Lewandowska Dominika, Zhang Runxuan, Colas Isabelle, Uzrek Nicola, Waugh Robbie
Cell and Molecular Sciences, The James Hutton Institute, Dundee, United Kingdom.
Information and Computational Sciences, The James Hutton Institute, Dundee, United Kingdom.
Front Plant Sci. 2019 Apr 2;10:393. doi: 10.3389/fpls.2019.00393. eCollection 2019.
Meiosis is a highly dynamic and precisely regulated process of cell division, leading to the production of haploid gametes from one diploid parental cell. In the crop plant barley (), male meiosis occurs in anthers, in specialized cells called meiocytes. Barley meiotic tissue is scarce and not easily accessible, making meiosis study a challenging task. We describe here a new micro-proteomics workflow that allows sensitive and reproducible genome-wide label-free proteomic analysis of individual staged barley anthers. This micro-proteomic approach detects more than 4,000 proteins from such small amounts of material as two individual anthers, covering a dynamic range of protein relative abundance levels across five orders of magnitude. We applied our micro-proteomics workflow to investigate the proteome of the developing barley anther containing pollen mother cells in the early stages of meiosis and we successfully identified 57 known and putative meiosis-related proteins. Meiotic proteins identified in our study were found to be key players of many steps and processes in early prophase such as: chromosome condensation, synapsis, DNA double-strand breaks or crossover formation. Considering the small amount of starting material, this work demonstrates an important technological advance in plant proteomics and can be applied for proteomic examination of many size-limited plant specimens. Moreover, it is the first insight into the proteome of individual barley anther at early meiosis. The proteomic data have been deposited to the ProteomeXchange with the accession number PXD010887.
减数分裂是一个高度动态且受到精确调控的细胞分裂过程，可使一个二倍体亲代细胞产生单倍体配子。在农作物大麦中，雄性减数分裂发生在花药中，即在称为减数分裂细胞的特化细胞中。大麦的减数分裂组织稀少且不易获取，这使得减数分裂研究成为一项具有挑战性的任务。我们在此描述一种新的微量蛋白质组学工作流程，该流程能够对单个发育阶段的大麦花药进行灵敏且可重复的全基因组无标记蛋白质组分析。这种微量蛋白质组学方法能够从两个单个花药这么少量的材料中检测到4000多种蛋白质，涵盖了五个数量级的蛋白质相对丰度水平的动态范围。我们应用微量蛋白质组学工作流程来研究处于减数分裂早期、含有花粉母细胞的发育中大麦花药的蛋白质组，并成功鉴定出57种已知的和推测的与减数分裂相关的蛋白质。我们研究中鉴定出的减数分裂蛋白是减数分裂前期许多步骤和过程的关键参与者，例如：染色体浓缩、联会、DNA双链断裂或交叉形成。考虑到起始材料量少，这项工作展示了植物蛋白质组学的一项重要技术进步，可应用于对许多大小受限的植物标本进行蛋白质组学检测。此外，这是对减数分裂早期单个大麦花药蛋白质组的首次深入研究。蛋白质组学数据已存入ProteomeXchange，登录号为PXD010887。
