Development of new bioremediation strategy through next generation biotechnology
The development of next generation sequencing technology grow rapidly in recent years. we can obtain lot of biological information by NGS technology (Next generation sequencing, NGS) because of its high throughput and precise sequencing ability , and its large-scale sequencing , the average price is much lower than the cost of sanger sequencing . Microbes are the most diverse populations in nature , have been used in human society and in many aspects of life. To take this technology（NGS） to explore the wildlife population information comprehensive; such as food fermentation, industrial production , etc. However, there is a microorganism known only one percent less than the number of inhabitation in nature .we explore more information by NGS applications to understand microbial physiology . The use of biological metabolism or adsorption of the environmental pollutant to remove contaminant substances technique called bioremediation technology ; bioremediation can eliminate pollutants permanently , rather than transfer contaminant to other types of particle to the environment. and it can be moderately remediation processes with physical or chemical combination . Because of this technique is relatively inexpensive and environmentally friendly , it has become widely used in recent years. This project targets Anshun factory in Tainan, Sinopec , which is heavily contaminated sites for many years, we plane to make a series of soil samples respectively, in the sea pool, phytoremediation area ( formerly PCP area ) and industrial factory zone soil sample, it would be to screen out a high dioxin degradation efficiency and tolerability of microorganisms ; Our research framework mainly based on the concept of train bioremediation strategy , bioremediation will be divided into three parts: on the one hand to direct remediation of high contamination soil ; conjugate with growing plants in contaminated sites and inoculated plants endophyte strategies. The other hand will combine with the sieved high tolerance and degradation dioxin bacteria to form complex bio-agents , co-bioremediate in the contaminant sites, not only increase the number of microbial populations in soil ,but improve soil organisms richness, and should profit to the metabolize of dioxin contamination . The project timeline will be divided into two phases . The first stage is to confirm that we apply the new generation of microbial sequencing technologies for genome and transcriptome technologies by Rhodococcus. we isolated Rhodococcus erythropolis B11 from sea pool , with OCDF eight chlorine dioxin to test degradation ability ; after three weeks culture ,its dioxin degradation rate of 30% , which also successfully sequenced the complete genome sequence : there are many different candidate genes in its genome relate with some aromatic cyclic compounds metabolism , but also contain dioxin decomposition path -related genes ; particular dioxygenase gene , the candidate genes which dioxin benzene ring opening key genes , but we need more information to prove the gene biosynthesis ( such as transcriptome sequencing confirmed ) , and this high chlorine degradation bacteria directly applied in soil dioxin contaminant remediation ; In addition to Rhodococcus, we also sieve Burkholderia cenocepacia a endophyte with plants and benefit with plant growth and high dioxin, mercury tolerance bacteria, our team also put the strain to whole genome sequencing , and discovery some degradation dioxin potential genes. because of its versatility, we take it to transcriptomic sequencing , the exact RNA information compared to microbe RNA blast information , such as (tonB family, osmC gene ) , estimated to be involved in the intracellular transport of dioxin , and the decomposition of some aromatic benzene metabolic pathway genes . This two bacterial fit for remediation train concept of dioxin contamination catabolism. The second stage we also screened and isolated B.subtilis, Rhodococcus, Providencia, Pseudomonas, Micrococcus, B.licheniformis, Serratia Plymuthica, Shigella boydii, Burkholderia cenocepacia, Termitomyces, Phytobecteria, ; and confirm dioxin tests and found very good tolerability , 2ppm TCDD is still alive , 2ppm OCDF many bacteria have been found tolerance and resistance , is already having considerable potential for bacterial dioxin decomposition tests. we try to combine this bio-agents approach with phytoremediation and do auxiliary remediation of the train concept . Mixing all parts , the role Rhodococcus erythropolis B11 as a bioreactor , using its highly toxic dioxin ability to decompose the soil contaminant , and plant grow on the soil with Burkholderia cenocepacia as endophyte accelerate for vegetation restoration ; supplemented by these bio-agents , improve soil ecology diversity, increase soil fertility to achieve real field restoration goals.
next generation sequencing;dioxin degradation;microbe isolation