成果報告書詳細
管理番号20100000000372
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) エネルギー植物の形質転換技術及び組換え植物栽培施設での栽培技術の研究開発
公開日2010/6/2
報告書年度2009 - 2009
委託先名国立大学法人筑波大学 国立大学法人千葉大学
プロジェクト番号P07015
部署名新エネルギー技術開発部 バイオマスグループ
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
バイオマス転換エネルギーの効率的生産のため、非食糧系エネルギー植物の遺伝子組換えを利用した改良に大きな期待が寄せられている。しかし、これらの非食糧系エネルギー植物の形質転換技術に関する研究は乏しく、形質転換技術の活用による改良が困難な状況にある。我々は、これまでに難形質転換植物の克服を可能にすると期待されるスーパーアグロバクテリウムを開発してきた。スーパーアグロバクテリウムとは、アグロバクテリウムを介した植物への遺伝子導入の際、感染時に植物体が生成する制御因子として知られているエチレン生成を抑制する能力が付与されたアグロバクテリウムのことをいう。これによって、植物への遺伝子導入能力を向上させることに成功した。本研究開発では、このスーパーアグロバクテリウムを活用した非食糧系エネルギー植物の形質転換技術の開発及び組換え植物栽培施設での栽培技術の開発を目指す。
具体的には、
1)スーパーアグロバクテリウムの高度化に関する研究開発(担当:筑波大学

2)スーパーアグロバクテリウムの効果的利用技術に関する研究開発(担当:千葉大学)
3)エネルギー植物の組換え植物栽培施設での栽培技術の研究開発(担当:筑波大学)
を分担して遂行した。
英文要約Title: R&D of Genetic Transformation and Cultivation Technology for Energy Plants (FY2009-FY2011) FY 2010 Annual Report
In this R&D, genetic transformation methods for non-food energy plants using super-Agrobacterium and cultivation technologies of non-food energy plants in GMP facilities will be developed.
1) Advancement of super-Agrobacterium
In utilization of antibiotics to maintain plasmid carrying ACC deaminase, the encoding gene is integrated into Agrobacterium genome. However, homologous recombination method has been reported in only a few cases. We aim to establish a recombination protocol in the first year. Considering that IAA can induce ethylene accumulation, regions containing genes which are down-regulated by IAA were chosen as marker. Selection of the marker gene and the electroporation condition were modified and colonies were isolated from the medium containing proper antibiotics. The inserted marker gene extracted from the genome was confirmed by PCR. The ACC deaminase gene is scheduled to be introduced into Agrobacterium by using this method in the second year. Moreover, a next generation super-Agrobacterium with high capability of ethylene reduction activity will be developed by modification of ACC deaminase.
To determine the effect of super-Agrobacterium producing ACC deaminase, both immature zygotic embryos of Sorghum bicolor (L.) Moench reported before and proliferating leaf segments of Erianthus ravennae have been transformed. Currently, these plants are being cultured in medium for co-culture and callus induction. There is need to examine the transformation efficiency using the reporter gene, E. coli uidA, which encodes the beta-glucuronidase in the second year.
2) Effective utilization technology of super-Agrobacterium
We have developed the culture systems of potential energy plants, Erianthus ravannae, Miscanthus giganteus, Pennisetum purpurem, Panicum maximum, and Saccharum officinarum for genetic transformation. In this year, somatic embryogenesis and plant regeneration were formed from calli induced utilizing proliferating leaf segments of Erianthus ravannae and seeds of Saccharum officinarum. In the second year, we will develop the culture systems for the other energy plants and genetic transformation methods using the next generation super-Agrobacterium.
3) Cultivation technology of energy plants in GMP cultivation facilities
To develop cultivation technology of energy plants in GMP facilities, non-transformed plants of Sorghum bicolor and Erianthus ravannae have been cultivated in screen greenhouse for generation promotion between summer and winter. After growth in sterile culture medium, plants were successfully transfered to soil and grown at 23 degrees Celsius in long-day condition. Afterwards, they were translocated to screen greenhouse in short-day condition for the purpose of flowering and seed harvest. Plants of Sorghum bicolor but not Erianthus ravannae were grown to maturity. There is need to change the culture condition such as temperature or light quality.
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