Thermal cycler for plant molecular genetics

该提案要求提供资金，用于购买一台热循环仪，以支持PCR（聚合酶链式反应），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和这种类型的仪器在分子生物学和遗传学实验室中对于许多方法和实验方法是必不可少的，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正在进行的研究的核心。该仪器将支持与植物细胞壁生物合成有关的三个领域的研究，并为实用目的改进木本物种的细胞壁。一组研究涉及次生细胞壁生物合成的调节（其形成树木和木质纤维素生物能源原料中的大部分木质生物质）。我们已经确定了一个负反馈回路中的监管网络管理次生壁形成的模式植物拟南芥，并正在调查的功能，在这种调控工作中涉及的转录因子使用各种方法，依赖于PCR。在另一组研究中，我们正在将这些信息和其他信息转化为白杨，这是一种快速生长的树木，是生物能源原料的有力候选者。利用PCR技术研究白杨次生细胞壁（木材）形成的调控，并利用重要细胞壁基因的天然变异体来改良原料。研究的最后一个领域是研究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一个模型系统，以调查基因编码所需的外壁生物合成，这是知之甚少，并将在很大程度上依赖于PCR和所要求的热循环仪进行这些研究。

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.