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폐기물매립장 침출수내 미생물군집 구조 해석을 위한 T-RFLP의 활용
유재철(Jae Cheul Yu),Tomonori Ishigaki,Yoichi Kamagata,이태호(Tae Ho Lee) 大韓環境工學會 2010 대한환경공학회지 Vol.32 No.4
폐기물매립장의 안정화에는 미생물이 중요한 역할을 수행한다. 폐기물매립장에서 미생물군집 변화 모니터링에 말단 제한절편다형성(Terminal Restriction Fragment Length Polymorphism; T-RFLP)법의 활용 가능성을 평가하고자 박테리아의 16S rDNA 서열에 기초한 T-RFLP법으로 4개의 폐기물매립장 내부에서 채취한 침출수의 미생물군집 구조를 조사하였다. T-RFLP법을 사용하여 해석한 침출수 내 우점 미생물군집 구조와 일반적으로 널리 사용되고 있는 16S rDNA 클론 해석법에 의한 우점 미생물군집구조는 유사하였다. 또한, T-RFLP법을 이용하여 폐기물매립장의 구조, 매립 폐기물 종류, 운영기간이 다른 폐기물매립장 침출수의 우점 미생물군집 구조가 서로 다르게 나타나는 것을 확인 할 수 있었다. 따라서 T-RFLP법을 사용하여 폐기물매립장 침출수내 미생물군집 구조를 장기적으로 모니터링 한다면 많은 비용과 시간이 소요되는 클론해석법의 반복적인 수행 없이도 비교적 간단하게 폐기물매립장의 안정화 정도를 평가할 수 있을 것으로 기대한다. Microorganisms are key-role player for stabilization of landfill sites. In order to evaluate the availability of T-RFLP(Terminal Restriction Fragment Length Polymorphism) for monitoring microbial community variations during stabilization of landfill sites, the phylogenic diversity of microbial community in the leachate from 4 different full-scale landfills was characterized by T-RFLP based on bacterial 16S rDNA. Main population of microbial community analyzed by T-RFLP was significantly similar with that of microbial community analyzed by clone library analysis. The results of T-RFLP analysis for main population of microbial community in the leachate from landfills with different landfill structures, waste types and landfill ages showed apparently different microbial diversity and structures. Therefore, long-term monitoring of microbial community in leachate from landfill sites by using T-RFLP is expected to be available for evaluation of landfill stability.
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Insect’s intestinal organ for symbiont sorting
Ohbayashi, Tsubasa,Takeshita, Kazutaka,Kitagawa, Wataru,Nikoh, Naruo,Koga, Ryuichi,Meng, Xian-Ying,Tago, Kanako,Hori, Tomoyuki,Hayatsu, Masahito,Asano, Kozo,Kamagata, Yoichi,Lee, Bok Luel,Fukatsu, Tak National Academy of Sciences 2015 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.112 No.37
<P><B>Significance</B></P><P>In general, animals have a mouth for feeding, an anus for defecation, and a gut connecting them for digestion and absorption. However, we discovered that the stinkbug’s gut is functionally disconnected in the middle by a previously unrecognized organ for symbiont sorting, which blocks food fluid and nonsymbiotic bacteria but selectively allows passing of a specific bacterial symbiont. Though very tiny and inconspicuous, the organ governs the configuration and specificity of stinkbug gut symbiosis, wherein the posterior gut region is devoid of food flow, populated by a specific bacterial symbiont, and transformed into an isolated organ for symbiosis. Mutant analyses showed that the symbiont’s flagellar motility is needed for passing the host organ, highlighting intricate host–symbiont interactions underpinning the symbiont sorting process.</P><P>Symbiosis has significantly contributed to organismal adaptation and diversification. For establishment and maintenance of such host–symbiont associations, host organisms must have evolved mechanisms for selective incorporation, accommodation, and maintenance of their specific microbial partners. Here we report the discovery of a previously unrecognized type of animal organ for symbiont sorting. In the bean bug <I>Riptortus pedestris</I>, the posterior midgut is morphologically differentiated for harboring specific symbiotic bacteria of a beneficial nature. The sorting organ lies in the middle of the intestine as a constricted region, which partitions the midgut into an anterior nonsymbiotic region and a posterior symbiotic region. Oral administration of GFP-labeled <I>Burkholderia</I> symbionts to nymphal stinkbugs showed that the symbionts pass through the constricted region and colonize the posterior midgut. However, administration of food colorings revealed that food fluid enters neither the constricted region nor the posterior midgut, indicating selective symbiont passage at the constricted region and functional isolation of the posterior midgut for symbiosis. Coadministration of the GFP-labeled symbiont and red fluorescent protein-labeled <I>Escherichia coli</I> unveiled selective passage of the symbiont and blockage of <I>E. coli</I> at the constricted region, demonstrating the organ’s ability to discriminate the specific bacterial symbiont from nonsymbiotic bacteria. Transposon mutagenesis and screening revealed that symbiont mutants in flagella-related genes fail to pass through the constricted region, highlighting that both host’s control and symbiont’s motility are involved in the sorting process. The blocking of food flow at the constricted region is conserved among diverse stinkbug groups, suggesting the evolutionary origin of the intestinal organ in their common ancestor.</P>
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