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Functional verification of TaPDCD5 via BSMV-VIGS. (A) Leaf characteristics of positive control (PDS gene-silenced) plants. (A), Early stage of positive control plants. (B), Metaphase of positive control plants. (C), Late stage of positive control plants. (D-F) Fertility of ears. (D), Negative control. E, Positive control. F, TaPDCD5-silenced plants. (G-I) Anther morphology. (G), Negative control. (H), Positive control. L, TaPDCD5-silenced plants. (J) Relative expression of gene-silenced plants. Significant differences were determined by Student's t-test (* p < 0.05).
Source publication
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 the...
Contexts in source publication
Context 1
... positive control group showed an obvious whitening of leaves. The silencing of the TaPDCD5 (TraesCS1A02G368100) led to a significant decrease in the expression level of TaPDCD5 and seed setting rate of YS3038 under fertile conditions (Figure 9 and Supplementary Materials Table S10). In the TaPDCD5-silencing plant, the anther showed an abnormal shape. ...
Context 2
... positive control group showed an obvious whitening of leaves. The silencing of the TaPDCD5 (TraesCS1A02G368100) led to a significant decrease in the expression level of TaPDCD5 and seed setting rate of YS3038 under fertile conditions (Figure 9 and Supplementary Materials Table S10). In the TaPDCD5-silencing plant, the anther showed an abnormal shape. ...
Citations
... 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][50][51]. ...
... 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). ...
Background
Cytoplasmic male sterility (CMS) is the basis of heterosis exploitation. CMS has been used to hybrid production in cotton, but its molecular mechanism remains unclear. CMS is associated with advanced or delayed tapetal programmed cell death (PCD), and reactive oxygen species (ROS) may mediate this process. In this study, we obtained Jin A and Yamian A, two CMS lines with different cytoplasmic sources.
Results
Compared with maintainer Jin B, Jin A anthers showed advanced tapetal PCD with DNA fragmentation, producing excessive ROS which accumulated around the cell membrane, intercellular space and mitochondrial membrane. The activities of peroxidase (POD) and catalase (CAT) enzymes which can scavenge ROS were significantly decreased. However, Yamian A tapetal PCD was delayed with lower ROS content, and the activities of superoxide dismutase (SOD) and POD were higher than its maintainer. These differences in ROS scavenging enzyme activities may be caused by isoenzyme gene expressions. In addition, we found the excess ROS generated in Jin A mitochondria and ROS overflow from complex III might be the source in parallel with the reduction of ATP content.
Conclusion
ROS accumulation or abrogation were mainly caused by the joint action of ROS generation and scavenging enzyme activities transformation, which led to the abnormal progression of tapetal PCD, affected the development of microspores, and eventually contributed to male sterility. In Jin A, tapetal PCD in advance might be caused by mitochondrial ROS overproduction, accompanied by energy deficiency. The above studies will provide new insights into the cotton CMS and guide the follow-up research ideas.
... 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). ...
Xanthomonas axonopodis pv. citri (Xac) belongs to the Gram-negative species, causing citrus canker that seriously affects the fruit yield and quality of many rutaceae plants. Herein, we found that compound 2-(butyldisulfanyl) quinazolin-4(3H)-one exhibited remarkable anti-Xac activity in vitro with a half effective concentration (EC50) of 2.6 μg/mL, while the positive controls thiodiazole-copper with 57 μg/mL and bismerthiazol with 68 μg/mL and this compound showed great anti-citrus canker activity in vivo. This active compound also was confirmed to reduce biofilm formation, increase the level of reactive oxygen species, damage the morphological structure of the bacteria, and cause bacterial death. Proteomics and RT-qPCR analysis results indicated that this compound down-regulated the expression of enzymes in the MEP (2-methyl-D-erythritol 4-phosphate) pathway and might achieve destructive ability of Xac. Overall, this study indicates that such derivatives could be a promising scaffold to develop novel bactericides to control citrus canker.
Cytoplasmic male sterility (CMS) plays a crucial role in the utilization of heterosis. The petaloid anther abortion in oil tea (Camellia oleifera Abel.) constitutes a CMS phenomenon, which is of great value for the hybrid breeding of oil tea. However, as the mechanism of its CMS is still poorly understood, it is necessary to study the cytology and physiological characteristics of anther abortion. In this study, a C. oleifera cultivar, Huashuo (HS), and its petalized CMS mutant (HSP) were used as materials to explore this mechanism. Compared with HS, cytological analysis demonstrated that HSP showed early-onset tapetum programmed cell death (PCD) and an organelle disorder phenotype during the tetrad stage. In HSP, anthers exhibited elevated levels of calcium deposition in anther wall tissues, tapetum layers, and microspores, and yet calcium accumulation was abnormal at the later stage. The contents of hydrogen peroxide and MDA in HSP anthers were higher, and the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were lower than those of HS, which resulted in an excessive accumulation of reactive oxygen species (ROS). Real-time quantitative PCR confirmed that the transcription levels of CoPOD and CoCAT genes encoding key antioxidant enzymes in HSP were downregulated compared with HS in early pollen development; the gene CoCPK, which encodes a calcium-dependent protein kinase associated with antioxidase, was upregulated during the critical period. Thus, we suggest that excessive ROS as a signal breaks the balance of the antioxidant system, and along with an abnormal distribution of calcium ions, leads to the early initiation of PCD in the tapetum, and ultimately leads to pollen abortion for HSP. These results lay a cytological and physiological foundation for further studies on the CMS mechanism, and provide information for breeding male-sterile lines of C. oleifera.
Background
Chinese wheat mosaic virus (CWMV) often causes severe damage to wheat (Triticum aestivum L.) growth and yield. It is well known that a successful infection in plants depends on a complex interaction between the host plant and the pathogen. Post-translational modification (PTM) of proteins is considered to be one of the main processes that decides the outcome of the plant-pathogen arms race during this interaction. Although numerous studies have investigated PTM in various organisms, there has been no large-scale phosphoproteomic analysis of virus-infected wheat plants. We therefore aimed to investigate the CWMV infection-induced phosphoproteomics changes in wheat by high-resolution liquid chromatography-tandem mass spectroscopy (LC–MS/MS) using affinity-enriched peptides followed by comprehensive bioinformatics analysis.
Results
Through this study, a total of 4095 phosphorylation sites have been identified in 1968 proteins, and 11.6% of the phosphorylated proteins exhibited significant changes (PSPCs) in their phosphorylation levels upon CWMV infection. The result of Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that most of the PSPCs were associated with photosynthesis, plant-pathogen interactions, and MAPK signaling pathways. The protein–protein interaction (PPI) network analysis result showed that these PSPCs were mainly participated in the regulation of biosynthesis and metabolism, protein kinase activities, and transcription factors. Furthermore, the phosphorylation levels of TaChi1 and TaP5CS, two plant immunity-related enzymes, were significantly changed upon CWMV infection, resulting in a significant decrease in CWMV accumulation in the infected plants.
Conclusions
Our results indicate that phosphorylation modification of protein plays a critical role in wheat resistance to CWMV infection. Upon CWMV infection, wheat plants will regulate the levels of extra- and intra-cellular signals and modifications of enzyme activities via protein phosphorylation. This novel information about the strategies used by wheat to resist CWMV infection will help researchers to breed new CWMV-resistant cultivars and to better understand the arms race between wheat and CWMV.
