Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.)
| العنوان: | Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.) |
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| المؤلفون: | Hina Ali, Syed Muhammad Azam, Simon Peter Ojolo, Bingyan Hu, S. V. G. N. Priyadarshani, Weimin Li, Maokai Yan, Haifeng Jia, Lihua Zhao, Zia ur Rahman, Junjie Xiong, Yuan Qin, Qingsong Wu |
| المصدر: | Plant Methods, Vol 14, Iss 1, Pp 1-12 (2018) Plant Methods |
| بيانات النشر: | BMC, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0106 biological sciences, 0301 basic medicine, Plant Science, Cellulase, Biology, lcsh:Plant culture, Transfection, 01 natural sciences, 03 medical and health sciences, Tissue culture, Genetics, lcsh:SB1-1110, Ananas, lcsh:QH301-705.5, NAA, Protoplast, Research, Pineapple, fungi, food and beverages, Subcellular localization, biology.organism_classification, BAP, Protein subcellular localization prediction, Transformation (genetics), 030104 developmental biology, Biochemistry, lcsh:Biology (General), biology.protein, Organelle localization, 010606 plant biology & botany, Biotechnology |
| الوصف: | Background An efficient transformation protocol is a primary requisite to study and utilize the genetic potential of any plant species. A quick transformation system is also crucial for the functional analysis of genes along with the study of proteins and their interactions in vivo. Presently, however, quick and effective transformation systems are still lacking for many plant species including pineapple. This has limited the full exploration of the genetic repository of pineapple as well as the study of its genes, protein localization and protein interactions. Results To address the above limitations, we have developed an efficient system for protoplast isolation and subcellular localization of desired proteins using pineapple plants derived from tissue culture. A cocktail of 1.5% (W/V) Cellulase R-10 and 0.5% (W/V) Macerozyme R-10 resulted in 51% viable protoplasts with 3 h digestion. Compared to previously reported protocols, our protoplast isolation method is markedly faster (saving 4.5 h), requires only a small quantity of tissue sample (1 g of leaves) and has high yield (6.5 × 105). The quality of the isolated protoplasts was verified using organelle localization in protoplasts with different organelle markers. Additionally, colocalization analysis of two pineapple Mg2+ transporter genes in pineapple protoplasts was consistent with the results in a tobacco transient expression system, confirming that the protoplast isolation method can be used to study subcellular localization. Further findings showed that the system is also suitable for protein–protein interaction studies. Conclusion Based on our findings, the presently described method is an efficient and effective strategy for pineapple protoplast isolation and transformation; it is convenient and time saving and provides a greater platform for transformation studies. |
| اللغة: | English |
| تدمد: | 1746-4811 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::37cd12e87aacdb7361d015c81b90ba46Test http://link.springer.com/article/10.1186/s13007-018-0365-9Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....37cd12e87aacdb7361d015c81b90ba46 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 17464811 |
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