A High Through-put Screen to Identify and Clone Maize and Arabidopsis Cell-wall Biogenesis-related Genes

傅里叶变换红外光谱（FTIR）的判别分析已被开发为一种特异性高通量的方法，以识别影响植物细胞壁组分和结构的突变。 该方法将用于证明在玉米和拟南芥的诱变群体中鉴定在生长和发育期间植物细胞壁结构的生物发生和动态改变中的广泛的突变体所需的通量率。 将采用正向和反向遗传筛选。 FTIR“正向”屏幕将检测由DNA“标签”插入引起的壁结构和结构的特定改变，而不管改变的遗传基础如何。 在这方面，屏幕将在最终产品层面上具有强大的选择性。 待鉴定的突变可能包括在生长期间引起壁基质形成、壁组分分泌和靶向、壁结构和动力学缺陷的那些突变。 随后可以基于标签的插入位置来鉴定这些突变体中被破坏的基因。 “反向”遗传学方法将检查将DNA标签插入已知与细胞壁生物合成有关的基因的影响。 预计第二种方法将提供一组广泛的细胞壁突变体，这将有助于进一步开发FTIR光谱库。 这些文库将有助于诊断FTIR选择的未知遗传缺陷的突变体。 一个主要的实际目标是产生在细胞壁组成和结构中具有遗传限定的变异的植物，以允许评估这些修饰对植物发育的影响。 随着FTIR突变得到确认，植物生物学社区将通过网站了解这些突变，种子和克隆将通过已建立的库存中心提供给社区。

Discriminate analysis of Fourier Transform Infrared Spectra (FTIR) has been developed as a specific, high throughput method to identify mutations that affect plant cell wall components and architecture. This method will be employed to demonstrate the throughput rates needed to identify, in mutagenized populations of maize and Arabidopsis, a broad range of mutants in the biogenesis and dynamic alteration of plant cell wall architecture during growth and development. Both forward and reverse genetic screens will be employed. The FTIR "forward" screen will detect specific alterations in wall structure and architecture caused by the insertion of DNA 'tags", regardless of the genetic basis of the alteration. In this respect, the screen will be powerfully selective at the end-product level. Mutations to be identified could include those causing defects in wall substrate formation, wall component secretion and targeting, wall architecture and dynamics during growth. Genes disrupted in these mutants may subsequently be identified based on the insertion position of the tags. The "reverse" genetics approach will examine the effects of inserting DNA tags into genes that are already known to be related to cell wall biosynthesis. It is anticipated that this second approach will provide a broad set of cell wall mutants that will serve to further the development of FTIR spectrum libraries. These libraries will be useful to diagnose FTIR-selected mutants of unknown genetic deficiency. A major practical goal is to generate plants with genetically defined variation in cell wall composition and architecture to permit assessment of these modifications on plant development. As FTIR mutations are confirmed, the plant biology community will be informed of them through a web site and seeds and clones will be provided to the community through established stock centers.