• Asian Journal of Turfgrass Science
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• v.18 no.4
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• pp.211-219
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• 2004

Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

• Asian Journal of Turfgrass Science
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• v.18 no.3
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• pp.141-148
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• 2004

In this report, several factors such as infection time and concentration of bacterial suspension, influencing on transient gene expression in Agrobacterium-mediated transformation were evaluated. An appropriate concentration (O.D 600nm = 1.0-1.2) of bateria and 30 min of infection time showed a higher level of GUS expression. To improve transformation efficiency (TE), friable embryogenic calli (FEC) were bombarded by tungsten particles without plasmid DNA, and then co-cultivated with A. tumefaciens LBA4404 which contains pTOK233 super binary vector, carrying neomycin phosphotransferase (NPTII), hygromycin phosphotransferase (hpt) and$\beta-glucuronidase$ (GUS) genes. Three days after co-cultivation with A. tumefaciens and particle bombardment, FEC cultures were transferred to the selection medium (SM: MS medium supplemented with BA 1mg/l, hygromycin 100mg/l, cefotaxime 250 mg/l and vancomycin 200mg/l). They were cultured for 2 weeks and then transferred to the second SM containing hygromycin 50mg/l, cefotaxime 200 mg/l and vancomycin 100mg/l. Later, stable GUS expression was detected 4 to 6 weeks after transfer to the SM. Further, TE from Agrobacterium-mediated transformation after particle bombardment increased to about 3-folds compared with Agrobacterium-mediated transformation without particle bombardment. In the present study, we established an efficient transformation protocol of zoysiagrass by using A. tumefaciens in the combination with particle bombardment for the first time.

• Asian Journal of Turfgrass Science
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• v.15 no.1
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• pp.9-14
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• 2001

Callus formation and plant regeneration from the seeds of zoysiagrass cv. Zenith was tested on MS basal medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid(2,4-D) and of several cytokinins. A concentration of 1mg/L 2,4-D on medium stimulated callus formation. In the presence of 5mg/L 2,4-D, addition of 1mg/L kinetin significantly enhanced callus formation and plant regeneration over 2,4-D alone. To transfer foreign DNA into turfgrass, parameters for the bombardment of embryogenic callus with the particle bombardment were partially optimized using transient expression assay of a $chimeric \beta$-glucuronidase(GUS) gene driven by the CaMV 35S promoter. GUS gene was strongly expressed at helium pressure 1,100 psi and 6~9cm target distance.

• Proceedings of the Turfgrass Society of Korea Conference
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• 2005.04a
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• pp.24-25
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• 2005

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.327-335
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• 2009

Creeping bentgrass (Agrostis stolonifera L.) is economically important as the principal turfgrass species for golf course greens and fairways in temperate climates around the world. As the utilization area of the turfgrass species increases recently, the demand for new and improved cultivars increases. Thus, substantial progress has been made in applying modern biotechnology to develop genetically engineered (i.e., biotech) creeping bentgrass with new traits that eluded the breeders. This review article addresses the advances made in developing biotech creeping bentgrass, which are categorized in the following topics: (i) genetic transformation of creeping bentgrass, (ii) development of various biotech creeping bentgrasses by genetic engineering, and (iii) progresses in the deregulation of herbicideresistant creeping bentgrass.

• Asian Journal of Turfgrass Science
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• v.7 no.1
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• pp.31-34
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• 1993

There is a growing concern in the United Stares over the environmental and human health implications associated with heavy use of water, pesticides, and inorganic ferilizers in maintaining picture perfect golf courses. There is also a growing awareness that a beautiful course is not necessarily a healthy course. The following discussion reviews the interrelationship of turfgrass and the soil that supports it and provides basic information on currently available alternatives to turf management practices that feature intensive application of inorganic fertilizers. water and pesticides. Soil is a dynamic natural environment in which microorganisms play an important role. Soil contains a large mass of microorganisms which produce thousands of enzymes that can catalyze the transformation and degradation of many organic molecules. (In top soil under optimum conditions may contain 10 billion cells per gram of soil.). Turfgrass and the soil which supports it are interdependent. The natural organic cycle as applied to turf and soil begins with healthy vigorous grass plants storing up the sun's energy in green plant tissues as chemical energy. Animals obtain energy by eating plants and when plants and animals die, their wastes are returned to the soil and provide "food" for soil microorganisms. In the next step of the organic cycle soil microorganisms break down complex plant tissues into more basic forms and make the nutrients available to grass roots. Finally, growing plants extract the available nutrients from the soil. By free operation of this organic cycle, natural grasslands have some of the most fertile soils on earths.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.10a
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• pp.103-103
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• 2004

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.400-407
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• 2010

Zoysiagrass (Zoysia japonica Steud.), also called Korean or Japanese lawngrass, is the most popular warm-season turfgrass in Korea and is widely used for home lawns, parks, roadsides, golf courses and athletic fields. Its use is rapidly expanding in Korea and the other countries, due to its excellent characteristics which include tolerance to heat, drought and salinity. As the utilization area of this turfgrass increases, there is an increase in the demand for improved cultivars with disease and insect tolerance or with herbicide-tolerance or with extended greening periods. Conventional breeding methods have been used to improve the traits described above with limited success. However, with the advances in biotechnology, genetic transformation can be utilized for turfgrass improvement. In this paper, we review recent progress in biotechnological improvement of zoysiagrass and discuss future molecular breeding of this species.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.4
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• pp.427-432
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• 2009

Seashore Paspalum (Paspalum vaginatum Swartz) is a warm season grass and indigenous to tropical and subtropical regions of coastal areas worldwide. The species is used as feed for cattle and horses and has been very successful for golf courses worldwide. One of the most outstanding characteristics of seashore paspalum is its tolerance to saline soils compared to other warm season turfgrasses. The development of new seashore paspalum cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to product for herbicide resistant seashore paspalum using Arobacterium-mediated transformation and this study is the first report on transformation and herbicideresistant transgenic plants in seashore paspalum. Embryogenic calli were induced from the seeded variety of pseashore paspalum. Embryogenic calli were transformed with Agrobacterium tumefaciens strain EHA105 carrying the binary vector pCAMBIA3301 with two genes encoding gusA and bar. Transformed calli and plants were selected on medium containing 3 mg/l PPT. PCR detected the presence of the gusA and bar gene, indicating both genes are integrated into the genome of seashore paspalum. A chlorophenol red assay was used to confirm that the bar gene was expressed. By application of herbicide BASTA, the herbicide resistance in the transgenic seashore paspalum plants was confirmed.

• Asian Journal of Turfgrass Science
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• v.12 no.4
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• pp.195-202
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• 1998

The development of a protocol for high efficiency of embryogenic callus separation, maintenance and plant regeneration from the seeds of zoysiagrass cv. Zenith was studied. Embryogenic callus ratio is absolutely determined by genotype, but by adding high concentration of copper into medium, changing light condition and maintaining callus on initial induction medium for 8∼10 weeks, embryogenic callus can be easily distinguished and its growth can be promoted. There were significant differences among selected callus lines (each from one seed) according to their growth rates and regeneration percentages. Callus pre-treatment with activated charcoal inhibited callus growth, increased the level of precocious germination during culture and promoted shoot cluster formation after transfer to regeneration medium. For long-term callus maintenance, N6AA medium was better than MS medium, because the former inhibited non-embryogenic callus formation and kept vigor of embryogenic callus. The best callus lines Z-(5) has been successfully used for transformation and somaclonal variation selection in our laboratory.