Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small (2.71 - 5.50㎛)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

사시나무의 클론간 생장 특성(전체 건중량, 줄기 건중량, 잎의 전체 건중량)과 대사물질간의 관계를 구명하여 조기선발을 위한 생장 관련 주요 인자를 구명하고자 사시나무 삽목묘(유묘)를 대상으로 우량클론과 일반클론간의 생장특성을 조사하였고, 수체내의 대사물질을 분석하였다. 사시나무 유묘의 줄기 건중량과 잎 내의 glucose, sucrose, inositol 등 당 함량과 정의 상관관계(p<0.05)가 있었으며, 잎의 건중량과는 줄기 내의 glutamic acid, alanine, threonine등과 같은 아미노산류의 함량과 정의 상관관계가 있었다. 사시나무 잎 내의 대사물질은 우량클론에서 glucose, sucrose, inositol 등과 같은 당류가 일반클론보다 고도의 유의 차이(p<0.01)가 있게 높게(1.3배 이상) 함유한 것으로 나타났으며,rythronic acid, malic acid, pentanedioic acid 및 proline도높게 함유한 것으로 나타났다. 또한 우량클론의 줄기내의 대사물질은 glutamic acid, alanine, threonine등과 같은 아미노산류의 함량이 일반클론에 비해 유의적으로 높게(1.2배 이상)함유하고 있는 것으로 나타났고, 더불어 succinic acid, maleicacid의 함량도 고도의 유의차이(p<0.01)로 우량클론에서 높게 함유하고 있었다. 생장이 우수한 사시나무 클론은 유묘의잎에서는 당류가 줄기에서는 아미노산류가 높게 함유하여 이러한 복합적 인자가 사시나무 우량클론 선발을 위한 대사 표지자의 개발에 필요한 기초 자료로 이용될 수 있을 것이다.

Gastrodia elata Blume is a achlorophyllous orchid plant that has been used in traditional medicine to treat many human illnesses, such as vertigo, hemiplegia, and epilepsy. It contains three major pharmacological substances; gastrodin, ergothioneine, and vanillyl alcohol. Here we investigated the quantitative changes in those functional components by cultivation conditions and harvest times. The content of ergothioneine was different with cultivation areas, while gastrodin accumulated the higher levels than vanillyl alcohol regardless of cultivation areas. The smaller tubers contained the higher concentrations of gastrodin and the lower concentrations of ergothioneine than did the larger tubers. The cultivation under sun shade screen increased the concentrations of both gastrodin and vanillyl alcohol but decreased the levels of ergothioneine. We also determined the contents of functional substances depending on the harvest times. The concentration of gastrodin was higher in October than in April, while the level of ergothioneine was higher in April than in October. Interestingly, the use of sexually propagated seed tubers produced 1.9 times higher levels of gastrodin compared to vegetatively propagated seed tubers. In this study, we found that three major functional substances in G. elata tubers altered depending on various cultivation practices, which might be important to increase the economic values of G. elata tubers.

Gastrodia elata has been cultivated as an important medicinal resources to treat various human diseases. One of the major problems associated with its field production is the degeneration of seed tubers, which is mainly caused by soil-borne pathogens. This study was conducted to produce disease-free seed tubers by the development of in vitro micropropagation method. First, tubers of G. elata were treated with HgCl2 prior to culturing in vitro. Among various culture medium tested, water agar (WA) and WPM medium were the most effective on the induction and growth of vegetative stems. NAA (0.1mg/l) or TDZ (1.0mg/l) in WA medium showed better growth of vegetative stems compared to other plant hormones. Finally the induction and growth of vegetative stems were better in the dark compared to the light condition. In this study, we established an in vitro micropropagation system of G. elata, which might be an efficient way to increase the yield and quality of G. elata tubers in the field production.

We have established adventitious root culture systems of Codonopsis lanceolata, Codonopsis pilosula and Codonopsis ussuriensis. Root segments of C. lanceolata were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 18.8±1.9 roots per explant. Root segments of C. pilosula were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 1.0mg/l NAA and produced 8.5±1.8 roots per explant. Leaf segments of C. ussuriensis were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 7.8±0.4 roots per explant. In liquid culture, the best production of adventitious root (fresh weight) was obtained in 1/2 MS medium with 1.0mg/l NAA. This study demonstrated for the first time to produce adventitious roots in C. pilosula and C. ussuriensis.

Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon's information Index (S.I.) indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.

신속하게 생장하는 소나무의 조기 선발인자를 구명하고자 32년생 소나무의 풍매차대 검정결과로 얻어진 재적지수를 기준으로 상, 중, 하로 구분하여 선발된 12가계의 생장특성과 동일한 가계의 유묘 생장특성과의 상관관계를 분석하였다. 선발된 12가계의 종자 특성과 동일 가계의 성목 생장특성과는 상관관계가 없었으며 다만 소나무 유묘의 초기 생장과는 관련이 있는 것으로 나타났다. 가계별 1년생의 유묘 건물중과 성목의 재적지수(32년생)와는 1%의 유의적 정의 상관