• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• Korean Journal of Pharmacognosy
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• v.53 no.4
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• pp.226-233
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• 2022

Ephedra herbs are defined as stem of Ephedra sinica , Ephedra intermedia and Ephedra equisetina in the Korean Pharmacopoeia. It is important to use pure herbs to derive the safety and efficacy of herbal medicine. However, the identification of these herbs by conventional taxonomic methods is difficult. Recently, many studies have applied these DNA barcoding for the identification of herbal medicinal species using standard DNA markers. In this study, we report a case study in which the identification of Ephedra species was done by DNA barcoding. For identification of Ephedra species, 17 samples were collected, and a reference DNA barcode library was developed using 6 markers (rbcL, matK, ITS2, ycf1, ycf3, and rpoC2). To develop KASP-SNP markers, we selected 4 markers (ycf1, ycf3, rpl2, and rbcL), which were able to distinguish three Ephedra species. In the result, the specific markers for each of the three Ephedra were clustered into FAM-positive section, whereas non-targeted plants were clustered either HEX-positive or negative section. Therefore, we have developed KASP assay that allow rapid and easy Ephedra species identification using three KASP markers.

• Korean Journal of Plant Taxonomy
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• v.51 no.1
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• pp.109-114
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• 2021

Veronica nakaiana Ohwi (Plantaginaceae) is an endemic taxon on Ulleungdo Island, Korea. We report the second complete chloroplast genome sequence of V. nakaiana. Its genome size is 152,319 bp in length, comprising a large single-copy of 83,195 bp, a small single-copy of 17,702 bp, and a pair of inverted repeat regions of 25,711 bp. The complete genome contains 115 genes, including 51 protein-coding genes, four rRNA genes, and 31 tRNA genes. When comparing the two chloroplast genomes of V. nakaiana, 11 variable sites are recognized: seven SNPs and four indels. Two substitutions in the coding regions are recognized: rpoC2 (synonymous substitution) and rpl22 (nonsynonymous substitution). In nine noncoding regions, one is in the tRNA gene (trnK-UUU), one is in the intron of atpF, and seven are in the intergenic spacers (trnH-GUG~psbA, trnK-UUU, rps16~trnQ-UUG, trnC-GCA~petN, psbZ~trnG-GCC, ycf3~trnS-GGA, ycf4~cemA, and psbB~psbT). The data provide the level of genetic variation in V. nakaiana. This result will be a useful resource to formulate conservation strategies for V. nakaiana, which is a rare endemic species in Korea.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.98-98
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• 2019

제니(薺苨, Adenophorae Remotiflori Radix)는 "대한민국약전외한약(생약)규격집(KHP)"에 모시대(Adenophora remotiflorus Miquel)의 뿌리로 수재되어있으나, 형태학적으로 유사한 잔대(A. triphylla), 당잔대(A. stricta) 및 더덕(Codonopsis lanceolata)과 오 혼용 우려가 있어 이들을 구별하기 위한 정확하고 객관적인 종 감별법이 필요하다. 본 연구에서는 '제니'의 기원인 모시대와 오 혼용 우려가 있는 종들을 구별 할 수 있는 유전자 마커를 개발하기 위하여 Genbank에 등록된 ycf2 구간을 활요하여 모시대와 잔대, 당잔대를 구분 할 수 있는 INDEL (insertion/deletion) 마커를 개발하였다. 또한, 보다 정확한 종감별을 위해 DNA 바코드로 활용되고 있는 유전자 부위의 염기서열을 분석하여 ITS (25%), atpB-rbcL (15%), atpF-atpH (14%), rpl16 (13%), trnL-F (10%), matK (9%), rpoC1 (7%)에서 변이율(percent of variable sites)을 확인하였다. 향후, 본 연구에서 개발된 INDEL 마커와 더불어 추가적으로 개발을 진행 중인 분자 마커는 한약재 '제니'의 품질관리에 활용 가능할 것으로 사료된다.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.19-19
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• 2023

제비꽃속 34분류군의 61개체를 대상으로 엽록체 DNA 게놈 특이 유전자의 특징을 알아보고자 하였다. 61개체의 엽록체 게놈 전체 길이는 155,535~158,940 bp 로 모두 전형적인 사분할 구조였다. 지역별로는 LSC 지역이 84,826~87,250 bp, SSC 지역이 16,338~18,654 bp, 그리고 IR 지역이 26,029~27,192 bp 였다. 유전자 개수는 131개로 84개 protein coding-gene, 37개 tRNA 유전자, 8개 rRNA유전자, 그리고 2개의 유사유전자인 𝜓rps19, 𝜓ycf1으로 구성되어 있었다. LSC/IRa 경계에 위치한 rps19 유전자 길이는 279 bp로 모든 분류군에서 동일하였으며, 𝜓rps19의 길이는 다양했으나 유전자 개수에는 영향을 미치지 않았다. SSC/IRb 경계에 위치한 ycf1 유전자 길이는 약 5,600 bp 였으나, V. japonica (MZ151699) 1개체에서는 다른 종에 비해 약 1,000 bp 위치에서 발생한 점돌연변이로 인해 종결 코돈이 나타나는 특징을 보였다. 한편 13분류군의 23개체에서는 𝜓ycf1의 길이가 650 bp 정도 짧은 것을 확인하였는데, 이 종류들은 원예종인 V. tricolor (ON262802) 이외에는 모두 줄기가 없는 분류군들로 IR 지역의 확장과 SSC 지역의 수축에 의한 것으로 판단된다. ndhF는 대체로 SSC 지역에 위치하나, V. inconspicua (MZ065354), V. mongolica (MW802534, ON548135), V. yunnanfuensis (MW802541) 등 4개체에서는 IRa/SSC 경계에 위치하면서 유사유전자가 발생하였고, 그 결과 다른 제비꽃 종류에 비해 유전자 개수가 132개로 차이를 보였다. 또한, V. collina (OP271831), V. mirabilis (MH256000), V. tricolor (ON262802) 등 3분류군에서는 SSC 지역이 inversion 되어 엽록체 이성질체가 존재함을 확인하였다. 이상의 결과를 종합하면, 제비 꽃속 엽록체 게놈 61개체의 ycf1, 𝜓ycf1, ndhF, 𝜓ndhF 등은 유전자 길이와 개수 등에 차이를 보이는 것으로 나타났으며, 제비꽃속에서도 엽록체 이성질체가 존재함을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1266-1271
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• 2017

Lactobacillus fermentum strain JDFM216, isolated from a Korean infant feces sample, possesses the ability to enhance the longevity and immune response of a Caenorhabditis elegans host. To explore the characteristics of strain JDFM216 at the genetic level, we performed whole-genome sequencing using the PacBio system. The circular draft genome has a total length of 2,076,427 bp and a total of 2,682 encoding sequences were identified. Five phylogenetically featured genes possibly related to the longevity and immune response of the host were identified in L. fermentum strain JDFM216. These genes encode UDP-N-acetylglucosamine 1-carboxyvinyltransferase (E.C. 2.5.1.7), ErfK/YbiS/YcfS/YnhG family protein, site-specific recombinase XerD, homocysteine S-methyltransferase (E.C. 2.1.1.10), and aspartate-ammonia ligase (E.C. 6.3.1.1), which are involved in peptidoglycan synthesis and amino acid metabolism in the gut environment. Our findings on the genetic background of L. fermentum strain JDFM216 and its potential candidate genes for host longevity and immune response provide new insight for the application of this strain in the food industry as newly isolated functional probiotic.

• Journal of Species Research
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• v.4 no.2
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• pp.152-158
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• 2015

Dendranthema boreale and D. indicum are easily distinguished from other Korean Dendranthema spp. by having yellow flowers. We have found a putative new taxon of Dendranthema having white flowers, except for sharing most characters with Dendranthema boreale. The chloroplast (cp) genome of the putative new taxon of Dendranthema, Dendranthema sp. K247003, registered in National Agro-Biodiversity Center (ABC), was completely characterized as a genetic barcode. The cp-genome of Dendranthema sp. K247003 was 151,175-bp in size: LSC was 82,886-bp, IR 24,971-bp, SSC 18,347-bp. The cp-genome of Dendranthema sp. K247003 contains 113 genes and 21 introns consisted of 79 protein coding genes, 4 RNA genes, and 30 tRNA genes, with 20 group II introns and one group I intron. Some of the genes and there introns were duplicated in IR. The cp-DNA of Dendranthema sp. K247003 is distinguished from that of D. boreale IT121002 by 67 SNPs in genic regions of 24 protein coding genes and by a 9-bp INDEL in ycf1. Further cp-DNA study will give us better information on genetic markers of Dendranthema species.

• BMB Reports
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• v.40 no.4
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• pp.577-587
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• 2007

Single-copy chloroplast loci are used widely to infer phylogenetic relationship at different taxonomic levels among various groups of plants. To test the utility of chloroplast loci and to provide additional data applicable to hybrid evolution in Musa, we sequenced two introns, rpl16 and ndhA, and two intergenic spacers, psaA-ycf3 and petA-psbJ-psbL-psbF and combined these data. Using these four regions, Musa acuminata Cola(A)- and M. balbisiana Colla (B)-containing genomes were clearly distinguished. Some triploid interspecific hybrids contain A-type chloroplasts (the AAB/ABB) while others contain B-type chloroplasts (the BBA/BBB). The chloroplasts of all cultivars in 'Namwa' (BBA) group came from the same wild maternal origin, but the specific parents are still unrevealed. Though, average sequence divergences in each region were little (less than 2%), we propose that petA-psbJ intergenic spacer could be developed for diversity assessment within each genome. This segment contains three single nucleotide polymorphisms (SNPs) and two indels which could distinguish diversity within A genome whereas this same region also contains one SNP and an indel which could categorize B genome. However, an inverted repeat region which could form hairpin structure was detected in this spacer and thus was omitted from the analyses due to their incongruence to other regions. Until thoroughly identified in other members of Musaceae and Zingiberales clade, utility of this inverted repeat as phylogenetic marker in these taxa are cautioned.