• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.3
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• pp.237-242
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• 2016

Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.641-644
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• 2000

In vitro true digestibility of cup-plant (Silphium perfoliatum L.) is higher than other alternative forages and comparative to alfalfa (Medicago sativa L.) even at the high neutral detergent fiber (NDF) concentration. This study was conducted to determine whether the digestion kinetic parameters of cup-plant could explain high in vitro true digestibility of cup-plant at the several NDF levels. Cup-plant and alfalfa were both collected in Arlington and Lancaster, Wisconsin to meet the NDF content within 40 to 50% range. The collected samples were incubated with rumen juice to investigate the digestion kinetics at 3, 6, 9, 14, 20, 28, 36, 48, and 72 h. Kinetics was estimated by the model $R=D_0\;e-k(t-L)+U$ where R is residue remaining at time t, and $D_0$ is digestible fraction, k is digestion rate constant, L is discrete lag time, and U is indigestible fraction. Parameters of the model were estimated by the direct nonlinear least squares (DNLS) method. Digestion rate and potential extent of digestion were not statistically different in either forage. However, alfalfa had shorter lag time (p<0.05). The indigestible fraction increased with maturation in alfalfa and in cup-plant (p<0.05). The ratio of indigestible fraction to acid detergent lignin (ADL) was higher in cup-plant than in alfalfa (p<0.05). From the results, alfalfa is probably digested more rapidly than cup-plant, however, cup-plant maintains higher digestibility with maturation due to a relatively slower increase of indigestible fraction in NDF.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.630-635
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• 2000

Previous research has not evaluated the effects of various rates and frequencies of manure application and frequencies of cutting on yield and stand persistence of cool-season grasses and alfalfa-grass mixtures. The primary objective of this study was to compare the effects of cutting management systems on herbage yield and stand persistence of orchardgrass (Dactylis glomerata L.) and an alfalfa (Medicago sativa L.)-orchardgrass mixture from various rates and frequencies of dairy slurry application. A randomized complete block design with treatments in a sub-subplot arrangement with four replicates was used. The main plot consisted of 2 cutting management systems (4 and 5 annual cuttings). The subplots were 9 fertility treatments: 7 slurry rate and frequency of application treatments, one inorganic fertilizer treatment, and an unfertilized control. The split-split-plots were the two forage species: orchardgrass and alfalfa-orchardgrass mixture. The study was initiated after 1st cutting in 1995. Cumulative yields of the 2nd and subsequent cuttings of both orchardgrass and alfalfa-orchardgrass in 1995 were higher for the 5-cutting system than the 4-cutting system. The 1995 growing season was abnormally dry. In 1996, an abnormally wet year, the reverse was true, total herbage yields being higher for the 4-cutting system than the 5-cutting system. Species response to fertility rate/frequency treatments was different in both years. Higher application rates early in the season and carryover of nutrients from late season applications the previous year appear to be responsible for the yield increases of those fertility treatments having significant yield differences between the cutting management systems. The stand ratings of orchardgrass were not affected by cutting management. In the spring of 1997, however, the stand ratings of alfalfa-orchardgrass in the 4-cutting management system were significantly greater than the 5-cutting management system. The very high manure application rate significantly reduced the stand ratings of alfalfa-orchardgrass in the 5-cutting system.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.4
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• pp.430-434
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• 1989

Alfalfa saponin was isolated from the roots of alfalfa(Medicage sativa L.) and incoporated at the level of 0.2% and 0.4% cholesterol-free diets and cholesterol enriched-diets. Wistar male rats weighing approximately 80g were fed with for 21 days. The serum lipids and fecal excretion lipids of the rats were determined and the result obtained are as follows. 1. Alfalfa saponin contained 36.4% medicagenic acid. 2. The feeding of alfalfa saponin increased cholesterol excretion but not bile acid excretion. 3. Alfalfa saponin showed hypocholesterolemic activity at 0.4% level in both the diets with and without cholesterol but the effect was not observed at 0.2% level in a diet. 4. It was indicated that the hypocholesterolemic effect of alfalfa saponin was associated with the decreased absorption of cholesterol from alimentary canal.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.2
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• pp.192-197
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• 1981

Alfalfa (Medicago sativa L. cv. Luna) seeded in agar was inoculated with two strains of Rhizobium meliloti isolated from root nodules of alfalfa for assessment of nodulation. The seedling growth after six weeks was remarkably increased by adding each rhizobia strains into agar media and also by nitrate application (70ug N/ml), but there was no significant difference among them. Nodulations started one week after inoculation and increased its numbers and sizes as seedling grew. Therefore, the two strains isolated from alfalfa root were concluded to be effective strains. For determining seed inoculation method the same cultivar was inoculated with both rhizobia strains using different inoculation methods such as broth-vacuum, peat-adhesive, peat & lime pelleting. They were seeded in pots of river sand and supplied with culture solution excluded nitrogen. The peat & lime pelleting was recognized the best method in both of nodulation and seedling growth after eight weeks growth. There were significant correlations between the weight of nodules and the shoot or root dry weight of alfalfa in both rhizobia strains.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.1
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• pp.56-64
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• 2011

This study was conducted to evaluate the use of exogenous enzymes as a potential means of improving the ruminal digestion (i.e., degradability) of alfalfa hay and rice straw. Twenty six enzyme-additives were examined in terms of protein concentration and enzymic activities on model substrates. The exogenous enzymes contained ranges of endoglucanase, xylanase, ${\beta}$-glucanase, ${\alpha}$-amylase, and protease activities. Six of the enzyme additives were chosen for further investigation. The enzyme additives and a control without enzyme were applied to mature quality alfalfa hay substrate and subsequently incubated in rumen batch cultures. Five of the enzyme additives (CE2, CE13, CE14, CE19, and CE24) increased total gas production (GP) at 48 h of incubation compared to the control (p<0.05). The two additives (CE14 and CE24) having the greatest positive effects on alfalfa hay dry matter, neutral detergent fibre (NDF) and acid detergent fibre (ADF) degradability were further characterized for their ability to enhance degradation of low quality forages. The treatments CE14, CE24, a 50:50 combination of CE14 and CE24 (CE14+24), and control (no enzyme) were applied to mature alfalfa hay and rice straw. For alfalfa hay, application of the two enzyme additives, alone and in combination, increased GP compared to the control at 48 h fermentation (p<0.05), whereas only CE14 and CE14+24 treatments improved GP from rice straw (p<0.05). Rumen fluid volatile fatty acid concentrations throughout the incubation of rice straw were analyzed. Acetate concentration was slightly lower (p<0.05) for CE14${\times}$CE24 compared to the control, although individually, CE14 and CE24 acetate concentrations were not different from the control. Increases (p<0.05) in alfalfa hay NDF degradability measured at 12 and 48 h of incubation occurred only for CE14 (at 12 h) and for CE14+24 (at 12 and 48 h). Similarly, ADF degradability increased (p<0.05) with CE14 and CE14+24. As for rice straw, increased DM degradability was observed at 12 and 48 h of incubation for all enzyme treatments with an exception for CE14 at 12 h. The degradability of NDF was improved by all the enzyme treatments at either incubation time, while ADF degradability was only enhanced at 48 h. Overall, the enzymes led to enhanced digestion of mature alfalfa and there was evidence of improved digestibility of rice straw, an even lower quality forage.

• Journal of The Korean Society of Grassland and Forage Science
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• v.11 no.2
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• pp.84-89
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• 1991

This experiment was carried out to determine the effects of oversowing and other seeding methods (oversowing+ raking, oversowing+ raking+compaction, tillage+ broadcasting+ compaction and tillage + drilling+ compaction) on growth, dry matter and crude protein yield of alfalfa (Medicago satiua L.). The results obtained are summarized as follows:1. Soil pH and soil properties were improved by tillage.2. Establishment of alfalfa was increased with raking and compaction, and more by tillage than by oversowing(P

• Asian-Australasian Journal of Animal Sciences
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• v.2 no.2
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• pp.115-121
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• 1989

A comparative slaughter feeding experiment with steer calves weighing 280 kg and a concurrent digestion trial with wether lambs was conducted to study the energy value of rice straw as influenced by ammonia treatment and mixing with alfalfa hay. Steers were ad libitum fed one of nine completely mixed experimental diets: basal (high concentrate); 25 or 50% of untreated rice straw (URS) or ammoniated (plastic covered bales, 4.6% $NH_3$ by weight) rice straw (ARS) proportionately replacing part of the basal; a 50:50 % mixture of URS or ARS and alfalfa replacing a proportion of the basal at 25 and 50%. Digestibility of the nine complete diets (pelleted to prevent sorting) was determined with four ad libitum fed lambs. Ammoniation increased crude protein level (from 3.6 to 10.8%) and in vitro dry matter digestibility of the rice straw by 15%. The improvement in DE, NEm and NEg by ammoniation of rice straw was 20, 52 and 117%, respectively. Ammoniation of rice straw fed as 50% of the diet improved gains over the diet containing 50% URS, but no significant influence on animal performance was observed when rice straw was fed at the 25% level. Each 10% addition of URS to basal diet decreased empty body gain of steers by 116 gram per day compared with a decrease of 70 gram per day when rice straw had been ammoniated. There was no beneficial effect of ammoniation when the roughage component of the diet was a 50:50 mixture of rice straw and alfalfa. Ammoniation of straw and inclusion of alfalfa generally increased the concentration of total volatile fatty acids in rumen fluid. Ammoniation resulted in reduced concentrations of acetic and propionic acid, but increased concentration of butyric acid. Digestibility of URS was improved by mixing with alfalfa. However, alfalfa hay did not influence digestibility of ARS. Diets in which ARS replaced the basal mixture at 25 and 50% had higher NEm and NEg values than comparable URS diets. The same pattern was observed in the straw: alfalfa mixtures, but differences between URS and ARS were significant only for the 50% roughage diets.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.12
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• pp.1701-1704
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• 2001

The evaluation of structural carbohydrate losses and its effect on silages digestibility in alfalfa (Medicago sativa L.) and orchardgrass (Dactylis glomerata L.) was studied during 5, 21 and 56 days ensiling. About 70 and 60 kg fresh matter of the two forages were ensiled in 9 silos of 120 L capacity. The digestion trials were conducted in two phases using the two grasses in two $4{\times}4$ Latin square design according to the four treatments being the grass and the three silages. There were no differences in the DM and CP contents resulting from 5 to 56 days ensiling in both forages. The water-soluble carbohydrates (WSC), hemicelllose, pectin, and energy were slightly reduced and appeared lower in 56 days silage. The ether extract and cellulose contents slightly increased as the ensiling process advanced in the two species. Hemicellulose losses of 29 and 41 g/kg DM were obtained in alfalfa and orchardgrass, respectively, 56 days after ensiling. While the cellulose losses in both species were very little, compared to that for hemicellulose, the pectin losses, 56 days after ensiling were 15 and 12 g/kg DM in alfalfa and orchardgras respectively. The total structural carbohydrates lost (ie., hemicellulose + cellulose + pectin) in g/kg DM of fresh material forage ensiled, is about four fifths the amount lost by WSC, in alfalfa and about two thirds, in orchardgrass, by 21 days ensiling after the activity of microorganism terminated, indicating that appreciable amount was used as substrate for silage fermentation. Ensiling alfalfa and orchardgrass for 0, 5, 21 and 56 days maintained a decreasing trend of 83.8, 82.5, 79.3 and 78.9% digestibility in alfalfa and 80.5, 77.0, 77.1 and 76.4% digestibility in orchardgrass. While the digestibility of cellulose and ether extract increased in silage in both species, the digestible energy values in silage were reduced from 2.6 to 2.3 and 2.9 to 2.7 Mcal/kg DM respectively in alfalfa and orchard during 5-56 days ensiling.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.3
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• pp.243-247
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• 2016

Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.