Limits...
iTRAQ-based quantitative proteome and phosphoprotein characterization reveals the central metabolism changes involved in wheat grain development.

Ma C, Zhou J, Chen G, Bian Y, Lv D, Li X, Wang Z, Yan Y - BMC Genomics (2014)

Bottom Line: Hierarchical clustering analysis indicated that the DEPs involved in starch biosynthesis, storage proteins, and defense/stress-related proteins significantly accumulated at the late grain development stages, while those related to protein synthesis/assembly/degradation and photosynthesis showed an opposite expression model during grain development.Numerous DEPs are involved in grain starch and protein syntheses as well as adverse defense, which set an important basis for wheat yield and quality.Particularly, some key DEPs involved in starch biosynthesis and stress/defense were phosphorylated, suggesting their roles in wheat grain development.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China. zhimin206@263.net.

ABSTRACT

Background: Wheat (Triticum aestivum L.) is an economically important grain crop. Two-dimensional gel-based approaches are limited by the low identification rate of proteins and lack of accurate protein quantitation. The recently developed isobaric tag for relative and absolute quantitation (iTRAQ) method allows sensitive and accurate protein quantification. Here, we performed the first iTRAQ-based quantitative proteome and phosphorylated proteins analyses during wheat grain development.

Results: The proteome profiles and phosphoprotein characterization of the metabolic proteins during grain development of the elite Chinese bread wheat cultivar Yanyou 361 were studied using the iTRAQ-based quantitative proteome approach, TiO2 microcolumns, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Among 1,146 non-redundant proteins identified, 421 showed at least 2-fold differences in abundance, and they were identified as differentially expressed proteins (DEPs), including 256 upregulated and 165 downregulated proteins. Of the 421 DEPs, six protein expression patterns were identified, most of which were up, down, and up-down expression patterns. The 421 DEPs were classified into nine functional categories mainly involved in different metabolic processes and located in the membrane and cytoplasm. Hierarchical clustering analysis indicated that the DEPs involved in starch biosynthesis, storage proteins, and defense/stress-related proteins significantly accumulated at the late grain development stages, while those related to protein synthesis/assembly/degradation and photosynthesis showed an opposite expression model during grain development. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 12 representative genes encoding different metabolic proteins showed certain transcriptional and translational expression differences during grain development. Phosphorylated proteins analyses demonstrated that 23 DEPs such as AGPase, sucrose synthase, Hsp90, and serpins were phosphorylated in the developing grains and were mainly involved in starch biosynthesis and stress/defense.

Conclusions: Our results revealed a complex quantitative proteome and phosphorylation profile during wheat grain development. Numerous DEPs are involved in grain starch and protein syntheses as well as adverse defense, which set an important basis for wheat yield and quality. Particularly, some key DEPs involved in starch biosynthesis and stress/defense were phosphorylated, suggesting their roles in wheat grain development.

Show MeSH

Related in: MedlinePlus

An overlook of stress/defense related proteins and ROS scavenging system involved in wheat grain development. LEA, TLP, SLP, CAP, CRP Emp and wail17 have coordinating role in various osmotic stresses during grain development. αAI and XI are mainly involved in protection of starch and cell wall, respectively. Purothionin, antifungal protein R (APR), chltinase, rRNA N-glycosidase (rRNA N-g) and wheatwin2 were mainly involved in resistance to pathogen and pest infection. Serpin, CTI and Hsps are involved in protein protection processes, serpin and CTI prevent protein from degradation by protese, Hsps prevent degraded protein from aggregation and help misfolded protein refolding, these processes also involved in resistance to pathogen/insect and osmotic stress. All defense process can produce reactive oxygen and which can be scavenged by ROS scavenging system. The protein levels are shown in coloured squares, indicating the change of expression for each developmental stage. The phosphorylated DEPs are marked.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4301063&req=5

Fig9: An overlook of stress/defense related proteins and ROS scavenging system involved in wheat grain development. LEA, TLP, SLP, CAP, CRP Emp and wail17 have coordinating role in various osmotic stresses during grain development. αAI and XI are mainly involved in protection of starch and cell wall, respectively. Purothionin, antifungal protein R (APR), chltinase, rRNA N-glycosidase (rRNA N-g) and wheatwin2 were mainly involved in resistance to pathogen and pest infection. Serpin, CTI and Hsps are involved in protein protection processes, serpin and CTI prevent protein from degradation by protese, Hsps prevent degraded protein from aggregation and help misfolded protein refolding, these processes also involved in resistance to pathogen/insect and osmotic stress. All defense process can produce reactive oxygen and which can be scavenged by ROS scavenging system. The protein levels are shown in coloured squares, indicating the change of expression for each developmental stage. The phosphorylated DEPs are marked.

Mentions: Based on our results, an overview of DEPs involved in stress/defense could be drawn (Figure 9), which depicts a coordinated adverse response and defense mechanism during grain development. Since the grain development is liable to affected by adverse environments such as various biotic and abiotic stresses, various defense proteins were recruited to resist and adapt to the adverse environments. Considerable DEPs involved in defense/stress processes were activated and significantly up-regulated during grain development. Moreover, some DEPs such as LEA, serpins, Hsp90, and wail7 were phosphorylated, which could enhance the activity of stress/defense-related proteins and their interactions with other proteins. The coordinated functions of these stress/defense-related proteins could protect grain development from adverse harm, and benefit wheat yield and quality.Figure 9


iTRAQ-based quantitative proteome and phosphoprotein characterization reveals the central metabolism changes involved in wheat grain development.

Ma C, Zhou J, Chen G, Bian Y, Lv D, Li X, Wang Z, Yan Y - BMC Genomics (2014)

An overlook of stress/defense related proteins and ROS scavenging system involved in wheat grain development. LEA, TLP, SLP, CAP, CRP Emp and wail17 have coordinating role in various osmotic stresses during grain development. αAI and XI are mainly involved in protection of starch and cell wall, respectively. Purothionin, antifungal protein R (APR), chltinase, rRNA N-glycosidase (rRNA N-g) and wheatwin2 were mainly involved in resistance to pathogen and pest infection. Serpin, CTI and Hsps are involved in protein protection processes, serpin and CTI prevent protein from degradation by protese, Hsps prevent degraded protein from aggregation and help misfolded protein refolding, these processes also involved in resistance to pathogen/insect and osmotic stress. All defense process can produce reactive oxygen and which can be scavenged by ROS scavenging system. The protein levels are shown in coloured squares, indicating the change of expression for each developmental stage. The phosphorylated DEPs are marked.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4301063&req=5

Fig9: An overlook of stress/defense related proteins and ROS scavenging system involved in wheat grain development. LEA, TLP, SLP, CAP, CRP Emp and wail17 have coordinating role in various osmotic stresses during grain development. αAI and XI are mainly involved in protection of starch and cell wall, respectively. Purothionin, antifungal protein R (APR), chltinase, rRNA N-glycosidase (rRNA N-g) and wheatwin2 were mainly involved in resistance to pathogen and pest infection. Serpin, CTI and Hsps are involved in protein protection processes, serpin and CTI prevent protein from degradation by protese, Hsps prevent degraded protein from aggregation and help misfolded protein refolding, these processes also involved in resistance to pathogen/insect and osmotic stress. All defense process can produce reactive oxygen and which can be scavenged by ROS scavenging system. The protein levels are shown in coloured squares, indicating the change of expression for each developmental stage. The phosphorylated DEPs are marked.
Mentions: Based on our results, an overview of DEPs involved in stress/defense could be drawn (Figure 9), which depicts a coordinated adverse response and defense mechanism during grain development. Since the grain development is liable to affected by adverse environments such as various biotic and abiotic stresses, various defense proteins were recruited to resist and adapt to the adverse environments. Considerable DEPs involved in defense/stress processes were activated and significantly up-regulated during grain development. Moreover, some DEPs such as LEA, serpins, Hsp90, and wail7 were phosphorylated, which could enhance the activity of stress/defense-related proteins and their interactions with other proteins. The coordinated functions of these stress/defense-related proteins could protect grain development from adverse harm, and benefit wheat yield and quality.Figure 9

Bottom Line: Hierarchical clustering analysis indicated that the DEPs involved in starch biosynthesis, storage proteins, and defense/stress-related proteins significantly accumulated at the late grain development stages, while those related to protein synthesis/assembly/degradation and photosynthesis showed an opposite expression model during grain development.Numerous DEPs are involved in grain starch and protein syntheses as well as adverse defense, which set an important basis for wheat yield and quality.Particularly, some key DEPs involved in starch biosynthesis and stress/defense were phosphorylated, suggesting their roles in wheat grain development.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Capital Normal University, Beijing 100048, China. zhimin206@263.net.

ABSTRACT

Background: Wheat (Triticum aestivum L.) is an economically important grain crop. Two-dimensional gel-based approaches are limited by the low identification rate of proteins and lack of accurate protein quantitation. The recently developed isobaric tag for relative and absolute quantitation (iTRAQ) method allows sensitive and accurate protein quantification. Here, we performed the first iTRAQ-based quantitative proteome and phosphorylated proteins analyses during wheat grain development.

Results: The proteome profiles and phosphoprotein characterization of the metabolic proteins during grain development of the elite Chinese bread wheat cultivar Yanyou 361 were studied using the iTRAQ-based quantitative proteome approach, TiO2 microcolumns, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Among 1,146 non-redundant proteins identified, 421 showed at least 2-fold differences in abundance, and they were identified as differentially expressed proteins (DEPs), including 256 upregulated and 165 downregulated proteins. Of the 421 DEPs, six protein expression patterns were identified, most of which were up, down, and up-down expression patterns. The 421 DEPs were classified into nine functional categories mainly involved in different metabolic processes and located in the membrane and cytoplasm. Hierarchical clustering analysis indicated that the DEPs involved in starch biosynthesis, storage proteins, and defense/stress-related proteins significantly accumulated at the late grain development stages, while those related to protein synthesis/assembly/degradation and photosynthesis showed an opposite expression model during grain development. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 12 representative genes encoding different metabolic proteins showed certain transcriptional and translational expression differences during grain development. Phosphorylated proteins analyses demonstrated that 23 DEPs such as AGPase, sucrose synthase, Hsp90, and serpins were phosphorylated in the developing grains and were mainly involved in starch biosynthesis and stress/defense.

Conclusions: Our results revealed a complex quantitative proteome and phosphorylation profile during wheat grain development. Numerous DEPs are involved in grain starch and protein syntheses as well as adverse defense, which set an important basis for wheat yield and quality. Particularly, some key DEPs involved in starch biosynthesis and stress/defense were phosphorylated, suggesting their roles in wheat grain development.

Show MeSH
Related in: MedlinePlus