• 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.17 no.8
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• pp.1098-1106
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• 2004

In a split-unit design, 12 Javan rusa (Cervus timorensis russa) stags (6 castrated and 6 entire) were used to investigate seasonal (winter, spring and summer) effects on intake, digestibility of food constituents, volatile fatty acid profile, and nitrogen retention, when given lucerne (Medicago sativa) or rhodes grass (Chloris guyana) hays. Entire stags ate 9% more dry matter (DM) than castrates (p<0.05). Both castrated and entire stags exhibited seasonal changes in DM intake, these being higher (p<0.05) in winter (62.3 g/kg $W^{0.75}$) than spring (56.9 g/kg $W^{0.75}$) or summer (55.3 g/kg $W^{0.75}$). Intakes of lucerne hay (211 g protein/kg DM) were significantly higher than of rhodes grass hay (49 g protein/kg DM) in all seasons (p<0.05). Digestibilities of DM (0.58), neutral detergent fibre (0.59) and acid detergent fibre (0.47) were similar between castrates and entires. DM digestibility was higher (p<0.0001) for lucerne (0.66) than rhodes grass (0.55), and was higher (p<0.05) in winter (0.60) than spring (0.58) or summer (0.57). The ruminal acetate concentrations were higher (p<0.001) in spring than summer (78 and 73 molar % respectively). Apparent digestibility of protein was significantly less (p<0.0001) for rhodes grass (0.37) than lucerne (0.75). N retention was positive for lucerne (15.2 g/d) but negative for rhodes grass (-2.8 g/d) (p<0.0001), and was higher (p<0.001) in summer (12.0 g/d) than spring (4.3 g/d) or winter (2.4 g/d). The tropical rusa deer exhibits seasonal variations in feed intake, food constituent digestibilities, VFA profile and N retention. Castration did not alter these traits. The results do not support the view that rusa deer can thrive on low-quality pastures. The productivity and commercial exploitation of rusa deer could be optimised if they are given high-protein feed during spring and summer.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.929-934
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• 2000

Optimum stand density of alfalfa (Medicago sativa L.) varies with locations and climates. Stand density is one of the factors that determines herbage yield, forage quality and persistence of alfalfa. As establishment costs increase, the question arises whether present population densities are optimum for obtaining maximum herbage yield and forage quality. The objectives of this study were: 1) to determine the optimum plant density for highest herbage yield and forage quality for the dehydrated alfalfa industry under Edmontons climatic conditions in Alberta, Canada; 2) to compare herbage yield and forage quality of the cultivars 'Algonquin' and 'Vernal' grown at a range of stand densities. Alfalfa seedlings of both cultivars were either transplanted at spacings of 6, 10, 15 and 25 cm or direct seeded at the 4.5 cm plant spacings, providing population densities of 494, 278, 100, 45 and $16plants/m^2$. The experimental design was a randomized complete block with a split-plot arrangement having three replicates; the main plots consisted of two alfalfa cultivars Algonquin and Vernal, and the sub-plots were the five population densities. The cultivar Vernal had significantly higher annual yield than did the cultivar Algonquin. There was no significant effect of plant density on herbage yield. There was no difference in crude protein (CP) between the two cultivars. At the first cut, there was a significant quadratic effect of plant density on CP content and the greatest CP occurred at the 100 plants/m2 density. Crude protein was not affected by plant density at the second cut. Acid detergent fiber (ADF) and neutral detergent fiber (NDF) were not affected by plant density. The cultivar Algonquin usually had a lower ADF and NDF than cultivar Vernal. In conclusion, high population densities ($278plants/m^2$ or more) of alfalfa did not improve herbage yield and forage quality compared with low plant population densities ($100plants/m^2$ or less) of alfalfa.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.1
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• pp.5-11
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• 1993

First crop of alfalfa (Medicago sativa L.) was harvested, wilted and ensiled with or without NaOH or $NH_3$, and fed to three rumen fistulated goats in a $3{\times}3$ Latin-square design. Each alkali treatment (2.44% of alfalfa dry matter) was made by spraying its solution prior to ensiling. Silage pH, $NH_3-N$ and butyric acid concentration were increased with each alkali addition, and NaOH-treated silage showed the lowest chemical quality. Compared with untreated silage, digestibilities of organic matter, ADF and cellulose were depressed by both alkali treatments, and the reductions in NaOH-treated silage were significant. Crude protein digestibility was also significantly decreased in NaOH-treated silage, but the goats receiving the silage excreted less nitrogen in urine than those on the other two silages. Nitrogen retention of goats was not different among the treatments. Ruminal solubility and potential degradability of dry matter and nitrogen determined with the in situ bag technique were reduced, and rate of degradation of the two components were increased by the NaOH treatment. Addition of $NH_3$ provided ruminal soluble nitrogen to the silage, but the rate of degradation was similar to that of untreated silage. These results suggest that NaOH treatment would denature the protein and reduce the susceptibility to microbial degradation in the rumen, while no positive effect of alkali treatment on fiber digestion and nitrogen utilization was observed in this study.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.6
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• pp.818-823
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• 2008

The interaction of fibre degrading microbes and methanogens was studied using two forages, lucerne (Medicago sativa) hay and maize (Zea mays) hay, as substrate and 2-bromoethanesulphonic acid (BES) as an additive in an in vitro gas production test. Gas and methane production (ml/g dry matter) were significantly higher (p<0.05) on lucerne as compared to maize hay. Inclusion of BES in the incubation medium significantly suppressed methane emission irrespective of substrate. The population density of total bacteria, fungi, Ruminococcus flavefaciens and Fibrobacter succinogenes was higher, whereas that of methanogens was lower with maize hay as compared to lucerne as substrate. BES suppressed methanogen population by 7 fold on lucerene and by 8.5 fold on maize at 24 h incubation as estimated by real time-PCR. This suppression was accompanied by almost complete (>98% of control) inhibition of methanogenesis. The proportion of acetate decreased, whereas that of propionate increased significantly by inclusion of BES, resulting in narrowing of acetate to propionate ratio. In vitro true digestibility (IVTD) of lucerne was significantly higher as compared to maize but BES inclusion did not affect IVTD.

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.3
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• pp.264-268
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• 2015

The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to $250{\mu}M$ of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.4
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• pp.371-378
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• 1997

The purpose of this research was to investigate the effect of P fertilization and cutting fkquency without N fertilizer on nutritive value and nitrogen fixation activity of alfalfa. A field experiment was conducted on the experimental farm of Kongju National University at Yesan from 1994 to 1996. The experimental design was a split plot design with two P levels as main plot and two cutting frequencies as the sub-plot. The results obtained are summarized as follows. Available phosphate, exchangeable Ca and Na in the soil were slightly increased with P fertilizer and basal treatment of lime. Average DM yield was 1.5 ton and 1.4 ton in second and third year. There was no significant difference within P level and cutting kquency. The first cut showed higest CP value as cutting times increased the value tend to be lowered. Crude protein content in 3 and 4 cutting regimes were from 16.3 to 16.6% 17.4 to 17.9% respectively, but no significant effect were found among the main and sub-plot. ADF and NDF in 1995 were ranged from 38.2 to 39.9, 45.1 to 58.1% respectively. ADF was 2% more higher in 1996 compared to previous year and NDF was slightly higher in 1996. There was significant difference of ADF between main plot in 1995. .P, Ca, K and Mg of 200kg P level with 3 times cutting treatment in 1995 and 1996 were 0.32 and 0.23, 0.95 and 0.84, 3.24 and 2.87, and 0.36 and 0.26. Significant difference of K and Mg between cutting frequency was observed in 1995. Nitrogen fixing activity of 400kg P fertilizer plots were higher than 200kg P level and 3 times cutting plot also showed higher acetylene reduction activity than that of 4 cutting per year. Phosphate level and cutting frequency treatment of did not significantly affect nitrogen fixation activity.

• 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.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.6
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• pp.821-832
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• 1997

The aim of this study was to separate or purify some bioactive compounds from flowers of alfalfa(Medicago sativa L.) and to test of the isolated compounds on alfalfa for their autotoxicity and on Echinochloa crus-galli for their allelopathy for seed germination and seedling weight. Using thin layer chromatography(TLC) of $CHCl_3$ extracts, the most inhibitory band to alfalfa seed germination was determined. Germination inhibition of this extract suggested a complex chemical interaction. Separation and purification of compounds with CHCl$_3$ extract of fresh alfalfa flowers were conducted by a silica gel TLC, and microcrystalline cellulose TLC(MCTLC), followed by droplet countercurrent chromatography(DCCC) bioassay. Preliminary identification was done by high perfomance liquid chromatography(HPLC) on the most inhibitory fractions in DCCC. Ferulic acid, caffeic acid, vanillic acid, rutin, narringin were identified in fraction 5 and ferulic acid, caffeic acid, vanillic acid, rutin, coumarin in fraction 6. The phytotoxicity of their individual compound was tested on alfalfa and Echinochloa crus-galli seed germination and seedling weight. Coumarin and ferulic acid showed the most inhibitory effect on alfalfa seed germination and Echinochloa crus-galli seedling fresh and dry weight. These compounds may be, at least in part, involved in autotoxicity and allelopathy.

• Journal of Animal Science and Technology
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• v.63 no.5
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• pp.1098-1113
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• 2021

Maturation process of alfalfa (Medicago sativa L.) could be prevented by the reduction of lignin content in terms of conventional breeding or transgenic technology. Alfalfa could exhibit higher leaf/stem ratio, with a concern of yield loss. The objective of this study was to compare forage yield and nutritive value of low lignin alfalfa and two reference varieties subjecting to two harvest intervals and three seeding rates. The experimental design was a randomized complete block in a split-split plot arrangement with four replicates, where harvest intervals (28-day and 35-day) were assigned to whole plots, seeding rates were subplots, and varieties were sub-subplots. The weighted mean nutritive value was applied to two production years of 2016 and 2017. Hi-Gest 360 (low lignin alfalfa) provided similar yield potential and increased nutritive value compared to two reference varieties. Over a two-year production period, alfalfa harvested at every 28-day interval provided more economic returns than those at 35-day interval. For the seeding year and first production year, five cuts made by the 28-day interval produced more yield than four cuts by the 35-day interval. Due to limited rainfall in May 2017, a sharp drop of the first cutting overturned the advantage of the five-cut system. Shorter intervals between harvests generally increased crude protein (CP) concentrations. The differences of relative feed value (RFV) between two harvest intervals tended to be great during the first and second cuttings. Overall, harvest interval had a large effect on nutritive value and a more significant effect on alfalfa dry matter yield than variety selection. Seeding rate did not affect alfalfa yield and nutritive value.