Kolanchi, P and Sanivarapu, H and Marimuthu, M and Yogendra, K and Onkarappa, D and Jaba, J and Venkatasamy, B and Muthu, A (2025) Combining complementary multiplex extraction chemistries enhances proteome coverage and analytical insights in tiny insects: A study on sweet potato whitefly Bemisia tabaci. International Journal of Biological Macromolecules (TSI), 333 (1). ISSN 0141-8130
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Advancements in protein extraction methodologies are essential for enhancing insect proteomics, especially in sap-feeding small insects, where high lipid content and limited tissue volume impede efficient protein recovery. This study systematically assessed seven extraction chemistries, lysis buffer (LB), dissolution buffer (DB), TRIzol®, trifluoroacetic acid (TFA), trichloroacetic acid (TCA), TCA-phenol, and phenol— under varying pH conditions to determine the most effective workflow for comprehensive proteomic profiling of Bemisia tabaci. Qualitative two-dimensional electrophoresis (2-DE) analysis indicated that phenol and TCA-phenol methods maintained protein integrity across a wide molecular weight range, while acidic conditions enhanced the resolution of low-abundance proteins. Quantitative liquid chromatography-tandem mass spectrometry (LC–MS/MS) analyses demonstrated that the TCA-phenol method yielded the highest protein recovery and reproducibility (coefficient of variation = 12.6 %), identifying 1010 proteins with balanced representation of soluble and membrane fractions. Principal component and overlap analyses confirmed distinct, method-specific proteomic signatures, with TCA-phenol and TFA achieving the most extensive proteome coverage. Functional annotation revealed that DB and LB enriched primary metabolic and biosynthetic processes, whereas the TCA-phenol method provided the broadest functional spectrum, capturing detoxification, energy metabolism, and stress-related proteins. Overall, this research illustrates that no single extraction chemistry is universally optimal; rather, combining complementary methods enhances proteome coverage. These findings establish a standardized comparative framework for small-insect proteomics and offer valuable methodological insights to advance research in insect physiology, stress adaptation, and ecological interactions.
| Item Type: | Article |
|---|---|
| Divisions: | Global Research Program - Accelerated Crop Improvement |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | whitefly, sweet potato, proteome coverage, tiny insects |
| Subjects: | Others > Genetics and Genomics Others > Entomology |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 09 Dec 2025 04:28 |
| Last Modified: | 09 Dec 2025 04:28 |
| URI: | http://oar.icrisat.org/id/eprint/13405 |
| Official URL: | https://www.sciencedirect.com/science/article/abs/... |
| Projects: | UNSPECIFIED |
| Funders: | UNSPECIFIED |
| Acknowledgement: | The authors sincerely thank the Department of Agricultural Entomology, Tamil Nadu Agricultural University (TNAU), Coimbatore, Tamil Nadu, India, and the Cell & Molecular Biology and Trait Engineering, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India, for providing instrumental facilities. I also extend my thanks to NF-OBC (UGC-CSIR; 202324-231610061191) for providing a fellowship... |
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