ (δab - δad), was used to calculate the leaf dorsiventral CO2 gradient. Leaf-side-specific epicuticular wax deposition (amphiwaxy level) was estimated and related to ASL. In HS species, the CO2 concentration in the adaxial epidermis was lower than in the abaxial one, independently of light conditions. In AS leaves grown in high-light and low-light conditions, the isotopic polarity and CO2 gradient varied in parallel with ASL. The AS leaves grown in high-light conditions increased ASL compared with low light, and δab - δad approached near-zero values. Changes in ASL occurred concomitantly with changes in amphiwaxy level. Leaf wax isotopic polarity is a newly identified leaf trait, distinguishing between hypo- and amphistomatous species and indicating that increased ASL in sun-exposed AS leaves reduces the CO2 gradient across the leaf mesophyll. Stomata and epicuticular wax deposition follow similar leaf-side patterning.

Response of photosynthesis, the xanthophyll cycle, and wax in Japanese yew (Taxus cuspidata L.) seedlings and saplings under high light conditions.

In order to understand the adaptative changes of the Japanese yew (Taxus cuspidate L.) to high light conditions, this study investigated gas-exchange, chlorophyll fluorescence, chlorophyll, and the impact of epicuticular wax on the gas-exchange and photoinhibition of Japanese yew seedlings and saplings. The chlorophyll content per unit area and photosynthetic rate in seedling leaves were significantly lower than in sapling leaves. When leaves from seedlings and saplings were exposed to 1,200 μmol·m-2·s-1 photon flux density (PFD) for 2 h, seedling leaves exhibited a greater down-regulation of maximum quantum yield (Fv/Fm) and actual photosystem II efficiency ( PSII). Non-photochemical quenching (NPQ) and high energy quenching (qE) in sapling leaves were much higher than in seedling leaves when both were exposed to 1,200 μmol·m-2·s-1 PFD for 2 h. At a low level of O2, the photorespiration rate (Pr) and the ratio of photorespiration/gross photosynthetic rate (Pr/Pg) in seedling leaves were lower than in sapling leaves when both were exposed to 1,200 μmol·m-2·s-1 PFD, but this difference did not reach statistical significance (P < 0.05). Compared with sapling leaves, seedling leaves exhibited lower levels of xanthophyll pool. Epicuticular wax content on seedling leaves was significantly lower than on sapling leaves. The results of this study showed that wax coverage on the leaf surface decreased the photosynthetic rate in sapling leaves as a consequence of decreased stomatal conductance. Epicuticular wax is related to tree age and photoinhibition prevention in the Japanese yew. It is possible that lower photosynthetic rate, lower NPQ depending on the xanthophyll cycle, and lower deposition of epicuticular wax results in seedling plants that are not adapted to high light conditions.

How Different Na+ Concentrations Affect Anatomical, Nutritional Physiological, Biochemical, and Morphological Aspects in Soybean Plants: A Multidisciplinary and Comparative Approach

Soybean is a legume widely cultivated globally for its seeds, which are rich in oil and protein suitable for animal and human nutrition, and as a biofuel source. One of the main factors that limits production is soil salinity; currently there are an estimated 800 million hectares of agricultural land affected by salt stress worldwide. The aim of this research was to determine whether anatomical, morphological, nutritional, physiological, and biochemical parameters are negatively affected in soybean plants cultivated under different levels of salt stress. The experiment was randomized into five treatments (0, 50, 100, 150, and 200 mM Na+). Plants subjected to concentrations of 50 to 200 mM Na+ exhibited reductions in K (range 21% to 57%), Ca (range 38% to 63%), and Mg (range 20% to 41%) compared to controls (without Na+). Na+ stress progressively produced negative effects on photosynthetic machinery, gas exchange, and photosynthetic pigments, results clearly related to oxidative stress generated by the saline growth conditions. Interestingly, our study revealed that at concentrations up to 100 mM Na+ deposition of epicuticular wax occurred, the quantity and shape of the stomata changed, and the thickness of the leaf epidermis increased. Our broad-based, multidisciplinary, and comparative study proved that soybean plants suffer significant deleterious effects modulated by Na+ stress, mainly at concentrations above 100 mM Na+.

Micro-morpho-anatomical transitions at various stages of in vitro development of Crinum malabaricum Lekhak and Yadav: A critically endangered medicinal plant.

Crinum malabaricum Lekhak & Yadav is a recently discovered and critically endangered aquatic bulbous plant of the family Amaryllidaceae. It gained attention as a wild source of the acetylcholinesterase inhibiting alkaloid 'galanthamine' used to treat Alzheimer and Parkinson diseases. The bulbs of this plant contain the highest amount of galanthamine among Crinum species. In vitro regeneration systems were developed to produce quality uniform plantlets of C. malabaricum. Bright field light microscopy was used to analyse micro-morpho-anatomical developments taking place in the leaves and roots during in vitro, ex vitro and in vivo transitions of plantlets. Leaves and roots of plants raised in vitro possessed a higher degree of microscopic structural anomalies, such as underdeveloped epicuticular wax deposition, immature and non-functional stomata, more aquiferous parenchyma with a reduced lumen. Roots developed in vitro were characterized by extremely large, uneven cortical cells and reduced intercellular spaces. The vascular tissues were under-developed and only primary vascular tissues were observed. As a result of ex vitro acclimation, there was a significant acceleration in the improvement of tissue systems in leaves and roots. Such plantlets can tolerate elevated temperatures and light under in vivo conditions. Thus, the microscopic evaluation of the structural trajectory in different stages of plantlet development provides an understanding of the acclimation process and structural adaptations, which could help enhance survival of in vitro raised plantlets under ex vitro and in vivo conditions.

NUCLEAR TRANSPORT FACTOR 2-LIKE improves drought tolerance by modulating leaf water loss in alfalfa (Medicago sativa L.).

Drought is a major environmental factor that limits the production of alfalfa (Medicago sativa). In the present study, M. sativa NUCLEAR TRANSPORT FACTOR 2-LIKE (MsNTF2L) was identified as a nucleus-, cytoplasm-, and plasma membrane-localized protein. Its transcriptional expression was highly induced by ABA and drought stress. Overexpression of MsNTF2L in Arabidopsis resulted in hypersensitivity to ABA during both the seed germination and seedling growth stages. However, transgenic Arabidopsis plants exhibited enhanced tolerance to drought stress by reducing the levels of reactive oxygen species (ROS) and increasing the expression of stress/ABA-inducible genes. Consistently, analysis of MsNTF2L overexpression (OE) and RNA interference (RNAi) alfalfa plants revealed that MsNTF2L confers drought tolerance through promoting ROS scavenging, a decrease in stomatal density, ABA-induced stomatal closure, and epicuticular wax crystal accumulation. MsNTF2L highly affected epicuticular wax deposition, as a large group of wax biosynthesis and transport genes were influenced in the alfalfa OE and RNAi lines. Furthermore, transcript profiling of drought-treated alfalfa WT, OE, and RNAi plants showed a differential drought response for genes related to stress/ABA signaling, antioxidant defense, and photosynthesis. Taken together, these results reveal that MsNTF2L confers drought tolerance in alfalfa via modulation of leaf water loss (by regulating both stomata and wax deposition), antioxidant defense, and photosynthesis.

Abscisic Acid Deficiency Alters Epicuticular Wax Metabolism and Morphology That Leads to Increased Cuticle Permeability During Sweet Orange (Citrus sinensis) Fruit Ripening.

Citrus fruit ripening is coupled with the synthesis and deposition of epicuticular waxes, which reduces water loss during fruit postharvest storage. Although abscisic acid (ABA) is a major regulator of citrus fruit ripening, whether ABA mediates epicuticular wax formation during this process remains poorly understood. We investigated the implication of ABA in cuticle properties and epicuticular wax metabolism, composition, and morphology by comparing the Navelate orange [Citrus sinensis (L.) Osbeck] and its ABA biosynthesis-impaired mutant Pinalate in four ripening stages. ABA deficiency had minor effects on cuticle thickness and epicuticular wax load, but correlated with cuticle permeability. ABA content aligned with mostly fatty acids accumulation in both cultivars, and also with specific alkane, terpenoid, and aldehyde constituents in the parental fruit. In turn, cuticle permeability correlated with the fatty acid profile during fruit ripening in the Navelate and Pinalate, and with primary alcohols, terpenoids, and aldehydes, but only in the mutant fruit. Low ABA levels increased the susceptibility of waxes to crack and were lost from the epicuticular layer. The RNA-seq analysis highlighted the differential regulation of a list of 87 cuticle-related genes between genotypes and ripening stages. Changes in the gene expression of the selected genes in both cultivars were consistent with the content of the aliphatics and terpenoid fractions during ripening. The results suggest a role for ABA in the regulation of fatty acid content and primary alcohol composition, and point out the importance of alkane and triterpenoid for controlling water permeance through fruit cuticles.

The Molecular Priming of Defense Responses is Differently Regulated in Grapevine Genotypes Following Elicitor Application against Powdery Mildew.

Molecular changes associated with response to powdery mildew (PM) caused by Erysiphe necator have been largely explored in Vitis vinifera cultivars, but little is known on transcriptional and metabolic modifications following application of resistance elicitors against this disease. In this study, the whole transcriptome sequencing, and hormone and metabolite analyses were combined to dissect long-term defense mechanisms induced by molecular reprogramming events in PM-infected ‘Moscato’ and ‘Nebbiolo’ leaves treated with three resistance inducers: acibenzolar-S-methyl, potassium phosphonate, and laminarin. Although all compounds were effective in counteracting the disease, acibenzolar-S-methyl caused the most intense transcriptional modifications in both cultivars. These involved a strong down-regulation of photosynthesis and energy metabolism and changes in carbohydrate accumulation and partitioning that most likely shifted the plant growth-defense trade-off towards the establishment of disease resistance processes. It was also shown that genotype-associated metabolic signals significantly affected the cultivar defense machinery. Indeed, ‘Nebbiolo’ and ‘Moscato’ built up different defense strategies, often enhanced by the application of a specific elicitor, which resulted in either reinforcement of early defense mechanisms (e.g., epicuticular wax deposition and overexpression of pathogenesis-related genes in ‘Nebbiolo’), or accumulation of endogenous hormones and antimicrobial compounds (e.g., high content of abscisic acid, jasmonic acid, and viniferin in ‘Moscato’).

Efficiency of glyphosate and carfentrazone-ethyl in the control of Macroptilium atropurpureum (DC.) Urb. under different light intensities

Variation in environmental conditions, such as light availability, can affect herbicide efficacy by altering leaf morphoanatomy. Accordingly, the aim of the present study was to evaluate the influence of light level and herbicide application on leaf anatomy and herbicide efficacy, using Macroptilium atropurpureum as a model weed species. The experiment involved a factorial design (3 × 4), using three light levels (0, 50, or 70% shadow) and four herbicide treatments (no herbicide, glyphosate, carfentrazone-ethyl, or a combination of glyphosate and carfentrazone-ethyl). When combined, the herbicides were efficient in controlling M. atropurpureum under all light levels. However, when applied independently, the herbicides failed to satisfactorily control plants cultivated in full sunlight, and plants cultivated in shaded environments were more sensitive to glyphosate application than those cultivated in full sunlight. The greater efficacy of the herbicides on plants grown under shaded conditions can likely be attributed to changes in leaf morphoanatomy. Plants grown under restricted light conditions exhibit greater average leaf area, lower trichome density, reduced epicuticular wax deposition, thinner leaves, and lower dry mass accumulation, characteristics which are closely correlated to chemical control. The combination of the herbicides presents an additional effect in the control of M. atropurpureum. Also, changes in leaf anatomy caused by light restriction reduce the tolerance of M. atropurpureum to the herbicides.

The semidominant mutation w5 impairs epicuticular wax deposition in common wheat (Triticum aestivum L.).

The semidominant EMS-induced mutant w5 affects epicuticular wax deposition and mapped to an approximately 194-kb region on chromosome 7DL. Epicuticular wax is responsible for the glaucous appearance of plants and protects against many biotic and abiotic stresses. In wheat (Triticum aestivum L.), β-diketone is a major component of epicuticular wax in adult plants and contributes to the glaucousness of the aerial organs. In the present study, we identified an ethyl methanesulfonate-induced epicuticular wax-deficient mutant from the elite wheat cultivar Jimai22. Compared to wild-type Jimai22, the mutant lacked β-diketone and failed to form the glaucous coating on all aerial organs. The mutant also had significantly increased in cuticle permeability, based on water loss and chlorophyll efflux. Genetic analysis indicated that the mutant phenotype is controlled by a single, semidominant gene on the long arm of chromosome 7D, which was not allelic to the known wax gene loci W1-W4, and was therefore designated W5. W5 was finely mapped to an ~ 194-kb region (flanked by the molecular markers SSR2 and STARP11) that harbored four annotated genes according to the reference genome of Chinese Spring (RefSeq v1.0). Collectively, these data will broaden the knowledge of the genetic basis underlying epicuticular wax deposition in wheat.

Application of Nano-Silicon Dioxide Improves Salt Stress Tolerance in Strawberry Plants

Silicon application can improve productivity outcomes for salt stressed plants. Here, we describe how strawberry plants respond to treatments including various combinations of salt stress and nano-silicon dioxide, and assess whether nano-silicon dioxide improves strawberry plant tolerance to salt stress. Strawberry plants were treated with salt (0, 25 or 50 mM NaCl), and the nano-silicon dioxide treatments were applied to the strawberry plants before (0, 50 and 100 mg L−1) or after (0 and 50 mg L−1) flowering. The salt stress treatments reduced plant biomass, chlorophyll content, and leaf relative water content (RWC) as expected. Relative to control (no NaCl) plants the salt treated plants had 10% lower membrane stability index (MSI), 81% greater proline content, and 54% greater cuticular transpiration; as well as increased canopy temperature and changes in the structure of the epicuticular wax layer. The plants treated with nano-silicon dioxide were better able to maintain epicuticular wax structure, chlorophyll content, and carotenoid content and accumulated less proline relative to plants treated only with salt and no nano-silicon dioxide. Analysis of scanning electron microscopic (SEM) images revealed that the salt treatments resulted in changes in epicuticular wax type and thickness, and that the application of nano-silicon dioxide suppressed the adverse effects of salinity on the epicuticular wax layer. Nano-silicon dioxide treated salt stressed plants had increased irregular (smoother) crystal wax deposits in their epicuticular layer. Together these observations indicate that application of nano-silicon dioxide can limit the adverse anatomical and biochemical changes related to salt stress impacts on strawberry plants and that this is, in part, associated with epicuticular wax deposition.

Ultrastructural and histochemical analysis of the trichomes of Cyclea peltata (Menispermaceae)

Cyclea peltata is a medicinal climber belonging to the family Menispermaceae. The global distribution of this species is recorded from the tropical Asia regions. Trichome phenology studies of C. peltata utilizing Scanning Electron Microscopy (SEM), light microscopy and fluorescent microscopy demonstrated the structural and organizational heterogeneity of trichomes. All the trichomes observed were non-glandular. C. peltata possessed three types of non-glandular trichomes such as long trichomes, short trichomes and ribbon-like trichomes. Of these, ribbon-like trichomes are limited to the surface of the leaves. The majority of the trichomes observed were unicellular, long and hook-shaped. Auto fluorescence analysis revealed a large amount of cell wall materials on the basal cell. Additional morphological features of the trichomes included the occurrence of grooves and deposition of epicuticular wax. The trichome morphology of C. peltata explains the possible influence of environmental cues.

&lt;b&gt;Leaf histology of greenhouse plants, in vitro cultured and somatic embryo-derived plants of &lt;i&gt;Bactris gasipaes&lt;/i&gt; Kunth

In vitro cultures of peach palm ( Bactris gasipaes Kunth) were established by somatic embryogenesis but some improvements in maturation and conversion steps are still needed. The aim of this study was to analyze morpho-anatomical differences in peach palm leaves from greenhouse cultured plants, in vitro plants developed from in vitro germinated seeds and somatic embryo-derived plants . Expanded leaves were prepared for histological analyses and scanning electron microscopy. No significant difference was found between ex vitro and in vitro cultured plants, but the somatic embryo-derived plants showed structural alterations of the leaves. The epidermal cells were elongated in shape, the mesophyll cells were thicker and the vascular bundle was not very developed. In somatic embryo-derived leaves the cuticle was thinner than in other leaves and epicuticular wax was present but poorly deposited. In in vitro cultured plants, the deposition of epicuticular wax on the leaves was irregular while in the greenhouse plants it was regular and abundant. These alterations in somatic embryo-derived leaves could hinder the acclimatization and development of peach palm plants so it is necessary to improve the protocol for somatic embryogenesis to produce better plants.

The maize lilliputian1 (lil1) gene, encoding a brassinosteroid cytochrome P450 C-6 oxidase, is involved in plant growth and drought response.

Brassinosteroids (BRs) are plant hormones involved in many developmental processes as well as in plant-environment interactions. Their role was investigated in this study through the analysis of lilliputian1-1 (lil1-1), a dwarf mutant impaired in BR biosynthesis in maize (Zea mays). We isolated lil1-1 through transposon tagging in maize. The action of lil1 was investigated through morphological and genetic analysis. Moreover, by comparing lil1-1 mutant and wild-type individuals grown under drought stress, the effect of BR reduction on the response to drought stress was examined. lil1-1 is a novel allele of the brassinosteroid-deficient dwarf1 (brd1) gene, encoding a brassinosteroid C-6 oxidase. We show in this study that lil1 is epistatic to nana plant1 (na1), a BR gene involved in earlier steps of the pathway. The lill-1 mutation causes alteration in the root gravitropic response, leaf epidermal cell density, epicuticular wax deposition and seedling adaptation to water scarcity conditions. Lack of active BR molecules in maize causes a pleiotropic effect on plant development and improves seedling tolerance of drought. BR-deficient maize mutants can thus be instrumental in unravelling novel mechanisms on which plant adaptations to abiotic stress are based.

Electron microscopic observations of stomata, epicuticular waxes, and papillae in Chamaecyparis obtusa: Reconsidering the traditional concept of Y-shaped white stomatal bands.

The foliar morphological characters of hinoki (Chamaecyparis obtusa) were revisited using optical and scanning electron microscopy. In C. obtusa, typical Y-shaped white stomatal bands were evident on the abaxial leaf surfaces. Two facial leaves and two lateral leaves were observed at the same node. Waxy papillae and oval stomata were arranged in two or three rows with protuberant rims on the abaxial leaf surfaces. Higher magnifications revealed the deposition of epicuticular waxes (tubules) on the Y-shaped white stomatal bands. Given the absence of stomatal bands after dewaxing with organic solvents, the white stomatal bands in C. obtusa were related to the epicuticular waxes rather than the presence of aggregated stomata alone. In contrast to C. obtusa, a single median leaf and two lateral leaves were observed at the same node of oriental arborvitae (Platycladus koraiensis). Neither stomatal bands nor papillae were observed on P. koraiensis leaves. The stomatal density and epicuticular waxes in the stomatal regions of C. obtusa were higher than those of P. koraiensis. This study suggests that the traditional concept of Y-shaped white stomatal bands in C. obtusa should be revised to describe the arrangement of the aggregated waxy stomata that occur in rows.

Molecular mapping and candidate gene analysis of a new epicuticular wax locus in sorghum (Sorghum bicolor L. Moench).

A new epicuticular wax (bloom) locus has been identified and fine mapped to the 207.89kb genomic region on chromosome 1. A putative candidate gene, Sobic.001G269200, annotated as GDSL-like lipase/acylhydrolase, is proposed as the most probable candidate gene involved in bloom synthesis/deposition. Deposition of epicuticular wax on plant aerial surface is one strategy that plants adapt to reduce non-transpiration water loss. Epicuticular wax (bloom)-less mutants in sorghum with their glossy phenotypes exhibit changes in the accumulation of epicuticular wax on leaf and culm surfaces. We report molecular mapping of a new sorghum locus, bloomless mutant (bm39), involved in epicuticular wax biosynthesis in sorghum. Inheritance studies involving a profusely bloom parent (BTx623) and a spontaneous bloomless mutant (RS647) indicated that the parents differed in a single gene for bloom synthesis. Bloomless was recessive to bloom deposition. Genetic mapping involving F2 and F7 mapping populations in diverse genetic backgrounds (BTx623×RS647; 296A×RS647 and 27A×RS647) identified and validated the map location of bm39 to a region of 207.89kb on chromosome 1. SSR markers, Sblm13 and Sblm16, flanked the bm39 locus to a map interval of 0.3cM on either side. Nine candidate genes were identified, of which Sobic.001G269200 annotated for GDSL-like lipase/acylhydrolase is the most likely gene associated with epicuticular wax deposition. Gene expression analysis in parents, isogenic lines and sets of near isogenic lines also confirmed the reduced expression of the putative candidate gene. The study opens possibilities for a detailed molecular analysis of the gene, its role in epicuticular wax synthesis and deposition, and may help to understand its function in moisture stress tolerance and insect and pathogen resistance in sorghum.

Morphophysiological, ultrastructural, and nutritional changes induced by Cu toxicity in young Erythrina fusca plants

ABSTRACTErythrina fusca is an important legume used for shade cover in cacao plantations in Brazil. Cacao plantations receive large quantities of copper (Cu)-containing agrochemicals, mainly for control of diseases. Therefore, Cu toxicity was investigated in seedlings grown in hydroponics with increasing concentrations of Cu (0.005–32 mg L−1) in a greenhouse. Ultrastructural analyses showed cell plasmolysis in the root cortical area and changes in thylakoid membranes at 8 mg Cu L−1 and higher. There were changes in epicuticular wax deposition on the leaf surface at the 16 and 32 mg Cu L−1 treatments. Leaf gas exchanges were highly affected 24 hours after application of treatments beginning at 8 mg Cu L−1 and higher Cu concentrations. Chemical analyses showed that Cu content in E. fusca roots increased as Cu concentration in the nutrient solution increased, whereas the shoot did not show significant changes. It is also observed that excess Cu interfered with Zn, Fe, Mn, Mg, K, P, and Ca content in the different E. fusca organs. Investigation of Cu toxicity symptoms focusing on morphophysiological, ultrastructural, gas exchange, and nutritional changes would be useful to alleviate Cu toxicity in E. fusca under field conditions, an important agroforestry species in cacao plantation.

The Genes Bm2 and Blmc that Affect Epicuticular Wax Deposition in Sorghum are Allelic

Epicuticular waxes (EW) coat the aerial surfaces of the sorghum [Sorghum bicolor (L.) Moench] plant and have a role in the plant's resistance to abiotic and biotic stresses, such as drought, ultraviolet light, insects, and fungal pathogens. Mutants of the Bloomless2 (Bm2) locus, such as bm2‐3, lack visible EW and have inhibited synthesis of the cuticle. The sorghum mutant bloom‐cuticle (blmc) also lacks visible EW and has inhibited synthesis of the cuticle, as well as many other effects. In this study, an allelism test was performed, which revealed that blmc is an allele of the Bm2 locus and is hereby designated as bm2‐7. This finding allows the data that have been generated separately from each of these mutations to be merged. Thus, the results suggest that the Bm2 locus is on sorghum chromosome 10 and could have pleiotropic effects, including cuticle proper synthesis, leaf water retention, greenbug biotype C (Schizaphis graminum Rondani) susceptibility, and resistance to the fungal pathogen Exserohilum turcicum Pass., which causes northern corn leaf blight. Further examination of the Bm2 locus can be accomplished by the use of only one mutant line.

Physiological and leaf ultrastructural characteristics of perennial ryegrass (Lolium perenne L.) biotypes from Tunisia under sulfonylurea herbicide application

During the last decade, reduced efficacy and failure to control perennial ryegrass (Lolium perenne L.) with labeled rates of Acetolactate Synthase (ALS) inhibitor herbicide (MSM-IDS) were reported. In this context, the effects of MSM-IDS (Sulfonylurea) on suspected resistant (Mateur) and susceptible (Tinja) ryegrass biotypes, prospected in Tunisian wheat fields, were investigated in comparison with those of Pinoxaden, an Acetyl CoA Carboxylase (ACCase) inhibitor herbicide, based on physiological and ultrastructural studies. At their labeled rates, both herbicides significantly affected biomass and gas exchange of Tinja biotype. In Mateur biotype, similar responses as control were observed after ALS inhibitor application. At the molecular level, the most common mutations (P197 and W574) of the ALS gene were not identified in Mateur biotype. Microscopically, a marked collapse of Tinja biotype mesophyll cells was observed by Scanning Electron Microscopy (SEM) after both herbicide applications. However, Mateur biotype mesophyll surface exhibited abundant epicuticular wax deposits and was not affected by ALS inhibitor herbicide. By Transmission Electron Microscopy (TEM), cuticle in Mateur biotype presented a continuous thick lamellate region, while in Tinja biotype, the lamellate region was discontinuous indicating an easy diffusion of MSM-IDS through the cuticle. MSM-IDS resistant Tunisian Mateur ryegrass biotype seems to have developed efficient micromorphological and ultrastructural barriers against sulfonylurea absorption, which in turn allowed a photosynthetic capacity similar to control.

Structural effects on Cattleya xanthina leaves cultivated in vitro and acclimatized ex vitro

In vitro orchid micropropagation is efficient biotechnological strategy for conservation and commercial plantlet production. However, micropropagated plantlets generally need to adapt to survive severe changes in humidity, irradiance, and growing medium that accompany the transfer to ex vitro conditions. Such adaptive cellular changes would give insights into the phenotypic plasticity of the model plant Cattleya xanthina (L.) Van den Berg. Therefore, we aimed to evaluate structural changes in the leaves of C. xanthina cultivated in vitro and acclimatized ex vitro using qualitative and quantitative analyses. During acclimatization, we observed a higher accumulation of dry mass, a greater convexity of the outer surface of epidermal cells, an increased deposition of epicuticular waxes, a greater elongation of mesophyll parenchymatic cells, and finally, the presence of chloroplasts with organized thylakoids and well-developed grana. Stomatal density was not changed. Furthermore, a gradual acclimatization allows this species the best adaptation to a new environment.

The maize fused leaves1 (fdl1) gene controls organ separation in the embryo and seedling shoot and promotes coleoptile opening.

The fdl1-1 mutation, caused by an Enhancer/Suppressor mutator (En/Spm) element insertion located in the third exon of the gene, identifies a novel gene encoding ZmMYB94, a transcription factor of the R2R3-MYB subfamily. The fdl1 gene was isolated through co-segregation analysis, whereas proof of gene identity was obtained using an RNAi strategy that conferred less severe, but clearly recognizable specific mutant traits on seedlings. Fdl1 is involved in the regulation of cuticle deposition in young seedlings as well as in the establishment of a regular pattern of epicuticular wax deposition on the epidermis of young leaves. Lack of Fdl1 action also correlates with developmental defects, such as delayed germination and seedling growth, abnormal coleoptile opening and presence of curly leaves showing areas of fusion between the coleoptile and the first leaf or between the first and the second leaf. The expression profile of ZmMYB94 mRNA-determined by quantitative RT-PCR-overlaps the pattern of mutant phenotypic expression and is confined to a narrow developmental window. High expression was observed in the embryo, in the seedling coleoptile and in the first two leaves, whereas RNA level, as well as phenotypic defects, decreases at the third leaf stage. Interestingly several of the Arabidopsis MYB genes most closely related to ZmMYB94 are also involved in the activation of cuticular wax biosynthesis, suggesting deep conservation of regulatory processes related to cuticular wax deposition between monocots and dicots.