New insights into the role of the root system of epiphytic bromeliads: comparison of root and leaf trichome functions in acquisition of water and nutrients.

BACKGROUND: In epiphytic bromeliads, the roots used to be considered poorly functional organs in the processes of absorption and metabolization of water and nutrients, while the leaves always acted as protagonists in both functions. More recent discoveries have been changing this old view of the root system. SCOPE: In this review, we will address the old thoughts of the scientific community regarding the function performed by the roots of epiphytic bromeliads (mere holdfast structures with low physiological activity) and the importance of a reduced or lack of root system for the emergence of epiphytism. We will present indirect and direct evidence that contradicts this older hypothesis. Furthermore, the importance of the root absorptive function mainly for juvenile tankless epiphytic bromeliads and the characteristics of the root absorption process of adult epiphytic tank bromeliads will be thoroughly discussed in physiological aspects. Finally, some factors (species, substrate, environmental conditions) that influence the absorptive capability of the roots of epiphytic tank bromeliads will also be considered in this review, highlighting the importance that the absorptive role of the roots have for the plasticity of bromeliads that live on trees, which is an environment characterized by the intermittent availability of water and nutrients. CONCLUSIONS: The roots of tank-forming epiphytic bromeliads play important roles in the absorption and metabolization of nutrients and water. The importance of roots stands out mainly for juvenile tankless bromeliads since the root is the main absorptive organ. In larger plants with tank, although the leaves become the protagonists in the resource acquisition process, the roots complement the absorptive function of the leaf trichomes, resulting in a better growth of the bromeliad. The physiological and biochemical properties of the processes of absorption and distribution of resources in the tissues seem to differ between absorption by trichomes and roots.

Simultaneous Impact of Rhizobacteria Inoculation and Leaf-Chewing Insect Herbivory on Essential Oil Production and VOC Emissions in Ocimum basilicum.

Inoculation with rhizobacteria and feeding by herbivores, two types of abiotic stress, have been shown to increase the production of secondary metabolites in plants as part of the defense response. This study explored the simultaneous effects of inoculation with Bacillus amyloliquefaciens GB03 (a PGPR species) and herbivory by third-instar Spodoptera frugiperda larvae on essential oil (EO) yield and volatile organic compound (VOC) emissions in Ocimum basilicum plants. The density of glandular trichomes was also examined, given that they are linked to EO production and VOC emission. Herbivory increased EO content, but inoculation on its own did not. When combined, however, the two treatments led to a 10-fold rise in EO content with respect to non-inoculated plants. VOC emissions did not significantly differ between inoculated and non-inoculated plants, but they doubled in plants chewed by the larvae with respect to their undamaged counterparts. Interestingly, no changes were observed in VOC emissions when the treatments were tested together. In short, the two biotic stressors elicited differing plant defense responses, mainly when EO was concerned. PGPR did not stimulate EO production, while herbivory significantly enhanced it and increased VOC emissions. The combined treatment acted synergistically, and in this case, PGPR inoculation may have had a priming effect that amplified plant response to herbivory. Peltate trichome density was higher in inoculated plants, those damaged by larvae, and those subjected to the combination of both treatments. The findings highlight the intricate nature of plant defense mechanisms against various stressors and hint at a potential strategy to produce essential oil through the combined application of the two stressors tested here.

Leaf secretory structures in Rosa lucieae (Rosaceae): two times of secretion-two ecological functions?

Secretory trichomes and colleters are two of the secretory structures whose exudates may cover the body of the plant. Such secretions comprise resins or mucilages which are associated with an array of ecological roles. In Rosaceae, secretory trichomes have been reported for the leaves while colleters associated with leaf teeth. Our study aimed to identify the secretory structures of Rosa lucieae and understand the ecological role played by these glands as interpreted by morphoanatomical and histochemical studies. Samples from developing and fully mature leaves were collected, fixed, and processed according to usual techniques for light and scanning electron microscopy. In R. lucieae, colleters are restricted to the leaf and stipular margins and are associated with the teeth. They present a parenchymatous axis surrounded by a secretory palisade epidermis and usually fall off after the secretory activity is finished. Different from colleters, secretory trichomes are persistent. They present a multicellular secretory head and stalk. They are found at the base of the leaflet, petiolule, rachis, and petiole and occasionally on the stipular and leaf margins. The colleters predominantly secrete mucilages while the secretory trichomes secrete lipids and terpenes, both via cuticle rupture. The secretory activity of colleters is predominant in the leaf primordia, holding leaflets together and protecting meristems and leaves from desiccation, while the secretory trichomes maintain their secretory activity at different stages of leaf development, protecting different regions of the leaf against pathogens and herbivores.

Epidermal and subepidermal changes during the formation of hairy galls induced by Eriophyidae on Avicennia schaueriana leaves.

Leaf-galling Eriophyidae (Acarina) may promote simple or complex alterations in the organs of their host plants, such as an increase in indumentum density or the reorganization of epidermis and ground system tissue patterns. To test if hairy galls of Eriophyidae on Avicennia schaueriana (Acanthaceae) are related to complex changes, leaf galls in distinct developmental phases were compared to non-galled leaves using anatomical, histochemical, and histometric analyses. Quantitative comparisons of preferential gall induction sites and gall area according to distinct leaf portions were made to evaluate if the impacts of gall formation can be related to the distinct potentialities of leaf microsites. The apical portion of the leaves and leaf margins were the sites with the highest occurrence of galls, but no relationship was detected between gall area and induction site. The gall anatomy revealed that epidermal features are influenced the most with the development of abnormal stomata and projected or sunken salt glands. The most striking change is the neoformation of elongated filiform trichomes on the abaxial surface (where the mites occur) that accumulate reducing sugars and proteins. The filiform trichomes may protect the inducers against abiotic stressors and enemies, and the primary metabolites that accumulate are important foods for mites. The mesophyll has simple alterations, only in the spongy parenchyma. Complex alterations occur only in abaxial epidermal cells close to feeding sites of the inducer. The number of inducers per gall seems to be the most important influence on gall size, since gall area is not related to the position in the leaves.

Differential levels of soybean resistance to the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) under controlled and uncontrolled environments are associated with plant age, damage intensity, and trichome density

Whiteflies are a severe threat to soybean production in the tropics. This study aimed to evaluate the soybean resistance level of the whitefly Bemisia tabaci in controlled and uncontrolled environments that is associated with plant age, damage intensity, and trichome density. The research was conducted under two conditions: non-sprayed (NS) and sprayed (SP). This study used 50 soybean genotypes arranged in a randomized block design with three replicates. The whitefly population was derived from natural infestations. The results showed that the highest wild population of B. tabaci occurred at 40 days after planting (DAP), i.e., 126.08 adults/plant in the NS environment and 22.57 adults/plant in the SP environment. The peak damage intensity occurred at 50 DAP, 20.71% in the NS environment, and 17.15% in the SP environment. In the NS environment, there were six resistant genotypes (including the resistant control G100H), 25 moderate, and 19 susceptible genotypes. In the SP environment, 19 genotypes were resistant, 22 genotypes were moderate, and nine genotypes were susceptible, respectively. Six soybean genotypes showed consistent resistance to B. tabaci in NS and SP environments. The low density of leaf trichomes in soybean may influence the high resistance to B. tabaci. The resistant genotypes identified in this study could be utilized in breeding programs for B. tabaci resistance.

Population Dynamics of Heliothrips haemorrhoidalis (Thysanoptera: Thripidae) in Strawberry Cultivars in Southern Brazil.

Heliothrips haemorrhoidalis is a polyphagous thrips species that has been reported as a potential pest in strawberry cultivation in Brazil. Since it has only recently emerged as a pest species in production areas, understanding seasonal trends in the species' activity is particularly important for designing management strategies. Therefore, this study aims to investigate the influence of climate factors (temperature and relative humidity) and six strawberry cultivars ('Albion', 'Aromas', 'Camino Real', 'Monterey', 'Portola', and 'San Andreas') on the population dynamics of H. haemorrhoidalis during the 2018-2019 and 2019-2020 growing seasons in southern Brazil. The greatest peak in thrips populations in the cultivars assessed occurred between January and March (summer) in both growing seasons. The relationship between the incidence of H. haemorrhoidalis and temperature was observed to be negative during the autumn, spring, and summer months but positive during winter. Relative humidity levels above 70% had a negative influence on the abundance of insects. Populations of H. haemorrhoidalis varied between the cultivars as follows: 'Aromas' > 'Monterey' > 'San Andreas' > 'Albion' > 'Camino Real' > 'Portola'. Cross-sections of strawberry leaves showed that 'Monterey' had the highest density of nonglandular trichomes/mm2 (7.26), while 'Portola' cultivar, the greatest predominance of glandular trichomes/mm2 (12.67), which may have contributed to the incidence of H. haemorrhoidalis. The demographic information presented on H. haemorrhoidalis in strawberry cultivars provides a starting point to better understand the insect's population dynamics. These results also indicate that climatic conditions are a limiting factor for the development of H. haemorrhoidalis.

Classification of Various Marijuana Varieties by Raman Microscopy and Chemometrics.

The Raman analysis of marijuana is challenging because of the sample's easy photo-degradation caused by the laser intensity. In this study, optimization of collection parameters and laser focusing on marijuana trichome heads allowed collecting Raman spectra without damaging the samples. The Raman spectra of Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) standard cannabinoids were compared with Raman spectra of five different types of marijuana: four Sativa varieties (Amnesia Haze, Amnesia Hy-Pro, Original Amnesia, and Y Griega) and one Indica variety (Black Domina). The results verified the presence of several common spectral bands that are useful for marijuana characterization. Results were corroborated by the quantum chemical simulated Raman spectra of their acid-form (tetrahydrocannabinol acid (THCA), cannabidiol acid (CBDA)) and decarboxylated cannabinoids (THC, CBD, and CBN). A chemometrics-assisted method based on Raman microscopy and OPLS-DA offered good classification among the different marijuana varieties allowing identification of the most significant spectral bands.

Introgression of the sesquiterpene biosynthesis from Solanum habrochaites to cultivated tomato offers insights into trichome morphology and arthropod resistance.

MAIN CONCLUSION: Cultivated tomatoes harboring the plastid-derived sesquiterpenes from S. habrochaites have altered type-VI trichome morphology and unveil additional genetic components necessary for piercing-sucking pest resistance. Arthropod resistance in the tomato wild relative Solanum habrochaites LA1777 is linked to specific sesquiterpene biosynthesis. The Sesquiterpene synthase 2 (SsT2) gene cluster on LA1777 chromosome 8 controls plastid-derived sesquiterpene synthesis. The main genes at SsT2 are Z-prenyltransferase (zFPS) and Santalene and Bergamotene Synthase (SBS), which produce α-santalene, ß-bergamotene, and α-bergamotene in LA1777 round-shaped type-VI glandular trichomes. Cultivated tomatoes have mushroom-shaped type-VI trichomes with much smaller glands that contain low levels of monoterpenes and cytosolic-derived sesquiterpenes, not presenting the same pest resistance as in LA1777. We successfully transferred zFPS and SBS from LA1777 to cultivated tomato (cv. Micro-Tom, MT) by a backcrossing approach. The trichomes of the MT-Sst2 introgressed line produced high levels of the plastid-derived sesquiterpenes. The type-VI trichome internal storage-cavity size increased in MT-Sst2, probably as an effect of the increased amount of sesquiterpenes, although it was not enough to mimic the round-shaped LA1777 trichomes. The presence of high amounts of plastid-derived sesquiterpenes was also not sufficient to confer resistance to various tomato piercing-sucking pests, indicating that the effect of the sesquiterpenes found in the wild S. habrochaites can be insect specific. Our results provide for a better understanding of the morphology of S. habrochaites type-VI trichomes and paves the way to obtain insect-resistant tomatoes.

Epicormic bud protection traits vary along a latitudinal gradient in a neotropical savanna.

Regrowth via production of epicormic shoots is an important strategy for many woody plants after environmental disturbances such as fire, drought, and herbivory. Populations spreading across a broad latitudinal gradient offer opportunities to investigate if essential traits vary with heterogenous environmental conditions, such as in savanna ecosystems. This information can help us predict plant responses to climate change. Here, we evaluated if epicormic bud protection traits varied among populations of three focal savanna species (Miconia albicans, Solanum lycocarpum, and Zeyheria montana) that have a wide distribution and grow under variable climatic conditions. We randomly sampled 225 individuals over five spatially independent sites (7°, 10°, 15°, 18°, and 24° S) in Brazil, totaling 15 individuals per species per area. We analyzed anatomical transverse sections of five buds per species per area to assess the relative area occupied by crystal and phenolic idioblasts, the thickness of the trichome boundary layer, and to test if these traits were associated with climatic conditions. The buds were protected by cataphylls and composed of a variable number of undeveloped leaves enveloping the shoot apex. For M. albicans, we found an association between maximum temperature and both phenolic idioblasts and trichome boundary layer, but no association with crystal idioblasts. In S. lycocarpum, only the trichome boundary layer was associated with maximum temperature plus high radiation. Z. montana showed no variation. Combination of two or more traits can lead to the development of adaptative strategies to different climatic conditions. We present for the first time an analysis of epicormic bud traits in plant populations occurring in an extensive latitudinal gradient and shed light on how maximum temperature is associated with these traits, contributing to a better understanding of plant resprouting capabilities in widespread savanna plant species.

Do food trichomes occur in Pinguicula (Lentibulariaceae) flowers?

BACKGROUND AND AIMS: Floral food bodies (including edible trichomes) are a form of floral reward for pollinators. This type of nutritive reward has been recorded in several angiosperm families: Annonaceae, Araceae, Calycanthaceae, Eupomatiaceae, Himantandraceae, Nymphaeaceae, Orchidaceae, Pandanaceae and Winteraceae. Although these bodies are very diverse in their structure, their cells contain food material: starch grains, protein bodies or lipid droplets. In Pinguicula flowers, there are numerous multicellular clavate trichomes. Previous authors have proposed that these trichomes in the Pinguicula flower play the role of 'futterhaare' ('feeding hairs') and are eaten by pollinators. The main aim of this study was to investigate whether the floral non-glandular trichomes of Pinguicula contain food reserves and thus are a reward for pollinators. The trichomes from the Pinguicula groups, which differ in their taxonomy (species from the subgenera: Temnoceras, Pinguicula and Isoloba) as well as the types of their pollinators (butterflies/flies and bees/hummingbirds), were examined. Thus, it was determined whether there are any connections between the occurrence of food trichomes and phylogeny position or pollination biology. Additionally, we determined the phylogenetic history of edible trichomes and pollinator evolution in the Pinguicula species. METHODS: The species that were sampled were: Pinguicula moctezumae, P. esseriana, P. moranensis, P. emarginata, P. rectifolia, P. mesophytica, P. hemiepiphytica, P. agnata, P. albida, P. ibarrae, P. martinezii, P. filifolia, P. gigantea, P. lusitanica, P. alpina and P. vulgaris. Light microscopy, histochemistry, and scanning and transmission electron microscopy were used to address our aims with a phylogenetic perspective based on matK/trnK DNA sequences. KEY RESULTS: No accumulation of protein bodies or lipid droplets was recorded in the floral non-glandular trichomes of any of the analysed species. Starch grains occurred in the cells of the trichomes of the bee-/fly-pollinated species: P. agnata, P. albida, P. ibarrae, P. martinezii, P. filifolia and P. gigantea, but not in P. alpina or P. vulgaris. Moreover, starch grains were not recorded in the cells of the trichomes of the Pinguicula species that have long spurs, which are pollinated by Lepidoptera (P. moctezumae, P. esseriana, P. moranensis, P. emarginata and P. rectifolia) or birds (P. mesophytica and P. hemiepihytica), or in species with a small and whitish corolla that self-pollinate (P. lusitanica). The results on the occurrence of edible trichomes and pollinator syndromes were mapped onto a phylogenetic reconstruction of the genus. CONCLUSION: Floral non-glandular trichomes play the role of edible trichomes in some Pinguicula species (P. agnata, P. albida, P. ibarrae, P. martinezii, P. filifolia and P. gigantea), which are mainly classified as bee-pollinated species that had originated from Central and South America. It seems that in the Pinguicula that are pollinated by other pollinator groups (Lepidoptera and hummingbirds), the non-glandular trichomes in the flowers play a role other than that of a floral reward for their pollinators. Edible trichomes are symplesiomorphic for the Pinguicula species, and thus do not support a monophyletic group such as a synapomorphy. Nevertheless, edible trichomes are derived and are possibly a specialization for fly and bee pollinators by acting as a food reward for these visitors.

Class I TCP transcription factors regulate trichome branching and cuticle development in Arabidopsis.

Trichomes and the cuticle are two specialized structures of the aerial epidermis that are important for plant organ development and interaction with the environment. In this study, we report that Arabidopsis thaliana plants affected in the function of the class I TEOSINTE BRANCHED 1, CYCLOIDEA, PCF (TCP) transcription factors TCP14 and TCP15 show overbranched trichomes in leaves and stems and increased cuticle permeability. We found that TCP15 regulates the expression of MYB106, a MIXTA-like transcription factor involved in epidermal cell and cuticle development, and overexpression of MYB106 in a tcp14 tcp15 mutant reduces trichome branch number. TCP14 and TCP15 are also required for the expression of the cuticle biosynthesis genes CYP86A4, GPAT6, and CUS2, and of SHN1 and SHN2, two AP2/EREBP transcription factors required for cutin and wax biosynthesis. SHN1 and CUS2 are also targets of TCP15, indicating that class I TCPs influence cuticle formation acting at different levels, through the regulation of MIXTA-like and SHN transcription factors and of cuticle biosynthesis genes. Our study indicates that class I TCPs are coordinators of the regulatory network involved in trichome and cuticle development.

A Comprehensive Analysis of Wheat Resistance to Rhopalosiphum padi (Hemiptera: Aphididae) in Brazilian Wheat Cultivars.

Rhopalosiphum padi L. is one of the predominant aphids affecting wheat crops worldwide. Therefore, the identification of resistant genotypes and the understanding of molecular response mechanisms involved in wheat resistance to this aphid may contribute to the development of new breeding strategies. In this study, we evaluated the resistance of 15 wheat cultivars to R. padi and performed morpho-histological and gene expression analyses of two wheat cultivars (BRS Timbaúva, resistant and Embrapa 16, susceptible) challenged and unchallenged by R. padi. The main findings of our work are as follows: 1) most Brazilian wheat cultivars recently released are resistant to R. padi; 2) Green leaf volatiles are probably involved in the resistance of the BRS Timbaúva cultivar to the aphid; 3) trichomes were more abundant and larger in the resistant cultivar; 4) the internal morphology did not show differences between cultivars; 5) the lipoxygenase-encoding gene was downregulated in the susceptible cultivar and basal expression remained level in the resistant cultivar; and 6) the expression of resistance-related proteins was induced in the resistant but not in the susceptible cultivar. Lipoxygenase is the first enzyme in the octadecanoic pathway, a well-known route for the synthesis of signaling molecules involved in the activation of plant defense. The overall analyses suggest that the key steps in BRS Timbaúva resistance to R. padi may be presence or absence of green leaf volatiles decreasing the aphid preference and the action of nonglandular trichome as a physical barrier, which allows continuous lipoxygenase-encoding gene expression.

Artemisia annua L. and photoresponse: from artemisinin accumulation, volatile profile and anatomical modifications to gene expression.

KEY MESSAGE: Blue and yellow light affected metabolism and the morphology. Blue and red promote the DOXP/MEP pathway. ADS gene expression was increased in plants cultivated under blue, promoting artemisinin content. Artemisinin-based combination therapies are the most effective treatment for highly lethal malaria. Artemisinin is produced in small quantities in the glandular trichomes of Artemisia annua L. Our aim was to evaluate the effect of light quality in A. annua cultivated in vitro under different light qualities, considering anatomical and morphological changes, the volatile composition, artemisinin content and the expression of two key enzymes for artemisinin biosynthesis. Yellow light is related to the increase in the number of glandular trichomes and this seemed to positively affect the molecular diversity in A. annua. Yellow light-stimulated glandular trichome frequency without triggered area enhancement, whereas blue light stimulated both parameters. Blue light enhanced the thickness of the leaf epidermis. The B-promoting effect was due to increased cell size and not to increased cell numbers. Green and yellow light positively influenced the volatile diversity in the plantlets. Nevertheless, blue and red light seemed to promote the DOXP/MEP pathway, while red light stimulates MVA pathway. Amorpha-4,11-diene synthase gene expression was significantly increased in plants cultivated under blue light, and not red light, promoting artemisinin content. Our results showed that light quality, more specifically blue and yellow light, positively affected secondary metabolism and the morphology of plantlets. It seemed that steps prior to the last one in the artemisinin biosynthesis pathway could be strongly influenced by blue light. Our work provides an alternative method to increase the amount of artemisinin production in A. annua without the use of transgenic plants, by the employment of blue light.

Induction of essential oil production in Mentha x piperita by plant growth promoting bacteria was correlated with an increase in jasmonate and salicylate levels and a higher density of glandular trichomes.

Plant growth promoting bacteria (PGPB) are agriculturally important soil bacteria that increase plant growth. We subjected peppermint to inoculation with three species of PGPB. After inoculation, the plants were sprayed with methyl jasmonate solution (MeJA) or SA (salicylic acid). Then, the plants were harvested and the plant growth parameters, trichome density, EO content and endogenous phytohormones were measured. Shoot fresh weight was reduced in plants inoculated and treated with MeJA whereas EO content varied depending on the MeJA concentration applied. Plants inoculated and treated with MeJA 2 mM showed the maximum increase in EO production, revealing a synergism between PGPB and MeJA. SA treatments also enhanced EO yield. The increased growth and EO production observed upon PGPB application were at least partly due to an increase in the JA and SA concentrations in the plant, as well as to an associated rise in the glandular trichome density.

Triticum aestivum in open skin wounds: cytotoxicity and collagen histopathology / Triticum aestivum em feridas cutâneas abertas: citotoxicidade e histopatologia de colágeno

Phytoterapic compounds have been used in wound healing for many centuries. Nowadays, scientific evidences of phytotherapeutics is a requirement of the legislation. The scientific literature notes the need for healing topics yielding scars that are both aesthetically appealing and resistant. We aimed to evaluate the cytotoxicity of several doses of T. aestivum extract (2 mg mL-1, 4 mg mL-1, 6 mg mL-1, 8 mg mL-1 and 10 mg mL-1) in a fibroblast cell line and the healing process in an in vivo experimental model (New Zealand rabbits). For this, MTT test in 3T6 cells was performed in duplicates using MEM (0 mg ml-1) as negative control. Cell viability was calculated as: absorbance average in treatments/absorbance average in controls x 100. In vivo test was performed in 78 skin wounds in rabbits that were treated with 2 mg ml-1and 10 mg ml-1 of T. aestivum and non-ionic cream for 21 days. After this period, it was evaluated the histology using picrosorius and Gomoris trichrome staining. Statistical analysis was evaluated using T test (Graphpad) for cytotoxicity assay, Fischer test for the gomori trichrome test (Grahpad) and Kruskal-Wallis (Statistic 9.0) for picrosirius test. The in vitro test resulted in cytotoxicity observed at 2mg mL-1 whereas cells were viable at higher doses. On the other hand, it was observed that collagen formation of wounds was more uniform with this dose than with 10mg mL-1 extract in the in vivo study. Thus, we conclude that the 2mg mL-1 T. aestivum aqueous extract dose was more efficient in the in vivo wound healing study, despite its cytotoxic effects in vitro.(AU)

Os extratos vegetais têm sido utilizados na cicatrização de feridas a muitos séculos. No entanto nos dias atuais a comprovação científica dos fitoterápicos é uma exigência da legislação. Na literatura científica se observa a necessidade de cicatrizantes tópicos que proporcionem uma cicatriz estética e resistente. Devido a isso objetivou-se avaliar a citotoxicidade de diversas doses de T. aestivum (2 mg mL1, 4 mg mL-1, 6 mg mL-1, 8 mg mL-1 e 10 mg mL-1) em linhagem celular de fibroblasto, e o processo cicatricial em modelo experimental (New Zealand rabbits) in vivo. Para isso foi realizado o teste de MTT em linhagem celular 3T. Tests were performed in duplicates, using MEM (0 mg mL-1) as negative control. Cell viability was calculated as: absorbance average in treatments/absorbance average in controls x 100. No ensaio in vivo foram realizadas 78 feridas experimentais em coelhos que foram tratadas T. aestivum 2mg mL-1, T. aestivum 10 mg mL-1e creme não iônico por 21 dias, após foi avaliado a histologia do tricrômico de golmori e de picrosirius. Na análise estatística do ensaio de citotoxicidade foi realizado o teste de t (Graphpad), para avaliação do tricomico de golmori foi realizado o teste de fischer (Graphpad) e no picrosirius foi avaliado através de Kruskal - Wallis (Statistic 9.0). O resultado in vitro demonstrou que a dose de 2mg mL-1 foi citotóxica para as células e que doses maiores a essa apresentavam viabilidade celular. No entanto no estudo in vivo foi constatado que as feridas tratadas com essa dose apresentaram a formação de colágeno mais uniforme que as tratadas com 10 mg mL-1. Concluí-se que a dose de 2mg mL-1 do extrato aquoso de T. aestivum é eficiente no ensaio in vivo com as feridas experimentais, o que não foi observado no estudo in vitro.(AU)

Triticum aestivum in open skin wounds: cytotoxicity and collagen histopathology / Triticum aestivum em feridas cutâneas abertas: citotoxicidade e histopatologia de colágeno

Phytoterapic compounds have been used in wound healing for many centuries. Nowadays, scientific evidences of phytotherapeutics is a requirement of the legislation. The scientific literature notes the need for healing topics yielding scars that are both aesthetically appealing and resistant. We aimed to evaluate the cytotoxicity of several doses of T. aestivum extract (2 mg mL-1, 4 mg mL-1, 6 mg mL-1, 8 mg mL-1 and 10 mg mL-1) in a fibroblast cell line and the healing process in an in vivo experimental model (New Zealand rabbits). For this, MTT test in 3T6 cells was performed in duplicates using MEM (0 mg ml-1) as negative control. Cell viability was calculated as: absorbance average in treatments/absorbance average in controls x 100. In vivo test was performed in 78 skin wounds in rabbits that were treated with 2 mg ml-1and 10 mg ml-1 of T. aestivum and non-ionic cream for 21 days. After this period, it was evaluated the histology using picrosorius and Gomori’s trichrome staining. Statistical analysis was evaluated using T test (Graphpad) for cytotoxicity assay, Fischer test for the gomori trichrome test (Grahpad) and Kruskal-Wallis (Statistic 9.0) for picrosirius test. The in vitro test resulted in cytotoxicity observed at 2mg mL-1 whereas cells were viable at higher doses. On the other hand, it was observed that collagen formation of wounds was more uniform with this dose than with 10mg mL-1 extract in the in vivo study. Thus, we conclude that the 2mg mL-1 T. aestivum aqueous extract dose was more efficient in the in vivo wound healing study, despite its cytotoxic effects in vitro.

Os extratos vegetais têm sido utilizados na cicatrização de feridas a muitos séculos. No entanto nos dias atuais a comprovação científica dos fitoterápicos é uma exigência da legislação. Na literatura científica se observa a necessidade de cicatrizantes tópicos que proporcionem uma cicatriz estética e resistente. Devido a isso objetivou-se avaliar a citotoxicidade de diversas doses de T. aestivum (2 mg mL1, 4 mg mL-1, 6 mg mL-1, 8 mg mL-1 e 10 mg mL-1) em linhagem celular de fibroblasto, e o processo cicatricial em modelo experimental (New Zealand rabbits) in vivo. Para isso foi realizado o teste de MTT em linhagem celular 3T. Tests were performed in duplicates, using MEM (0 mg mL-1) as negative control. Cell viability was calculated as: absorbance average in treatments/absorbance average in controls x 100. No ensaio in vivo foram realizadas 78 feridas experimentais em coelhos que foram tratadas T. aestivum 2mg mL-1, T. aestivum 10 mg mL-1e creme não iônico por 21 dias, após foi avaliado a histologia do tricrômico de golmori e de picrosirius. Na análise estatística do ensaio de citotoxicidade foi realizado o teste de t (Graphpad), para avaliação do tricomico de golmori foi realizado o teste de fischer (Graphpad) e no picrosirius foi avaliado através de Kruskal - Wallis (Statistic 9.0). O resultado in vitro demonstrou que a dose de 2mg mL-1 foi citotóxica para as células e que doses maiores a essa apresentavam viabilidade celular. No entanto no estudo in vivo foi constatado que as feridas tratadas com essa dose apresentaram a formação de colágeno mais uniforme que as tratadas com 10 mg mL-1. Concluí-se que a dose de 2mg mL-1 do extrato aquoso de T. aestivum é eficiente no ensaio in vivo com as feridas experimentais, o que não foi observado no estudo in vitro.

Leaf surface traits and water storage retention affect photosynthetic responses to leaf surface wetness among wet tropical forest and semiarid savanna plants.

While it is reasonable to predict that photosynthetic rates are inhibited while leaves are wet, leaf gas exchange measurements during wet conditions are challenging to obtain due to equipment limitations and the complexity of canopy-atmosphere interactions in forested environments. Thus, the objective of this study was to evaluate responses of seven tropical and three semiarid savanna plant species to simulated leaf wetness and test the hypotheses that (i) leaf wetness reduces photosynthetic rates (Anet), (ii) leaf traits explain different responses among species and (iii) leaves from wet environments are better adapted for wet leaf conditions than those from drier environments. The two sites were a tropical rainforest in northern Costa Rica with ~4200 mm annual rainfall and a savanna in central Texas with ~1100 mm. Gas exchange measurements were collected under dry and wet conditions on five sun-exposed leaf replicates from each species. Additional measurements included leaf wetness duration and stomatal density. We found that Anet responses varied greatly among species, but all plants maintained a baseline of activity under wet leaf conditions, suggesting that abaxial leaf Anet was a significant percentage of total leaf Anet for amphistomatous species. Among tropical species, Anet responses immediately after wetting ranged from -31% (Senna alata (L.) Roxb.) to +21% (Zamia skinneri Warsz. Ex. A. Dietr.), while all savanna species declined (up to -48%). After 10 min of drying, most species recovered Anet towards the observed status prior to wetting or surpassed it, with the exception of Quercus stellata Wangenh., a savanna species, which remained 13% below Anet dry. The combination of leaf wetness duration and leaf traits, such as stomatal density, trichomes or wax, most likely influenced Anet responses positively or negatively. There was also overlap between leaf traits and Anet responses of savanna and tropical plants. It is possible that these species converge on a relatively conservative response to wetness, each for divergent purposes (cooling, avoiding stomatal occlusion, or by several unique means of rapid drying). A better understanding of leaf wetness inhibiting photosynthesis is vital for accurate modeling of growth in forested environments; however, species adapted for wet environments may possess compensatory traits that mitigate these effects.

Characterization of Antixenosis in Soybean Genotypes to Bemisia tabaci (Hemiptera: Aleyrodidae) Biotype B.

Bemisia tabaci biotype B (Gennadius) is one of the most important soybean pest worldwide. Herein, 15 soybean genotypes were evaluated, to characterize the occurrence of antixenosis to B. tabaci biotype B. Initially, a multiple-choice test with all genotypes was carried out, evaluating the settling and oviposition preference at 3 d after infestation, and the colonization by nymphs after 48 d of infestation. Subsequently, a no-choice test, using 14 genotypes, was conducted with infested plants individually, and the number of eggs was counted after 72 h. Then, 10 genotypes were selected (indicative of resistance and susceptibility), which were evaluated for whitefly settling 24, 48, and 72 h after infestation and for oviposition 72 h after infestation. The trichomes of the leaflets were characterized for density, size, and inclination to establish possible correlations with the settling and oviposition in the genotypes. In the first multiple-choice test, involving 15 genotypes, 'IAC-17,' 'IAC-19,' and UX-2569-159 expressed antixenosis against B. tabaci. 'Jackson,' 'P98Y11,' and PI-229358 exhibited the same behavior in the no-choice test. In the multiple-choice test, 'Jackson,' 'P98Y11,' and 'TMG1176 RR' were the least attractive and least used for oviposition. The antixenosis shown by 'Jackson,' 'P98Y11,' and PI-229358 may be related to the characteristics of the trichomes (lower density and inclined). Based on the experiments carried out, 'IAC-17,' 'IAC-19,' 'Jackson,' 'P98Y11,' PI-229358, TMG1176 RR, and UX-2569-159 are considered promising for resistance to B. tabaci biotype B and may be exploited in soybean breeding programs for resistance to insects.

Direct Analyses of Secondary Metabolites by Mass Spectrometry Imaging (MSI) from Sunflower (Helianthus annuus L.) Trichomes.

Helianthus annuus (sunflower) displays non-glandular trichomes (NGT), capitate glandular trichomes (CGT), and linear glandular trichomes (LGT), which reveal different chemical compositions and locations in different plant tissues. With matrix-assisted laser desorption/ionization (MALDI) and laser desorption/ionization (LDI) mass spectrometry imaging (MSI) techniques, efficient methods were developed to analyze the tissue distribution of secondary metabolites (flavonoids and sesquiterpenes) and proteins inside of trichomes. Herein, we analyzed sesquiterpene lactones, present in CGT, from leaf transversal sections using the matrix 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA) (mixture 1:1) with sodium ions added to increase the ionization in positive ion mode. The results observed for sesquiterpenes and polymethoxylated flavones from LGT were similar. However, upon desiccation, LGT changed their shape in the ionization source, complicating analyses by MSI mainly after matrix application. An alternative method could be applied to LGT regions by employing LDI (without matrix) in negative ion mode. The polymethoxylated flavones were easily ionized by LDI, producing images with higher resolution, but the sesquiterpenes were not observed in spectra. Thus, the application and viability of MALDI imaging for the analyses of protein and secondary metabolites inside trichomes were confirmed, highlighting the importance of optimization parameters.

Functional Response of Aphidoletes aphidimyza Rondani (Diptera: Cecidomyiidae) to Aphis gossypii Glover (Hemiptera: Aphididae): Effects of Vermicompost and Host Plant Cultivar.

Interactions between natural enemies and herbivores may be affected by application of fertilizers and different cultivars. We investigated the functional response of the predatory gall midge, Aphidoletes aphidimyza Rondani (Diptera: Cecidomyiidae) larvae to the nymphs of the melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), reared on two commonly grown cucumber cultivars in Iran (Khasib and Karim) treated with different vermicompost/soil ratios (0:100, 10:90, 20:80, and 30:70%). Based on logistic regression analysis, A. aphidimyza revealed a type II functional response to the aphid in all treatments. Attack rates and handling times of A. aphidimyza larvae on different vermicompost/soil ratios ranged from 0.076 to 0.140 h(-1) and 0.969 to 1.164 h on Khasib and from 0.092 to 0.123 h(-1) and 0.905 to 1.229 h on Karim, respectively. Furthermore, increasing the density of the melon aphid on both cultivars amended with vermicompost/soil ratios resulted in increased prey consumption by the predator. Density of trichomes increased when plants received higher concentrations of vermicompost. So, trichomes may be responsible for different attack rates and handling times of A. aphidimyza on both cultivars.