Thermal cycler for plant molecular genetics

该提案要求提供资金购买热循环器以支持聚合酶链式反应，以支持对植物细胞壁的生物学和生物技术以及植物生物质生产的研究。在分子生物学和遗传学实验室中，包括基因克隆、基因决定、非定量或半定量基因表达和DNA测序在内的许多方法和实验手段都离不开这种类型的聚合酶链式反应和仪器。所有这些方法都是申请者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和改进木本植物细胞壁相关的三个领域的研究，用于实际目的。一组研究涉及对次生细胞壁生物合成的调控(这构成了树木和木质纤维素生物能源原料中木质生物量的大部分)。我们已经在模式植物拟南芥中发现了一个控制次生壁形成的调控网络中的负反馈环，并正在使用依赖于PCR的各种方法来研究参与这一调控工作的转录因子的功能。在另一组研究中，我们正在将这一信息和其他信息转化为杨树，这是一种生长迅速的树，是作为生物能源原料的有力候选者。利用聚合酶链式反应辅助的方法将研究杨树次生细胞壁(木材)形成的调节，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是调查已知的最坚固的植物细胞壁材料的形成，即花粉粒的外壁。我们正在使用拟南芥作为一个模型系统来研究外壁生物合成所需的基因编码，这一点人们知之甚少，并将严重依赖于PCR和所需的热循环仪来进行这些研究。

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.