Metroxylon sagu Rottb. or locally known as sago palm is a tropical starch crop grown for starch production in commercial plantations in Malaysia, especially in Sarawak, East Malaysia. This plant species accumulate the highest amount of edible starch compared to other starch-producing crops. However, the non-trunking phenomenon has been observed to be one of the major issues restricting the yield of sago palm starch. In this study, proteomics approach was utilised to discover differences between trunking and non-trunking proteomes in sago palm leaf tissues. Total protein from 16 years old trunking and non-trunking sago palm leaves from deep peat area were extracted with PEG fractionation extraction method and subjected to two-dimensional gel electrophoresis (2D PAGE). Differential protein spots were subjected to MALDI-ToF/ToF MS/MS. Proteomic analysis has identified 34 differentially expressed proteins between trunking and non-trunking sago samples. From these protein spots, all 19 proteins representing different enzymes and proteins have significantly increased in abundance in non-trunking sago plant when subjected to mass spectrometry. The identified proteins mostly function in metabolic pathways including photosynthesis, tricarboxylic acid cycle, glycolysis, carbon utilization and oxidative stress. The current study indicated that the several proteins identified through differentially expressed proteome contributed to physical differences in trunking and non-trunking sago palm.
Background Gel staining is a crucial step that allows the visualisation of proteins separated through SDS-PAGE. Colloidal Coomassie Brilliant Blue-G (CBB-G) staining is among the commonly used visualisation methods due to several factors such as compatibility with mass spectrometry (MS) analysis, sensitivity, reproducibility, and simplicity of the staining process. However, the standard colloidal CBB-G staining has a drawback: the resolution of protein bands is compromised because of diffusion of proteins during the washing step. Results A modification to an established colloidal CBB-G staining method, which greatly increases the resolution of protein bands, is described. The addition of a fixation step, which prevents the diffusion of proteins during the washing step, is shown to increase protein band resolution. Conclusion The fixation step is fast, flexible, and also retains all the advantages of the standard colloidal CBB-G staining methods. As there are no drawbacks, incorporating this fixation step into the standard colloidal CBB-G staining is an easy way to improve protein visualisation in SDS-PAGE. Graphical Abstract
Piper nigrum L., also known as the pepper plant, produces pepper berries, which is a spice used worldwide. Changes in the natural environment have forced the plant to undergo acclimatization for plant growth. Consequently, un-adapted plants developed unhealthy, stunted growth, affecting the production of pepper berries, and thus, causing loss to agriculture. Proteomic analysis using two-dimensional polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry was used to compare the differentially expressed proteins between healthy and unhealthy pepper plants. The proteins were successfully extracted from the stems and roots of P. nigrum and the two-dimensional protein profiles were compared. Two-dimensional analysis has indicated significant differences between the healthy and unhealthy parts with 41 and 25 differentially expressed proteins in the stems and roots respectively. A total of 10 differentially expressed proteins, characterized as unique (2) and over-expressed (8) proteins, from both plant parts were selected and sent for identification through matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry. The unique and over-expressed proteins in the unhealthy stems were identified as stress-related small heat shock proteins (sHSPs). These proteins act as a chaperone to protect other proteins against a stressful environment for plant defence and adaptation. Meanwhile, over-expressed proteins from the roots were found to be peroxidase 5, malate dehydrogenase, mitochondrial and gamma-glutamyl phosphate reductase which are involved in oxidative stress, photosynthetic activity, and plant development, respectively. The findings of the differentially expressed proteins in this study, therefore, may create opportunities for further investigations of stress management control and help to improve plantation practices in the future.
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