Comparativeproteomic analysis of the shoot apical meristem in maize between aZmCCT-associated near-isogenic line and its recurrent parent.
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Background: ZmCCT is a gene that is important for plant photoperiod response. For corn, long-term lighting delays the flowering time. This article explores the molecular mechanism of the light periodicity of corn species at the level of proteomics.
Source: apical meristem (SAM), derived from ZmCCT-positive strain NIL- cml vs. ZmCCT-negative strain H4
Technical route: iTRAQ , three biological replicates
Research result:
1. A total of 5259 proteins and 386 differential proteins were identified. These differential proteins are involved in energy production, photosynthesis, signal transduction, stress and defense, glucose metabolism, and protein metabolism, and also contain proteins of unknown function.
Identification of GO function analysis of proteins
Venn diagram of differential proteins
2. Under long-day conditions, the NIL-cml strain delayed vegetative growth and reproductive growth compared to the normal vegetative and reproductive growth of the H4 strain.
Analysis of protein expression from vegetative growth state to reproductive state
3. Select 52 differential proteins (directly or indirectly induced by ZmCCT, related to carbohydrate metabolism, signal transduction, defense response, etc.) for protein interaction analysis .
4. Validation at the protein level . Three proteins were selected for WB validation.
5. Conduct research from the transcriptional level . Six representative genes were selected: hisone H2B, ribonucleoprotein A, glycine-rich RNA-binding protein, calmodulin-binding protein, malate synthase, 14-3-3 protein for qPCR analysis.
The results of qPCR showed that the expression level of ZmCCT gene in the H4 strain (V3-V6 stage) was lower than that of the NIL-cml strain. At the same time, the expression patterns of protein level and mRNA level were not completely consistent, indicating that these proteins may have post-transcriptional modifications.
Xiaobian's experience: This is a typical literature published on the Scientific Report using proteomics technology (iTRAQ) to study the expression of plant tissues at the protein level under different stress conditions, through a series of common life letter analysis. ( GO, clustering, interaction ) and verification means ( WB, qPCR ) to obtain specific (or target) proteins with high credibility, laying the foundation for further research.
The structure of the article is clear, the framework is obvious, simple and easy to understand, it is worth learning!
Protein|modification|metabolism|lipid|structural confirmation
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