Molecular markers have been widely utilized in population genetics, diagnostic taxonomy, and genetic mapping, and can be applied to cultivar discrimination during field selection processes for alfalfa. In this study, whole-genome sequencing information was obtained for seven alfalfa lines and cultivars developed in Korea, including ‘Alfaone (MS001)’, using Next-Generation Sequencing (NGS). Single nucleotide polymorphism (SNP) analysis revealed that ‘Alfaone (MS001)’ could be distinguished from other lines and cultivars using six SNP loci. Specifically, only two SNP loci were sufficient to differentiate ‘Alfaone (MS001)’ from major lines and cultivars such as ‘MS002’ and ‘Alfaking (MSCB07)’. This set of SNP barcodes provides a reliable standard for alfalfa cultivar discrimination, contributing to domestic cultivar protection and the advancement of the Korea forage industry. Furthermore, the development of distinguishing markers across alfalfa cultivars will enhance genetic resource identification and support the breeding of high-quality new cultivars.

To study the effects of external ethanol pretreatment on growth and drought tolerance in alfalfa, 21-day-old plants were pretreated with ethanol at 0-1000 mM for 4 days and then subjected to drought stress for 9 days. Ethanol pretreatment significantly enhanced plant survival under drought conditions, with the highest survival rate observed in the 250 mM treatment. In contrast, fresh weight and dry weight were not significantly affected by ethanol pretreatment. Relative water content was increased 7.15% at 250 mM ethanol treatment compared to 0 mM ethanol treatment. Chlorophyll content decreased compared to the control, while carotenoid content remained unchanged. total soluble carbohydrate content was reduced by ethanol pretreatment. Hydrogen peroxide (H2O2) levels were maintained only in plants treated with 250 mM ethanol, whereas lower and higher concentrations resulted in significantly decreased H2O2 content. These results suggest that ethanol pretreatment does not affect alfalfa growth, but significantly improves survival rate under drought conditions by maintenance of relative water content. Furthermore, it suggests that chemical priming using ethanol pretreatment could be applied as a method to improve drought tolerance of alfalfa plants.

This study reports the development of a new alfalfa (Medicago sativa L.) variety, ‘Alfaone’, at the Forage Production Systems Division, National Institute of Animal Science, Rural Development Administration, Korea, from 2015 to 2023. The variety originated from an artificial cross between Xun Lu (maternal parent) and RadarⅡ Brand (paternal parent), followed by pedigree selection and performance testing. The elite line ‘MsCB01’ was subsequently released as ‘Alfaone’. Regional adaptability trials were conducted for two years (2022–2023) across four representative sites in Korea (Cheonan, Pyeongchang, Jeongeup, and Jinju) to evaluate agronomic traits, forage yield, and quality. Evaluated characteristics included plant height, regrowth ability, winter survival, and lodging resistance. The average dry matter yield of ‘Alfaone’ was 20,811 kg/ha, approximately by about 3% higher than that of the standard cultivar ‘Vernal’ (20,236 kg/ha). Yield superiority was particularly evident in Pyeongchang, suggesting excellent cold tolerance and winter hardiness. Assessment of forage nutritive traits indicated that ‘Alfaone’ was comparable to ‘Vernal’, demonstrating that its yield advantage did not come at the expense of quality. Overall, ‘Alfaone’ is a promising cultivar that combines high productivity with strong adaptability to unfavorable environments, particularly cold-prone regions. Its release is expected to promote the expansion of alfalfa cultivation, enhance forage self-sufficiency, and reduce dependence on imported hay in Korea.

Field drying remains the primary bottleneck to consistent alfalfa haymaking under Korea’s humid, rain-prone summers. This study evaluated how dry matter (DM) yield and tedding frequency shape seasonal drying trajectories and leaf shatter in Cheonan during 2024. The experiment was conducted in May, June, August, September, and October 2024 with three DM yield levels (3, 5, and 7 ton/ha) and three tedding regimes (no tedding, once daily, and twice daily). Favorable May–June weather enabled swaths to reach <20% moisture within 100 h, whereas in September–October, moisture commonly remained ≥50%, indicating conditions unsuitable for dry hay. Yield effects on drying were generally small except in August, when re-wetting events accentuated between-group differences. Tedding effects were season-dependent: treatment differences emerged after rainfall in August and were evident in September, with twice-daily tedding producing the lowest moisture among tedding treatments. Leaf dropout rose with greater tedding frequency and higher DM yield; significant tedding effects were detected in June and September, and twice-daily tedding often exceeded no-tedding for leaf losses, particularly at 7 ton/ha. Collectively, omitting tedding is a viable option for May–June hay (4 days to ~20% moisture), while for post-June harvests, tedding offers limited drying benefits and may elevate leaf shatter; ensiling is therefore recommended when drying conditions are suboptimal.

The objective of this study was to assess the effects of gypsum application on dry matter yield (DMY), mineral content of alfalfa (Medicago sativa L.), and soil properties in reclaimed tidal land in South Korea. The experiment was conducted in Seokmun, located on the west coast of South Korea, which is reclaimed with approximately 70 cm depth of degraded island soil. Treatments consisted of a control with no gypsum application (G0), 2 ton ha-1 (G2), and 4 ton ha-1 (G4) of gypsum application. The first harvest was carried out when the alfalfa reached 10% flowering, and subsequent harvests were conducted at 35-day intervals. Over the three-year experimental periods (2019-2021), the total DMY of G2 treatment was significantly higher than those of G0 and G4 (p<0.05). Although both G2 and G4 gypsum application treatments lowered soil pH, the G4 treatment increased the electrical conductivity (EC) content of the soil. Additionally, gypsum application affected the mineral contents of alfalfa, resulting in reduced concentration of sodium (Na) and Magnesium (Mg). Therefore, this present study suggests that a gypsum application rate of 2 ton ha-1 is optimal for improving alfalfa dry matter yield and mineral balance, as well as enhancing soil chemical properties in reclaimed tidal land in South Korea.

Dietary fiber plays a important role in improving gut micriobiome and reproductive performance in gestating sows. This study aimed to evaluate effects of dietary supplementation with domestically produced alfalfa, provided in pelleted or mashed form, on body weight, body condition, litter performance, fecal score and fecal microbiome in gestating sows. A total of nine crossbred F1 gilts (Yorkshire × Landrace) with an average body weight of 230.33 ± 12.63 kg at 50 days of gestation were randomly assigned to three dietary treatments (n = 3 per treatment): (1) control (CON, basal diet), (2) Pellet (basal diet + 10% pelleted alfalfa), and (3) Mash (basal diet + 10% mashed alfalfa). Results indicated that alfalfa supplementation had no significant effect on weight gain during gestation (p>0.05), although sows in both alfalfa-supplemented groups showed numerically reduced weight loss during lactation compared to the control. The mashed alfalfa improved average daily weight gain of piglets during lactation (p<0.05). Moreover, the pelleted alfalfa alleviated constipation and numerically increased the relative abundance of positive microbes such as Firmicutes (phylum level), Lactobacillaceae and Ruminococcaceae (family level), and Clostridium_sensu_stricto_1, Lactobacillus, and Ruminococcaceae_UCG_002 (genus level) (p<0.05). On the other hands, it numerically decreased the relative abundance of negative microbes such as Proteobacteria in phylum level and Escherichia_Shigella in genus level. These findings suggest that domestically grown alfalfa, particularly in pellet or mash form, may be a valuable dietary fiber source for gestating sows to support piglet performance, gut health, and welfare. However, further studies with larger sample sizes are warranted to confirm these effects.

Near infrared reflectance spectroscopy (NIRS) is widely used to assess the nutrient composition of forages. In forage, the leaf to stem ratio of alfalfa greatly affects its forage quality, with a high ratio of leaf indicated as high quality. This study aimed to evaluate the predictability of the alfalfa leaf-to-stem ratio and feed value using NIRS. Alfalfa hay was manually separated into leaves and stems by hand and the analysis samples were then made in the controlled range between 0 and 100%. Calibration models (n=320) were developed using modified partial least squares regression (MPLS) based on cross-validation. The optimal calibrations were selected based on the highest coefficients of determination in cross-validation (R2) and the lowest standard error of cross-validation (SECV). The prediction accuracy for the leaf-to-stem ratio (SECV, 5.95 vs. 5.71%; R2, 0.91 vs. 0.91) in alfalfa hay was comparable. For leaves, the standard error of calibration (SEC) was 4.94% (R2=0.94), and for stems, it was 4.81% (R2=0.94). The leaves and stems of the SEC were 4.94% (R2=0.94) and 4.81% (R2=0.94), respectively. The prediction accuracy for feed value, based on the leaf-to-stem ratio, predicted SECV values of 0.92% (R2=0.88) for crude protein (CP) content, 1.92% (R2=0.91) for neutral detergent fiber (NDF) content, 1.36% (R2=0.91) for total digestibility nutrient (TDN) content, and 9.86 (R2=0.81) for relative feed value (RFV). The results of this study demonstrate the potential of the NIRS method as a reliable tool for predicting the leaf-to-stem ratio of alfalfa hay, and show available techniques for routine feed value evaluation.

Alfalfa hay varieties have been developed and cultivated in Korea to enhance the self-sufficiency rate of domestic roughage. The objective of this study was to compare the effects of feeding domestic and imported alfalfa on the growth performance and economic feasibility of Hanwoo bulls. This study was conducted using eight Hanwoo bulls, aged 6 to 7 months, during their growing period. Hanwoo bulls were divided into two groups: one group fed imported alfalfa and the other group fed domestic alfalfa. Dry matter intake, weight gain, and blood metabolites were analyzed for two months of the growing stage. There was no significant difference between the two groups in dry matter intake and body weight. However, the price of domestic alfalfa is lower than that of imported alfalfa, so feeding domestic alfalfa is expected to have a positive effect on reducing production costs. Further study is needed to compare not only the growth performance but also carcass characteristics after slaughter by feeding domestic alfalfa to Hanwoo for a longer period during their growing stage.

Drought stress is one of the major factors that reduce plant growth and productivity. This study was conducted to investigate the effect of exogenous acetic acid pretreatment on drought stress tolerance response in plants. Fourteen-day-old alfalfa plants were pretreated with 15 mM acetic acid, and then subsequently subjected to drought stress for 6 days. The fresh weight and relative water content in the leaves of acetic acid pretreated alfalfa plants were increased compared to the control group. The chlorophyll and carotenoid contents were slightly decreased in the acetic acid treatment. The H2O2 and proline contents were also significantly decreased in the acetic acid treatment. These results suggest that the scavenging mechanism of reactive oxygen species in alfalfa activated by acetic acid pretreatment is involved in conferring tolerance to drought stress.

A new variety of Alfalfa (Medicago sativa L.), named 'Alfaking' was developed between 2015 and 2023 at the Grassland and Forages Division, National Institute of Animal Science, Rural Development Administration, Cheonan, Republic of Korea. The variety was produced through artificial hybridization, with ‘Paravivo’ serving as the maternal line and ‘WL514’ as the paternal line. ‘Alfaking’ underwent field tests across four regions (Cheonan, Pyeongchang, Jeongeup, and Jinju) to evaluate its agronomic characteristics and forage production over two years (2022-2023). The dry matter yield of ‘Alfaking’ reached 22,516 kg/ha, which is 11% higher than the control variety, ‘Vernal.’ ‘Alfaking’ exhibited 2.1% higher crude protein content than ‘Vernal’ in forage nutritive value. The development of this new alfalfa variety, which exhibits excellent adaptability to challenging environmental conditions, is expected to enhance forage cultivation and productivity in Korea.

The alfalfa weevil (Hypera postica) is an important pest that causes significant damages to alfalfa crops, reducing yield and quality, but there’s a solution. This research had two main goals to evaluate the efficacy of insecticides available in Korea for alfalfa weevil control and to provide data for pest management studies. The experiment, conducted from 2023 to 2024 at the National Institute of Animal Science in Cheonan, Republic of Korea, included four treatment plots: control (CON), early-occurrence (EAR), mid-occurrence (MID), and late-occurrence (LAT). It also included two frequency plots with primary and secondary insecticide using 50% fenitrothion emulsion, which made it truly comprehensive study. The primary insecticide was applied at the first observation of the alfalfa weevil larvae, with subsequent secondary applications at specified intervals. The results showed that two-times insecticide applications significantly reduced larvae populations and increased yield and nutrient content compared to a single application. Specifically, control rates ranged from 94 to 94.7% on the third day after treatment and from 72.2 to 93.4% on the seventh day. Plots with two applications had higher yields and crude protein content. The study concluded that the timing and frequency of insecticide applications are critical to maximizing alfalfa yield and quality, emphasizing the importance of optimized application strategies for effective pest control.

This study aims to contribute to resolving the critical issue of weed management in newly established alfalfa fields, study has been conducted on effective herbicide use. The study evaluated the impact of various domestically available foliar herbicides on alfalfa phytotoxicity, weed control, yield, and nutritive value. The experiment was designed in a randomized complete block design with four treatments. Alfalfa ‘SW 5615’ seeded in the spring of 2024 on a 1 ha field (March 18), with herbicide treatments including fluazifop-P-butyl (FPB), bentazone (BEN), and a mixture of these herbicides (MIX). Herbicide efficacy, alfalfa yield, and nutritive value were assessed 30 days post-application. Results indicated that the MIX treatment achieved superior weed control comparable to hand weeding (HW), although it exhibited higher phytotoxicity, requiring extended recovery periods. While MIX led to lower overall yield, it enhanced alfalfa purity, resulting in higher crude protein (CP) content and relative feed value (RFV) compared to other treatments. The study concludes that despite the potential for increased phytotoxicity, mixed herbicide treatments could offer a strategic advantage in enhancing the quality of alfalfa feed through effective weed management, thereby improving CP and RFV, critical factors for the nutritional value of alfalfa. These findings provide valuable insights for optimizing weed management practices in alfalfa cultivation, suggesting that mixed herbicide application, although associated with increased phytotoxicity on the plants, could improve the overall feed quality by reducing weed competition.

The objective of this study was to investigate the comparison of feeding a total mixed ration (TMR) containing imported alfalfa hay and TMR containing hot-air dried domestic alfalfa on rumination activity, milk production, and composition in lactating dairy cows. Ten Holstein dairy cows were divided into two groups: control (imported alfalfa hay + TMR) and treatment (hot-air dried domestic alfalfa + TMR) groups. The study was conducted over a total period of 18 days, including a 13-days adaptation period and a 5-days main experimental period. The results indicated no differences in total digestible nutrient and net energy intake between the imported and hot-air dried alfalfa. Body weight was not significantly different between the two groups (p>0.05), however, total feed intake and rumination time were significantly and tendentially higher in the treatment group compared with control group, respectively (p<0.001; p=0.075). Milk yield was not significantly different between the two groups (p>0.05), however, milk fat (kg) and lactose (%) concentration were significantly higher in the treatment group compared with control group (p=0.016; p=0.02). This study confirms that feeding TMR with hot-air dried domestic alfalfa results in no differences of feed intake, rumination activity, and milk productivity. Therefore, it is considered that hot-air dried domestic alfalfa can be used as a substitute for imported alfalfa on lactating dairy cows.

It would be advantageous to grow legume forage crops in order to increase the productivity and sustainability of sloped croplands in Hamkyongbukdo. In particular, the identification of potential cultivation areas for alfalfa in the given region could aid decision-making on policies and management related to forage crop production in the future. This study aimed to analyze the climate suitability of alfalfa in Hamkyongbukdo under current and future climate conditions using the Fuzzy Union model. The climate suitability predicted by the Fuzzy Union model was compared with the actual alfalfa cultivation area in the northern United States. Climate data obtained from 11 global climate models were used as input data for calculation of climate suitability in the study region to examine the uncertainty of projections under future climate conditions. The area where the climate suitability index was greater than a threshold value (22.6) explained about 44% of the variation in actual alfalfa cultivation areas by state in the northern United States. The climatic suitability of alfalfa was projected to decrease in most areas of Hamkyongbukdo under future climate scenarios. The climatic suitability in Onseong and Gyeongwon County was analyzed to be over 88 in the current climate conditions. However, it was projected to decrease by about 66% in the given areas by the 2090s. Our study illustrated that the impact of climate change on suitable cultivation areas was highly variable when different climate data were used as inputs to the Fuzzy Union model. Still, the ensemble of the climate suitability projections for alfalfa was projected to decrease considerably due to summer depression in Hamkyongbukdo. It would be advantageous to predict suitable cultivation areas by adding soil conditions or to predict the climate suitability of other leguminous crops such as hairy vetch, which merits further studies.

This study was conducted to determine the optimal spring seeding dates for alfalfa yield and feed value. The experiment was conducted annually for three years (2021~2023) at the field in the Department of Animal Resources Development, NIAS, located in Cheonan. The treatments involved six seeding dates ranging from February 24 to April 14, with 10days intervals. Alfalfa was harvested four times a year at the early flowering stage. Dry matter yield showed a tendency to decrease with delayed the seeding date. However, depending on the climatidc condisions in the seeding year, the dry matter yield on March 14 or 24 was comparable to that on February 24. Annual dry matter yield varied, influenced by the daylight conditions each year. The average feed value did not significantly differ within in the same year with delayed seeding dates (p>0.05). Therefore, the most stable period for alfalfa spring seeding in the central area of South Korea is considered to be from February 24 to April 4, with February 24 indentified as the optimal date.

In this study, we examined the antagonistic effects of sprout-borne lactic acid bacteria (LAB) on Salmonella enterica serovar Enteritidis. This antagonism is promoted as a means of controlling contamination during sprout production and provides additional LAB for consumers. We isolated a total of 24 LAB isolates in nine species and five genera from seven popular vegetable sprouts: alfalfa (Medicago sativa), clover (Trifolium pratense), broccoli (Brassica oleracea ssp. italica), vitamin (B. rapa ssp. narinosa), red radish (Raphanus sativus), red kohlrabi (B. oleracea var. gongylodes), and Kimchi cabbage (B. campestris var. pekinensis). Based on 16S rRNA gene sequences, the LAB species were identified as Enterococcus casseliflavus, E. faecium, E. gallinarum, E. mundtii, Lactococcus taiwanensis, Leuconostoc mesenteroides, Pediococcus pentosaceus, and Weissella cibaria, and W. confusa. A total of 16 LAB isolates in seven species including E. faecium, E. gallinarum, E. mundtii, L. taiwanensis, L. mesenteroides, P. pentosaceus, and W. cibaria showed antagonistic activity toward S. enterica. The growth inhibition of sprout LAB on S. enterica was confirmed by co-culture. Unexpectedly, sprout LAB failed to suppress the growth of S. enterica in alfalfa sprouts, whereas all LAB strains stimulate S. enterica growth even if it is not significant in some strains. The findings of this study indicate that S. enterica-antagonistic LAB are detrimental to food hygiene and will contribute to further LAB research and improved vegetable sprout production.

본 연구는 수분함량과 미생물 첨가제가 알팔파 사일리지의 발효특성과 사료가치에 미치는 영향을 알아보기 위해 수행되었다. 알팔파는 개화 10% 시기에서 수확되었으며 수분 함량(M60, M50, M40 및 M30)이 60, 50, 40 및 30%일 때 각각 이용하였다. 1500g을 샘플링한 후, 증류수 10mL을 첨가한 미첨가구(NAD)와 Lactococcus lactis 와 Pediococcus pentosaceus의 혼합물을 증류수(0.1g/10mL)에 1.5 x 1010cfu/g 농도로 희석한 후 접종한 미생물 균주 첨가구(ADD)를 3개월 및 6개월 발효하였다. 수분 및 미생물 첨가제에 따른 연평균 알팔파의 조단백, 중성세제 불용성 섬유 및 산성세제 불용성 섬유에서 차이가 없었다(p>0.05). 모든 발효기간에서 pH는 ADD의 M40에서 가장 낮았다(p<0.05). NAD 처리구에서는 젖산이 M50에서 가장 높았고(p<0.05), ADD 처리군에서는 M40 시험구에서 젖산이 가장 높았다(p<0.05). NAD와 ADD의 M60은 젖산 함량이 다른 수분 함량들에 비해 가장 낮으며(p<0.05) 낙산이 유일하게 검출되었다. 미생물군집의 상대적 풍부도는 ADD 처리구의 M40과 M50에서 Homo LAB (Enterococcus, Lactiplantibacillus, Lacticaseibacillus, Lactococcus, Pediococcus)의 비율이 가장 높았고, Clostridium은 M60에서 가장 높았다.

Iron (Fe) is a vital element for plants and other organisms, involving in several physiological processes including respiration, chlorophyll biosynthesis, and photosynthesis. Unfortunately, how Fe accumulation regulates in response to light quality has not been well established in plants. Therefore, the aim of the study was to explore the mechanism of Fe homeostasis by light quality. In this study, we found morpho-physiological attributes were significantly improved in response to blue (λmax: 450) compared to white (λ max: 500) and red (λmax: 660) light. The root-shoot length, plant biomass, photosynthesis efficiency (Fv/Fm) and leafgreen (SPAD) significantly declined in response to white and red light. However, these parameters were improved and iron deficiency was substantially alleviated by blue light exposure in alfalfa seedlings. This study might be useful to the forage breeders and farmers for improving alfalfa yield and nutritional benefits.

Silicon (Si) has the potential to improve plant growth and stress tolerance. The study aimed to explore Si-involving plant responses and molecular characterization of different Si-responsive genes in alfalfa. In this study, the exogenous supplementation of Si enhanced plant growth, and biomass yield. Si-acquisition in alfalfa root and shoot was higher in Si-supplemented compared to silicon deficient (-Si) plants, implying Si-acquisition has beneficial on alfalfa plants. As a consequence, the quantum efficiency of photosystem II (Fv/Fm) was significantly increased in silicon-sufficient (+Si) plants. The quantitative gene expression analysis exhibited a significant upregulation of the Lsi1, Lsi2, Lsi3, NIP5;1, and NIP6;1 genes in alfalfa roots, while BOR1, BOR4, NIP2, and NIP3 showed no significant variation in their expression. The MEME results further noticed the association of four motifs related to the major intrinsic protein (MIP). The interaction analysis revealed that NIP5;1 and Lsi1 showed a shared gene network with NIP2, BOR1, and BOR4, and Lsi2, Lsi3 and NIP3-1, respectively. These results suggest that members of the major intrinsic proteins (MIPs) family especially Lsi1, Lsi2, Lsi3, NIP5;1, and NIP6;1 genes helped to pass water and other neutral solutes through the cell membrane and those played significant roles in Si uptake and transport in plants. Together, these insights might be useful for alfalfa breeding and genome editing approaches for alfalfa improvement.