Establishment of a Protoplasts-Based Transient Expression System in Banana (Musa spp.)

文献类型： 外文期刊

第一作者： Zhao, Chunhui

作者： Zhao, Chunhui;Li, Shuyu;Gao, Hui;Cui, Haitao;Du, Chanjuan;Yang, Di;Fu, Gang

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关键词： banana; protoplast isolation; Musa spp.; transient expression; transformation

期刊名称：AGRONOMY-BASEL （ 影响因子：3.949； 五年影响因子：4.117 ）

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年卷期： 2022 年 12 卷 11 期

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收录情况： SCI

摘要： The breeding of triploid banana cultivars with improved traits, such as yield and disease resistance, remains a major challenge for breeders. One reason is that the molecular study and functional gene analysis in bananas fall behind due to the difficulties of its genetic manipulation. The plant protoplast-based transient transformation has been documented and widely used as a versatile and convenient system for functional gene analysis in many plant species. However, an efficient high-quality protoplast isolation and transformation system is still lacking for bananas. Here, we established an efficient protoplast isolation and transformation method for bananas by selecting proper source materials, optimizing conditions for enzymatic hydrolysis and PEG-mediated transfection. We found the best source materials for banana protoplasts' isolation are young suckers, which give a yield of protoplasts ranging from 2.5 x 10(6) to 10.1 x 10(7) g(-1) fresh weight after 5 to 6 h of enzymolysis. The yield is sufficient for most assays that have been established in protoplasts-based systems, such as protein subcellular localization and protein interaction assays. Moreover, using the established transient gene expression system in banana protoplasts, we validated the subcellular localization of Arabidopsis VESICLE SORTING RECEPTOR 1 (VSR1) and the protein self-interaction of Arabidopsis CNGC20 on the cell membrane. The results indicated this system works well and could be routinely used for the functional characterization of banana genes.

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