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Comparative studies of mitochondrial proteomics reveal an intimate protein network of male sterility in wheat (Triticum aestivum L.).

Wang S, Zhang G, Zhang Y, Song Q, Chen Z, Wang J, Guo J, Niu N, Wang J, Ma S - J. Exp. Bot. (2015)

Bottom Line: A total of 71 differentially expressed mitochondrial proteins were found to be involved in pollen abortion and further identified by MALDI-TOF/TOF MS (matrix-assisted laser desorption/ionization-time of fight/time of flight mass spectrometry).Interactions between identified proteins were demonstrated by bioinformatics analysis, enabling a more complete insight into biological pathways involved in anther abortion and pollen defects.The results provide intriguing insights into the metabolic pathway of anther abortion induced by CHA-SQ-1 and also give useful clues to identify the crucial proteins of PHYMS and CMS in wheat.

View Article: PubMed Central - PubMed

Affiliation: College of Agronomy, Northwest A&F University, National Yangling Agricultural Biotechnology & Breeding Center, Yangling Branch of State Wheat Improvement Centre, Wheat Breeding Engineering Research Center, Ministry of Education, Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling, Shaanxi 712100, P. R. China.

No MeSH data available.


Related in: MedlinePlus

ROS burst induces premature tapetal PCD. (A–F) The O2– production rate (A), H2O2 (B), and MDA (C) contents, and the activities of SOD (D), POD (E), and CAT (F) in developing anthers. Data are means ±SD of three independent experiments. The significance of differences was assessed by Student’s t-test (*P<0.05, **P<0.01). Euns, the early uninucleate stage; Bns, the binucleate stage; Tns, the trinucleate stage. (G) Analysis of oxidation stress and energy supply genes by RT-PCR assay. Primers are listed in Supplementary Table S3 at JXB online. ACO, aconitase; APX, ascorbate peroxidase; PNDOR, pyridine nucleotide-disulphide oxidoreductase; GAPDH, glyceraldehydes-3-phosphate dehydrogenase; NFUDP4, nifU-like domain protein 4. (H) Detection of premature PCD by TUNEL assay in the anthers at different developmental stages. Red fluorescence represents staining with propidium iodide, and green fluorescence indicates TUNEL-positive nuclei of PCD cells. E, epidermis; En, endothecium; ML, middle layer; T, tapetum; Msp, microspore; P, pollen; dP, degenerated pollen. Scale bar=100 μm.
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Figure 4: ROS burst induces premature tapetal PCD. (A–F) The O2– production rate (A), H2O2 (B), and MDA (C) contents, and the activities of SOD (D), POD (E), and CAT (F) in developing anthers. Data are means ±SD of three independent experiments. The significance of differences was assessed by Student’s t-test (*P<0.05, **P<0.01). Euns, the early uninucleate stage; Bns, the binucleate stage; Tns, the trinucleate stage. (G) Analysis of oxidation stress and energy supply genes by RT-PCR assay. Primers are listed in Supplementary Table S3 at JXB online. ACO, aconitase; APX, ascorbate peroxidase; PNDOR, pyridine nucleotide-disulphide oxidoreductase; GAPDH, glyceraldehydes-3-phosphate dehydrogenase; NFUDP4, nifU-like domain protein 4. (H) Detection of premature PCD by TUNEL assay in the anthers at different developmental stages. Red fluorescence represents staining with propidium iodide, and green fluorescence indicates TUNEL-positive nuclei of PCD cells. E, epidermis; En, endothecium; ML, middle layer; T, tapetum; Msp, microspore; P, pollen; dP, degenerated pollen. Scale bar=100 μm.

Mentions: ROS contents and antioxidant enzyme activities were first measured (Fig. 4A–C). The levels of O2– in PHYMS-XN1376 and CMS-XN1376 anthers were significantly higher than in MF-XN1376 from the early uninucleate stage to the trinucleate stage (Fig. 4A); the levels increased rapidly and remained 20% higher in PHYMS-XN1376, and reached a maximum value of 133% in CMS-XN1376. Excess O2– was catalysed to form H2O2. The change of H2O2 content was similar to that of O2–; it apparently increased with anther development in PHYMS-XN1376 and reached a maximum value of 190% at the trinucleate stage, and remained 35% higher in CMS-XN1376 (Fig. 4B). ROS scavenging depends on antioxidant enzymes, such as SOD, CAT, and POD. However, in this experiment, the activities of these enzymes were significantly decreased in PHYMS-XN1376 and CMS-XN1376 anthers when excess ROS were generated (Fig. 4D–F), and this further disrupted the oxidative/antioxidative balance. Subsequently, excess ROS degraded polyunsaturated lipids to form MDA, and its level was significantly increased in anthers of PHYMS-XN1376 (maximum value of 147%) and CMS-XN1376 (maximum value of 432%) (Fig. 4C). Moreover, RT-PCR analysis showed that two oxidation stress-related genes [ascorbate peroxidase (APX) and pyridine nucleotide-disulphide oxidoreductase (PNDOR)] were down-regulated in PHYMS-XN1376 and CMS-XN1376 (Fig. 4G). The above results indicate that the production of ROS overwhelmed the antioxidant capacity, which disrupted the balance between ROS production and clearance in male-sterile anthers; therefore oxidative stress occurred. Therefore, these results suggest that the oxidative stress of the anther in PHYMS-XN1376 was the same as that in CMS-XN1376.


Comparative studies of mitochondrial proteomics reveal an intimate protein network of male sterility in wheat (Triticum aestivum L.).

Wang S, Zhang G, Zhang Y, Song Q, Chen Z, Wang J, Guo J, Niu N, Wang J, Ma S - J. Exp. Bot. (2015)

ROS burst induces premature tapetal PCD. (A–F) The O2– production rate (A), H2O2 (B), and MDA (C) contents, and the activities of SOD (D), POD (E), and CAT (F) in developing anthers. Data are means ±SD of three independent experiments. The significance of differences was assessed by Student’s t-test (*P<0.05, **P<0.01). Euns, the early uninucleate stage; Bns, the binucleate stage; Tns, the trinucleate stage. (G) Analysis of oxidation stress and energy supply genes by RT-PCR assay. Primers are listed in Supplementary Table S3 at JXB online. ACO, aconitase; APX, ascorbate peroxidase; PNDOR, pyridine nucleotide-disulphide oxidoreductase; GAPDH, glyceraldehydes-3-phosphate dehydrogenase; NFUDP4, nifU-like domain protein 4. (H) Detection of premature PCD by TUNEL assay in the anthers at different developmental stages. Red fluorescence represents staining with propidium iodide, and green fluorescence indicates TUNEL-positive nuclei of PCD cells. E, epidermis; En, endothecium; ML, middle layer; T, tapetum; Msp, microspore; P, pollen; dP, degenerated pollen. Scale bar=100 μm.
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Related In: Results  -  Collection

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Figure 4: ROS burst induces premature tapetal PCD. (A–F) The O2– production rate (A), H2O2 (B), and MDA (C) contents, and the activities of SOD (D), POD (E), and CAT (F) in developing anthers. Data are means ±SD of three independent experiments. The significance of differences was assessed by Student’s t-test (*P<0.05, **P<0.01). Euns, the early uninucleate stage; Bns, the binucleate stage; Tns, the trinucleate stage. (G) Analysis of oxidation stress and energy supply genes by RT-PCR assay. Primers are listed in Supplementary Table S3 at JXB online. ACO, aconitase; APX, ascorbate peroxidase; PNDOR, pyridine nucleotide-disulphide oxidoreductase; GAPDH, glyceraldehydes-3-phosphate dehydrogenase; NFUDP4, nifU-like domain protein 4. (H) Detection of premature PCD by TUNEL assay in the anthers at different developmental stages. Red fluorescence represents staining with propidium iodide, and green fluorescence indicates TUNEL-positive nuclei of PCD cells. E, epidermis; En, endothecium; ML, middle layer; T, tapetum; Msp, microspore; P, pollen; dP, degenerated pollen. Scale bar=100 μm.
Mentions: ROS contents and antioxidant enzyme activities were first measured (Fig. 4A–C). The levels of O2– in PHYMS-XN1376 and CMS-XN1376 anthers were significantly higher than in MF-XN1376 from the early uninucleate stage to the trinucleate stage (Fig. 4A); the levels increased rapidly and remained 20% higher in PHYMS-XN1376, and reached a maximum value of 133% in CMS-XN1376. Excess O2– was catalysed to form H2O2. The change of H2O2 content was similar to that of O2–; it apparently increased with anther development in PHYMS-XN1376 and reached a maximum value of 190% at the trinucleate stage, and remained 35% higher in CMS-XN1376 (Fig. 4B). ROS scavenging depends on antioxidant enzymes, such as SOD, CAT, and POD. However, in this experiment, the activities of these enzymes were significantly decreased in PHYMS-XN1376 and CMS-XN1376 anthers when excess ROS were generated (Fig. 4D–F), and this further disrupted the oxidative/antioxidative balance. Subsequently, excess ROS degraded polyunsaturated lipids to form MDA, and its level was significantly increased in anthers of PHYMS-XN1376 (maximum value of 147%) and CMS-XN1376 (maximum value of 432%) (Fig. 4C). Moreover, RT-PCR analysis showed that two oxidation stress-related genes [ascorbate peroxidase (APX) and pyridine nucleotide-disulphide oxidoreductase (PNDOR)] were down-regulated in PHYMS-XN1376 and CMS-XN1376 (Fig. 4G). The above results indicate that the production of ROS overwhelmed the antioxidant capacity, which disrupted the balance between ROS production and clearance in male-sterile anthers; therefore oxidative stress occurred. Therefore, these results suggest that the oxidative stress of the anther in PHYMS-XN1376 was the same as that in CMS-XN1376.

Bottom Line: A total of 71 differentially expressed mitochondrial proteins were found to be involved in pollen abortion and further identified by MALDI-TOF/TOF MS (matrix-assisted laser desorption/ionization-time of fight/time of flight mass spectrometry).Interactions between identified proteins were demonstrated by bioinformatics analysis, enabling a more complete insight into biological pathways involved in anther abortion and pollen defects.The results provide intriguing insights into the metabolic pathway of anther abortion induced by CHA-SQ-1 and also give useful clues to identify the crucial proteins of PHYMS and CMS in wheat.

View Article: PubMed Central - PubMed

Affiliation: College of Agronomy, Northwest A&F University, National Yangling Agricultural Biotechnology & Breeding Center, Yangling Branch of State Wheat Improvement Centre, Wheat Breeding Engineering Research Center, Ministry of Education, Key Laboratory of Crop Heterosis of Shaanxi Province, Yangling, Shaanxi 712100, P. R. China.

No MeSH data available.


Related in: MedlinePlus