N supply in stockless organic cereal production under northern temperate conditions. Undersown legumes, or whole-season green manure?

Two systems for nitrogen (N) supply to organic spring cereals were compared under Norwegian conditions. Repeated undersowing of clover in the cereals in four growing seasons was compared to a whole-season green manure in the second year. Cereal yields were higher in the treatments with clover than in the controls. The yield increasing effect of undersown clover was residual. One year of whole-season green manure increased subsequent cereal yields significantly, but not enough to compensate for the loss of yield over the total four-year period. If phytopathological problems can be avoided, repeated undersowing of legumes seems to be more profitable than green manure each fourth year in stockless organic cereal production systems. The soil mineral N decreased during the study, demonstrating a negative N balance. Hence, additional N sources should be found for stockless organic cereal systems under Norwegian conditions

Repeated use of green-manure catch crops in organic cereal production - grain yields and nitrogen supply

By restricted access to manure, nitrogen (N) supply in organic agriculture relies on biological N-fixation. This study compares grain yields after one full-season green manure (FSGM) to yields with repeated use of a green-manure catch crop. At two sites in south-eastern Norway, in a simple 4-year rotation (oats/wheat/oats/wheat), the repeated use of ryegrass, clover, or a mixture of ryegrass and clover as catch crops was compared with an FSGM established as a catch crop in year 1. The FSGM treatments had no subsequent catch crops. In year 5, the final residual effects were measured in barley. The yield levels were about equal for grains with no catch crop and a ryegrass catch crop. On average, the green-manure catch crops increased subsequent cereal yields close to 30%. The FSGM increased subsequent cereal yields significantly in two years, but across the rotation the yields were comparable to those of the treatments without green-manure catch crop. To achieve acceptable yields under Norwegian conditions, more than 25% of the land should be used for full-season green manure, or this method combined with green-manure catch crops. The accumulated amount of N in aboveground biomass in late autumn did not compensate for the N removed by cereal yields. To account for the deficiency, the roots of the green-manure catch crops would have to contain about 60% of the total N (tot-N) required to balance the cereal yields. Such high average values for root N are likely not realistic to achieve. However, measurement of biomass in late autumn may not reflect all N made available to concurrent or subsequent main crop

Quality of fish sludge as fertiliser to spring cereals: Nitrogen effects and environmental pollutants

The aim of this study was to contribute to development of organic fertiliser products based on fish sludge (i.e. feed residues and faeces) from farmed smolt. Four dried fish sludge products, one liquid digestate after anaerobic digestion and one dried digestate were collected at Norwegian smolt hatcheries in 2019 and 2020. Their quality as fertilisers was studied by chemical analyses, two 2-year field experiments with spring cereals and soil incubation combined with a first-order kinetics N release model. Cadmium (Cd) and zinc (Zn) concentrations were below European Union maximum limits for organic fertilisers in all products except one (liquid digestate). Relevant organic pollutants (PCB7, PBDE7, PCDD/F + DL-PCB) were analysed for the first time and detected in all fish sludge products. Nutrient composition was unbalanced, with low nitrogen/phosphorus (N/P) ratio and low potassium (K) content relative to crop requirements. Nitrogen concentration in the dried fish sludge products varied (27–70 g N kg-1 dry matter), even when treated by the same technology but sampled at different locations and/or times. In the dried fish sludge products, N was mainly present as recalcitrant organic N, resulting in lower grain yield than with mineral N fertiliser. Digestate showed equally good N fertilisation effect as mineral N fertiliser, but drying reduced N quality. Soil incubation in combination with modelling is a relatively cheap tool that can give a good indication of N quality in fish sludge products with unknown fertilisation effects. Carbon/N ratio in dried fish sludge can also be used as an indicator of N quality.publishedVersio

Direktesåing i mye planterester

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Tang som gjødsel

Denne rapporten er en sammenstilling av ulike utprøvinger av tang som gjødsel, utført av Norsk senter for økologisk landbruk (NORSØK). Disse ble gjennomført som en del av et større prosjekt med tittelen «Alternative gjødselkilder i økologisk drift med lite eller ikke noe husdyrgjødsel». Dette prosjektet utførte NORSØK i perioden 1998–2000. Tangforsøket ble avsluttet i 2001. Sommeren 2000 ble det gjennomført en studietur rettet mot bruk av alternative næringskilder i økologisk landbruk og bruk av tang i landbruket. Erfaringene fra studieturen er oppsummert i en egen rapport(Holm & McKinnon 2000). De delene av rapporten som berører bruk av tang og tare er gjengitt i denne rapporten

Delt gjødsling til vårraps

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Vårhvetesorter og soppbekjempelse

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Comparison of RNAi efficiency mediated by tetracycline-responsive H1 and U6 promoter variants in mammalian cell lines

Conditional expression of short hairpin RNAs (shRNAs) to knock down target genes is a powerful tool to study gene function. The most common inducible expression systems are based on tetracycline-regulated RNA polymerase III promoters. During the last years, several tetracycline-inducible U6 and H1 promoter variants have been reported in different experimental settings showing variable efficiencies. In this study, we compare the most common variants of these promoters in several mammalian cell lines. For all cell lines tested, we find that several inducible U6 and H1 promoters containing single tetracycline operator (tetO) sequences show high-transcriptional background in the non-induced state. Promoter variants containing two tetO sequences show tight suppression of transcription in the non-induced state, and high tet responsiveness and high gene knockdown efficiency upon induction in all cell lines tested. We report a variant of the H1 promoter containing two O2-type tetO sequences flanking the TATA box that shows little transcriptional background in the non-induced state and up to 90% target knockdown when the inducer molecule (dox–doxycycline) is added. This inducible system for RNAi-based gene silencing is a good candidate for use both in basic research on gene function and for potential therapeutic applications

Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma

BACKGROUND: MicroRNAs (miRNAs) regulate expression of many cancer-related genes through posttranscriptional repression of their mRNAs. In this study we investigate the proto-oncogene MYCN as a target for miRNA regulation. METHODS: A luciferase reporter assay was used to investigate software-predicted miRNA target sites in the 30 -untranslated region (30 UTR) of MYCN. The miRNAs were overexpressed in cell lines by transfection of miRNA mimics or miRNA-expressing plasmids. Mutation of the target sites was used to validate MYCN 30 UTR as a direct target of several miRNAs. To measure miRNA-mediated suppression of endogenous N-myc protein, inhibition of proliferation and inhibition of clonogenic growth, miRNAs were overexpressed in a MYCN-amplified neuroblastoma cell line. RESULTS: The results from this study show that MYCN is targeted by several miRNAs. In addition to the previously shown mir-34a/c, we experimentally validate mir-449, mir-19a/b, mir-29a/b/c, mir-101 and let-7e/mir-202 as direct MYCN-targeting miRNAs. These miRNAs were able to suppress endogenous N-myc protein in a MYCN-amplified neuroblastoma cell line. The let-7e and mir-202 were strong negative regulators of MYCN expression. The mir-101 and the let-7 family miRNAs let-7e and mir-202 inhibited proliferation and clonogenic growth when overexpressed in Kelly cells. CONCLUSION: The tumour-suppressor miRNAs let-7 and mir-101 target MYCN and inhibit proliferation and clonogenic growth of MYCN-amplified neuroblastoma cells

Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma

Background: Neuroblastoma is a childhood cancer derived from immature cells of the sympathetic nervous system. The disease is clinically heterogeneous, ranging from neuronal differentiated benign ganglioneuromas to aggressive metastatic tumours with poor prognosis. Amplification of the MYCN oncogene is a well established poor prognostic factor found in up to 40% of high risk neuroblastomas. Using neuroblastoma cell lines to study neuronal differentiation in vitro is now well established. Several protocols, including exposure to various agents and growth factors, will differentiate neuroblastoma cell lines into neuron-like cells. These cells are characterized by a neuronal morphology with long extensively branched neurites and expression of several neurospecific markers. Results: In this study we use retrovirally delivered inducible short-hairpin RNA (shRNA) modules to knock down MYCN expression in MYCN-amplified (MNA) neuroblastoma cell lines. By addition of the inducer doxycycline, we show that the Kelly and SK-N-BE(2) neuroblastoma cell lines efficiently differentiate into neuron-like cells with an extensive network of neurites. These cells are further characterized by increased expression of the neuronal differentiation markers NFL and GAP43. In addition, we show that induced expression of retrovirally delivered anti- MYCN shRNA inhibits cell proliferation by increasing the fraction of MNA neuroblastoma cells in the G1 phase of the cell cycle and that the clonogenic growth potential of these cells was also dramatically reduced. Conclusion: We have developed an efficient MYCN-knockdown in vitro model system to study neuronal differentiation in MNA neuroblastomas