How does plant leaf senescence of grassland species influence decomposition kinetics and litter compounds dynamics?

The influence of litter quality on plant litter decomposition rates is a crucial aspect of the soils C cycle. In grassland ecosystems, leaf litter, which is not removed either by herbivores or by mowing, returns to soil after the senescence process (brown litter). In grassland managed by mowing, another significant proportion of litter returns to the soil before senescence through harvesting losses (green litter). We hypothesized that changes in leaf tissue quality due to the senescence process would lead to contrasting decomposition dynamics of brown litter compared to green litter. Our conceptual approach included the monitoring of decomposition of green (fresh leaves) and brown litter (dead leaves, still attached to the plant) of three different grassland species (Lolium perenne, Festuca arundinacea and Dactylis glomerata) during a 1 year field incubation. After 0, 2, 4, 20 and 44 weeks, we retrieved the litterbags and analysed the remaining material for carbon and nitrogen content and stable isotope composition. Additionally, we determined the lignin content and composition by CuO oxidation and the non-cellulosic neutral carbohydrate content and composition after TFA hydrolysis. As expected, green litter, being higher in N and soluble compounds, while showing a lower C:N ratio and lower lignin contents compared to brown litter, was degraded at a higher rate. Carbon decomposition kinetics suggests that both leaf litter types consist of two pools with contrasting turnover times. The size of the active pool was related to the initial content of soluble plant litter compounds and the size of the recalcitrant pool was related to the lignin to N ratio of initial plant material. More lignin was lost from green litter compared to brown litter. P-coumaryl-type lignin units were decomposed at a higher rate than vanillyl and syringyl units. Total non cellulosic polysaccharide content showed little changes for both litter types. In contrast, the ratios of hexoses/pentoses (C6/C5) and desoxy sugars/pentoses (desoxy/C5) increased during decomposition of green litter only. This is an indication for an increasing contribution of microbial derived compounds being consistant with the higher decomposition rate of this material. Our results showed that grassland management (grazing versus mowing) could influence soil carbon sequestration through different proportions of green and brown litter returned to soil.     

Color characteristics of Beijing's regional woody vegetation based on Natural Color System

Seasonality is a typical characteristic of Beijing's regional vegetation, and plant color is one of the most prominent visual factors of vegetation dynamic. In this research, we explored the composition and dynamic characteristics of plant color in Beijing's urban vegetation, involving the analysis of overall characteristics and respective features of leaf, flower, and fruit colors. Color data was collected from 177 woody plant species in Beijing Botanical Garden, spanning their annual life cycle, and identified with the colorimetry of the Natural Color System (NCS). Correlation and regression analyses were applied to reveal the temporal dynamic features of overall plant color richness. Cluster analysis was applied to categorize tree species based on typical colors of various plant organs. Color richness and color dispersion were introduced as two factors to measure color diversity of various tree species, applied in species evaluation by sorting and principal component analysis (PCA). Color dispersion of three‐dimensional NCS data was measured with a modified SD based on the calculation of mean spatial distance in the NCS space. Main results are as follows. The first part is plant color composition. The composition of all plant colors contains 862 NCS color species, 20 blackness species ranging from 3 to 90, 20 chromaticness species ranging from 0 to 90, 35 hue species ranging from G10Y‐B90G, and N. The second part is temporal dynamic of overall color richness. Leaf color richness and total color richness are significantly positively correlated with pentad (5‐day) sequence; flower color richness is significantly negatively correlated with pentad sequence; and fruit color richness first increases and then decreases over time. The third part is cluster analysis of tree species. Based on typical growing‐leaf color, various tree species were clustered into 6 categories; based on typical senescent‐leaf color, various tree species were clustered into 6 categories; based on typical flower color, various tree species were clustered into 15 categories; based on typical fruit color, various tree species were clustered into 7 categories. The fourth part is color diversity evaluation of various tree species with PCA. According to the PCA of flower‐leaf color diversity, the species with higher leaf color diversity and higher flower color diversity include Cotinus coggygria, Lagerstroemia indica, and Amygdalus triloba; the species with higher flower color diversity and lower leaf color diversity include Campsis radicans and Tamarix chinensis; the species with higher leaf color diversity and lower flower color diversity include Acer ginnala and Crataegus pinnatifida; the species with lower color diversity both for flower and leaf colors include Fontanesia fortune and Gleditsia sinensis. According to the PCA of leaf color diversity, the species with higher leaf color diversity in both leaf growth period and leaf senescence period include Diospyros kaki, Lagerstroemia indica and Paeonia suffruticosa; the species with higher leaf color diversity in leaf growth period and lower leaf color diversity in leaf senescence period include Amygdalus persica ‘Atropurpurea’ and Prunus virginiana ‘Canada Red’; the species with higher leaf color diversity in leaf senescent period and lower color diversity in leaf growth period include Quercus palustris, Armeniaca sibirica, and Metasequoia glyptostroboides; the species with lower leaf color diversity for the whole leaf development period include Gleditsia sinensis and Swida walteri.     

Role of the Lipoxygenase/lyase Pathway of Host-food Plants in the Host Searching Behavior of Two Parasitoid Species, Cotesia glomerata and Cotesia plutellae

To elucidate the role of the plant lipoxygenase (LOX)/lyase pathway for host search behavior of two parasitic wasps attacking herbivorous larvae, an Arabidopsis mutant (all84) was isolated with a mutation somewhere in the LOX/lyase pathway. Detached leaves of the mutant were shown to release less (Z)-3-hexenal, a first green leaf volatile (GLV) product of the LOX/lyase pathway. The braconid larval parasitoids studied, Cotesia glomerata and Cotesia plutella, differ in their ability to discriminate among plant volatiles induced by feeding of lepidopteran hosts and nonhosts: C. plutella only responds to plant volatiles induced by hosts (Plutella larvae), whereas the response by the more generalist C. glomerata is not host specific. The Arabidopsis mutant all84 infested by Pieris larvae was less attractive to C. glomerata than Arabidopsis wild type (wt) infested by the host larvae. C. glomerata was attracted by two of the GLV biosynthesized through the LOX/lyase pathway, (E)-2-hexenal and (Z)-3-hexenyl acetate. However, attraction of C. plutellae to volatiles from Plutella-infested all84 plants did not differ from attraction to host-infested wt Arabidopsis. Both wasp species were arrested to the respective host-infested edge of the wt leaf by showing characteristic antennal searching behavior on the edge. In C. glomerata, the duration of this searching behavior at the infested leaf edge was significantly shorter on all84 plants than on wt plants. By contrast, the duration of the searching behavior of C. plutellae on the host-infested leaf edge of all84 was not significantly different from that on the wt leaf. These data suggest that the LOX/lyase pathway is directly involved in the production of attractants and arrestants important for host search behavior of the more generalist C. glomerata, but not for the specialist C. plutellae.     

Growth and yield response of glasshouse- and field-grown sweetpotato to nitrogen supply

Nitrogen (N) is an essential element for producing optimum crop yields, but negative responses to high N supply are commonly reported in sweetpotato (Ipomoea batatas) production. This study assessed contrasting responses of sweetpotato yield as a result of N application rates of 0, 30, 60, 90, 130, 160 and 230 kg ha^?1 in a glasshouse trial, and rates of 0, 50, 100, 150, 200 and 250 kg ha^?1, equivalent to 160, 210, 260, 310, 360 and 410 kg ha^?1 when soil N supply is included. The glasshouse-grown sweetpotato produced a maximum number and dry-biomass of storage roots, aboveground biomass and leaf area at 130 kg N ha^?1, while leaf N concentration peaked at 90 kg N ha^?1. Further increasing N application to 230 kg ha^?1 did not result in significant change in any of these attributes. In field-grown sweetpotato, leaf and storage root N concentrations increased with increasing N supply. Although N supply had no effect on the number of storage roots, total yield peaked at 260 kg ha^?1. Further increase of N supply reduced the total yield by up to 14% of the maximum yield. With increasing N supply, the glasshouse-grown sweetpotato yield linearly increased with leaf area; the arrangement of the trial permitting light interception to exceed the pot surface area. The yield reduction in field-grown plants was attributed to excess growth of aboveground parts, beyond that needed for efficient light capture. Respirational demand of the aboveground growth occurred at the expense of storage root yields.     

Damage-induced alkaloids in tobacco: Pot-bound plants are not inducible

Field-grown wild tobacco plants (Nicotiana sylvestris) were subjected to a defoliation regime designed to mimic the rate and amount of leaf mass removed by one tobacco hornworm per plant. Undamaged leaves on these plants undergo a dramatic (457% for leaf position 5, 410% for leaf position 8) increase in total leaf alkaloids compared to same-age and positioned control leaves on undamaged control plants. However, potted greenhouse-grown plants fail to exhibit the same damage-induced increase in alkaloid content. The greenhouse environment differs from the field environment in factors known to influence leaf alkaloid content, particularly soil N, P, K, near-UV radiation, and relative humidity. However, altering these environmental factors does not make potted plants able to increase their leaf alkaloid levels in response to defoliation. Transplanting plants into larger pots with more soil does allow the plants to respond to defoliation. Thirty days after transplanting, the plants are again unresponsive to damage, probably as a result of becoming pot-bound. This result suggests a mechanism for the induction response, specifically that leaf damage triggers synthesis of these alkaloids in the roots, and offers a potentially valuable experimental tool for the study of induced-plant defenses in tobacco and other plants that synthesize alkaloids in their root tissues.     

Flavonoid pigments in marbled white butterfly (Melanargia galathea) are dependent on flavonoid content of larval diet

Analyses of two-dimensional chromatographic flavonoid patterns of butterflies reared on different grass species have shown that the flavonoid pattern ofMelanargia galathea is dependent on the flavonoid content of the larval diet. This confirms the dietary origin of flavonoid pigments inM. galathea. The flavonoid patterns of butterflies reared on different grass species differ from each other and from the larval food plants;M. galathea reared on the same grass species have identical flavonoid patterns. Differences in the butterfly and larval food plant flavonoid patterns indicate that the ingested flavonoids are metabolized byM. galathea or its gut flora before sequestration. The distinct flavonoid patterns of butterflies reared on different larval food plants have been defined as the flavonoid fingerprint profiles for each grass species. Similarity between theFestuca rubra flavonoid fingerprint profile and the constant flavonoid pattern characteristic of wild-capturedMelanargia suggests thatMelanargia larvae are not generalist grass feeders, but are specific toF. rubra or toF. rubra and a few closely related grass species in the wild.     

Allelopathic Potential of <Emphasis Type="Italic">Robinia pseudo-acacia</Emphasis> L.

Robinia pseudo-acacia L. (black locust) is a nonindigenous species currently invading the central part of Japanese grasslands. Several allelochemicals were identified and characterized from the leaf tissue. The growth of both radicle and hypocotyl in the tested species (barnyard grass, white clover, lettuce, and Chinese cabbage) was reduced when grown in soil mixed with the leaves of R. pseudo-acacia at various concentrations. Aqueous leaf extracts, when bioassayed, exhibited a significant suppression of radicle growth. Chromatographic separation of an ethanolic extract of R. pseudo-acacia leaves resulted in isolation of three compounds, identified as robinetin (1), myricetin (2), and quercetin (3) by nuclear magnetic resonance and mass spectroscopy. All inhibited root and shoot growth of lettuce. Robinetin, found in a large amount, caused 50% suppression of the root and shoot growth of lettuce at 100 ppm. The presence of these bioactive substances in leaf tissue suggests a potential role for flavonoids in R. pseudo-acacia invasion in introduced habitats.     

Rumex obtusifolius L: Release of allelochemical agents and their influence on small-scale spatial distribution of meadow species

DecomposingRumex obtusifolius L. leaves and their extracts were most toxic for germination and root growth of meadow speciesLolium perenne, Trifolium repens, Poa pratensis, andDactylis glomerata after seven days' decomposition, although the toxicity level was, in some cases, still high after 21 days. The hypothesis thatR. obtusifolius exerts allelopathic control over meadow species is supported by small-scale distribution of meadow species in the neighborhood ofR. obtusifolius plants. The area affected and the intensity of the effect both increase with the size of the individualR. obtusifolius plant.     

Late season nitrogen fertilization of soybeans: effects on leaf senescence, yield and environment

Nitrogen demand from soybean seeds during seed filling is very high and has been proposed as the cause of nitrogen remobilization and leaf senescence. Previous research has not shown consistent effects of late season fertilization on seed yield, while its effects on leaf senescence have not been evaluated. Two field experiments were performed to determine the effects of a late season N fertilization on leaf senescence and fall, seed yield and its components, and residual soil nitrate, and to evaluate the potential risk of groundwater contamination. Two rates of nitrogen (50 and 100 kg N ha^–1) were applied at the R3 and R5 development stages. Nitrogen fertilization, either at R3 or R5, increased soil nitrate availability during the seed-filling period. Seed yield, seed number and protein content were not affected by N fertilization. The addition of 100 kg N ha^–1 produced a small delay of 1–2 days in the leaf fall, and slightly increased seed size (3.6%). Our results suggest that increasing soil N availability during the seed-filling period is not an effective way to delay leaf senescence or to increase seed growth and yield of soybean. Nitrogen fertilization increased the level of residual nitrate in the top soil at one site (the one with lowest seed yield), increasing the risk of nitrate leaching during subsequent fallow.     

Mutation of Leaf Senescence 1 Encoding a C2H2 Zinc Finger Protein Induces ROS Accumulation and Accelerates Leaf Senescence in Rice

Premature senescence of leaves causes a reduced yield and quality of rice by affecting plant growth and development. The regulatory mechanisms underlying early leaf senescence are still unclear. The Leaf senescence 1 (LS1) gene encodes a C2H2-type zinc finger protein that is localized to both the nucleus and cytoplasm. In this study, we constructed a rice mutant named leaf senescence 1 (ls1) with a premature leaf senescence phenotype using CRISPR/Cas9-mediated editing of the LS1 gene. The ls1 mutants exhibited premature leaf senescence and reduced chlorophyll content. The expression levels of LS1 were higher in mature or senescent leaves than that in young leaves. The contents of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were significantly increased and catalase (CAT) activity was remarkably reduced in the ls1 plants. Furthermore, a faster decrease in pigment content was detected in mutants than that in WT upon induction of complete darkness. TUNEL and staining experiments indicated severe DNA degradation and programmed cell death in the ls1 mutants, which suggested that excessive ROS may lead to leaf senescence and cell death in ls1 plants. Additionally, an RT-qPCR analysis revealed that most senescence-associated and ROS-scavenging genes were upregulated in the ls1 mutants compared with the WT. Collectively, our findings revealed that LS1 might regulate leaf development and function, and that disruption of LS1 function promotes ROS accumulation and accelerates leaf senescence and cell death in rice.     

Investigations on the growth ofRhodococcus rubra in relation to the formation of stable biological foams

The growth characteristics of a foam-forming species,Rhodococcus rubra were studied on different substrates. The basic medium contained Czapek (3.34%), yeast extract (0.2%), potassium dihydrogen phosphate (0.12%), dipotassium hydrogen phosphate (0.25%) and ammonium chloride (0.1%). This was supplemented with varying concentrations of glucose (0–2%). The same basic medium was also used to examine the growth ofR. rubra in combination with varying concentration of n-hexadecane (0.0–0.5%) as a source of energy while varying the concentration of ammonium chloride in the range 1–3 gl ⁻¹. Studies based on determining the biomass concentration, the surface activity related to the cell suspensions and measuring the variations in broth pH revealed that glucose encouraged the growth ofR. rubra, compared to the control. However, increasing the glucose concentration from 0.1 to 2.0% had no further effect on growth. The surface activity of the cell suspensions increased with increasing glucose concentration. Results similar to glucose were exhibited by the addition of n-hexadecane, suggesting same degree of growth among different concentrations with higher surface activity increasing with increase in substrate concentration. Results have also shown that the pH of all the culture broths decreased as the ammonium chloride concentration increased, suggesting that there was a production of hydrogen ions during the course of its metabolism.     

Relationship of Tarbush Leaf Surface Terpene Profile with Livestock Herbivory

Tarbush (Flourensia cernua DC.) is a Chihuahuan Desert shrub with a resinous leaf surface containing terpenes that may affect livestock herbivory. Cattle, sheep, and goats were densely stocked in paddocks containing tarbush in two consecutive years for six to nine days and defoliation of 160 plants was recorded daily. Plants were categorized as exhibiting high or low defoliation. Leaves were collected from these plants the third year for chemical analysis. A selection procedure was used to generate two variable sets closely related to defoliation category. One set contained 14 variables (dry matter, ash, -pinene, sabinene, 3-carene, p-cymene, limonene, camphor, borneol, cis-jasmone, -caryophyllene, -humulene, ledene, and flourensadiol) and the other set contained 14 unidentified compounds. When subjected to multivariate analysis, each group distinguished between the two defoliation categories (P < 0.001 and P < 0.0019 for known and unknown variable sets, respectively). These data support the hypothesis that leaf surface chemistry of individual tarbush plants is related to extent of defoliation by livestock.     

Phytotoxicity of <Emphasis Type="Italic">Phytolacca americana</Emphasis> Leaf Extracts on the Growth,and Physiological Response of <Emphasis Type="Italic">Cassia mimosoides</Emphasis>

We examined the allelochemical effects of control soil, native soil (treated soil), and leaf extracts of Phytolacca americana (pokeweed) on the germination rate and seedling growth of Cassia mimosoides var. nomame. We also studied the resulting changes in root-tip ultrastructure and peroxidase isozyme biochemistry. P. americana leaf extract inhibited seed germination, seedling growth, and biomass when compared to control and treated soil. Root and shoot growth in treated soil was stimulated relative to control soil, but root growth was inhibited by 50% in the leaf extract treatment. Biomass of C. mimosoides seedlings grown on leaf extract was reduced sevenfold when compared to the control seedlings. The amounts of total phenolic compounds in the leaf extract, treated soil, and control soil were 0.77, 0.14, and 0.03 mg l⁻¹, respectively. The root tips of C. mimsoides treated with leaf extracts of P. americana showed amyloplasts and large central vacuoles with electron-dense deposits inside them when compared to control root tips. The activity of guaiacol peroxidase (GuPOX) in whole plant, roots, and shoots of C. mimosoides increased as leaf extract increased; maximum activity was observed in extract concentrations of 75% and higher. Root GuPOX activity was three times higher than in shoots. Therefore, we conclude that inhibition of C. mimosoides growth is related to the phenolic compounds in the P. americana leaf extract and the ultrastructure changes in root-tip cells and increased GuPOX activity is a response to these allelochemicals.     

Leaf and root litter of a legume tree as nitrogen sources for cacaos with different root colonisation by arbuscular mycorrhizae

Traditionally cacao (Theobroma cacao L.) is cultivated under legume shade trees, which produce N-rich litter that improves soil organic matter content, microbial activity, and recycles N to the crop. Arbuscular mycorrhiza forming fungi (AMF) are known to play an important role in plant nutrient uptake, yet their role in plant N uptake from organic residues in tropical agroforestry systems is not clear. We studied root and leaf litter of the legume shade tree Inga edulis Mart. as a source of N for cacao and the importance of AMF colonisation in the uptake of litter N under controlled conditions. Leaf and root litter of I. edulis enriched with ¹⁵N was added to cacao pots filled with field soil. Half of the cacao saplings were AMF-inoculated and the soil of non-inoculated saplings was treated with fungicide to suppress AMF. During the 10-week experiment, young cacao leaves were sampled for ¹⁵N analyses and at the end of the experiment whole plants were harvested. Microbial populations in the soil were determined using phospholipid fatty acid (PLFA) analysis, and AMF structures in the roots were quantified. Fungicide treatment decreased AMF structures in roots and increased bacterial populations, but did not affect the decomposition rate of either litter type. Inoculated and non-inoculated cacao saplings used 2.6 and 2.1%, respectively, of N added to the pots in leaf litter and 12.1 and 7.1% of N available in root litter indicating that root litter of I. edulis may be a more efficient N source than leaf litter for cacao. Although the fungicide treatment did not completely suppress AMF in non-inoculated pots, it created sufficient contrast in root AMF colonisation for concluding that AMF significantly enhanced cacao N use from both litter types. The role of root litter of shade trees as a N source in agroforestry should not be neglected.     

LPS1, Encoding Iron-Sulfur Subunit SDH2-1 of Succinate Dehydrogenase,Affects Leaf Senescence and Grain Yield in Rice

The iron-sulfur subunit (SDH2) of succinate dehydrogenase plays a key role in electron transport in plant mitochondria. However, it is yet unknown whether SDH2 genes are involved in leaf senescence and yield formation. In this study, we isolated a late premature senescence mutant, lps1, in rice (Oryza sativa). The mutant leaves exhibited brown spots at late tillering stage and wilted at the late grain-filling stage and mature stage. In its premature senescence leaves, photosynthetic pigment contents and net photosynthetic rate were reduced; chloroplasts and mitochondria were degraded. Meanwhile, lps1 displayed small panicles, low seed-setting rate and dramatically reduced grain yield. Gene cloning and complementation analysis suggested that the causal gene for the mutant phenotype was OsSDH2-1 (LOC_Os08g02640), in which single nucleotide mutation resulted in an amino acid substitution in the encoded protein. OsSDH2-1 gene was expressed in all organs tested, with higher expression in leaves, root tips, ovary and anthers. OsSDH2-1 protein was targeted to mitochondria. Furthermore, reactive oxygen species (ROS), mainly H₂O₂, was excessively accumulated in leaves and young panicles of lps1, which could cause premature leaf senescence and affect panicle development and pollen function. Taken together, OsSDH2-1 plays a crucial role in leaf senescence and yield formation in rice.     

Allelopathic properties ofPolygonella myriophylla

Polygonella myriophylla is a perennial shrub endemic to the Florida scrub. Striking bare zones surround maturePolygonella stands. Quantitative measurements of root distribution show that fewPolygonella roots extend into the bare zones, supporting the hypothesis that the bare zones result from chemical interference byPolygonella with the growth of other species. Bioassays of soils collected biweekly for one year from beneathPolygonella, the bare zone, and adjacent grassed areas confirm that the germination and growth of grasses is reduced inPolygonella soil and soil from the surrounding bare zone. Compared to adjacent grassed area soil, the average germination of bahiagrass (Paspalum notatum) was 71% inPolygonella soil and 81% in bare zone soil, and average shoot dry weight was 48% inPolygonella soil and 81% in bare zone soil. Seasonal variation in the inhibition of grass germination and growth was not pronounced.     

Transgenerationally Transmitted DNA Demethylation of a Spontaneous Epialleles Using CRISPR/dCas9-TET1cd Targeted Epigenetic Editing in Arabidopsis

CRISPR/dCas9 is an important DNA modification tool in which a disarmed Cas9 protein with no nuclease activity is fused with a specific DNA modifying enzyme. A previous study reported that overexpression of the TET1 catalytic domain (TET1cd) reduces genome-wide methylation in Arabidopsis. A spontaneous naturally occurring methylation region (NMR19-4) was identified in the promoter region of the PPH (Pheophytin Pheophorbide Hydrolase) gene, which encodes an enzyme that can degrade chlorophyll and accelerate leaf senescence. The methylation status of NMR19-4 is associated with PPH expression and leaf senescence in Arabidopsis natural accessions. In this study, we show that the CRISPR/dCas9-TET1cd system can be used to target the methylation of hypermethylated NMR19-4 region to reduce the level of methylation, thereby increasing the expression of PPH and accelerating leaf senescence. Furthermore, hybridization between transgenic demethylated plants and hypermethylated ecotypes showed that the demethylation status of edited NMR19-4, along with the enhanced PPH expression and accelerated leaf senescence, showed Mendelian inheritance in F1 and F2 progeny, indicating that spontaneous epialleles are stably transmitted trans-generationally after demethylation editing. Our results provide a rational approach for future editing of spontaneously mutated epialleles and provide insights into the epigenetic mechanisms that control plant leaf senescence.     

In vivo synthesis of nanomaterials in plants: location of silver nanoparticles and plant metabolism

Metallic nanoparticles (MeNPs) can be formed in living plants by reduction of the metal ions absorbed as soluble salts. It is very likely that plant metabolism has an important role in MeNP biosynthesis. The in vivo formation of silver nanoparticles (AgNPs) was observed in Brassica juncea, Festuca rubra and Medicago sativa. Plants were grown in Hoagland''s solution for 30 days and then exposed for 24 h to a solution of 1,000 ppm AgNO₃. In the leaf extracts of control plants, the concentrations of glucose, fructose, ascorbic acid, citric acid and total polyphenols were determined. Total Ag content in plant fractions was determined by inductively coupled plasma atomic emission spectroscopy. Despite the short exposure time, the Ag uptake and translocation to plant leaves was very high, reaching 6,156 and 2,459 mg kg⁻¹ in B. juncea and F. rubra, respectively. Ultrastructural analysis was performed by transmission electron microscopy (TEM), and AgNPs were detected by TEM X-ray microanalysis. TEM images of plant fractions showed the in vivo formation of AgNPs in the roots, stems and leaves of the plants. In the roots, AgNPs were present in the cortical parenchymal cells, on the cell wall of the xylem vessels and in regions corresponding to the pits. In leaf tissues, AgNPs of different sizes and shapes were located close to the cell wall, as well as in the cytoplasm and within chloroplasts. AgNPs were not observed in the phloem of the three plant species. This is the first report of AgNP synthesis in living plants of F. rubra. The contents of reducing sugars and antioxidant compounds, proposed as being involved in the biosynthesis of AgNPs, were quite different between the species, thus suggesting that it is unlikely that a single substance is responsible for this process.

MSC 2010

92 Biology and other natural sciences; 92Cxx Physiological, cellular and medical topics; 92C80 Plant biology