Grass Tissues Research Articles

The effect of dead standing (marcescent) biomass on litter decomposition in herbaceous flora is governed by plant functional group

Abstract In autumn, temperate herbs begin to senesce and gradually shed their litter. However, surprisingly large amounts of dead biomass remain standing, that is, marcescent. The consequences of marcescence for the decomposition of biomass once it finally reaches the soil are largely unknown. Here, we aimed to determine whether marcescence affects subsequent litter decomposition in the organic layer to such an extent that its mass loss and chemistry are distinguishable from those of directly shed biomass. We further aimed to disentangle the role of plant functional traits and groups (forbs vs. grasses) concerning the marcescence effect on decomposition. To this end, we sampled the living, marcescent and shed senescent biomass of 39 herbaceous plant species grown in a common garden experiment, determined plant functional traits and incubated the marcescent and shed plant tissues in the field in an allochthonous organic layer for 6 months. We determined the mass loss, C and N contents, chemical composition and microbial community structure of the decomposed tissues. Our results show that marcescent tissues decomposed more slowly than directly shed tissues (mass loss 37.3% vs. 63.2% for forbs, 43.5% vs. 45.5% for grasses), likely due to more favourable conditions for decomposition in the organic layer. These were reflected in a significantly higher microbial colonization of shed (~333 and 708 μg biomass C g−1 for forbs and grasses, respectively) than marcescent tissue (~189 and 543 μg biomass C g−1 for forbs and grasses, respectively) even after 6 months in the organic layer. Moreover, higher relative contributions of aliphatics and polyphenolics in shed tissues indicated a more advanced stage of decomposition. Notably, marcescent tissues of forbs, with a more complex growth architecture (being composed of stems [marcescent] and leaves [shed]), decomposed substantially more slowly than directly shed tissues. In contrast, differences in decomposition between marcescent and shed tissues of grasses, with a more uniform growth architecture, were substantially less pronounced. These findings highlight that marcescence in the temperate herbaceous flora can strongly affect litter decomposition and thus C and nutrient cycling through temperate ecosystems, but that the extent to which marcescence affects decomposition depends on plant functional group.

The Teleost CXCL13-CXCR5 Axis Induces Inflammatory Cytokine Expression through the Akt-NF-κB, p38-AP-1, and p38-NF-κB Pathways.

The ancestors of chemokines originate in the most primitive of vertebrates, which has recently attracted great interest in the immune functions and the underlying mechanisms of fish chemokines. In the current study, we identified an evolutionarily conserved chemokine, CiCXCL13, from a teleost fish, grass carp. CiCXCL13 was characterized by a typical SCY (small cytokine CXC) domain and four cysteine residues (C34, C36, C61, C77), with the first two cysteines separated by a random amino acid residue, although it shared 24.2-54.8% identity with the counterparts from other vertebrates. CiCXCL13 was an inducible chemokine, whose expression was significantly upregulated in the immune tissues of grass carps after grass carp reovirus infection. CiCXCL13 could bind to the membrane of grass carp head kidney leukocytes and promote cell migration, NO release, and the expression of >15 inflammatory cytokines, including IL-1β, TNF-α, IL-10 and TGF-β1, thus regulating the inflammatory response. Mechanistically, CiCXCL13 interacted with its evolutionarily conserved receptor CiCXCR5 and activated the Akt-NF-κB and p38-AP-1 pathways, as well as a previously unrevealed p38-NF-κB pathway, to efficiently induce inflammatory cytokine expression, which was distinct from that reported in mammals. Zebrafish CXCL13 induced inflammatory cytokine expression through Akt, p38, NF-κB, and AP-1 as CiCXCL13. Meanwhile, the CiCXCL13-CiCXCR5 axis-mediated inflammatory activity was negatively shaped by grass carp atypical chemokine receptor 2 (CiACKR2). The present study is, to our knowledge, the first to comprehensively define the immune function of CXCL13 in inflammatory regulation and the underlying mechanism in teleosts, and it provides a valuable perspective on the evolution and biology of fish chemokines.

Morphological analysis of <i>Polemonium Coeruleum</i> L. herb using stereo- and luminescent microscopy

Aim to establish the morphological features of the ground part of Polemonium coeruleum L.
 Material and methods. The material of the study was the samples of Greek-valerian polemonium grass (Polemonium coeruleum L.), cultivated and harvested in the Voronezh region, the city of Bobrov, during the flowering period in the second year of life in 2021. The State Pharmacopoeia XIV standard ed. OFS.1.5.3.0003.15 "Technique for microscopic and microchemical examination of medicinal plant materials and medicinal plant preparations" was used for macroscopic analysis.
 Results. When studying the Greek-valerian polemonium herb, its external features were described, which can be used for "Authenticity" section when developing regulatory documentation for this type of plant material. The difference in the pubescence of Greek-valerian polemonium leaves depending on their topography was established. The presence of simple multicellular trichomes on all parts of the plant, as well as glandular trichomes on some elements of the flower, was revealed. For the first time, the nature of the autoluminescence of tissues of the Greek-valerian polemonium grass was described.

Spatial expression patterns of genes encoding sugar sensors in leaves of C4 and C3 grasses.

The mechanisms of sugar sensing in grasses remain elusive, especially those using C4 photosynthesis even though a large proportion of the world's agricultural crops utilize this pathway. We addressed this gap by comparing the expression of genes encoding components of sugar sensors in C3 and C4 grasses, with a focus on source tissues of C4 grasses. Given C4 plants evolved into a two-cell carbon fixation system, it was hypothesized this may have also changed how sugars were sensed. For six C3 and eight C4 grasses, putative sugar sensor genes were identified for target of rapamycin (TOR), SNF1-related kinase 1 (SnRK1), hexokinase (HXK) and those involved in the metabolism of the sugar sensing metabolite trehalose-6-phosphate (T6P) using publicly available RNA deep sequencing data. For several of these grasses, expression was compared in three ways: source (leaf) versus sink (seed), along the gradient of the leaf, and bundle sheath versus mesophyll cells. No positive selection of codons associated with the evolution of C4 photosynthesis was identified in sugar sensor proteins here. Expressions of genes encoding sugar sensors were relatively ubiquitous between source and sink tissues as well as along the leaf gradient of both C4 and C3 grasses. Across C4 grasses, SnRK1β1 and TPS1 were preferentially expressed in the mesophyll and bundle sheath cells, respectively. Species-specific differences of gene expression between the two cell types were also apparent. This comprehensive transcriptomic study provides an initial foundation for elucidating sugar-sensing genes within major C4 and C3 crops. This study provides some evidence that C4 and C3 grasses do not differ in how sugars are sensed. While sugar sensor gene expression has a degree of stability along the leaf, there are some contrasts between the mesophyll and bundle sheath cells.

Fibulorhizoctonia psychrophila, causal agent of lenticel spot in pome fruit, rarely detected in Dutch pear and apple orchards

Fibulorhizoctonia psychrophila causes lenticel spot in stored pome fruit. A Dutch survey revealed that damage can be observed regularly in individual lots at low incidence to occasionally incidences above 25% after storage for five months or longer under specific controlled atmosphere storage conditions. The source of inoculum of the psychrophilic pathogen is not known. A species-specific TaqMan PCR assay was developed to quantify the pathogen DNA in orchard litter considered as potential inoculum source. 2008 samples of various litter types such as dead leaves, fallen fruits and mummies and necrotic tissues of grasses and weeds, collected in 2009–2012 in ten apple and twenty pear orchards, were analyzed. Although other fruit rot pathogens such as Neofabraea alba, Cadophora luteo-olivacea and Stemphylium vesicarium were detected in the samples at high incidences in earlier studies, F. psychrophila was only detected in three samples. The results show that F. psychrophila is not ubiquitously present in Dutch orchards at traceable concentrations. The main sources of the disease remain unknown. Possibly, the pathogen is present on storage bins or dust in storage facilities.

Water stress protection by the arbuscular mycorrhizal fungus Rhizoglomus irregulare involves physiological and hormonal responses in an organ‐specific manner

Arbuscular mycorrhizal fungi may alleviate water stress in plants. Although several protection mechanisms have already been described, little information is available on how these fungi influence the hormonal response to water stress at an organ-specific level. In this study, we evaluated the physiological and hormonal responses to water stress in above and below-ground tissues of the legume grass Trifolium repens colonized by the arbuscular mycorrhizal fungus Rhizoglomus irregulare. Plants were subjected to progressive water stress and recovery. Different leaf and root physiological parameters, as well as phytohormone levels, were quantified. Water-stressed mycorrhizal plants showed an improved water status and no photoinhibition compared to uncolonized individuals, while some stress markers like α-tocopherol and malondialdehyde content, an indicator of the extent of lipid peroxidation, transiently increased in roots, but not in leaves. Water stress protection exerted by mycorrhiza appeared to be related to a differential root-to-shoot redox signaling, probably mediated by jasmonates, and mycorrhization enhanced the production of the cytokinin trans-zeatin in both roots and leaves. Overall, our results suggest that mycorrhization affects physiological, redox and hormonal responses to water stress at an organ-specific level, which may eventually modulate the final protection of the host from water stress.

Evaluation of colonization and mutualistic endophytic symbiosis of Escherichia coli with tomato and Bermuda grass seedlings.

Escherichia coli is generally considered a bacterium associated with animal microbiomes. However, we present evidence that E. coli may also mutualistically colonize roots of plant species, even to the extent that it may become endophytic in plants. In this study we used GFP tagged E. coli to observe its colonization and effects on tomato (Solanum lycopersicum) and Bermuda grass (Cynodon dactylon) seedling development and growth. Inoculation with the bacterium significantly improved root development of both seedlings tested. Treatment also increased the photosynthetic pigments in Bermuda grass seedlings. However, effects on shoot length in both seedlings were not significant. This bacterium was found to produce indole acetic acid (IAA) up to 8.68 ± 0.43 µg ml−1 in the broth medium amended with tryptophan. Effects on seedling root growth could, in part, be explained by IAA production. Bacteria successfully colonized the root surfaces and interiors of both seedlings. Tagged bacteria expressing the GFP were observed in the vascular tissues of Bermuda grass seedling roots. Seedlings with bacteria showed greater survival and were healthier than seedlings without bacteria, indicating that E. coli set up a successful mutualistic symbiosis with seedlings. E. coli is not commonly considered to be a plant endophyte but is more generally considered to be a crop contaminant. In this study we show that E. coli may also be an endophyte in plant tissues.

Differentiating grazing-induced rangeland grass nutrition status in situ using near infrared reflectance

Assessing the nutrient content of rangeland grass using earth observation technologies presents opportunities for non-destructive, rapid and repetitive appraisals in support of rangeland management. The biochemical-related concentrations of macronutrients (N, P, K, Ca, Mg) are among the factors that influence near infrared reflectance (ρNIR) by a turgid leaf. High grazing intensity (GI) enhances nutrient recycling in rangelands, resulting in high concentrations in aboveground grass tissue. This study sought to establish whether grazing-induced macronutrient abundance in rangeland grass can be remotely sensed in situ through ρNIR in the multiple species natural settings. Sampling was conducted at peak grass growth stage in large savannah rangelands with similar grass species mixtures, in low moisture stress conditions. The sampling was widely distributed in GI-contrasted livestock and wild grazer rangelands, and in grazing-restricted control sites. Two species representing high and low nutrient grass species, respectively, were sampled. At each sampling site (N = 116), reflectance was measured in the 350–2500 nm range from leaf and stem tissue of healthy-looking specimens of the two species (≤10 m apart), using a 1.1 nm – 1.4 nm resolution spectroradiometer. Macronutrient concentrations (MNC) in the tissue were then determined in the laboratory. For each species, the hyperspectral NIR (λ700 – 1100 nm) reflectance was averaged per sampling site. Individual and total MNC, respectively, were correlated with the hyperspectral ρNIR, and then with atmospheric effect-corrected ρNIR from 10 m-resolution, field sampling-concurrent Sentinel-2 MSI multispectral images. For each sampling site, the two species’ hyperspectral ρNIR values, individual and total MNC, respectively, were then averaged. Respective models relating the paired hyperspectral ρNIR and averaged MNC were developed, by linear regression and partial least squares regression (PLSR). Tissue MNC were lowest in the control sites and higher in the grazed rangelands. Covariance in MNC in the two species suggested location-context similarities in all grasses. Individual and total MNC, respectively, had statistically significant (p < 0.01) negative correlations with both tissue level hyperspectral and multispectral ρNIR. The negative relationship was detected on empirical line-retrieved ρNIR on historical Hyperion hyperspectral images. PLSR yielded a more accurate predictive model (R2 = 0.760) than linear regression (R2 = 0.601, F = 138.415, p < 0.0001). The study shows that there are possibilities to remotely sense grazing-induced MNC in aboveground grass tissue through changes in ρNIR.

Different response of perennial ryegrass\u2014Epichlo\xeb endophyte symbiota to the elevated concentration of heavy metals in soil

The phenomenon of plant mutualistic symbiosis with microbes may have a positive effect on the improvement of plant tolerance to environmental stresses as well as on the ability of plants to accumulate heavy metal (HM) ions from soil. The influence of Epichloë fungal endophyte (Ascomycota, Clavicipitaceae) on perennial ryegrass (Lolium perenne L.) plants grown in the presence of elevated concentrations of HM ions (Cd2+, Pb2+, and Cu2+) in soil was studied. The presence of Epichloë in the host grass tissues resulted in different accumulation of HM ions in the aboveground parts of the plants. In some cases, endophyte infection positively affected ryegrass ability to accumulate HM ions from soil. In plants with (E +) and without (E −) endophytes, the hormesis effect was induced by the elevated concentration of Cu2+ ions, resulting in better growth and photosynthesis, as examined by measurements of Chl a fluorescence. The obtained results indicate that based on the laboratory evaluation of the efficiency of HM accumulation, we were able to choose the best associations of perennial ryegrass with endophytes for HM phytoremediation.

Gene analysis reveals that leaf litter from Epichloë endophyte-infected perennial ryegrass alters diversity and abundance of soil microbes involved in nitrification and denitrification

Although Epichloë endophytes are present only in aboveground tissues of grasses they indirectly influence soil biological characteristics through increased litter incorporation and root exudation. Epichloë endophytes have been reported to affect the decomposition rates of litter by altering litter quality and microbial decomposers to affect soil characteristics. However, it is not well-studied that underlying effects on functional genes involved in nitrification and denitrification after incorporation of litter containing Epichloë endophytes into soil. We collected soil samples at S0 (first litter incorporation), S1 (second litter incorporation), S2 (third litter incorporation) and S3 (120 days after the third litter incorporation) times after incorporation of litter that was either infected with the fungal endophyte Epichloë festucae var. Lolii or free of Epichloë endophyte, respectively. . We sampled the soil fororganic carbon (OC), total nitrogen (TN), total phosphorus (TP), ammonium nitrogen (NH4+-N: AN), nitrate nitrogen (NO3−-N: NN), soil microbial biomass carbon (MBC), nitrogen (MBN), ammonia-oxidizing bacteria (AOB-amoA) and ammonia-oxidizing archaea (AOA-amoA) functional genes for nitrification, nitrite reduction (nirS and nirK) and nitrous oxide reduction (nosZ) functional genes for denitrification. We found that Epichloë-infected plant increased the soil properties (OC, TN, AN, NN) and microbial biomass C in S1 to S3 times, and microbial biomass N in S2 and S3 times, but reduced soil pH in S1 times and the C_N ratio in S2 and S3 times by litter incorporation. The absolute abundance of the AOB-amoA functional gene at S1 and S3 times and relative abundances of Planctomycetes, Methylobacterium, Nitosomonas, Gemmata, Geodermatophilus and Sinorhizobium in the nirK functional gene community were significantly higher in soil with Epichloë-infected litter compared to soil with Epichloë-free litter at S1 and S3 times. While the absolute abundance at S1 and S3 times and diversity at S3 times of the nirK functional gene, and the absolute abundance at S1 to S3 times and diversity at S1 times of the nosZ functional gene, and relative abundance of Proteobacteria in the AOB-amoA functional gene were significantly lower in soil with litter containing Epichloë endophyte than that of soil with litter without the endophyte. The soil moisture, and the contents of TP, OC, AN and MBN were the best explanatory variables for the altered microbial community. Our results provide a new understanding of the responses of soil nitrification and denitrification genes through incorporation of litter containing Epichloë endophytes into soil.

Root decomposition in silvopastures is influenced by grazing, fertility, and grass species

AbstractGrass root production and decomposition is a major source of C entering soils, although rates are largely unknown based on edaphic and management factors. Therefore, study objectives were to evaluate four explanatory variables including forage species (native and nonnative), fertility (poultry litter and a control), soil moisture (udic and aquic), and pasture management (grazed and an ungrazed control) in order to evaluate driving factors for root turnover and subsequent soil organic matter formation in silvopastoral systems using the root litter bag technique. Native grass root decomposition was accelerated relative to the nonnative forage based on root mass balance, as well as the exponential decay function, likely owing to greater five‐ and six‐C sugars and more digestible root tissues of native grasses. These physiochemical results suggest more favorable microbial food sources, which culminate in faster decomposition and greater microbially derived organic matter. Overall, there was greater root sloughing and subsequent soil organic matter formation potential with native grass species and poultry litter applications, with soil moisture affecting decomposition to a lesser extent. This study contributes to the understanding of complex interactions of grass species, soil moisture, nutrients, and grazing, which controls primary productivity, as well as nutrient cycling and C sequestration in silvopastures.

Improved analysis of arabinoxylan-bound hydroxycinnamate conjugates in grass cell walls

BackgroundArabinoxylan in grass cell walls is acylated to varying extents by ferulate and p-coumarate at the 5-hydroxy position of arabinosyl residues branching off the xylan backbone. Some of these hydroxycinnamate units may then become involved in cell wall radical coupling reactions, resulting in ether and other linkages amongst themselves or to monolignols or oligolignols, thereby crosslinking arabinoxylan chains with each other and/or with lignin polymers. This crosslinking is assumed to increase the strength of the cell wall, and impedes the utilization of grass biomass in natural and industrial processes. A method for quantifying the degree of acylation in various grass tissues is, therefore, essential. We sought to reduce the incidence of hydroxycinnamate ester hydrolysis in our recently introduced method by utilizing more anhydrous conditions.ResultsThe improved methanolysis method minimizes the undesirable ester-cleavage of arabinose from ferulate and p-coumarate esters, and from diferulate dehydrodimers, and produces more methanolysis vs. hydrolysis of xylan-arabinosides, improving the yields of the desired feruloylated and p-coumaroylated methyl arabinosides and their diferulate analogs. Free ferulate and p-coumarate produced by ester-cleavage were reduced by 78% and 68%, respectively, and 21% and 39% more feruloyl and p-coumaroyl methyl arabinosides were detected in the more anhydrous method. The new protocol resulted in an estimated 56% less combined diferulate isomers in which only one acylated arabinosyl unit remained, and 170% more combined diferulate isomers conjugated to two arabinosyl units.ConclusionsOverall, the new protocol for mild acidolysis of grass cell walls is both recovering more ferulate- and p-coumarate-arabinose conjugates from the arabinoxylan and cleaving less of them down to free ferulic acid, p-coumaric acid, and dehydrodiferulates with just one arabinosyl ester. This cleaner method, especially when coupled with the orthogonal method for measuring monolignol hydroxycinnamate conjugates that have been incorporated into lignin, provides an enhanced tool to measure the extent of crosslinking in grass arabinoxylan chains, assisting in identification of useful grasses for biomass applications.

Nutrient concentrations in two savannah grass species and their implications for grazers

The diversity of grazers on savannahs exerts pressure on the grass to provide nutrition. In this study the concentrations of major and trace animal nutrients in above ground grass tissue as influenced by grazing were examined. Two palatable grass species grazed by both livestock and wild grazers were sampled: Brachiaria nigropedata and Eragrostis lehmanniana. Samples of grass tissue were collected from widely spread sampling sites in both livestock and wildlife grazing sites, at the end of the growing season. Although B. nigropedata had higher nutrient concentrations, there was general covariance in nutrient levels in the two grasses. The covariance indicated that the respective sites generally had location context-dependent low or high nutrient concentrations in all grasses. The adequacy of the nutrient concentrations for grazer nutrition was assessed using literature-sourced minimum and maximum tolerable limit requirements of cattle and the high population gazelle-like species as illustration. The results showed that, in these nitrogen-limited savannahs, the grass generally met grazer requirements in terms of crude protein, potassium, molybdenum, nickel and zinc but that it had deficiencies in phosphorous, sodium, and boron, as well as grazing intensity-related localised toxic levels of potassium, phosphorous, magnesium, iron and manganese. The deficiencies and toxicities were inferred to result in abnormalities in grazer physical condition and reproduction. Given the importance of livestock in semiarid savannahs, these effects on livestock grazers were inferred as threatening human food security. The study shows that high grazing intensity can raise the concentrations of some animal nutrients in above ground grass tissue to levels above grazer tolerable limits. Grazing rotation to reduce grazing intensity is recommended as a proactive grazing management strategy to limit the effects.

Tissue-specific Transcriptome analysis reveals lignocellulose synthesis regulation in elephant grass (Pennisetum purpureum Schum)

BackgroundThe characteristics of elephant grass, especially its stem lignocellulose, are of great significance for its quality as feed or other industrial raw materials. However, the research on lignocellulose biosynthesis pathway and key genes is limited because the genome of elephant grass has not been deciphered.ResultsIn this study, RNA sequencing (RNA-seq) combined with lignocellulose content analysis and cell wall morphology observation using elephant grass stems from different development stages as materials were applied to reveal the genes that regulate the synthesis of cellulose and lignin. A total of 3852 differentially expressed genes (DEGs) were identified in three periods of T1, T2, and T3 through RNA-seq analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of all DEGs showed that the two most abundant metabolic pathways were phenylpropane metabolism, starch and sucrose metabolism, which were closely related to cell wall development, hemicellulose, lignin and cellulose synthesis. Through weighted gene co-expression network analysis (WGCNA) of DEGs, a ‘blue’ module highly associated with cellulose synthesis and a ‘turquoise’ module highly correlated with lignin synthesis were exhibited. A total of 43 candidate genes were screened, of which 17 had function annotations in other species. Besides, by analyzing the content of lignocellulose in the stem tissues of elephant grass at different developmental stages and the expression levels of genes such as CesA, PAL, CAD, C4H, COMT, CCoAMT, F5H and CCR, it was found that the content of lignocellulose was related to the expression level of these structural genes.ConclusionsThis study provides a basis for further understanding the molecular mechanisms of cellulose and lignin synthesis pathways of elephant grass, and offers a unique and extensive list of candidate genes for future specialized functional studies which may promote the development of high-quality elephant grass varieties with high cellulose and low lignin content.

A fungal endophyte of an annual weed reduces host competitive ability and confers associational protection to wheat

Epichloid fungal endophytes (Epichloë spp., Ascomycota: Clavicipitaceae) inhabit aerial tissues of several cool-season grasses, and enhance host growth and defence against herbivores. The presence of these symbionts can also affect interactions between the host and other non-epichloid plants. The role of an epichloid endophyte on interspecific competition has been tested using perennial grasses with contrasting results, but it has been scarcely tested using annual species in agroecosystems. We evaluated the impact of Epichloë-grass symbiosis on the competitive interaction between a non-host cereal crop (Triticum aestivum, wheat) and a host weed (Lolium multiflorum, ryegrass), growing in the presence of invertebrate herbivores (aphids) under no resource limitation. We conducted an outdoor mesocosm experiment with wheat plants growing in monoculture or in mixture with low or high proportions of ryegrass plants. Ryegrass plants presented either low (E-) or high (E+) incidence of Epichloë occultans (i.e. frequency of epichloid endophytic plants). We measured wheat vegetative and reproductive yield and its natural aphid infestation. Although epichloid endophyte incidence did not affect ryegrass biomass, wheat reproductive yield in mixtures (relative to wheat monocultures) was 45% higher when grown with E+ ryegrass plants than E- conspecific plants. Aphids preferred wheat plants grown with E- plants rather than wheat plants grown with E+ plants, but only in mixtures with high proportion of ryegrass. Our results demonstrate that epichloid endophyte incidence can decrease host competitive ability and confers associational protection to the non-endophytic neighbouring plants. Thus, ryegrass-endophyte symbiosis can increase crop yield by positive neighbourhood effects through different mechanisms probably related to the density of the weed. The benefits of this endosymbiont cannot be considered host-exclusive since they can be disseminated to non-endophytic plants. Furthermore, our results suggest that the epichloid endophyte incidence on annual weeds can contribute to agroecosystem sustainability by influencing pest management and increasing crop yield.

Citizen science campaign reveals widespread fallout of contaminated dust from mining activities in the central Peruvian Andes

Abstract Inadequate management of mine tailings at Cerro de Pasco, one of Peru’s largest mining complexes, has resulted in elevated concentrations of Pb, As, Cu, Zn, and Ag in surface soil horizons across the Junín Plain, central Peru. During June 2016, in response to local concern over mine contamination, teams of local citizens armed with sample bags, plastic trowels, and GPS receivers acquired 385 surface soil samples and 9 plant samples from agricultural lands from an area ∼1000 km2 on the Junín Plain. Metal concentrations were determined by acid digestion and inductively coupled plasma–mass spectrometry, and results revealed elevated levels of Pb, As, Cu, Zn, and Ag in all samples within a 10 km radius of the center of mining activities, and measurable contamination at least 30 km to the south-southwest, in the direction of prevailing winds. Dust traps emplaced for a 12 month period confirmed that contamination is ongoing. High metal concentrations in grasses growing on contaminated soils revealed that a portion of the total metal contamination is removed from the soil and held in grass tissue, where it can be ingested by graminivores, especially llama, alpaca, and sheep, thereby entering the human food supply.

Evaluation of the content and bioaccessibility of selected metals from barley grass

The evaluation of the content of zinc and copper in plant tissues of barley grass growing on enriched in zinc/copper solutions (sulphate, glycine complex, rutin complex) were studied by inductively coupled plasma mass spectrometry. The highest concentrations of Zn were found in the roots and leaves, growing on solutions enriched in the rutin-zinc(II) and sulphate solutions. The highest concentrations of Cu were found in the leaves, growing on solutions enriched in the glycine-copper(II) complex solutions. The research showed that the barley accumulates the zinc/copper from the nutrient solution, therefore, in the next step of the investigation the evaluation of bioaccessibility was carried out. The bioaccessibility of copper and zinc were evaluated from the various enriched cultivation solutions by the inductively coupled mass spectrometry coupled with size exclusion chromatography. Plants growing on solutions enriched in the zinc/copper complexes with amino acids (glycine) and flavonoids (rutin) accumulated excessive amounts of these elements in tissues. Additionally, the bioaccessibility is significantly higher than from the plant’s tissues growing on zinc/copper salts solutions.

Accumulation of germanium (Ge) in plant tissues of grasses is not solely driven by its incorporation in phytoliths

Ge/Si ratios of plant phytoliths have been widely used to trace biogeochemical cycling of Si. However, until recently, information on how much of the Ge and Si transferred from soil to plants is actually stored in phytoliths was lacking. The aim of the present study is to (i) compare the uptake of Si and Ge in three grass species, (ii) localize Ge and Si stored in above-ground plant parts and (iii) evaluate the amounts of Ge and Si sequestrated in phytoliths and plant tissues. Mays (Zea mays), oat (Avena sativa) and reed canary grass (Phalaris arundinacea) were cultivated in the greenhouse on soil and sand to control element supply. Leaf phytoliths were extracted by dry ashing. Total elemental composition of leaves, phytoliths, stems and roots were measured by ICP-MS. For the localization of phytoliths and the determination of Ge and Si within leaf tissues and phytoliths scanning electron microscopy (SEM), energy dispersive x-ray spectroscopy (EDX) and laser ablation inductively coupled mass spectrometry (LA-ICP-MS) was used. The amounts of Si and Ge taken up by the species corresponded with biomass formation and decreased in the order Z. mays > P. arundinacea, A. sativa. Results from LA-ICP-MS revealed that Si was mostly localized in phytoliths, while Ge was disorderly distributed within the leaf tissue. In fact, from the total amounts of Ge accumulated in leaves only 10% was present in phytoliths highlighting the role of organic matter on biogeochemical cycling of Ge and the necessity for using bulk Ge/Si instead of Ge/Si in phytoliths to trace biogeochemical cycling of Si.

Linking a rapid throughput plate-assay with high-sensitivity stable-isotope label LCMS quantification permits the identification and characterisation of low \u03b2-L-ODAP grass pea lines

BackgroundGrass pea (Lathyrus sativus) is an underutilised crop with high tolerance to drought and flooding stress and potential for maintaining food and nutritional security in the face of climate change. The presence of the neurotoxin β-L-oxalyl-2,3-diaminopropionic acid (β-L-ODAP) in tissues of the plant has limited its adoption as a staple crop. To assist in the detection of material with very low neurotoxin toxin levels, we have developed two novel methods to assay ODAP. The first, a version of a widely used spectrophotometric assay, modified for increased throughput, permits rapid screening of large populations of germplasm for low toxin lines and the second is a novel, mass spectrometric procedure to detect very small quantities of ODAP for research purposes and characterisation of new varieties.ResultsA plate assay, based on an established spectrophotometric method enabling high-throughput ODAP measurements, is described. In addition, we describe a novel liquid chromatography mass spectrometry (LCMS)-based method for β-L-ODAP-quantification. This method utilises an internal standard (di-13C-labelled β-L-ODAP) allowing accurate quantification of β-L-ODAP in grass pea tissue samples. The synthesis of this standard is also described. The two methods are compared; the spectrophotometric assay lacked sensitivity and detected ODAP-like absorbance in chickpea and pea whereas the LCMS method did not detect any β-L-ODAP in these species. The LCMS method was also used to quantify β-L-ODAP accurately in different tissues of grass pea.ConclusionsThe plate-based spectrophotometric assay allows quantification of total ODAP in large numbers of samples, but its low sensitivity and inability to differentiate α- and β-L-ODAP limit its usefulness for accurate quantification in low-ODAP samples. Coupled to the use of a stable isotope internal standard with LCMS that allows accurate quantification of β-L-ODAP in grass pea samples with high sensitivity, these methods permit the identification and characterisation of grass pea lines with a very low ODAP content. The LCMS method is offered as a new ‘gold standard’ for β-L-ODAP quantification, especially for the validation of existing and novel low- and/or zero-β-L-ODAP genotypes.

A Brief History of Endophyte Detection Techniques in Grasses

Endophytes are the plant mutualists that live asymptomatically inside plant tissue and are found in nearly whole plant kingdom. Endophytic fungi receive shelter and nutrition from host plants and in return provide great advantages to the host. Grasses are a useful forage species and are of great agricultural and socio-economic value. The presence of endophytes in these grasses provide protection, persistence and improved yield against herbivores, insects, pathogens, drought and several other biotic and abiotic stresses. This review summarizes traditional and modern molecular techniques to identify endophytes from turf and forage grasses. Traditional approaches include direct observation, staining, laser micro dissection and pressure catapulting and cultivation-dependent methods that provide a morphological identification of endophytic mycobiota in grass tissues. Earlier studies on endophytes using these methods resulted in several technical implications which molecular approaches are able to solve now-a-days. Molecular approaches include DNA extraction, PCR based DNA Fingerprinting techniques, Denaturing Gradient Gel Electrophoresis, Sanger sequencing, Pyrosequencing, Immunoblot assay, Biosensors, DNA Barcoding and Molecular Phylogenetics etc. A comparison of these detection techniques will facilitate other researchers as well to develop new ways for the detection of endophytes that will contribute to the improvement of grassland in future.