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Publications

NIBIOs employees contribute to several hundred scientific articles and research reports every year. You can browse or search in our collection which contains references and links to these publications as well as other research and dissemination activities. The collection is continously updated with new and historical material.

2024

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Abstract

The aim of this study was to examine how silages from different grassland species and harvesting frequencies affect feed intake, milk production, and methane (CH4) emission in dairy cows. We hypothesized that cows consuming silages of more frequent harvest, grass species with greater organic matter digestibility and legumes with lower NDFom concentration would have greater silage dry matter intake and milk yield and thereby lower CH4 yield and intensity. Forty Norwegian Red cows were allocated to 5 treatments in a cyclic changeover design with 4 21-d periods (14 d of adaptation, 7 d of data collection). The 5 treatments evaluated were silages produced from timothy (Phleum pratense L.) in a 3-cut system (T3), timothy in a 2-cut system (T2), perennial ryegrass (Lolium perenne L.) in a 3-cut system (PR3), red clover (Trifolium pratense L.) in a 3-cut system (RC3) and a mix of T3 and RC3 (50:50 on DM basis) (T3/RC3). The treatments were prepared by mixing silages from each crop over the growing season, proportional to the harvested DM yield of each cut. Cows were offered the mixed silages ad libitum supplemented with a fixed level of concentrate. Gas emissions were measured using 2 Greenfeed units. Milk yield was recorded in the milking robot at each visit, and milk samples were collected at 3 consecutive milkings during the last 7 d of each period. Cows were weighed after each milking, and total-tract digestibility of each diet was estimated using acid insoluble ash as internal marker in fecal grab samples. The data were analyzed using the MIXED procedure of SAS with block, period and treatment as fixed effects and animal within block as random effect. Silage and total DMI did not differ between T3 and T2, but total DMI was lower for PR3 than for T3. There was a quadratic effect of increased proportion of red clover, with highest intakes of T3/RC3 and lower intakes of RC3 than of T3. Energy corrected milk (ECM) yield was lower for T2 than T3, and for PR3 than T3. There was a quadratic effect of increased proportion of red clover, with highest ECM yield in T3/RC3 and lower in RC3 than in T3. Organic matter digestibility was lower for T2 than T3, but it did not differ between T3 and PR3. Including red clover in the diet linearly decreased organic matter digestibility. Methane production (g/d) did not differ between T3 and T2, but CH4 intensity (g/kg ECM) was greater for T2 than for T3. There was no difference between T3 and PR3 for CH4 production but yield and intensity were greater for PR3 than T3. Including red clover in the diet linearly increased CH4 production, yield and intensity with greatest intensity in the 100% red clover diet. In conclusion, changing harvesting frequency for timothy from 2 to 3 harvests per year did not affect CH4 production or yield, but CH4 intensity was reduced. Replacing timothy with perennial ryegrass and increased inclusion rate of red clover both increased CH4 yield and intensity. Key words: enteric methane, timothy, red clover, perennial ryegrass, greenfeed system

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Abstract

In situ epitope tagging is crucial for probing gene expression, protein localization, and the dynamics of protein interactions within their natural cellular context. However, the practical application of this technique in plants presents considerable hurdles. Here, we comprehensively explored the potential of the CRISPR/Cas nuclease-mediated prime editing and different DNA repair pathways in epitope tagging of endogenous rice (Oryza sativa) genes. We found that a SpCas9 nuclease/microhomology-mediated end joining (MMEJ)-based prime editing (PE) strategy (termed NM-PE) facilitates more straightforward and efficient gene tagging compared to the conventional and other derivative PE methods. Furthermore, the PAM-flexible SpRY and ScCas9 nucleases-based prime editors have been engineered and implemented for the tagging of endogenous genes with diverse epitopes, significantly broadening the applicability of NM-PE in rice. Moreover, NM-PE has been successfully adopted in simultaneous tagging of the MAP kinase (MPK) genes OsMPK1 and OsMPK13 in rice plants with c-Myc and HA tags, respectively. Taken together, our results indicate great potential of the NM-PE toolkit in the targeted gene tagging for Rice Protein Tagging Project, gene function study and genetic improvement.