본 연구는 우리나라 해안에서 널리 서식 중인 해양 자원 중 하나인 전복(Haliotis discus hannai) 의 차세대염기서열분석 데이터 기반으로 선별한 신규 펩타이드의 항암 활성을 평가한 연구이 다. 펩타이드의 항암 활성은 교모세포종 세포주인 SNU-489에서 농도 의존적으로 처리 시간에 비례하여 증가하였으며, 200 μM로 48시간 처리하였을 때 암 세포 사멸율이 67%로 가장 높게 나타났다. 반면 정상 세포인 HaCaT에서 가장 높은 세포 사멸율은 18%로 농도 의존적이었으나 처리 시간과는 무관하였다. 또한 신규 펩타이드의 항암 메커니즘 과정을 밝히기 위해 세포자 멸괴사(Necroptosis) 관련 유전자의 발현 변화를 qRT-PCR 방법을 통해 검증하였다. RIPK3는 신 규 펩타이드 처리군에서 200 μM 처리 시 9배 이상 발현 증가, MLKL는 100 μM 처리군에서 대조군 대비 2배 이상 유의미하게 발현이 증가되었다. 이러한 결과로 미루어 볼 때, 전복 유래 신규 펩타이드는 암 세포 특이적으로 세포 독성을 가지며, 세포자멸괴사 메커니즘을 통해 암 세포 사멸을 일으키는 것으로 추측되므로 신규 펩타이드가 추후 교모세포종 치료제의 후보 물질로 활용될 수 있을 것으로 사료된다.

저어새의 먹이생물을 파악하기 위해 2010년 6월부터 2014년 6월까지 인천 남동유수지에서 저어새의 토사물 시료를 채집하여 현미경 관찰 및 차세대염기서열 (NGS) 기법으로 분석 하였다. 저어새의 먹이생물은 어류, 갑각류, 다모류, 곤충류로 구성되어 있었으며, 주로 저어새는 어류와 갑각류를 섭이하는 것으로 나타났다. 최우점 먹이생물은 풀망둑 (Acanthogobius hasta)이었으며, 이 외에도 길게 (Macrophthalmus abbreviates), 징거미새우류 (Macrobrachium sp.), 칠게 (Macrophthalmus japonicus), 각시흰새우 (Exopalaemon modestus), 참 갯지렁이 (Neanthes japonica)가 우점 먹이생물로 출현하였다. 이들 먹이생물은 번식지 인근지역인 송도갯벌과 시화호에서 흔히 발견되며, 저어새는 채식지로써 이들 지역에 대한 의존도가 높을 것으로 판단된다. 현미경과 NGS로 분석한 일부 먹이생물에서 차이를 보였는데, 이는 토사물 내 먹이생물은 저어새의 위 내에서 분해되어 현미경 분석을 통한 형태학적 분류 특징을 찾기 어려웠던 반면, NGS 분석은 유전자를 통해 분류가 가능하기 때문에 형태학적 분석의 결과보다 높은 종 다양성을 보인 결과이다.

스트레스 음파(5,000 Hz, 95 dB)는 아메리카잎굴파리(Liriomyza trifolii) 번데기의 성충 발육을 억제시켰다. 이러한 발육 교란에 대한 생리적 원인을 규명하고자 이 곤충의 스트레스 음파에 대한 발현 유전체를 무처리 곤충과 비교 분석하였다. 발현유전체는 전사체를 대상으로 하였으며, 이는 전체 RNA를 Illumina HiSeq2000을 이용한 차세대우전체 염기서열 분석 기술을 이용하였다. 처리구와 무처리구에서 전체 전사체의 분석염기수는 약 16 Gb였고, 이를 토대로 77,553개의 contig가 규명되었다. 이 contig들을 대상으로 스트레스 음파 처리에 따라 발현량 차이를 보인 비율은 약 62.8%를 차지하였다. 이 가운데 대조구에 비해 10배 이상 발현량이 감소한 contig 숫자는 3,994개이고, 증가한 contig 숫자는 1,120개를 나타냈다. 특별히 성장과 변태와 관련된 insulin 및 ecdysone 신호 유전자의 발현이 영향을 받았다. 또한 일반 물질대사와 관련된 유전자들의 발현이 스트레스 음파 처리에 따라 감소를 보였다. 그러나 스트레스 관련 유전자인 지질동원호르몬 신호전달과정 유전자들은 발현량이 증가하였다. 이상의 스트레스 음파에 따라 아메리카잎굴파리의 유전체 발현량 변화는 이 곤충의 정상적 성충 발육에 교란을 주었을 것으로 추정된다. 향후 스트레스 음파에 처리에 따라 발현에 교란을 받은 유전자들의 성충 발육 연관성에 대한 기능 분석이 필요하다.

Background : Atractylodes japonica koidz (AJ) is a perennial herb that belongs to Atractylodes genus. The dried rhizome of AJ is known as ‘Baek-chul’. The ‘Baek-chul’ is used as important traditional medicine in north-east Asia. It is considered to be effective for the treatment of stomach disorder, virus, diuresis, inflammation, arthritis. AJ is heavily depend on import from china and only few studies have been carried out. In this study, we develop SSR marker to build a foundation of breeding, to analyze genetic diversity and to construct core collection. Methods and Results : AJ resources was collected from each different place. To find simple sequence repeat (SSR) marker, we sequenced genomic DNA of AJ resources using Illumina HiSeq 2000 System. As a result of next generation sequencing (NGS), we obtained putative SSR loci. From these SSR primers, 553 SSR primer sets were designed successfully and confirmed polymorphism by in silico analysis. Nucleotide motifs ranged from tri- to penta-. Among these, 48 primer were tested in 4 individuals by capillary electrophoresis. Finally, selected 28 SSR marker were showed clear band and polymorphism by Electrophoresis. Conclusion : In this study, we developed 28 polymorphic SSR marker using NGS, and it could be used for analyzing genetic diversity of A. japonica. These marker would be useful for breeding of new cultivar in the future.

Background: Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth with radical leaves during the first year and shows reproductive growth with cauline leaves and bolting during the second year. In addition, the shape of the plant varies within the same species. For this reason, there are limitations to classifying the species by visual examination. However, there is not sufficient genetic information or molecular tools to analyze the genetic diversity of the plant. Methods and Results: Approximately 34.59 Gbp of raw data containing 342,487,502 reads was obtained from next generation sequencing (NGS) and these reads were assembled into 357,211 scaffolds. A total of 84,106 simple sequence repeat (SSR) regions were identified and 14,133 primer sets were designed. From the designed primer sets, 95 were randomly selected and were applied to the genomic DNA which was extracted from five plants and pooled. Thirty-nine primer sets showing more than two bands were finally selected as SSR markers, and were used for the genetic relationship analysis. Conclusions: The 39 novel SSR markers developed in this study could be used for the genetic diversity analysis, variety identification, new variety development and molecular breeding of A. triphylla.

Background : Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth by radical leaf until 1 year after sowing and shows reproductive growth during the second year and there is a characteristic of bolting by turning into cauline leaf. In addition, the phenotypes of plants varies even though they are belonging to the same species. For this reason, there is a limit for the classification of the species by the method of visual examination. Methods and Results : Simple sequence repeat (SSR) markers were developed based on the genomic sequence of A. triphylla using next generation sequencing to prepare the basis of molecular breeding and analyze the genetic diversity. Ninety-five primer sets including tri-, tetra- and penta-nucleotide motif types were randomly selected and they were applied to mixed genomic DNA and finally 39 primer sets showing from two to four bands were selected and used for genetic relationship analysis. Conclusions : Using the next generation sequencing, 39 polymorphic SSR markers were developed.

Background : Medicinal crop has been used in the traditional Asian medicinal methods. From ancient times, various kinds of medicinal crop are being cultivated in East Aisa including Korea, China and Japan. In Korea, they used a variety of medicinal plants in folk medicine and oriental medicine since ancient times. Molecular markers can be widely used in a variety of settings such as effective-loci estimation, genetic-diversity characterization, allelic-effect studies, gene-flow studies, quantitative-trait locus (QTL) mapping, and evolutionary studies. The genetic analyses of crops require large numbers of useful molecular markers for genetic or QTL identification, comparative mapping and breeding. Studying the genetic diversity and genetic relationship of crops can assist breeders. Crop genetics within a breeding program enable the economic and effective cultivar development. We tried to develop a variety of molecular markers from Angelica gigas genomic sequences for genetic studies and breeding. Methods and Results : A. gigas resources cultivated in Republic of Korea were collected. Fresh leaves were ground with liquid nitrogen and gDNA was extracted using a DNeasy Plant Mini kit (Qiagen, Valencia, CA, USA). We sequenced the whole genomes of five A. gigas accessions using Illumina HiSeq 2500 platform and identified genomic Simple Sequence Repeat (SSR) and InDel markers. DNA amplification was conducted using the PCR system (Bio-Rad T-100 Thermal Cycler). PCR products were separated by capillary electrophoresis on the ABI 3730 DNA analyzer (Applied Biosystems) and Fragment analyzer automated CE system (Advanced Analytical Technologies, Ankeny, IA, USA). Conclusion : We developed novel SSR and InDel markers from A. gigas genomic sequences for further genetic studies and breeding.

The transcriptomes of four ginseng accessions such as Cheonryang (Korean ginseng cultivar), Yunpoong (Korean ginseng cultivar), G03080 (breeding line of Korean ginseng), and P. quinquefolius (American ginseng) was characterized. As a result of sequencing, total lengths of the reads in each sample were 156.42 Mb (Cheonryang cultivar), 161.95 Mb (Yunpoong cultivar), 165.07 Mb (G03080 breeding line), and 166.48 Mb (P. quinquefolius). Using a BLAST search against the Phytozome databases with an arbitrary expectation value of 1E-10, over 20,000 unigenes were functionally annotated and classified using DAVID software, and were found in response to external stress in the G03080 breeding line, as well as in the Cheonryang cultivar, which was associated with the ion binding term. Finally, unigenes related to transmembrane transporter activity were observed in Cheonryang and P. quinquefolius, which involves controlling osmotic pressure and turgor pressure within the cell. The expression patterns were analyzed to identify dehydrin family genes that were abundantly detected in the Cheonryang cultivar and the G03080 breeding line. In addition, the Yunpoong cultivar and P. quinquefolius accession had higher expression of heat shock proteins expressed in Ricinus communis. These results will be a valuable resource for understanding the structure and function of the ginseng transcriptomes.