巨菌草叶片cDNA-AFLP体系建立初探
巨菌草(Pennisetum giganteum z.x.lin)是高产优质的菌草之一，不仅可作为食用菌高质量的培养料和食草动物的优质饲料经济草，还是可作为生物质发电、制造燃料乙醇的能源草，甚至可作为新材料用于生产纤维板和防弹衣等高科技领域。目前，阻碍巨菌草在北方大面积推广的主要的问题是无法自然过冬。因此，为了提高巨菌草的抗寒性，满足国内外低温地区发展菌草业的需要，本文首次以巨菌草为材料，通过对RNA提取、内切酶和引物筛选以及PCR扩增等关键环节的实验条件进行优化，试图建一套适用于巨菌草的cDNA-AFLP差异分析体系。实验结果表明：通过条件优化，初步建立起一套可用于菌草的低温胁迫下cDNA-AFLP差异分析的实验体系，这为转录组水平筛选获得巨菌草低温反应相关基因提供了技术支持，为进一步巨菌草抗寒分子育种研究和最终解决寒冷地区巨菌草推广奠定基础。
关键词：巨菌草；cDNA-AFLP；叶片；体系优化
First Exploration Establishment of the Pennisetum giganteum z.x.Lin blade cDNA - AFLP System
Abstract: Pennisetum giganteum z. x. Lin is one of the Juncao with high yield and quality. It can be used as high quality training material for edible mushroom and cultivate high-quality forage economy grass for herbivores. It also can be used as biomass power generation, energy grass of making fuel ethanol, and even can be used as a new material for the production of fiber board, bulletproof vests and other high-tech fields. At present, the main reason of hindering pennisetum giganteum z. x. Lin of the northern large area is not natural to spend the winter. Therefore, in order to improve its the cold resistance and to meet the need of the development of it in low temperature area at home and abroad, this paper chooses pennisetum giganteum z. x. Lin as material for the first time. Through the experimental conditions of RNA extraction, enzyme and primer screening and PCR amplification and other key aspects of the optimization, the paper try to build the difference analysis system of cDNA-AFLP that is suitable to pennisetum giganteum z. x. Lin. The experimental results showed that by optimizing the conditions, this difference analysis system of cDNA-AFLP provides technical support for the transcriptome selection getting the level of screening related genes to low temperature reaction of pennisetum giganteum z. x. Lin. It lays a foundation for further study on cold resistant molecular breeding of pennisetum giganteum z. x. Lin and the final settlement of promotion of pennisetum giganteum z. x. Lin in cold regions.
Key words: Pennisetum giganteum z.x.Lin; cDNA-AFLP; blade; system optimization
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