• Horticultural Science & Technology
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• v.35 no.3
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• pp.344-360
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• 2017

Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

• Journal of Plant Biotechnology
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• v.43 no.2
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• pp.213-222
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• 2016

This study aims to establish a system for the rapid discrimination of Zoysia species using metabolite fingerprinting of FT-IR spectroscopy combined with multivariate analysis. Whole cell extracts from leaves of 19 identified Zoysia japonica, 6 identified Zoysia sinica, and 38 different unidentified Zoysia species were subjected to Fourier transform infrared spectroscopy (FT-IR). PCA (principle component analysis) and PLS-DA (partial least square discriminant analysis) from FT-IR spectral data successfully divided the 25 identified turf grasses into two groups, representing good agreement with species identification using molecular markers. PC (principal component) loading values show that the $1,100{\sim}950cm^{-1}$ region of the FT-IR spectra are important for the discrimination of Zoysia species. A dendrogram based on hierarchical clustering analysis (HCA) from the PCA and PLS-DA data of turf grasses showed that turf grass samples were divided into Zoysia japonica and Zoysia sinica in a species-dependent manner. PCA and PLS-DA from FT-IR spectral data of Zoysia species identified and unidentified by molecular markers successfully divided the 49 turf grasses into Z. japonica and Z. sinica. In particular, PLS-DA and the HCA dendrogram could mostly discriminate the 47 Z. japonica grasses into two groups depending on their origins (mountainous areas and island area). Considering these results, we suggest that FT-IR fingerprinting combined with multivariate analysis could be applied to discriminate between Zoysia species as well as their geographical origins of various Zoysia species.

• Weed & Turfgrass Science
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• v.2 no.2
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• pp.191-197
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• 2013

Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

• Proceedings of the Turfgrass Society of Korea Conference
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• 2005.04a
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• pp.31-38
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• 2005

본 실험은 한국잔디 신품종 '세녹'(특허출원:10-2003-0072018) 개발에 관한 것이다. 한국에서 수집한 유전자원 중 갯잔디와 금잔디간에 인공교배를 수행해서 F1계통을 얻었다. 작성한 F1계통을 포장상태에서 방임수분을 통해 변이확대를 하였으며, 이들 방임수분 후대에서 선발한 것이 '세녹'이다. '세녹'은 유전적으로 잔디의 녹색도가 진하며, 낮게 자라는 특성이 있고 품질이 우수하며 잎의 강직성이 높다. 엽폭이 3.1mm로 중엽형이며, 초장이 14cm로 낮고, 잎각도가 67.3도로 넓다. 지면으로부터 최하위 엽의 잎 몸 기부까지의 길이가 2cm로 낮은 형태적 특성을 갖고 있어 낮은 잔디깍기에 적응할 수 있는 영양번식형 잔디이다. 또한 RAPD 밴드 특성에서 한국잔디 신품종 세녹(Senock)이 특이 밴드를 갖고있어 식별성이 높다.

• Asian Journal of Turfgrass Science
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• v.24 no.2
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• pp.88-96
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• 2010

Zoysiagrass, especially Jungji implicating medium-leaf type zoysiagrass has been widely used in South Korea recently. This study was carried out to classify 36 selected medium-leaf type zoysiagrass accessions compared to 5 basic zoysiagrass species using RAPDs. Morphological characteristics such as leaf width, leaf angle, leaf sheath length, existance of trichomes and stolon color were measured as useful characteristics for identification of species in Zoysia genus. Nineteen RAPD markers were identified using 8 selected random primers. The dissimilarity coefficient of variants ranged from 0 to 0.736. Three zoysiagrass groups were clustered by dissimilarity coefficient analysis. Group 1 consisted of Z. japonica and some US varieties including 'Zenith' and 'Meyer'. Group 2 consisted of Z. sinica, Z. macrostachya and Korean commercial varieties such as 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type grasses. Group 3 was genetically distinct from Group 1 and Group 2, and included Z. matrella and Z. tenuifolia. 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type zoysiagrasses showed very close genetic relationship with Z. sinica and Z. macrostachya.

• Asian Journal of Turfgrass Science
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• v.20 no.1
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• pp.1-10
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• 2006

This study was carried out to develope new zoysiagrass cultivar 'Millock'(Patent registration No. : 10-2005-0110051). Artificial selfing of collected line of MJ8 was conducted to develope F1 plant (MJ8S). Among the inbred progenies, MJ8S-9 (Millock) showed superior performance in color, density, and rust resistance. 'Millock' showed genetically dark green color, with medium-textured leaf ($4.2{\pm}0.44$ mm), short internode length ($3.5{\pm}0.28$ cm), and wide leaf angle ($52.5{\pm}10.8$ degree). Height to the lowest leaf blade of this cultivar was $1.9{\pm}0.91$ cm, which may allow low mowing height. 'Millock' has a yellowish green stolen. Also, specific bands with primer number OPB 8 by RAPD analysis can be used for a cultivar identification.

• Asian Journal of Turfgrass Science
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• v.18 no.4
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• pp.201-209
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• 2004

This study was carried out to develope new zoysiagrass cultivar 'Senock' (Patent registration: 10-2003-0072018). Artificial crossing between collected lines of Z. sinica and Z. matrella (collected in South Korea) was conducted to develope F1 plant (CSM) which was cultivated at field for open self-pollination. Among the open pollinated progenies, CSM8 (Senock) showed superior performance in color and density rating. 'Senock' showed genetically dark green color, with medium leaf width ($3.1\pm0.17mm$), low plant height ($14\pm5.67cm$), and wide leaf angle ($67.3\pm11.1degree$). Height to the lowest leaf blade of this cultivar was $2.0\pm0.14cm$, which may allow low mowing height. Specific bands with primer number OPB7 by RAPD analysis can be used for cultivar identification.

• Korean Journal of Environment and Ecology
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• v.23 no.6
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• pp.499-505
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• 2009

This study was carried out to investigate and evaluate the vegetations in the coastal dunes in the vicinity of swimming beaches on the East Sea, South Korea, and the vegetations that were investigated are as follows: Carex pumila community, Cynodon dactylon community, Zoysia macrostachya community, Rosa rugosa community, Vitex rotundifolia community and Carex kobomugi typical community. Some of these vegetations, such as Carex kobomugi, Calystegia soldanella, Ixeris repens, Elymus mollis, Lathyrus japonica, and Glehnia littoralis were found to be differential species which belongs to Caricion kobomugi of Glehnietea littoralis. It was also discovered that Vitex rotundifolia community was the most heterogeneous among vegetation communities of the coastal dunes surrounding swimming beaches, and Zoysia macrostachya community was most closely linked to the typical coastal dune vegetation community. Finally, Rosa rugosa community and Vitex rotundifolia community seemed to have formed near the hinterland of coastal dunes and swimming beaches respectively because of certain unnatural disturbances such as construction of buildings and facilities or natural disasters.

• Asian Journal of Turfgrass Science
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• v.19 no.2
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• pp.85-94
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• 2005

A fungal isolate was newly collected from Zoysiagrass 'Anyang-Joongji' in small circular patches on a fairway ofa golf course in Korea, which seriously occurred during the early summer period of 2005. The isolate presented on PDAmedium, named Scz1, was closely identical to Sclerotinia homoeocarpa, a casual fungus of dollar spot disease, in cool season turf grasses such as creeping bentgrass. Hereby, this study was accomplished to characterize the isolate and compare it with the fungus, named Scb1, isolated from dollar spot-infected creeping bentgrass (Agrostis palustris Huds. cv Penncross). On PDAmedium, individual mycelial appearance of three isolates was very similar except for the pigment. Mycelial pigments of Scz1 and Scz2 (another analogous isolate collected) were light pinkish on the reverse side of PDA medium but that of Scb1 was dark brownish. In a microscopic study, three isolates were barely distinguishable in the appearance of mycelia. As expected, in the temperaturesensitivity assay, all pathogens were very delicate to $32^{circ}C$ above but not to $30^{circ}C$ below, in which was explained to be one of typical characteristics in S. homoeocarpa. In an artificial inoculation assay, disease symptoms including leaf spots in Zoysiagrass were appeared within 6-7 days after inoculation through the hand inoculation method with the isolate-infested soil. Then the fungus was re-identified from the infected leaf tissues. Interestingly, inoculation of isolate Scz1 gave rise to distinct symptoms in only Zoysiagrass but not in creeping bentgrass 'Penncross' and Kentucky bluegrass 'Midnight'. The observation might be involved in host specific pathogenecity of S. homoeocarpa Scz1 to Zoysiagrass. In a chemical sensitivity assay for the isolate, Scz1, showed a high mycelial inhibition against two fungicides, iprodione and propiconazole. All results described above suggest that S. homoeocarpa Scz1 is a primary pathogen of Zoysia dollar spot disease.

• Korean Journal of Agricultural Science
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• v.40 no.2
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• pp.115-121
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• 2013

Polymorphic bands of two fine-leaf zoysiagrass mutants (CNU 70-1, CNU 70-2) induced via a gamma-ray irradiation on seeds of Zoysia japonica were obtained by using randomly amplified polymorphic DNA (RAPD) primers. The genotype-specific fragments were then converted into PCR-based sequence characterized amplified region (SCAR) markers, which are now amenable to detecting them among other zoysiagrass species widely noticeable in Korea. The CNU 70-1-specific primer set amplified about 900 bp successfully, while the CNU 70-6 marker produced the expected 1,500 bp band, by which those markers were nominated by CNU 70-1_900 and CNU 70-6_1500 SCARs, respectively. The developed SCAR markers can be an applicable tool in sod industry where illegal appropriation hampers breeder's right and profits due to the turfgrass plant vegetatively propagating.