成果報告書詳細
管理番号20110000000609
タイトル*平成22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2010 - 2010
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。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作出を目指している。
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, an environmental destruction and a depletion of fossil fuels have become major social problems on a global scale. Forestation is considered to be an effective tool to mitigate these problems because it can preserve the environment and supply raw materials at the same time, 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 that can grow on marginal lands.
1. Development of Harsh-environment-resistant Technology
To increase biomass production under environmental stress condition, fast-growing transgenic trees (hybrid aspen and eucalyptus) that introduced salt-tolerant genes (mangrin, coline oxidase [codA] and RNA binding protein) have been produced. As a first screening, existences of transgenes, expression levels and multiplication ability were investigated in each transgenic Eucalyptus camaldulensis. We also measured glycinebetain accumulation in transgenic plants with codA. Furthermore, these lines were evaluated by salt-tolerance in a growth room and several lines were used for further analysis. The selected lines are growing in a special netted-house in University of Tsukuba.
2. Studies on Developing Highly Functional Trees
It is important to select suitable promoter and terminator for desired gene expression in transgenic plants. Then, we compared two terminators (HSP and NOS). Four vectors using the GUS gene as a reporter were constructed and introduced into hybrid aspen and eucalyptus. Histochemical and fluorometric assays indicated that higher GUS expressions were observed in the hybrid aspen shoot with HSP terminator. To produce highly stress-tolerant trees, the vector containing salt-tolerant genes with HSP terminator introduced into hybrid aspen and eucalyptus. We produced over 30 transgenic lines and now analyzing expression levels.
3. Research on Mode of Action of Salt-tolerant Genes
Transgenic tobacco expressing SjRBP/SeFLA
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, a cDNA encoding chloroplast RNA binding protein (SjRBP) was isolated. In addition, SeFLA cDNA was isolated from Salconia europaea cDNA library by the functional screening method, previously. In this study, transgenic tobacco expressing SjRBP/SeFLA was constructed and the salt tolerance was investigated. The highest salt tolerance was observed in the transgenic tobacco expressing SjRBP/SeFLA. Coexpression of SjRBP/SeFLA may be useful method to construct transgenic plant for enhancement of salt tolerance.
4. Development of Evaluation Methods on GM Trees in Special Netted-house and Field Testing.
In this project, salt-tolerant transgenic trees (i.e. eucalyptus and hybrid aspen) are evaluated in special netted-houses to select some candidate lines that maintain some biomass production at unarable lands. In this fiscal year, we started cultivation trials of transgenic eucalyptus lines, which have coline oxydase gene (codA), and transgenic hybrid aspen lines, which have McRBP gene, in the special netted-house. On the results of analyses of colonization of rhizosphere microorganisms and results of analyses of allelopathic effects of eucalyptus leaves, no significant difference was found between transgenic eucalyptus lines and non-transgenic lines. Hybrid aspen is the deciduous tree. We observed a transgenic hybrid aspen line breaks winter dormancy earlier than other lines including a non-transgenic line. In addition, other transgenic eucalyptus lines, which have McRBP gene, are ready to start cultivation trial in the special netted-house.
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