국화과(Compositae)는 현화식물 중 세계에서 가장 넓게 분포하고, 쌍자엽식물 중 가장 진화된 식물분류군이며, 우리나라에는 약 300여종이 존재하는 것으로 알려져 있다. 구절초, 감국, 쑥, 쑥갓, 개미취, 참취, 곰취 등 국화과 식물들은 예로부터 민간에서 약용 및 식용 소재로써 다양하게 사용되어왔다. 본 연구는 국화 및 국화근연종 유용유전자원 선발을 통하여 육종 소재를 확대하고, 중간모본 및 신품종 육성기반을 구축하고자 DNA 마커시스템의 개발을 위해 수행되었다. 1. 화단국인 Smileball(Dendranthema grandiflorum) 품종을 사용하여 SSR-enriched library를 작성하였고, GS FLX 분석을 통해 18.83Mbp의 염기서열 결과를 얻었으며, read의 평균 길이는 280.06bp로 나타났다. 2. 단순반복염기서열(SSR) 부위를 포함하는 26,780개 clones 중 di-nucleotide motifs가 16,375개(61.5%)로 우세하였고, trinucleotide motifs(6,616개, 24.8%), tetra-nucleotide motifs(1,674개, 6.3%), penta-nucleotide motifs(1,283개, 4.8%), hexa-nucleotide motifs(693개, 2.6%) 순으로 나타났다. 3. 얻어진 di-nucleotide motifs들 중에서는, AC/CA class가 93.5%로 대부분이었고, tri-nucleotide motifs에서는 AAC class가 50.5%, tetra-nucleotide motifs는 ACGT class가 43.6%이고, pentanucleotide motif에서는 AACGT class 27.2%이며, hexa-nucleotide motif에서는 ACGATG class 21.8%였다. 4. 얻어진 염기서열 결과를 토대로 다양한 motif를 갖는 100개의 SSR 마커를 제작하였고, 차후 이를 활용하여 국화 유전자원의 다형성 및 유전자형 분석을 통해 분자유전학적 다양성 및 집단의 구조분석이 가능하고, 국화의 분자육종기반 구축을 위한 유용한 도구가 될 것 이다.

Methanogenic community shift and comparison were determined by 454 pryosequencing for two different full-scale anaerobic digesters treating municipal sludge. For monitoring long-term of microbial communities, samples were collected for two year at three-monthly basis. The two mesophilic AD bioreactor were operated at similar operating conditions, but different substrate streams. Methanospirillum were identified as the key drivers of methanogenesis in full-scale anaerobic digester treating municipal sludge. In Joongrang (JR) digester, Methanospirillum was dominant (48%±10.3) over almost all period, but the dominant genus move to Methanosaeta and Methanoculleus due to low acetate concentration (0.02 g/L), total ammonia nitrogen concentration, respectively. In Asan digester (AS), Methanospirillum also was dominant (41%±12.6) like JR digester, but methanogenic community shift was examined twice. One of those was from Methanospirillum to Methanophaerula due to pH sharply decrease (<5.5) and second shift was Methanosaeta increase due to low VFAs concentration (0.25 g/L).

For the development of SSR marker system in Vicia villosa Roth, an enriched library was constructed by using a modified biotin-streptavidin capture method and the selected clones were sequenced with GS-FLX(454). Of 37,794 sequenced reads, we found that 8,474 reads (22.4%) were redundant, leaving 29,320 unique ones (77.6%). Among the unique clones, 17,174 reads (58.6%) were having microsatellite repeating motifs. Sequence analysis of all SSR-containing reads revealed a predominance of the di-nucleotide SSRs (62.5%). The tri-nucleotide and the tetra-nucleotide SSRs were 5.7% and 22.5%, respectively. As the di-nucleotide type, the AG/GA class of repeat motif was most frequently identified (55.0% of the total di-nucleotide SSRs), followed by the CT/TC class (19.5%), and the TA/AT class (12.1%). Among the tri-nucleotide SSRs, the AGT/GTA/TAG class of repeat motifs was predominant (22.2%), followed by the ACT/CTA/TAC class (17.8%). Among the tetra-nucleotide SSRs, the CTTT/TTTC/TTCT/TCTT class of repeat motifs was predominant (31.2%), followed by the AAAG/AAGA/AGAA/ GAAA class (19.9%). Finally, we designed 779 primer pairs from the flanking sequences of SSR containing reads. We are undertaking the analysis of polymorphisms using the diverse collected accessions of Vicia villosa Roth now. This newly developed SSR marker set shall provide a very useful tool for implementing molecular diversity assessment and population structure studies of Vicia villosa Roth onward.