Thermal cycler for plant molecular genetics

本提案要求资金购买一个热循环器来支持PCR（聚合酶链反应），以支持植物细胞壁和植物生物质生产的生物学和生物技术研究。PCR和这种类型的仪器是必不可少的分子生物学和遗传学实验室的许多方法和实验方法，包括基因克隆，基因型测定，非定量或半定量基因表达，和DNA测序。所有这些方法都是申请人实验室进行的研究的核心。该仪器将支持与植物细胞壁生物合成和改善木本物种细胞壁有关的三个领域的实际研究。其中一组研究涉及次级细胞壁生物合成的调控（次级细胞壁构成了树木和木质纤维素生物能源原料的大部分木质生物质）。我们已经确定了模式植物拟南芥次生壁形成调控网络中的负反馈回路，并正在使用多种依赖于PCR的方法研究参与该调控工作的转录因子的功能。在另一组研究中，我们正在将这些和其他信息转化为杨树，一种生长迅速的树木，是生物能源原料的有力候选。pcr辅助方法将用于研究杨树次生细胞壁（木材）形成的调控，并利用重要细胞壁基因的自然变异来改善原料。研究的最后一个领域调查了已知最坚固的植物细胞壁材料的形成，即花粉粒的外壁（外壁）。我们正在使用拟南芥作为模型系统来研究外叶生物合成所需的基因编码，这是知之甚少的，并且将严重依赖于PCR和所需的热循环器来进行这些研究。

This proposal requests funds to purchase a thermal cycler to support for PCR (polymerase chain reaction) to support research on the biology and biotechnology of plant cell walls and plant biomass production. PCR and instruments of this type are essential in molecular biology and genetics labs for many methodologies and experimental approaches including gene cloning, genotype determination, non-quantitative or semi-quantitative gene expression, and DNA sequencing. All of these approaches are central to the research being carried out in the applicant's lab. The instrument will support research in three areas related to plant cell wall biosynthesis and the improvement of cell walls in woody species for practical purposes. One set of studies involves the regulation of secondary cell wall biosynthesis (which form the bulk of the woody biomass in trees and in lignocellulosic bioenergy feedstocks). We have identified a negative feedback loop in the regulatory network governing secondary wall formation in the model plant Arabidopsis, and are investigating the functions of the transcription factors involved in this regulation work using a variety of approaches that depend on PCR. In another set of studies, we are translating this and other information into poplar, a fast growing tree that is a strong candidate as a bioenergy feedstock. PCR-aided approaches will be used to investigate regulation of secondary cell wall (wood) formation in poplar, and the use natural variants in important cell wall genes to improve the feedstock. A final area of research study investigates the formation of the strongest plant cell wall material known, the outer wall (exine) of pollen grains. We are using Arabidopsis as a model system to investigate the genes encoding required for exine biosynthesis, which is poorly understood, and will rely heavily on PCR and the requested thermal cycler to carry out these studies.