Cytological and comparative proteomic analyses on male sterility in Brassica napus L. induced by the chemical hybridization agent monosulphuron ester sodium

Citation: Cheng Y, Wang Q, Li Z, Cui J, Hu S, et al. (2013) Cytological and Comparative Proteomic Analyses on Male Sterility in Brassica napus L. Induced by the Chemical Hybridization Agent Monosulphuron Ester Sodium. PLoS ONE 8(11): e80191. doi:10.1371/journal.pone.0080191Male sterility induced by a chemical hybridization agent (CHA) is an important tool for utilizing crop heterosis. Monosulphuron ester sodium (MES), a new acetolactate synthase-inhibitor herbicide belonging to the sulphonylurea family, has been developed as an effective CHA to induce male sterility in rapeseed (Brassica napus L.). To understand MES-induced male sterility in rapeseed better, comparative cytological and proteomic analyses were conducted in this study. Cytological analysis indicated that defective tapetal cells and abnormal microspores were gradually generated in the developing anthers of MES-treated plants at various development stages, resulting in unviable microspores and male sterility. A total of 141 differentially expressed proteins between the MES-treated and control plants were revealed, and 131 of them were further identified by MALDI-TOF/TOF MS. Most of these proteins decreased in abundance in tissues of MES-treated rapeseed plants, and only a few increased. Notably, some proteins were absent or induced in developing anthers after MES treatment. These proteins were involved in several processes that may be crucial for tapetum and microspore development. Down-regulation of these proteins may disrupt the coordination of developmental and metabolic processes, resulting in defective tapetum and abnormal microspores that lead to male sterility in MES-treated plants. Accordingly, a simple model of CHA-MES-induced male sterility in rapeseed was established. This study is the first cytological and dynamic proteomic investigation on CHA-MES-induced male sterility in rapeseed, and the results provide new insights into the molecular events of male sterility

Unravelling salt stress in plants through proteomics

Plants like other organisms are mostly under the threat of various stresses (both biotic as well as abiotic). Being sessile, plants lack the mechanisms to flee from these unfavourable situations. The development of exclusive and complicated responses to these environmental stresses in plants has fostered through evolution. Such alterations can influence plant growth, production and productivity in agriculture, plant nutritional potential and metabolic profile. Hence, plant abiotic stress has always been a matter of concern for the world economy and maintenance of human life on earth. Salinity stress, being one of the main abiotic stresses, may bring the morphological, anatomical, and physiological changes in plants. Distributed in both irrigated and non-irrigated areas of the world, around 6% of the world’s total land area is affected by salt stress. So, it is a major concern to adopt the strategies against this great challenge by unravelling the mechanisms to overcome salt stress. In order to meet the challenges for biotechnological improvement of crop productivity; various steps involving genes, transcripts, proteins and metabolites, controlling the stress resistance and/or architecture of crop plants in a wide array of environments needed to be recognized. Proteomics, the protein complement of genome, these days is one of the leading branches of research which enables the large-scale ­scanning of various substances, and offers great potential for post-genomics to elucidate the genotype-phenotype connections. The present chapter is an account of current knowledge in this regard. It focuses on effects of salt stress unrevealed by proteomics tools. It comprises information on recent advances in proteomics, which could be a new opportunity to comprehend abiotic responses and categorize genes responsible for significant crop traits

Universal sample preparation method integrating trichloroacetic acid/acetone precipitation with phenol extraction for crop proteomic analysis

Crop plants contain large amounts of secondary compounds that interfere with protein extraction and gel-based proteomic analysis. Thus, a protein extraction protocol that can be easily applied to various crop materials with minimal optimization is essential. Here we describe a universal protocol for total protein extraction involving trichloroacetic acid (TCA)/acetone precipitation followed by SDS and phenol extraction. Through SDS extraction, the proteins precipitated by the TCA/acetone treatment can be fully resolubilized and then further purified by phenol extraction. This protocol combines TCA/acetone precipitation, which aggressively removes nonprotein compounds, and phenol extraction, which selectively dissolves proteins, resulting in effective purification of proteins from crop tissues. This protocol can also produce high-quality protein preparations from various recalcitrant tissues, and therefore it has a wide range of applications in crop proteomic analysis. Designed to run on a small scale, this protocol can be completed within 5 h. © 2014 Nature America, Inc. All rights reserved

Separomics applied to the proteomics and peptidomics of low-abundance proteins: choice of methods and challenges - a review

The enrichment and isolation of proteins are considered limiting steps in proteomic studies. Identification of proteins whose expression is transient, those that are of low-abundance, and of natural peptides not described in databases, is still a great challenge. Plant extracts are in general complex, and contaminants interfere with the identification of proteins involved in important physiological processes, such as plant defense against pathogens. This review discusses the challenges and strategies of separomics applied to the identification of low-abundance proteins and peptides in plants, especially in plants challenged by pathogens. Separomics is described as a group of methodological strategies for the separation of protein molecules for proteomics. Several tools have been used to remove highly abundant proteins from samples and also non-protein contaminants. The use of chromatographic techniques, the partition of the proteome into subproteomes, and an effort to isolate proteins in their native form have allowed the isolation and identification of rare proteins involved in different processes