• Journal of Plant Biology
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• v.35 no.2
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• pp.165-171
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• 1992

In order to study regulation of rbcL gene expression, rbcL gene of chloroplast DNA (Cp DNA) from maize was cloned. Cp DNA was isolated from intact chloroplast and digested with BamHI. BamHI 9 fragment of Cp DNA containing rbcL gene was ligated to pUC19 and transformed into E. coli DH5a. This recombinant plasmid was named pRLYSl. pRLYSl was hybridized with a part of rbcL gene from rice and digested with restriction enzyme BamHI, HindIIl, and PstI. From these results, it was confirmed that pRLYS1 contains intact rbcL gene and orientation of BamHI 9 fragment of Cp DNA in pRLYS1 was determined.rmined.

• Journal of Ginseng Research
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• v.19 no.1
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• pp.51-55
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• 1995

The DNA fragment containing ginseng ribulose-1,5-bisphosphate carboxytase/oxygenase large subunit(rbcL) gene was cloned from the ginseng chloroplast EcoRl library by colony lift hybridization with tobacco rbcL gene probe. From the screened clone, the DNA fragment containing ginseng rbcL gene was digested with several restriction enzyme and analyzed by Southern blot hybridization for the construction of restriction map. The ginseng rbcL gene fragment was subcloned in pBluescript II SK + vector and sequence analysis was performed. The nucleotide sequence of ginseng rbcL gene was compared with those of petunia, tobacco, alfalfa, rice and barley, which showed a homology of 93.1%, 95.2%, 90.5%, 85.5% and 84.3%, respectively.

• Journal of Ginseng Research
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• v.27 no.3
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• pp.146-150
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• 2003

This study was carried out to obtain basic information on breeding using PCR-aided RFLP technology which can identify the variation inter- and intra-species of ginseng in the level of DNA. It was intended to investigate banding pattern on psbA and rbeL genes of chloroplast DNA in ginseng after treating with restriction enzymes. To isolate psbA and rbcL genes of chloroplast, both psbA-N, psbA-C primer and rbcL-N, PX-1 primer were used. As a result, 1,008 bp band of psbA gene and 1,336 bp band of rbcL gene were appeared, which was optimal and expected molecular weight. In addition, primers to isolate atpB, rpoB, trnL, and trnF genes were used, resulting in the expected 1366, 900, 1500 and 1008 bp bands. Genes of psbA and rbcL isolated by PCR were cut by restriction enzymes, Sau3A, TaqI, AluI, HaeIII, and RFLP pattern was investigated. KG line and other species of ginseng were cut by TaqI treatment, and bands were located in 800 bp. The treatment treated by AluI also showed the same 800 bp band in KG line and other species. In HaeIII treatment, 500 bp of faint bands were shown in case of KG line, whereas any bands were not observed in other species. All chloroplast genes formed bands by PCR amplification. However, it was not evident to distinguish intra-or inter-species of ginseng after restriction enzyme treatment. Therefore, more restriction enzyme treatment or sequence comparison method should be considered for further experiment.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.53-58
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• 2013

Objectives : Pinelliae Tuber has been used as a typical unauthentic herbal medicines. Due to the morphological similarity between Pinelliae Tuber and adulterants, the correct authentication is very difficult. Therefore, we introduced DNA barcode to establish a powerful tool for the authentication of Pinelliae Tuner from adulterants. Methods : To obtain DNA barcode regions, genomic DNA was extracted from nineteen specimens of Pinellia ternata, Pinellia pedatisecta, Pinellia tripartita, and Typhonium flagelliforme, and matK and rbcL genes were amplified. For identification of species specific sequences and analysis phylogenetic relationship, a comparative analysis were performed by the ClastalW and UPGMA based on entire sequences of matK and rbcL genes, respectively. Results : In comparison of two DNA barcode sequences, we elucidated the phylogenetic relationship showing distinct four groups depending on species and identified 40 and 20 species specific nucleotides enough to distinguish each species from matK and rbcL gene, respectively. The sequence differences at the corresponding positions were avaliable genetic marker nulceotides to discriminate the correct species among analyzed four species. These results indicated that phylogentic and comparative analysis of matK and rbcL genes are useful genetic markers to authenticate Pinelliae Tubers. Conclusions : The marker nucleotides enough to distinguish P. ternata, P. tripatrita, P. peditisecta, and T. flagelliform, were observed at 40 positions in matK gene and 20 positions in rbcL gene sequence, respectively. These differences can be used to authenticate Pinelliae Tuber from adulterants as well as discriminate each four species.

• ALGAE
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• v.21 no.4
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• pp.417-423
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• 2006

Morphology and molecular phylogeny of a red algal species, Hypnea flexicaulis that is recently described from Japan, were investigated based on 23 collections from Korea (21), Taiwan (1), and the Philippines (1). Hypnea flexicaulis has percurrent axes with flexuous, antler-like branches which have wide branching angles, and abaxially curved ultimate branchlets. In order to study DNA divergence and phylogenetic relationships of the species, we determined plastid rbcL and mitochondrial cox1 sequences from the 23 collections. All 21 specimens from five different locations in Korea were almost identical to H. flexicaulis from Japan in rbcL sequences. Although there was a difference of three to five base pairs (bp) between samples from Korea and the Philippines or between the Philippines and Taiwan, Bayesian analyses of the rbcL data showed that all specimens from Korea, Japan, the Philippines, and Taiwan were strongly monophyletic. However, it is interesting that specimens from the Philippines differed by 31-34 base pairs in mitochondrial cox1 gene from those of materials from Korea and Taiwan, which differed by one to seven bp in rbcL between them. Although H. boergesenii is different from H. flexicaulis in having many antler-like branchlets, both appeared as sisters in all analyses of the rbcL data. This is the first report of H. flexicaulis from Korea based on morphology, rbcL, and cox1 gene sequences.

• Journal of Plant Biology
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• v.36 no.3
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• pp.203-210
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• 1993

In order to investigate the effects of plant hormones on the quantitative changes in mRNA of maize (Zea mays L.) rbcL, we used GA3, IAA, ABA and BAP. GA3 at the concentration of 10-4M resulted in decrease in rbcL gene transcript to 62%. IAA decreased the amount of rbcL transcript to about 70% at all the hormone concentrations tested. ABA did not cause a noticeable change in the amount of rbcL transcript, but BAP increased the amount of rbcL transcript to 153% at 10-8M and 123% at 10-5M, respectively. Thus, it appears that BAP is related to the increase in the amount of rbcL transcript by light.

• ALGAE
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• v.20 no.1
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• pp.15-23
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• 2005

Polysiphonia pacifica is rhodomelaceous red algal species that includes five varieties in Pacific Ocean: P. pacifica var. delicatula, P. pacifica var. distans, P. pacifica var. determinata, P. pacifica var. disticha, and P. pacifica var. gracilis. We here report morphology and phylogeny of P. pacifica to confirm the relationships among previously described varieties as a loan of type specimens from US and to assess phylogenetic relationships of closely related species using plastid protein-coding rbcL gene. Polysiphonia pacifica is distinguished by having creeping filaments attached by unicellular rhizoids not cut off by cross walls, four pericentral cells, ecorticate, trichoblasts rare, ultimate branchlets attenuate at the tip but not pungent, and tetrasporangia in long straight series in the ultimate branchlets. The protein-coding plastid rbcL gene sequence data show that P. pacifica is distinctly different from the superficially similar species, P. morrowii and P. stricta. However, the rbcL sequences of P. pacifica var. pacifica and var. disticha are identical though they have morphological variation.

• Journal of Plant Biology
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• v.38 no.1
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• pp.107-113
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• 1995

pRLYS1 containing intact rbcL gene of maize (Zea mays L. cv Golden X Bantam T-51; Zm-A) was digested with several restriction enzymes to construct subcones carrying promoter region of rbcL. The DNA fragments of 0.20, 0.19, 0.92 and 1.55 kb among the EcoRI digests, the EcoRI-DdeI digests, the AvaI digests and the EcoRI-BamHI digests of pRLYS1 were subcloned into pBluscriptSK＋and named pRLPS2, pRLPS3, pRLPS14 and pRLPS35, respectively. Four subclones contain the 1.92 kb portion from 136 nucleotide downstream to 1780 nucleotide upstream from the ATG initiation codon of rbcL gene. pRLPS2 (-29 to -229) and pRLPS3 (-239 to -420 from the ATG) were sequenced. When nucleotide sequence of Zm-A was compared with sequence of rbcL promoter region of a different cultivar of maize (Zea mays L. cv WFG TMS X BS7; Zm-B), the difference rate between two cultivars was 4.3%. The mean of sequence divergence between Zm-A and three grass species in the same tribe, Andropogoneae, in the upstream region from 29 to 420 of ATG was 1.8%, whereas between Zm-B and above-mentioned three species was 5.4%. Therefore, Zm-A seems to evolutionarily closer to three other species in Andropogoneae tribe than Zm-B is.

• The Plant Pathology Journal
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• v.37 no.6
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• pp.512-520
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• 2021

In this study, we report genetic characterization of Orobanche cumana, the causal agent of sunflower wilting in Serbia. The genetic diversity of this parasitic plant in Serbia was not studied before. Random amplified polymorphic DNA (RAPD) markers and partial rbcL gene sequences analysis were used to characterize the O. cumana populations at the molecular level. While phylogenetic analyses of RAPD-PCR amplicons were performed using unweighted pair-group Method analyses, rbcL gene sequences were analyzed using neigbor joining method and minimum spanning tree. Molecular analyses of RAPD-PCR analysis revealed high genetic diversity of O. cumana populations which indicated high adaptive potential of this parasitic weed in Serbia. Further analyses of rbcL gene using minimum spanning tree revealed clear differences among diverse sections of Orobanche genus. Although this molecular marker lacked the resolution to display intrapopulation diversity it could be a useful tool for understanding the evolution of this parasitic plant. Our results suggested that O. cumana has great genetic potential which can lead to differentiation of more virulent races which is important for determining crop breeding strategies for their control.

• Korean Journal of Environment and Ecology
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• v.24 no.3
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• pp.318-324
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• 2010

The aim of this study is to examine feeding habits of the Korean water deer(Hydropotes inermis argyropus) from its rumen contents using a PCR-DGGE method. For this study, rumen contents were collected from water deer causalities by natural death or road-kill in two different sites(Cheorwon, Gangwon province and the Eastern part of Jeonnam province). DNA was extracted from rumen contents of a total of 44 individuals. Two primers, rbcLZ1aF(GC) and rbcL19bR, were used for PCR amplifications of ribulose-1,5-bisphosphate carboxylase large subunit (rbcL) gene. Among 44 samples, twenty-nine samples were successfully amplified by PCRs. The 29 PCR products of partial rbcL gene were applied for PCR-DGGE. Totally, six families of plants were detected from the diet analyses. Five families of plants were found in Cheorwon, Gangwon province, but only three families of plants were found in the Eastern part of Jeonnam province. The PCR-DGGE method will provide us with a potential tool to study feeding habits of ungulates including water deer, even though our results failed to identify the prey plants at the level of species.