Suspected intoxication by Kikuyu grass (Cenchrus clandestinus) of dairy cattle in the Azores, Portugal.

CASE HISTORY: An outbreak of suspected Kikuyu grass (Cenchrus clandestinus) intoxication among dairy cattle occurred on the island of Terceira in the Azores (Portugal), in October 2022. The animals affected were non-lactating dairy cows and heifers from five small farms grazing (free or tethered) a Kikuyu-dominant pasture. Of the 29 animals exposed, 17 were affected, and eight (five heifers and three adult cows) died, resulting in a morbidity rate of 58%, a mortality rate of 28% and a case mortality rate of 47%. CLINICAL FINDINGS AND TREATMENT: The clinical signs were non-specific and inconsistent, and included dry faeces, some with dark red blood; apathy and prostration; abdominal dilatation; tachycardia; tachypnoea; pale or jaundiced mucous membranes; sham drinking; sialorrhoea; and moderate to severe dehydration. Symptomatic treatment was provided but was ineffective. Haematology and serum biochemistry revealed an acute inflammatory leukogram, increased alkaline phosphatase (ALP) activity, decreased gamma-glutamyl transferase (GGT) activity, and azotaemia.The most consistent necropsy findings were haemorrhages in the epicardium and endocardium, an enlarged liver with rounded edges, non-perforated abomasal ulcers, and haemorrhagic lesions in the small and large intestines. Histopathology indicated myocarditis, hepatitis, interstitial nephritis, enteritis and colitis.Several fungal species were isolated from grass samples taken from affected pastures including several Fusarium spp., the genus implicated in Kikuyu toxicosis. Immediate removal of the animals from the pasture with Kikuyu was the only measure that prevented new cases and resulted in the recovery of some of the less affected animals. DIAGNOSIS: The epidemiological features of this outbreak and the clinical signs and micro- and macroscopic lesions observed were highly suggestive of Kikuyu grass poisoning. CLINICAL RELEVANCE: Although the weather conditions varied from other published cases, the grazing conditions (almost exclusive Kikuyu grass) and the post-mortem findings were very similar to those described in the literature, particularly the haemorrhages in the epicardium and endocardium. Kikuyu grass is very invasive and presents many desirable characteristics as cattle feed. Thus, an increase in cases of intoxication may be expected. Practitioners and farmers in areas where Kikuyu grass is abundant should be aware of the potential risks of grazing cattle exclusively on these pastures. They should also be aware of the early and subtle signs of Kikuyu intoxication to allow for timely removal of the animals from pasture.

Screening of Secondary Metabolites Produced by Nigrospora sphaerica Associated with the Invasive Weed Cenchrus ciliaris Reveals Two New Structurally Related Compounds.

In the search for new alternative biocontrol strategies, phytopathogenic fungi could represent a new frontier for weed management. In this respect, as part of our ongoing work aiming at using fungal pathogens as an alternative to common herbicides, the foliar pathogen Nigrospora sphaerica has been evaluated to control buffelgrass (Cenchrus ciliaris). In particular, in this work, the isolation and structural elucidation of two new biosynthetically related metabolites, named nigrosphaeritriol (3-(hydroxymethyl)-2-methylpentane-1,4-diol) and nigrosphaerilactol (3-(1-hydroxyethyl)-4-methyltetrahydrofuran-2-ol), from the phytotoxic culture filtrate extract were described, along with the identification of several known metabolites. Moreover, the absolute stereochemistry of (3R,4S,5S)-nigrosphaerilactone, previously reported as (3S,4R,5R)-4-hydroxymethyl-3,5-dimethyldihydro-2-furanone, was determined for the first time by X-ray diffraction analysis. Considering their structural relationship, the determination of the absolute stereochemistry of nigrosphaerilactone allowed us to hypothesize the absolute stereochemistry of nigrosphaeritriol and nigrosphaerilactol.

Analysis of global Napier grass (Cenchrus purpureus) collections reveals high genetic diversity among genotypes with some redundancy between collections.

Genetic diversity amongst genotypes of several Napier grass collections was analyzed and compared with the diversity in a set of open pollinated progeny plants. A total of 114,881 SNP and 46,293 SilicoDArT genome-wide markers were generated on 574 Napier grass genotypes. Of these, 86% of the SNP and 66% of the SilicoDArT markers were mapped onto the fourteen chromosomes of the Napier grass genome. For genetic diversity analysis, a subset of highly polymorphic and informative SNP markers was filtered using genomic position information, a maximum of 10% missing values, a minimum minor allele frequency of 5%, and a maximum linkage-disequilibrium value of 0.5. Extensive genetic variation, with an average Nei's genetic distance value of 0.23, was identified in the material. The genotypes clustered into three major and eleven sub-clusters with high levels of genetic variation contained both within (54%) and between (46%) clusters. However, we found that there was low to moderate genetic differentiation among the collections and that some overlap and redundancy occurred between collections. The progeny plants were genetically diverse and divergent from the germplasm collections, with an average FST value of 0.08. We also reported QTL regions associated with forage biomass yield based on field phenotype data measured on a subset of the Napier grass collections. The findings of this study offer useful information for Napier grass breeding strategies, enhancement of genetic diversity, and provide a guide for the management and conservation of the collections.

Urban sewage sludge stabilization by alkalization-composting-vermicomposting process: Crop-livestock residue use.

Waste management practices are vital for human health and the environment in a world where natural resources stress is expected to increase with the growth of population. Our study aimed to evaluate the potential use of crop-livestock residue as a bulking agent associated with the ideal level of hydrated lime for the stabilization and sanitization of urban sewage sludge through the alkalization-composting process. Therefore, we determined the alkalization efficiency on the heavy metal concentration in urban sewage sludge, quantified the viable eggs of helminths in pure and alkalized sludge, and measured the rate of earthworms (Eisenia fetida) surviving in the vermicomposting process using different levels of alkalized urban sewage sludge associated with crop-livestock residue. Four sequential trials were carried out in a completely randomized design with three replicates. The lime alkalization reduced the levels of Ba, As, Pb, Cu, Cr, Mo, Ni, and Zn compared to the pure urban sewage sludge. Using 30% w/w of lime in the urban sewage sludge (SS-30) for composting process reduced the viable helminth eggs by 71, 72, and 69% for sugarcane bagasse (Saccharum officinarum; SB), fresh chopped Napier-grass (Pennisetum purpureum; NG), and bovine ruminal content (BR), respectively. The ideal level of hydrated lime for stabilization and sanitization of urban sewage sludge was found to be 30%, which was able to reduce the heavy metals. The residues have the potential as a bulking agent for the composting of urban sewage sludge when associated with alkalization. The lime alkalization decreases the total number of helminth eggs and the number of viable eggs. The possibility of starting a vermicomposting using the mixtures is promising, evidenced by the earthworm survival in composting urban sewage sludge mixed with crop-livestock residues after 45 days of composting. The earthworm survival is maintained by an association of at least 80% of the crop-livestock residues.

A near-complete genome assembly of the allotetrapolyploid Cenchrus fungigraminus (JUJUNCAO) provides insights into its evolution and C4 photosynthesis.

JUJUNCAO (Cenchrus fungigraminus; 2n = 4x = 28) is a Cenchrus grass with the highest biomass production among cultivated plants, and it can be used for mushroom cultivation, animal feed, and biofuel production. Here, we report a nearly complete genome assembly of JUJUNCAO and reveal that JUJUNCAO is an allopolyploid that originated ∼2.7 million years ago (mya). Its genome consists of two subgenomes, and subgenome A shares high collinear synteny with pearl millet. We also investigated the genome evolution of JUJUNCAO and suggest that the ancestral karyotype of Cenchrus split into the A and B ancestral karyotypes of JUJUNCAO. Comparative transcriptome and DNA methylome analyses revealed functional divergence of homeologous gene pairs between the two subgenomes, which was a further indication of asymmetric DNA methylation. The three types of centromeric repeat in the JUJUNCAO genome (CEN137, CEN148, and CEN156) may have evolved independently within each subgenome, with some introgressions of CEN156 from the B to the A subgenome. We investigated the photosynthetic characteristics of JUJUNCAO, revealing its typical C4 Kranz anatomy and high photosynthetic efficiency. NADP-ME and PEPCK appear to cooperate in the major C4 decarboxylation reaction of JUJUNCAO, which is different from other C4 photosynthetic subtypes and may contribute to its high photosynthetic efficiency and biomass yield. Taken together, our results provide insights into the highly efficient photosynthetic mechanism of JUJUNCAO and provide a valuable reference genome for future genetic and evolutionary studies, as well as genetic improvement of Cenchrus grasses.

Insights into the Ecotoxicology of Radicinin and (10S,11S)-(-)-epi-Pyriculol, Fungal Metabolites with Potential Application for Buffelgrass (Cenchrus ciliaris) Biocontrol.

Buffelgrass (Cenchrus ciliaris L.) is an invasive C4 perennial grass species that substantially reduces native plant diversity of the Sonoran Desert through fire promotion and resource competition. Broad-spectrum herbicides are essentially used for its control, but they have a negative environmental and ecological impact. Recently, phytotoxicity on C. ciliaris has been discovered for two metabolites produced in vitro by the phytopathogenic fungi Cochliobolus australiensis and Pyricularia grisea. They were identified as (10S,11S)-(-)-epi-pyriculol and radicinin and resulted in being potential candidates for the development of bioherbicides for buffelgrass biocontrol. They have already shown promising results, but their ecotoxicological profiles and degradability have been poorly investigated. In this study, ecotoxicological tests against representative organisms from aquatic ecosystems (Aliivibrio fischeri bacterium, Raphidocelis subcapitata alga, and Daphnia magna crustacean) revealed relatively low toxicity for these compounds, supporting further studies for their practical application. The stability of these metabolites in International Organization for Standardization (ISO) 8692:2012 culture medium under different temperatures and light conditions was also evaluated, revealing that 98.90% of radicinin degraded after 3 days in sunlight. Significant degradation percentages (59.51-73.82%) were also obtained at room temperature, 30 °C or under ultraviolet (254 nm) light exposure. On the other hand, (10S,11S)-epi-pyriculol showed more stability under all the aforementioned conditions (49.26-65.32%). The sunlight treatment was also shown to be most effective for the degradation of this metabolite. These results suggest that radicinin could provide rapid degradability when used in agrochemical formulations, whereas (10S,11S)-epi-pyriculol stands as a notably more stable compound.

Root microbiome diversity and structure of the Sonoran desert buffelgrass (Pennisetum ciliare L.).

Buffelgrass (Pennisetum ciliare) is an invasive plant introduced into Mexico's Sonoran desert for cattle grazing and has converted large areas of native thorn scrub. One of the invasion mechanisms buffelgrass uses to invade is allelopathy, which consists of the production and secretion of allelochemicals that exert adverse effects on other plants' growth. The plant microbiome also plays a vital role in establishing invasive plants and host growth and development. However, little is known about the buffelgrass root-associated bacteria and the effects of allelochemicals on the microbiome. We used 16S rRNA gene amplicon sequencing to obtain the microbiome of buffelgrass and compare it between samples treated with root exacknudates and aqueous leachates as allelochemical exposure and samples without allelopathic exposure in two different periods. The Shannon diversity values were between H' = 5.1811-5.5709, with 2,164 reported bacterial Amplicon Sequence Variants (ASVs). A total of 24 phyla were found in the buffelgrass microbiome, predominantly Actinobacteria, Proteobacteria, and Acidobacteria. At the genus level, 30 different genera comprised the buffelgrass core microbiome. Our results show that buffelgrass recruits microorganisms capable of thriving under allelochemical conditions and may be able to metabolize them (e.g., Planctomicrobium, Aurantimonas, and Tellurimicrobium). We also found that the community composition of the microbiome changes depending on the developmental state of buffelgrass (p = 0.0366; ANOSIM). These findings provide new insights into the role of the microbiome in the establishment of invasive plant species and offer potential targets for developing strategies to control buffelgrass invasion.

(±)-3-Deoxyradicinin Induces Stomata Opening and Chloroplast Oxidative Stress in Tomato (Solanum lycopersicum L.).

Radicinin is a phytotoxic dihydropyranopyran-4,5-dione isolated from the culture filtrates of Cochliobolus australiensis, a phytopathogenic fungus of the invasive weed buffelgrass (Cenchrus ciliaris). Radicinin proved to have interesting potential as a natural herbicide. Being interested in elucidating the mechanism of action and considering radicinin is produced in small quantities by C. australiensis, we opted to use (±)-3-deoxyradicinin, a synthetic analogue of radicinin that is available in larger quantities and shows radicinin-like phytotoxic activities. To obtain information about subcellular targets and mechanism(s) of action of the toxin, the study was carried out by using tomato (Solanum lycopersicum L.), which, apart from its economic relevance, has become a model plant species for physiological and molecular studies. Results of biochemical assays showed that (±)-3-deoxyradicinin administration to leaves induced chlorosis, ion leakage, hydrogen peroxide production, and membrane lipid peroxidation. Remarkably, the compound determined the uncontrolled opening of stomata, which, in turn, resulted in plant wilting. Confocal microscopy analysis of protoplasts treated with (±)-3-deoxyradicinin ascertained that the toxin targeted chloroplasts, eliciting an overproduction of reactive singlet oxygen species. This oxidative stress status was related by qRT-PCR experiments to the activation of transcription of genes of a chloroplast-specific pathway of programmed cell death.

Anti-inflammatory, anti-nociceptive and anti-pyretic activities of Cenchrus ciliaris L.

ETHNOPHARMACOLOGICAL RELEVANCE: Cenchrus ciliaris L. belongs to the family Poaceae and is found all over the world. It is native to the Cholistan desert of Pakistan where it is locally known as 'Dhaman'. Owing to high nutritional value, C. ciliaris is used as fodder while seeds are used for bread making which are consumed by locals. It also possesses medicinal value and is extensively employed to treat pain, inflammation, urinary tract infection, and tumors. AIM OF STUDY: Studies on the pharmacological activities of C. ciliaris are scarce in spite of its several traditional uses. To the best of our knowledge, no comprehensive study has been conducted on anti-inflammatory, analgesic and anti-pyretic activity of C. ciliaris until now. Here we employed an integrative phytochemical and in - vivo framework to evaluate the potential biological activities of C. ciliaris against inflammation, nociception and pyrexia experimentally induced in rodents. MATERIAL AND METHODS: C. ciliaris was collected from the desert of Cholistan, Bahawalpur, Pakistan. Phytochemical profiling of C. ciliaris was done by employing GC-MS analysis. Anti-inflammatory activity of plant extract was initially determined by various in - vitro assays including albumin denaturation assay and RBC membrane stabilization assays. Finally, rodents were utilized to evaluate in - vivo anti-inflammatory, antipyretic and anti-nociceptive activities. RESULTS: Our data revealed the presence of 67 phytochemicals in methanolic extract of C. ciliaris. The methanolic extract of C. ciliaris provided RBC membrane stabilization by 65.89 ± 0.32% and protection against albumin denaturation by 71.91 ± 3.42% at 1 mg/ml concentration. In in - vivo acute inflammatory models, C. ciliaris exhibited 70.33 ± 1.03, 62.09 ± 8.98, 70.24 ± 0.95% anti-inflammatory activity at concentration of 300 mg/ml against carrageenan, histamine and serotonin induced inflammation. In CFA induced arthritis, inhibition of inflammation was found to be 48.85 ± 5.11% at 300 mg/ml dose after 28 days of treatment. In anti-nociceptive assays C. ciliaris exhibited significant analgesic activity in both peripheral and centrally mediated pain. The C. ciliaris also reduced the temperature by 75.26 ± 1.41% in yeast induced pyrexia. CONCLUSION: C. ciliaris exhibited anti-inflammatory effect against acute and chronic inflammation. It also showed significant anti-nociceptive and anti-pyretic activity which endorses its traditional use in the management of pain and inflammatory disorders.

Assessment of pure, mixed and diluted deep eutectic solvents on Napier grass (Cenchrus purpureus): Compositional and characterization studies of cellulose, hemicellulose and lignin.

Pretreatment with pure, mixed, and diluted deep eutectic solvents (DESs) was evaluated for its effect on Napier grass through compositional and characterization studies. The morphological changes of biomass caused by pretreatment were analyzed by FTIR and XRD. The cellulose and hemicellulose content after pretreatment using mixed DES increased and decreased 1.29- and 4.25-fold, respectively, when compared to untreated Napier grass. The crystallinity index (CrI. %) of mixed DES sample increased due to the maximum removal of hemicellulose (76 %) and delignification of 62 %. The material costs of ChCl/FA and ChCl/LA for a single run are ≈2.16 USD and ≈1.65 USD, respectively. Pure DES showed that ChCl/LA pretreatment enhanced delignification efficiency and that ChCl/FA increased hemicellulose removal. It was estimated that a single run using ChCl/LA:ChCl/FA to achieve maximum hemicellulose and lignin removal would cost approximately ≈1.89 USD. Future work will evaluate the effect of DES mixture on enzyme digestibility and ethanol production from Napier grass. HYPOTHESES: Deep eutectic solvent (DES) pretreatment studies on the fractionation of lignocellulosic biomass have grown exponentially. The use of pure and diluted DES has been reported to improve saccharification efficiency, delignification, and cellulose retention (Gundupalli et al., 2022). These studies have reported maximum lignin removal but also a lower effect on hemicellulose removal from lignocellulosic biomass. It was hypothesized that mixing two pure DESs could result in maximum removal of hemicellulose and lignin after pretreatment. To our knowledge, no studies have been performed to investigate the efficiency of pretreatment using a DES mixture and compared the outcome with pure and diluted DESs. Furthermore, it was hypothesized that using two pure DESs in a mixed form could lower the material cost for each experimental run. Process efficiency was determined by compositional, XRD, and FTIR analysis. Avenues for future research include determining glucose and ethanol yields during the enzymatic saccharification and fermentation processes.

Phytotoxic Interference of Culture Filtrates of Endophytic Bacteria Associated with Nerium oleander Leaf Against Seed Germination of the Invasive Noxious Weed Cenchrus echinatus.

Weeds cause destructive agricultural losses, so weed control is an urgent challenge facing agriculture. The extensive use of synthetic chemical herbicides has detrimental environmental impacts and promotes the emergence of resistant species. Therefore, in this study we tried to find a new natural weed control that can ensure biosafety and eco-sustainability. The phytotoxic potential of culture filtrates of the endophytes Bacillus inaquosorum NL1 and Bacillus safensis NL2 isolated from Nerium oleander leaf against the invasive harmful weed species Cenchrus echinatus was evaluated. Culture filtrates of both bacterial species exhibited potent phytotoxic activity, which resulted in 100% germination inhibition of C. echinatus. The chemical analysis of culture filtrates revealed high contents of total phenolics and n-alkanes that have phytotoxic effects against seed germination. According to the findings of this study the endophytic bacteria associated with N. oleander leaf can be used in the future to develop a sustainable bio-herbicide formulation.

Effects of exogenous Strigolactone on the physiological and ecological characteristics of Pennisetum purpureum Schum. Seedlings under drought stress.

BACKGROUND: Drought is one of the main environmental factors limiting plant growth and development. Pennisetum purpureum Schum. was used to explore the mitigation effects of exogenous strigolactone (SL) on drought stress during the seedling stage. The effects of different concentrations (1, 3, 5, and 7 µmol·L- 1) of SL on the photosynthesis characteristics, growth performance, and endogenous abscisic acid (ABA) of P. purpureum under drought stress were studied. RESULTS: Exogenous SL could effectively alleviate the inhibitory effect of drought stress on P. purpureum growth. Compared with drought stress, the net photosynthesis rate, stomatal conductance, transpiration rate, and water-use efficiency of the leaves of P. purpureum after SL treatment significantly increased, thereby exerting a significant mitigation effect on the decrease in photosystem II maximum photochemical efficiency and the performance index based on light absorption caused by drought. Moreover, the exogenous application of SL can effectively increase the fresh and dry weight of the leaves and roots and the main-root length. After applying SL for 120 h, the ABA content of P. purpureum decreased significantly. The activity of key enzymes of photosynthesis significantly increased after 48 h of external application of SL to P. purpureum. CONCLUSIONS: SL treatment can improve the photosynthesis performance of P. purpureum leaves under drought conditions and increase the antioxidant capacity of the leaves, thereby reducing the adverse effects of drought, promoting the growth of P. purpureum, and effectively improving the drought resistance of P. purpureum.

Effect of drought stress or soil pH on cesium accumulation in Napier grass.

Radio-cesium (Cs) decontamination efficiency by plants is supposedly affected by environmental conditions. To elucidate the factors influencing the unstable 137Cs-decontamination efficiency, we first examined the influence of drought or soil pH on Cs accumulation using cesium-133 (133Cs) using Napier grass (Pennisetum purpureum Schum.) grown under pot conditions. The experiment was performed on three different conditions with 150 µM 133Cs applied to soil: low pH (L-pH, pH = 5.6 ± 0.0), near-neutral pH (N-pH, pH = 6.6 ± 0.1), and the combination of low pH and drought stress (Drought). Drought stress had prominent negative effects on plant height, aboveground dry matter weight (DWabove), 133Cs concentrations in aboveground or root (Csabove or Csroot) parts, or 133Cs total content in the part aboveground (Cs-T). 133Cs concentration and total content in Drought conditions were reduced by 44.1% and 60.0% aboveground and 63.6% and 66.0% in root parts compared with counterpart normal soil moisture conditions (L-pH), respectively. Conversely, there were no significant effects of soil pH on Csabove, Csroot, or Cs-T in L-pH and N-pH conditions. Chlorophyll fluorescence parameters (Fv/Fm, Fv/F0) and the temperature in leaves were negatively affected by drought stress or soil pH conditions. From our results, drought strongly influenced plant growth and Cs accumulation in plants compared with soil acidity. Therefore, soil moisture appears to be a significant factor in maintaining 137Cs-decontamination efficiency by Napier grass.

Cloncurry buffel grass mitigated Cr(III) and Cr(VI) toxicity in tomato plant.

Contamination of agricultural soil with chromium (Cr) ions has threatened global crop, human and ecosystem health. Its two oxidation states viz. Cr(III) and Cr(VI) are most stable and readily available to the plants. The study explored the impact of increasing exposure (up to 500 ppm) of Cr(III) and Cr(VI) on bio-physical traits of 15-day-old seedlings (in vitro) as well as 60-day-old tomato plant (in vivo), and highlighted the importance of buffel grass (Cenchrus pennisetiformis) in mitigating Cr levels in the tomato plants. In vitro, Petri plate bioassays with 13 different concentrations (20-500 ppm) of Cr(III) and Cr(VI) depicted the highly toxic effect of metal ions ≥ 200 ppm on all bio-physical traits of tomato seedlings. In vivo, soil spiked with Cr(III) and Cr(VI) (200, 300, and 400 mg/kg) was amended with 1% and 2% dry biomass of buffel grass. Phytotoxicity was higher in Cr(VI)-spiked soil compared with Cr(III)-spiked soil. Cr was mainly accumulated in tomato roots, and more Cr was translocated from roots to shoots from Cr(VI)-spiked soil than Cr(III)-spiked soil. Soil amendments with 2% weed biomass reduced metal toxicity in plants, particularly at 200 and 300 mg/kg of Cr. Protein profiles through SDS-PAGE revealed 12-50 kDa (mainly PR proteins) as an important region in tomato leaf, where many new bands were expressed under different treatments, particularly in the treatments provided with buffel grass. PCA-based biplot clearly separated Cr tolerance treatments from highly sensitive treatments. For the cultivation of tomato plants in Cr(III) and Cr(VI) contaminated soil (200 and 300 mg/kg), the biomass of Cloncurry buffel grass should be considered an effective and easily available phyto-management option.

Climate change and land-use change impacts on future availability of forage grass species for Ethiopian dairy systems.

Forage grasses are central feed resources for livestock globally. In Ethiopian dairy systems, they serve as feed sources during both wet and dry seasons, yet escalating climate change could threaten forage supply. Here, we investigate projected climate change impacts on three forage grasses currently recommended for Ethiopian dairy systems. We determine areas of geographical suitability for each species using three climate projections generated by General Circulation Models (GCMs) and calculate their ability to meet predicted dry matter demand under four scenarios for livestock intensification and land availability. By 2050, Buffel grass (Cenchrus ciliaris) is likely to be negatively affected by climate change in regions such as Tigray, while Rhodes grass (Chloris gayana) and Napier grass (Cenchrus purpureus) may have improved suitability under future climates. Our findings suggest that feed demands could theoretically be met by production of these forage grasses under current and future climates. However, if land availability is reduced and herd composition shifts towards higher-productivity exotic breeds, forage resources will not meet cattle demand even with improved agronomic management.

In Vitro Effects of Fungal Phytotoxins on Cancer Cell Viability: First Insight into Structure Activity Relationship of a Potent Metabolite of Cochliobolus australiensis Radicinin.

Natural compounds have always represented an important source for new drugs. Although fungi represent one such viable source, to date, no fungal metabolite has been marketed as an anticancer drug. Based on our work with phytotoxins as potential chemical scaffolds and our recent findings involving three phytopathogenic fungi, i.e., Cochliobolus australiensis, Kalmusia variispora and Hymenoscyphus fraxineus, herein, we evaluate the in vitro anti-cancer activity of the metabolites of these fungi by MTT assays on three cancer cell models harboring various resistance levels to chemotherapeutic drugs. Radicinin, a phytotoxic dihydropyranopyran-4,5-dione produced by Cochliobolus australiensis, with great potential for the biocontrol of the invasive weed buffelgrass (Cenchrus ciliaris), showed significant anticancer activity in the micromolar range. Furthermore, a SAR study was carried out using radicinin, some natural analogues and hemisynthetic derivatives prepared by synthetic methods developed as part of work aimed at the potential application of these molecules as bioherbicides. This investigation opens new avenues for the design and synthesis of novel radicinin analogues as potential anticancer agents.

Determining the pre-grazing sward height of Kikuyu grass (Cenchrus clandestinus - Hochst. ex Chiov.) for optimizing nutrient intake rate of dairy heifers.

Understanding the grazing process and animal response to sward structures (e.g., sward height) is key to setting targets for efficient grazing management. We hypothesized that the short-term intake rate (STIR) of dry matter (DM) and digestible organic matter (OM) by dairy heifers is maximized with Kikuyu grass (Cenchrus clandestinus-Hochst. ex Chiov.) of intermediate sward heights. The treatments consisted of five pre-grazing sward heights (10, 15, 20, 25, and 30 cm) randomly assigned to two of ten paddocks. The experimental design included two measurements of each paddock at different periods and times of day. Three Holstein heifers (440 ± 42 kg body weight) were used to determine the STIR, which was estimated using the double-weighing technique with correction for insensible weight losses. The bite mass (BM), bite rate (BR), sward structural characteristics, and nutritional value of herbage samples were assessed. The data were analyzed using mixed models with a factorial arrangement of five sward heights, two times of day, and two evaluation periods. The sward height of Kikuyu grass that maximized both STIRs was approximately 20 cm. The STIR of the DM was 30% and 15% lower than the maximum in the shortest and tallest swards tested, respectively. In swards shorter than 20 cm, the STIR was lower because the BM decreased with sward height, whereas in those greater than 20 cm, the lower BM and STIR of DM was explained by a decrease in bulk density and bite volume. The top stratum was composed mainly of highly digestible leaf blades with similar nutrient content across sward heights; therefore the STIR of digestible OM was also maximized at 20 cm. Hence, the optimal pre-grazing sward height of Kikuyu grass should be managed at 20 cm under rotational stocking systems to maximize nutrient intake rate of dairy heifers.

Chromosome-scale genome assembly provides insights into speciation of allotetraploid and massive biomass accumulation of elephant grass (Pennisetum purpureum Schum.).

Elephant grass (Pennisetum purpureum Schum) is an important forage, biofuels and industrial plant widely distributed in tropical and subtropical areas globally. It is characterized with robust growth and high biomass. We sequenced its allopolyploid genome and assembled 2.07 Gb into A' and B subgenomes of 14 chromosomes with scaffold N50 of 8.47 Mb, yielding a total of 77,139 genes. The allotetraploid speciation occurred approximately 15 Ma after the divergence between Setaria italica and Pennisetum glaucum, according to a phylogenetic analysis of Pennisetum species. Double whole-genome duplication (WGD) and polyploidization events resulted in large-scale gene expansion, especially in the key steps of growth and biomass accumulation. Integrated transcriptome profiling revealed the functional divergence between subgenomes A' and B. A' subgenome mainly contributed to plant growth, development and photosynthesis, whereas the B subgenome was primarily responsible for effective transportation and resistance to stimulation. Some key gene families related to cellulose biosynthesis were expanded and highly expressed in stems, which could explain the high cellulose content in elephant grass. Our findings provide deep insights into genetic evolution of elephant grass and will aid future biological research and breeding, even for other grasses in the family Poaceae.

Habitat thermal quality for Gopherus evgoodei in tropical deciduous forest and consequences of habitat modification by buffelgrass.

Tortoises of the genus Gopherus evolved in North America and have survived major environmental challenges in the past 40 million years. However, this genus now faces multiple anthropogenic threats, such as the introduction of invasive plant species. Buffelgrass (Cenchrus ciliaris) is considered one of the greatest threats to arid and tropical ecosystems, where gopher tortoises inhabit, because the grass displaces native flora and fauna. Modification of the environment as a result of this invasive plant portends an alteration of the available thermal landscape. The aim of this paper is twofold: 1) to evaluate the thermal quality of the primary habitat of Gopherus evgoodei (tropical deciduous forest [TDF], and 2) determine the potential thermal changes due to habitat modification by buffelgrass. First, we obtained data on body temperature of active tortoises in semi-captivity. Second, we measured the operative environmental temperature during 5 years at three sites south of Sonora, Mexico that support G. evgoodei: a) a pristine TDF (Conserved-TDF); b) a forest patch surrounded by introduced buffelgrass pasture (Partial-TDF); and c) an introduced buffelgrass pasture area (Buffel-Pasture). Our results demonstrate that the intact microhabitats within the TDF provide G. evgoodei with high thermal quality at both spatial and temporal scales. However modified habitat by buffelgrass had higher operative temperatures for G. evgoodei than TDF. The thermal quality of the sites disturbed with buffelgrass can exceed the thermal requirements of G. evgoodei by up to 25 °C. Finally, we discussed potential collateral effects of habitat modification by invasion by buffelgrass.

Sandbur Drought Tolerance Reflects Phenotypic Plasticity Based on the Accumulation of Sugars, Lipids, and Flavonoid Intermediates and the Scavenging of Reactive Oxygen Species in the Root.

The perennial grass Cenchrus spinifex (common sandbur) is an invasive species that grows in arid and semi-arid regions due to its remarkable phenotypic plasticity, which confers the ability to withstand drought and other forms of abiotic stress. Exploring the molecular mechanisms of drought tolerance in common sandbur could lead to the development of new strategies for the protection of natural and agricultural environments from this weed. To determine the molecular basis of drought tolerance in C. spinifex, we used isobaric tags for relative and absolute quantitation (iTRAQ) to identify proteins differing in abundance between roots growing in normal soil and roots subjected to moderate or severe drought stress. The analysis of these proteins revealed that drought tolerance in C. spinifex primarily reflects the modulation of core physiological activities such as protein synthesis, transport and energy utilization as well as the accumulation of flavonoid intermediates and the scavenging of reactive oxygen species. Accordingly, plants subjected to drought stress accumulated sucrose, fatty acids, and ascorbate, shifted their redox potential (as determined by the NADH/NAD ratio), accumulated flavonoid intermediates at the expense of anthocyanins and lignin, and produced less actin, indicating fundamental reorganization of the cytoskeleton. Our results show that C. spinifex responds to drought stress by coordinating multiple metabolic pathways along with other adaptations. It is likely that the underlying metabolic plasticity of this species plays a key role in its invasive success, particularly in semi-arid and arid environments.