• Journal of The Korean Society of Grassland and Forage Science
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• v.27 no.4
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• pp.235-240
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• 2007

Optimum tissue culture conditions for an efficient induction of embryogenic callus from mature seeds of perennial ryegrass (Lolium perenne L.) and regeneration of plants from callus tissues were investigated. MS medium containing 3 mg/L 2,4-D and 0.1 mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency (58.3%) was observed when the embryogenic callus tissues were cultured on N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be helpful for molecular breeding of perennial ryegrass through Agrobacterium-mediated genetic transformation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.14 no.3
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• pp.177-185
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• 1994

Slow seedling growth rate and nodulation failure of white clover (Trifolium repens L.) has been limited its good establishment to pastures. The experiment was done to determine the effect of removal of cotyledon and unifoliolate on the shoot, root growth, and nodule formation of 4 white clover cultivars for 8 weeks after the treatment. Four white clover cv. Regal (large leaf), Louisiana S-I (medium-large leaf), Grasslands Huia (mediumsmall leaf), and Aberystwyth S184 (small leaf), were grown in IOcm plastic pot containing 2:l:l soi1:sand:peat moss mixture until grown to cotyledon or unifoliolate stage and then removed one (Cl) or two cotyledons (C2) at cotyledon stage, and unifoliolate only (U), unifoliolate and one cotyledon (UCl) or unifoliolate and two cotyledons (UC2) at the unifoliolate stage, and the plants were sampled at 4, 6 and 8 weeks after the treatments. The intact plants had greater shoot and root dry weights, and no. of nodules than removal-treated ones. Removal treatments at cotyledon stage, the dry weight and no. of nodules more decreased in C1 and C2 than that of unifoliolate stage. While the severer cotyledon removal, the more reduction. Although the dry weights and no. of nodules steadily inclined with regrowing period, the former were higher in Regal and La. S-1 than in the others since 6 weeks after removal treatment but the latter was more in S 184 than in the others 8 weeks after removal treatment. Relationship between no. of nodules and shoot or root dry weight was analysed as linear mode while the earlier and severe removal, the steeper slope. It was concluded that severer damage of cotyledon and unifoliolate had detrimental effects on the shoot and root growth, nodule formation, and aftermath establishment of white clover.

• Journal of The Korean Society of Grassland and Forage Science
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• v.7 no.2
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• pp.87-91
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• 1987

This study was carried out to investigate the effects of different cutting times at 0, 10 and 20 days after lodging on the percentage of dead plants and bare ground, live and dead matter build-up, and botanical composition of orchardgrass dominated pasture during the mid summer season. For the test, a randomized block design with 3 replications was treated with 3 different cutting times (July 17, July 27 and Aug. 6). The results obtained are summarized as follows: 1. The percentage of dead plants with cutting times at 0, 10 and 20 days after lodging was 26.3,41.7 and 46.7'%, respectively. 2. The percentage of bare ground with cutting times at 0, 10 and 20 days after lodging was 13.3,26.7 and 3 1.7%, respectively. 3. Dead matter build-up with cutting times at 0, 10 and 20 days after lodging was increased significantly with later cutting (p<0.05). The ratio of live and dead matter build-up with cutting times at 0, 10 and 20 days after lodging was 90.2:9.8, 47.7:52.3 and 39.1:60.9%, respectively. 4. Regrowth yield after lodging was decreased significantly (p<0.05) with later cutting time. 5. The percentage of legumes in botanical composition was increased greatly with later cutting time. The ratio of grasses and legumes with cutting times at 0, 10 and 20 days after lodging was 77.8:21.6,45.5: 50.3 and 36.5:61.7%, respectively.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.87-93
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• 2003

Brachiaria (Poaceae) is the most important forage genus for cattle production in Brazil. The genetic breeding of this genus is limited by the incompatibility among species, differences in ploidy level and the natural cloning of plants by apomixis (Valle and Miles 1992). However, plant regeneration via tissue culture methods and genetic engineering provide an opportunity to introduce new characteristics in plants of this genus. We have developed methods for the 'genetic modification of Brachiaria brizantha cv. Marandu via biolistic transformation. A higher number of shoots was obtained with 4 mg/L 2.4-diclorophenoxyacetic acid and 0.2 mg/L benzylaminopurine in calli induction medium and 0.1 mg/L naphtaleneacetic acid and 4.0 mg/L kinetin in shoot regeneration medium. A selection curve for mannose was determined to use phospho mannose isomerase (PMI) gene of Escherichia coli as a selection marker. Calli formation was inhibited from 5 g/L mannose, even in the presence of sucrose while calli that were formed in the presence of mannose failed to develop embryos showing that PMI gene can be used for selection of transformants of this grass. Different promoters were tested to evaluate the efficiency based on the detection of the GUS gene expression (Jefferson et al. 1987). The monocot promoters, act1-D and ubi-1, resulted in higher expression levels than dicot promoters, ubi-3 and act-2, or the CaMV35S and CVMV promoters.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.3
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• pp.231-236
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• 2017

Salt stress is one of the most limiting factors that reduce plant growth, development and yield. However, identification of salt-inducible genes is an initial step for understanding the adaptive response of plants to salt stress. In this study, we used an annealing control primer (ACP) based GeneFishing technique to identify differentially expressed genes (DEGs) in Italian ryegrass seedlings under salt stress. Ten-day-old seedlings were exposed to 100 mM NaCl for 6 h. Using 60 ACPs, a total 8 up-regulated genes were identified and sequenced. We identified several promising genes encoding alpha-glactosidase b, light harvesting chlorophyll a/b binding protein, metallothionein-like protein 3B-like, translation factor SUI, translation initiation factor eIF1, glyceraldehyde-3-phosphate dehydrogenase 2 and elongation factor 1-alpha. These genes were mostly involved in plant development, signaling, ROS detoxification and salt acclimation. However, this study provides new molecular information of several genes to understand the salt stress response. These genes would be useful for the enhancement of salt stress tolerance in plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.41 no.3
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• pp.176-182
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• 2021

Ensiling is the most preferred technology to preserve the silage quality with high nutrients by the presence of lactic acid bacteria. In this study, lactic acid bacteria RJ1 and S22 were used to make the silages from different leguminous plants such as alfalfa, hairy vetch and red clover. Experimental groups were divided into control and LAB inoculated groups. LAB inoculated group; all legumes treated with a mixture of RJ1 and S22 and made an anaerobic condition for 45d. Without the addition of LAB considered the control group. The results showed that the lactic acid content was higher in all silages in response to LAB treatment and acetic acid content was slightly increased except red clover by LAB compared to control silages. A poor silage quality marker butyric acid was reduced all legume silages in response to LAB inoculation than control silages. The organic acid is closely associated with microbial population experimental silages. We noted that higher LAB and lower yeast were found in the silage in response to LAB treatment. The contents of crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and total digestible nutrient (TDN) were not altered significantly between control and LAB treated silages. Overall data suggested that the inclusion of additional LAB potentially enhance the silage quality and preserved the nutrients for long period.

• The Plant Pathology Journal
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• v.39 no.2
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• pp.171-180
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• 2023

Spring black stem and leaf spot, caused by Phoma medicaginis, is an issue in annual Medicago species. Therefore, in this study, we analyzed the response to P. medicaginis infection in a collection of 46 lines of three annual Medicago species (M. truncatula, M. ciliaris, and M. polymorpha) showing different geographic distribution in Tunisia. The reaction in the host to the disease is explained by the effects based on plant species, lines nested within species, treatment, the interaction of species × treatment, and the interaction of lines nested within species × treatment. Medicago ciliaris was the least affected for aerial growth under infection. Furthermore, the largest variation within species was found for M. truncatula under both conditions. Principal component analysis and hierarchical classification showed that M. ciliaris lines formed a separate group under control treatment and P. medicaginis infection and they are the most vigorous in growth. These results indicate that M. ciliaris is the least susceptible in response to P. medicaginis infection among the three Medicago species investigated here, which can be used as a good candidate in crop rotation to reduce disease pressure in the field and as a source of P. medicaginis resistance for the improvement of forage legumes.

• Journal of The Korean Society of Grassland and Forage Science
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• v.38 no.3
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• pp.196-201
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• 2018

Soil is the main nitrogen (N) provider for plants but N in soil is not all available to advanced plants. Mineralization is a critical biological process for transferring organic N to inorganic N that can be used by plants directly. To investigate the effect of different levels of soil temperature and water content to soil mineralization, a field experiment was established on three different sites (A, B and C). We measured soil temperature, moisture and electrical conductivity once daily after swine slurry application. Average soil moisture and temperature in site A is the highest among three sites (40.9% and $9.7^{\circ}C$, respectively). Following is in site C (37.3% and $9.6^{\circ}C$) and the lowest is in site B (28.0% and $9.0^{\circ}C$). Ammonium N (NH4+-N) and nitrate N (NO3--N) were determined on the first and fifth day after treatment. Compared with site B and C, site A always had the highest soil total N content (1.54 g N kg-1 on day one; 1.22 g N kg-1 on day five) and highest NO3-- N content (93.18 mg N kg-1 on day one; 16.22 mg N kg-1 on day five) and a significant decrease on day five. Content of NH4+-N in site B and C reduced while in site A, it increased by 6.7%. Results revealed that net N mineralization positively correlated with soil temperature (P<0.5, $r=0.675^*$) and moisture (P<0.01, $r=0.770^{**}$), suggesting that to some extent, higher soil moisture and temperature contribute more to inorganic N that can be used by plants.

• Journal of The Korean Society of Grassland and Forage Science
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• v.17 no.1
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• pp.19-26
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• 1997

To find out the residual effect of phosphate fertilization on botanical composition, mineral contents and mineral nutrient deprivation of pasture plants, this experiment was arranged as a randomized complete block design with six treatments(0-0, 50-50, 100-65, 150-65, 200-65 and 250-65kg $P_2O_5$/ha), those were composed of three P,O, fertilization level(0, 50 and 65kg P,OJha) after phosphate fertilization trial with six treatments(0, 50, 100, 150, 200 and 250kg $P_2O_5$/ha) from 1989 to 1992, and conducted at hilly land in Kwangju, Kyonggi Province 60m 1993 to 1994. The results obtained are summarized as follows: The botanical composition of pasture plants without phosphate fertilization was very poor. With increasing available phosphate of soil, the percentage of grasses and legumes were remarkably increased and weeds was decreased. With 150-65kg $P_2O_5$/ha application for two years, the average DM yield was composed of 80.6% grasses, 15.4% legume and 4.0% weeds, the percentage was similar to that of 200-65kg $P_2O_5$/ha. As available phosphate of soil increase, P, K and Mg content of pasture plants were increased, but Ca/P ratio was lowed and the other mineral contents tended not to be regular. The mineral nutrient contents deprived by DM yield was increased as available phosphate of soil was increased, but those were not different between 150-65 and 250-65kg $P_2O_5$/ha. However, mineral nutrient of $P_2O_5$,$K_2O$ and MgO were higher in 200-65 and 250-65kg $P_2O_5$/ha. Most of applied mineral elements were accumulated at a subsurface(0-2.5cm) of grasslands, and mineral tended to be decrease than that of soil chemical analysis before the experiment of residual effect of phosphate fertilization.

• Journal of The Korean Society of Grassland and Forage Science
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• v.6 no.3
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• pp.151-156
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• 1986

There are differences in recommendations in the USA as to whether nitrogen fertilizer should be applied when establishing alfalfa (Medicago sativa L). The reason for not applying nitrogen is because some researchers found the addition of nitrogen reduced nodulation of alfalfa plants. A replicated experiment was conducted under controlled environmental conditions at the University of Nevada-Reno, Reno, Nevada, USA, to determine the effects of nitrogen application on seedling growth and nodulation of alfalfa when grown in a cool environment. A sterile sand was used in the growing media to which a complete nutrient solution minus nitrogen was applied volumetrically to each pot daily. Half of the pots received $NH_4NO_3$, at the rate of 11.2 kg/ha, at seeding and two and four weeks after planting, giving a total nitrogen application rate of 33.6 kg/ha. Rhizobia inoculant (R-12) consisted of a mixture of strains 171-15a. 1682c and 80 PI 265 of Rhizobium meliloti. Inoculant was applied to the seeds prior to planting and to the sand media at two and four weeks after seeding. Twenty seeds were planted in pots 14.0 cm in diameter and 11.5 cm deep. Seedlings were thinned after emergence to ten plants per pot. They were grown in a controlled environment chamber with a 16-hour light period. Soil temperatures at 6 cm depth ranged from $5.7^{\circ}C\;to\;21.5^{\circ}C$ and had a daily mean of $16.2^{\circ}C$ Plants were harvested at weekly intervals for seven weeks at which time root, shoot and total length, dry weight, volume and number of nodules per plant were determined. Root, shoot and total length were not affected by nitrogen fertilizer. However, application of nitrogen increased the size of the seedlings as determined by dry weight and volume when compared to plants which were not fertilized. This indicates that rhizobia did not fix enough atmospheric nitrogen to promote good growth. Nitrogen application resulted in significantly more nodules per plant. The effect of nitrogen fertilizer became more apparent as the plant became older. Results of this experiment show there are benefits from applying nitrogen at a low rate when establishing alfalfa under a cool environment.