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Title 建立結合新世代生物科技之污染場址監測與整治策略研發計畫(2/2)
Abstract 102年計畫目的在於結合生物復育與次世代定序技術，透過篩選及純化、並分析具分解汙染物能力的微生物之基因體與代謝轉錄體，找出相關的污染物代謝轉錄途徑，篩選出相關的功能性基因。於計畫中對微生物應用新世代定序技術在基因體與轉錄體技術，本團隊成功從台南中石化安順廠的海水池土壤中分離出Rhodococcus erythropolis B11，具OCDF八氯戴奧辛分解能力，也成功解序其完整基因體序列，又篩到Burkholderia cenocepacia 是一與植物共生具促進植物生長和高戴奧辛、金屬汞耐受性的細菌，我們團隊把這株菌進行了全基因體與轉錄體定序，確切的比對到了一些RNA的資訊可能參與了戴奧辛運輸到細胞內、及一些芳香苯環狀物分解的代謝途徑。103年計畫則期以微生物菌群在不同試驗環境下的差異分析，找出差異表達下的菌群及基因之代謝轉錄體，以將這些生物因子應用於環境鑑識上。於103年計畫中成功建立土壤微生物總基因體與總代謝轉錄體分析流程，並由總基因體分析中顯示雖然在不同土壤採樣點可能造成微生物的差異，但微生物因汙染物種類而影響的趨勢大致相同。本研究發現土壤微生物群在多環芳香烴影響下：Alcanivora菌屬族群大量生長，及Erythrobacteraceae科的微生物，族群有小幅上升趨勢，都有潛力成為檢測標的之一。六組實驗土壤交相比對，發現在具環境汙染物的實驗組放線菌Gaiellaceae科比例逐漸提高，可視為土壤的毒物壓力指標之一。此外在代謝轉錄體分析上，找到4個transcripts (mhpA、 dsrA、 MAP2K2、E2.8.3.)，可能可作為戴奧辛汙染的Biomarker，提供環境檢測的相關依據，冀望未來繼續更進一步的評估與研究。
EngTitle Development of new bioremediation strategy through next generation biotechnology
EngAbstract In the project of year 2013, we used both next generation sequencing technology and bioremediation approaches to discover the detailed mechanisms of dioxin degradation. The potential bacteria strains that can degrade dioxin were isolated and characterized via both genomic sequencing and transcriptomic analysis. The genome annotation and functional genes expressed under dioxin induction were analyzed and the specific metabolomic pathways were identified. We isolated Rhodococcus erythropolis strain B11 that can degrade OCDF (eight chlorine dioxin) and the whole genomic information was completed. Nevertheless, we also found an endophyte, Burkholderia cenocepacia, that can benefit plant growth and show tolerance to both highly chlorinated dioxin and mercury. Both whole genomic sequencing and transcriptomic analysis under dioxin inducing were finished. Using BLAST tool, important functional genes such as genes belong to tonB family that may involve in the intracellular dioxin transportation were identified. In the project of year 2014, the differential analysis of both meta-genome and meta-transcriptomic analysis were done under different contaminations to identify dominant bacteria and their functional transcripts. The identified biological factors identified from results were expected to further used in the field of environmental forensics. In addition, we established the overall pipeline of soil samples for meta-genome and meta-transcript analysis. Based on metagenomic data, dominant bacteria still survived, suggesting that specific contaminations may not interfere substantial tendency although different sample locations may affect soil microbe constitutions. We also found that soil microbial population in aromatic polycyclic, hydrocarbons condition: Alcanivora genusis is dominant while Erythrobacteraceae family population is slightly rising. Both of them have potentials to become detectable targets. Furthermore, compared all six experimental samples, we found the proportion of Gaiellaceae family of Actinomyces gradually increased, and it could be regarded as one of the pressure indicators in the contaminated soil. On the other hand, based on the transtriptomic data, we found four specific transcripts (mhpA, dsrA, MAP2K2, E2.8.3) that may be used as dioxin contamination biomarker. It provides the indication of environmental testing that can be further investigated and evaluated.
ProjectYear 103
SponsorOrg 環檢所
ExecutingOrg 中興大學生科中心
PublicFullVersionURL http://epq.epa.gov.tw/project/filedownload.aspx?fid=74147