成果報告書詳細
管理番号20110000000524
タイトル*平成21年度中間年報 新エネルギー技術研究開発 バイオマスエネルギー等高効率転換技術開発(先導技術開発) 遺伝子組換えによるバイオマスエネルギー高生産樹木の創生に関する研究開発
公開日2011/6/7
報告書年度2009 - 2009
委託先名日本製紙株式会社 国立大学法人東京農工大学 国立大学法人筑波大学
プロジェクト番号P07015
部署名新エネルギー技術開発部
和文要約和文要約等以下本編抜粋:1.研究開発の内容及び成果等 本プロジェクトにおいては、遺伝子組換えの手法を用いて不良環境においても生産性の高い樹木を創生する。樹木に環境ストレス耐性等を付与することにより、不良環境においてもバイオマスの生産力が維持できる樹木の選抜、評価を目指す。耕作不適の原因として乾燥が挙げられ、塩類の集積による塩害が問題となっていることから、まず耐塩性遺伝子に注目し、その作用機構の解明を実施すると共に、作成した耐塩性遺伝子組換え樹木から、耐塩性が高く実用化に適した系統の選抜、特性評価試験を実施し、劣悪環境下でバイオマス生産を可能とする樹木の第一種使用申請を目指している。平成21年度はこれまでに作成していた各種組換え体の評価試験と、新たなベクターの構築及び導入を実施した。
英文要約Title: New Energy Technology Development. Development of Technology for High-efficiency Conversion of Biomass and Other Energy. R&D of Genetic Engineering to Produce Trees with High Biomass Energy Productivity
Recently, environmental destruction and the depletion of fossil fuels have become major social problems on a global scale. Because it can preserve the environment and supply raw materials at the same time, reforestation is considered to be an effective tool to mitigate these problems. In response, it is important to breed environmental stress-tolerant and fast-growing trees. Our purpose is to produce trees with high biomass energy productivity even if they plant on marginal lands. 1. Development of Harsh-environment-resistant Technology To increase biomass production under environmental stress condition, fast-growing GM trees (hybrid aspen, eucalyptus) that introduced a salt-tolerant gene (mangrin, coline oxidase (codA), RNA binding protein) have been produced. The transgenic Eucalyptus camaldulensis witch have the codA gene form a soil bacterium were investigated. By biochemical analysis, higher levels of glycinebetain accumulation were found in high expression lines. Some plantlet were grown in a growth room and evaluated with 300mM NaCl. The high expression lines indicated salt-tolerance compered with a non-transgenic control line. 2. Studies on Developing Highly Functional Trees  To improvement the desired gene expression, studies on a promoter and a terminator were carried out. The four vectors using the GUS gene as a reporter were constructed and introduced into a hybrid aspen and a eucalyptus. Histochemical GUS expression could be found in all kinds of transgenic callus and regenerated buds from the explants.
To produce highly stress-tolerant trees, the vector having two salt-tolerant genes were constructed and introduced into a hybrid aspen. The transgenic shoots will be evaluated with NaCl and compared to shoots having a salt-tolerant gene. 3. Research on Mode of Action of Salt-tolerant Genes  Mesembryanthemum crystallium is a highly salt-tolerant halophyte. In order to isolate genes for the salt tolerance, functional screening was performed using E. coli MG1655 as a host organism. In this screening, cDNAs encoding chloroplast RNA binding protein and DNA binding protein homologue was isolated. In this study, chloroplast RNA binding protein (McRBP) in ice plant was investigated. Overexpression of McRBP in E. coli and tobacco plant displayed enhanced salt tolerance. In addition, we constructed a method to measure RNA chaperone activity. McRBP showed RNA chaperone property in vivo. Therefore, it can be postulate that expression of chloroplast RNA binding protein enhances salt tolerance in E. coli and tobacco plant. 4. Development of Method to Evaluate GM Trees Cultivated in a Special Netted-houses and Field Testing Procedures In this project, salt-tolerant transgenic eucalyptus lines are evaluated in special netted-house to select some candidate lines that maintain some biomass production at unarable land. The salt-tolerant transgenic eucalyptus lines, which have mangrin gene, were evaluated in special netted-house. The tolerance level was estimated by using over one meter height plants. The tolerant lines maintained the water contents in leaves and were controlling the inflow of salt. Those high tolerant lines are candidates for field trial. On the other hand, from the analysis of colonization of mycorrhizal fungus, it was found that infection of mycorrhizal fungus might lead to give higher salt tolerance to transgenic eucalyptus.
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