Laser weeding of common weed species.

The massive use of herbicides since the 1950s has resulted in increasing problems with herbicideresistant weeds and pollution of the environment, including food, feed, and water. These side effects have resulted in political pressures to reduce herbicide application. The European Commission aims to reduce the use and risk of chemicals and more hazardous pesticides in the EU. Therefore, new weed control methods are in demand. Laser weeding might be an alternative to replace or supplement herbicides and other weed control methods in an Integrated Weed Management (IPM) strategy. This work aimed to investigate how increasing laser energy affected common weeds when the apical meristem was exposed to irradiation at the early stages of development. A 50 W thulium-doped fibre laser with a diameter of 2 mm and a wavelength of 2 µm was used. The highest efficacy of laser irradiation was achieved when the grass weed (Alopecurus myosuroides) had one leaf and the dicot species were at the cotyledon stage. There was a large difference between the species' susceptibility to irradiation probably caused by differences in morphology and growth habit. At the 4-leaf stage, most of the species regrew after irradiation. Laser weeding may be a solution to replace or supplement other weed control methods in some crops, but in general the weeds must be irradiated when they are at the cotyledon to 2-leaf stage to avoid regrowth.

Laser weeding: opportunities and challenges for couch grass (Elymus repens (L.) Gould) control.

Laser weeding may contribute to less dependency on herbicides and soil tillage. Several research and commercial projects are underway to develop robots equipped with lasers to control weeds. Artificial intelligence can be used to locate and identify weed plants, and mirrors can be used to direct a laser beam towards the target to kill it with heat. Unlike chemical and mechanical weed control, laser weeding only exposes a tiny part of the field for treatment. Laser weeding leaves behind only ashes from the burned plants and does not disturb the soil. Therefore, it is an eco-friendly method to control weed seedlings. However, perennial weeds regrow from the belowground parts after the laser destroys the aerial shoots. Depletion of the belowground parts for resources might be possible if the laser continuously kills new shoots, but it may require many laser treatments. We studied how laser could be used to destroy the widespread and aggressive perennial weed Elymus repens after the rhizomes were cut into fragments. Plants were killed with even small dosages of laser energy and stopped regrowing. Generally, the highest efficacy was achieved when the plants from small rhizomes were treated at the 3-leaf stage.

Chenopodium quinoa, a New Host for Alternaria Section Alternata and Alternaria Section Infectoriae Causing Yellow Leaf Blotch Disease.

Quinoa (Chenopodium quinoa Willd.) is a native American crop mainly grown in the Andes of Bolivia and Peru. During the last decades, the cultivation of quinoa has expanded to more than 125 countries. Since then, several diseases of quinoa have been characterized. A leaf disease was observed on quinoa plants growing in an experimental plot in Eastern Denmark in 2018. The symptoms produced by the associated fungi consisted of small yellow blotches on the upper surface of leaves with a pale chlorotic halo surrounding the lesion. These studies used a combination of morphology, molecular diagnostics, and pathogenicity tests to identify two different Alternaria species belonging to Alternaria sections Infectoriae and Alternata as the causal agent of observed disease symptoms. To the best of our knowledge, this is the first report of Alternaria spp. as foliar pathogens of quinoa. Our findings indicate the need for additional studies to determine potential risks to quinoa production.

Utilization of the halophytic shrubs Atriplex nummularia Lindl and Rhagodia preissii Moq as crops in salt-affected semi-arid regions: How temperature, salinity, seed weight and size affect seed germination.

The perennial halophytic shrubs Atriplex nummularia and Rhagodia preissii are native to Australia and can be planted on saline land to produce sheep and cattle feed during the autumn. However, an impediment to the adoption of the species on saline land has been the challenges in achieving successful establishment by direct seeding due to a lack of knowledge of the optimal conditions for germination. Therefore, the optimal germination requirements in relation to temperature, salinity level and seed size was assessed for each species to ascertain the optimal conditions for successful establishment by direct seeding on saline land. Seeds of both species showed optimal germination temperature at 10°C. Atriplex nummularia seeds were more tolerant to temperatures above or below 10°C than R. preissii. The germination percentage of A. nummularia was unchanged at 0-200 mM NaCl2. The germination percentage of Rhagodia preissii declined when the NaCl2 content exceeded 50 mM. There was no correlation between seed size, germination and emergence for any of the species. Based on the study, we suggest that sowing operations are performed during the cold winter months in subtropical areas or autumn and spring in temperate areas, to improve the successful establishment of these shrubs by direct seeding.

Eco-friendly approach to improve traits of winter wheat by combining cold plasma treatments and carbonization of subtropical biomass waste.

This study aims to improve the quality and quantity of winter wheat by using the potential of combining the use of cold plasma and waste biorefinery products for improving wheat yield. Plasma was applied by a radio frequency (RF) plasma reactor operated with air for 180 s and 50 W. The waste biorefinery products, including pyroligneous acid, biochar, and azolla compost, were used as plant nutrition. The effects of cold plasma treatment and waste biorefinery products were determined by measuring plant photosynthesis, grain yield, and content of chlorophyll, carotenoids, anthocyanin, protein, and starch. The experiment was conducted during the cropping seasons 2016-18 in a randomized complete block design with four replications. The combination of cold plasma and pyroligneous acid increased the grain yield up to 40.0%. The photosynthesis rate was improved up to 39.3%, and total chlorophyll content up to 48.3% in both years. Seed plasma treatment combined with biochar application increased the starch content by 36.8%. Adding azolla compost increased the protein content by 35.4%. Using seed plasma treatment with biochar increased the microbial biomass carbon by 16.0%. The application of plasma and azolla compost increased the microbial biomass nitrogen by 29.0%.

Quinoa Phenotyping Methodologies: An International Consensus.

Quinoa is a crop originating in the Andes but grown more widely and with the genetic potential for significant further expansion. Due to the phenotypic plasticity of quinoa, varieties need to be assessed across years and multiple locations. To improve comparability among field trials across the globe and to facilitate collaborations, components of the trials need to be kept consistent, including the type and methods of data collected. Here, an internationally open-access framework for phenotyping a wide range of quinoa features is proposed to facilitate the systematic agronomic, physiological and genetic characterization of quinoa for crop adaptation and improvement. Mature plant phenotyping is a central aspect of this paper, including detailed descriptions and the provision of phenotyping cards to facilitate consistency in data collection. High-throughput methods for multi-temporal phenotyping based on remote sensing technologies are described. Tools for higher-throughput post-harvest phenotyping of seeds are presented. A guideline for approaching quinoa field trials including the collection of environmental data and designing layouts with statistical robustness is suggested. To move towards developing resources for quinoa in line with major cereal crops, a database was created. The Quinoa Germinate Platform will serve as a central repository of data for quinoa researchers globally.

A Review of Chenopodium quinoa (Willd.) Diseases-An Updated Perspective.

The journey of the Andean crop quinoa (Chenopodium quinoa Willd.) to unfamiliar environments and the combination of higher temperatures, sudden changes in weather, intense precipitation, and reduced water in the soil has increased the risk of observing new and emerging diseases associated with this crop. Several diseases of quinoa have been reported in the last decade. These include Ascochyta caulina, Cercospora cf. chenopodii, Colletotrichum nigrum, C. truncatum, and Pseudomonas syringae. The taxonomy of other diseases remains unclear or is characterized primarily at the genus level. Symptoms, microscopy, and pathogenicity, supported by molecular tools, constitute accurate plant disease diagnostics in the 21st century. Scientists and farmers will benefit from an update on the phytopathological research regarding a crop that has been neglected for many years. This review aims to compile the existing information and make accurate associations between specific symptoms and causal agents of disease. In addition, we place an emphasis on downy mildew and its phenotyping, as it continues to be the most economically important and studied disease affecting quinoa worldwide. The information herein will allow for the appropriate execution of breeding programs and control measures.

Genetic variation for tolerance to the downy mildew pathogen Peronospora variabilis in genetic resources of quinoa (Chenopodium quinoa).

BACKGROUND: Quinoa (Chenopodium quinoa Willd.) is an ancient grain crop that is tolerant to abiotic stress and has favorable nutritional properties. Downy mildew is the main disease of quinoa and is caused by infections of the biotrophic oomycete Peronospora variabilis Gaüm. Since the disease causes major yield losses, identifying sources of downy mildew tolerance in genetic resources and understanding its genetic basis are important goals in quinoa breeding. RESULTS: We infected 132 South American genotypes, three Danish cultivars and the weedy relative C. album with a single isolate of P. variabilis under greenhouse conditions and observed a large variation in disease traits like severity of infection, which ranged from 5 to 83%. Linear mixed models revealed a significant effect of genotypes on disease traits with high heritabilities (0.72 to 0.81). Factors like altitude at site of origin or seed saponin content did not correlate with mildew tolerance, but stomatal width was weakly correlated with severity of infection. Despite the strong genotypic effects on mildew tolerance, genome-wide association mapping with 88 genotypes failed to identify significant marker-trait associations indicating a polygenic architecture of mildew tolerance. CONCLUSIONS: The strong genetic effects on mildew tolerance allow to identify genetic resources, which are valuable sources of resistance in future quinoa breeding.

Long-Term Storage and Longevity of Orthodox Seeds: A Systematic Review.

As part of conservation of plant genetic resources, long-term storage of seeds is highly relevant for genebanks. Here we present a systematic review and a meta-analysis of studies on seed longevity focusing on half-life (P50) under different storage conditions. Six studies were selected for the meta-analysis; in addition, a high number of additional references were included in the discussion of the results. The results show that under ambient conditions, half-life is short, from 5 to 10 years, while under more optimal conditions, which for orthodox seeds is at low humidity and low temperature, half-life is more in the 40-60 years range, although with large interspecies variation. Under long-term genebank conditions, with seeds dried to equilibrium and thereafter kept at minus 18-20°C in waterproof bags or jars, half-life can be twice or three times as long. In general, many of the grain legume seeds, as well as corn, common oat, and common barley are long-lived, while cereal rye, onion, garden lettuce, pepper, and some of the forage grasses are more short-lived. Conditions during maturation and harvesting influence longevity, and proper maturation and gentle handling are known to be of importance. Seed longevity models have been developed to predict final germination based on initial viability, temperature, humidity, storage time, and species information. We compared predicted germination to results from the long-term experiments. The predicted values were higher or much higher than the observed values, which demonstrate that something in the seed handling in the genebanks have not been optimal. Long-term studies are now available with data at least up to 60 years of storage. Our review shows that the knowledge and methodology developed for the conservation of plant genetic resources should also work for wild species of orthodox seed nature.

Seed Germination after 30 Years Storage in Permafrost.

More than 30 years ago, the Nordic Gene Bank established a long-term experiment on seeds stored under permafrost conditions in an abandoned mine corridor in Svalbard, as a tool to monitor storage life under these conditions. The study included seeds from 16 Nordic agricultural and horticultural crops, each represented by two or three cultivars (altogether 38 accessions). All seeds were ultra-dried to 3%-5% moisture before being sealed in glass tubes. Germination tests were performed in accordance with the International Seed Testing Association (ISTA) protocols. At the initiation of the experiment, the samples showed good germination with the median value at 92%. The overall picture remained stable over the first twenty to twenty-five years. However, the variation became larger over time and at 30 years, the median value had dropped to 80%. At the lower end, with a high drop in germination, we found rye, wheat, and English ryegrass. At the upper end, we found Kentucky bluegrass and cucumber. The lowest germination was found in samples with the highest initial seed moisture levels. Pre-storage conditions are likely to be of major importance for longevity.

Determination of Glyphosate in Dried Wheat by 1H-NMR Spectroscopy.

A wheat field was sprayed with a dosage of 1.1 kg a.i./ha Roundup PowerMax 10 days before harvest. The 1H Nuclear Magnetic Resonance (NMR) spectroscopy was used for the detection and quantification of the glyphosate (GLYP) in dried wheat spikelets, leaves, and stems. The quantification was done by the integration of the CH2-P groups doublet at 3.00 ppm with good linearity. The GLYP content varied between different samples and parts of the plant. On average, the largest content of herbicide was found in leaves (20.0 mg/kg), followed by stems (6.4 mg/kg) and spikelets (6.3 mg/kg). Our study shows that the 1H-NMR spectroscopy can be a rapid and reliable tool for GLYP detection and quantification in the field studies.

Assessing herbicide symptoms by using a logarithmic field sprayer.

BACKGROUND: In field experiments, assessment of herbicide selectivity and efficacy rarely takes advantage of dose-response regressions. The objective is to demonstrate that logarithmic sprayers, which automatically make a logarithmic dilution of a herbicide rate, can extract biologically relevant parameters describing the efficacy of herbicides in crops, and compare localities and time of assessment. RESULTS: In a conventional and an organic field, canola, white mustard, and no crop plots were sprayed with diflufenican and beflubutamid. A mixed effect log-logistic dose-response regression, with autoregressive correlation structure, estimated ED50 and ED90 for visual and Excess Green Index symptoms at various days after treatment (DAT). For visual assessment, ED50 differed within no crop between locations for beflubutamid at 12 DAT and 26 DAT. For diflufenican, the ED50 was different within crops at the two fields at 12 DAT, but not at 26 DAT. The Excess Green Indices at ED50 were not different among herbicides, locations, and corps; ED90 differed for white mustard and canola for beflubutamid but not for diflufenican. CONCLUSION: Suitable nonlinear regression models are now available for fitting dose-response data from a logarithmic sprayer in field experiments. The derived parameters (e.g. ED50 ) can compare selectivity and efficacy at numerous cropping systems. © 2018 Society of Chemical Industry.

Survival of weed seeds and animal parasites as affected by anaerobic digestion at meso- and thermophilic conditions.

Anaerobic digestion of residual materials from animals and crops offers an opportunity to simultaneously produce bioenergy and plant fertilizers at single farms and in farm communities where input substrate materials and resulting digested residues are shared among member farms. A surplus benefit from this practice may be the suppressing of propagules from harmful biological pests like weeds and animal pathogens (e.g. parasites). In the present work, batch experiments were performed, where survival of seeds of seven species of weeds and non-embryonated eggs of the large roundworm of pigs, Ascaris suum, was assessed under conditions similar to biogas plants managed at meso- (37°C) and thermophilic (55°C) conditions. Cattle manure was used as digestion substrate and experimental units were sampled destructively over time. Regarding weed seeds, the effect of thermophilic conditions (55°C) was very clear as complete mortality, irrespective of weed species, was reached after less than 2 days. At mesophilic conditions, seeds of Avena fatua, Sinapsis arvensis, Solidago canadensis had completely lost germination ability, while Brassica napus, Fallopia convolvulus and Amzinckia micrantha still maintained low levels (~1%) of germination ability after 1 week. Chenopodium album was the only weed species which survived 1 week at substantial levels (7%) although after 11 d germination ability was totally lost. Similarly, at 55°C, no Ascaris eggs survived more than 3h of incubation. Incubation at 37°C did not affect egg survival during the first 48 h and it took up to 10 days before total elimination was reached. In general, anaerobic digestion in biogas plants seems an efficient way (thermophilic more efficient than mesophilic) to treat organic farm wastes in a way that suppresses animal parasites and weeds so that the digestates can be applied without risking spread of these pests.

Dopaminergic differentiation of human neural stem cells mediated by co-cultured rat striatal brain slices.

Properly committed neural stem cells constitute a promising source of cells for transplantation in Parkinson's disease, but a protocol for controlled dopaminergic differentiation is not yet available. To establish a setting for identification of secreted neural compounds promoting dopaminergic differentiation, we co-cultured cells from a human neural forebrain-derived stem cell line (hNS1) with rat striatal brain slices. In brief, coronal slices of neonatal rat striatum were cultured on semiporous membrane inserts placed in six-well trays overlying monolayers of hNS1 cells. After 12 days of co-culture, large numbers of tyrosine hydroxylase (TH)-immunoreactive, catecholaminergic cells could be found underneath individual striatal slices. Cell counting revealed that up to 25.3% (average 16.1%) of the total number of cells in these areas were TH-positive, contrasting a few TH-positive cells (<1%) in non-induced areas. The presence of dopamine in the conditioned culture medium was confirmed by HPLC analysis. Interestingly, not all striatal slice cultures induced TH-expression in underlying hNS1 cells. Common to TH-inductive cultures was, however, the presence of degenerating, necrotic areas, suggesting that factors released during striatal degeneration were responsible for the dopaminergic induction of the hNS1 cells. Ongoing experiments aim to identify such factors by comparing protein profiles of media conditioned by degenerating (necrotic) versus healthy striatal slice cultures.

MALDI MS peptide mapping performance by in-gel digestion on a probe with prestructured sample supports.

Matrix-assisted laser desorption/ionization (tandem) mass spectrometry (MALDI MS) is widely used in protein chemistry and proteomics research for the identification and characterization of proteins isolated by polyacrylamide gel electrophoresis. In an effort to minimize sample handling and increase sample throughput, we have developed a novel in-gel digestion protocol where sample preparation is performed directly on a MALDI probe with prestructured sample support. The protocol consists of few sample-handling steps and has minimal consumption of reagents, making the protocol sensitive, timesaving, and cost-efficient. Performance of the on-probe sample preparation protocol was demonstrated by analysis of a set of rat liver proteins obtained from a fluorescently stained (Cy3 and SyproRuby) two-dimensional polyacrylamide gel. The success rate of protein identification by on-probe tryptic digestion and MALDI peptide mass mapping was 89%. The on-probe in-gel digestion procedure provided superior sensitivity and peptide mass mapping performance as compared to our standard in-gel digestion protocol. The on-probe digestion technique resulted in significantly improved amino acid sequence coverage of proteins, mainly due to efficient recovery and detection of large (>1.5 kDa) hydrophobic peptides. These observations indicate that numerous tryptic peptides are lost when using the standard in-gel digestion methods and sample preparation techniques for MALDI MS. This study also demonstrates that the on-probe digestion protocol combined with MALDI tandem mass spectrometry provides a robust platform for proteomics research, including protein identification and determination of posttranslational modifications.