Proteomic and Phosphoproteomic Analyses Reveal a Complex Network Regulating Pollen Abortion and Potential Candidate Proteins in TCMS Wheat

Abstract

Thermosensitive sterile lines are natural materials for exploring the effects of anther development on male fertility. To study the possible molecular mechanisms regulating protein activity during the induction of male sterility, proteomic and phosphoproteomic analyses with tandem mass tags (TMTs) were used to study the binucleate anther of the thermosensitive sterile wheat line YS3038. A total of 9072 proteins, including 5019 phosphoproteins, were identified. Enrichment analyses of differentially abundant proteins (DAPs) and phosphoproteins (DAPPs) in metabolic pathways showed that both were mainly related to energy metabolism. Soluble sugar and ATP content were significantly decreased, free fatty acid content was significantly increased, and ROS was abnormally accumulated in male sterile YS3038-A. In addition, 233 kinase–substrate pairs involved in potential phosphorylation control networks were predicted to regulate fertility. Candidate proteins were identified, and a quantitative real-time polymerase chain reaction (qRT-PCR) analysis was used to validate the TMT results. TaPDCD5 is likely to be involved in fertility conversion of YS3038 by barley stripe mosaic virus-induced gene silencing (BSMV-VIGS). Our data provide new insights into the mechanism of TCMS, which has value for identifying potential candidate proteins associated with the formation or abortion of pollen and promotion of wheat heterosis utilization.

ROS accumulation-induced tapetal PCD timing changes leads to microspore abortion in cotton CMS lines

“…NADPH oxidase was widely considered as the main source of ROS burst/generation in anther, especially RBOHE [ 48 , 55 , 56 ]. Therefore, we tested the expression of RBOHE (Additional file 4 : Table 2).…”

“…There are three main manifestations. Firstly, excessive ROS accumulation in sterile lines may affect the normal development of microspores and accompany with early degradation of tapetum, such as Kenaf CMS line 722HA and wheat YS3038-A [ 42 , 48 ]. Secondly, ROS remains at a continuously low level throughout microspore development, so that tapetal degradation is abolished or delayed, as shown in rice Defective Tapetum Death1 ( DTC1 ) anthers, Arabidopsis thaliana Respiratory burst oxidase homology ( RBOH ) mutant and Wheat male-sterile2 line [ 49 – 51 ].…”

Large-scale phosphoproteome analysis in wheat seedling leaves provides evidence for extensive phosphorylation of regulatory proteins during CWMV infection

“…Bioinformatics analysis was performed according to the previously described protocols [ 18 , 20 , 52 , 70 ]. The obtained proteins were further classified into three categories (biological process, cellular compartment, and molecular function) using the Uniprot-GOA database ( http://www.ebi.ac.uk/GOA/ , accessed on 20 May 2021) as a reference.…”

“…For example, Zeng and others found extensive phosphorylation of proteins associated with bast fiber growth in ramie using a phosphoproteomic analysis approach [ 17 ]. Ma and others also conducted proteomic and phosphoproteomic analyses and revealed a complex signaling network that is responsive to pollen abortion [ 18 ]. To better understand the global protein phosphorylation in the effector-triggered immunity (ETI), Yasuhiro and colleagues did a phosphoproteomic screen using Arabidopsis thaliana as the model plant and identified 109 differentially phosphorylated residues in the membrane-associated proteins [ 19 ].…”

Antibacterial Activity of Allicin-Inspired Disulfide Derivatives against Xanthomonas axonopodis pv. citri

“…A tandem mass tags (TMT)-labeled quantitative proteomics assay was carried out according to a method reported elsewhere [ 37 , 38 , 39 , 40 ]. For sample preparation, Xac cells (OD 595 ≈ 0.1) and compound 1 were co-incubated in NB medium in a shaker (250 rpm) at 28 °C until the negative control group had grown to OD 595 ≈ 0.6 (about 10 h).…”