Abstract Background Maize is one of the most important crops worldwide and has been a target of nuclear-based transformation biotechnology to improve it and satisfy the food demand of the ever-growing global population. However, the maize plastid transformation has not been accomplished due to the recalcitrant condition of the crop. Results In this study, we constructed two different vectors with homologous recombination sequences from maize (Zea mays var. LPC13) and grass (Bouteloua gracilis var. ex Steud) (pZmcpGFP and pBgcpGFP, respectively). Both vectors were designed to integrate into rrn23S/rrn16S from an inverted repeat region in the chloroplast genome. Moreover, the vector had the mgfp5 gene driven by Prrn, a leader sequence of the atpB gene and a terminator sequence from the rbcL gene. Also, constructs have an hph gene as a selection marker gene driven by Prrn, a leader sequence from rbcL gene and a terminator sequence from the rbcL gene. Explants of maize, tobacco and Escherichia coli cells were transformed with both vectors to evaluate the transitory expression–an exhibition of green and red fluorescent light under epifluorescence microscopy. These results showed that both vectors were expressed; the reporter gene in all three organisms confirmed the capacity of the vectors to express genes in the cell compartments. Conclusions This paper is the first report of transient expression of GFP in maize embryos and offers new information for genetically improving recalcitrant crops; it also opens new possibilities for the improvement in maize chloroplast transformation with these vectors. How to cite: Arevalo-Gallegos S, Varela-Rodriguez H, Lugo-Aguilar H, et al. Transient expression of a green fluorescent protein in tobacco and maize chloroplast. Electron J Biotechnol 2020;44. https://doi.org/10.1016/j.ejbt.2020.01.008

中文翻译：

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绿色荧光蛋白在烟草和玉米叶绿体中的瞬时表达

摘要背景玉米是世界上最重要的作物之一，一直是基于核转化生物技术的目标，以改进它并满足不断增长的全球人口的粮食需求。然而，由于作物的顽固条件，玉米质体转化尚未完成。结果 在本研究中，我们构建了两个不同的载体，它们具有来自玉米（Zea mays var. LPC13）和草（Bouteloua gracilis var. ex Steud）（分别为 pZmcpGFP 和 pBgcpGFP）的同源重组序列。两种载体都被设计为从叶绿体基因组中的反向重复区域整合到 rrn23S/rrn16S 中。此外，该载体具有由 Prrn 驱动的 mgfp5 基因、atpB 基因的前导序列和 rbcL 基因的终止子序列。还，构建体具有作为由 Prrn 驱动的选择标记基因的 hph 基因、来自 rbcL 基因的前导序列和来自 rbcL 基因的终止子序列。用两种载体转化玉米、烟草和大肠杆菌细胞的外植体以评估瞬时表达——在落射荧光显微镜下显示绿色和红色荧光。这些结果表明两种载体都表达了；所有三种生物体中的报告基因证实了载体在细胞区室中表达基因的能力。结论 本文首次报道了 GFP 在玉米胚中的瞬时表达，为遗传改良顽固作物提供了新信息；它还为使用这些载体改进玉米叶绿体转化开辟了新的可能性。如何引用：Arevalo-Gallegos S，Varela-Rodriguez H、Lugo-Aguilar H 等。绿色荧光蛋白在烟草和玉米叶绿体中的瞬时表达。电子 J 生物技术 2020；44。https://doi.org/10.1016/j.ejbt.2020.01.008