Banana is one of the most economically important fruit crops worldwide. Genetic improvement in banana is a challenging task due to its parthenocarpic nature and triploid genome. Genetic modification of crops via the CRISPR/Cas9 module has emerged as a promising tool to develop important traits. In the present work, a CRISPR/Cas9-based approach was used to develop the β-carotene-enriched Cavendish banana cultivar (cv.) Grand Naine (AAA genome). The fifth exon of the lycopene epsilon-cyclase (LCYε) gene was targeted. The targeting specificity of the designed guide-RNA was also tested by its ability to create indels in the LCYε gene at the A genome of cv. Rasthali (AAB genome). Sequence analysis revealed multiple types of indels in the genomic region of Grand Naine LCYε (GN-LCYε). Metabolic profiling of the fruit pulp of selected edited lines showed enhanced accumulation of β-carotene content up to 6-fold (~24 μg/g) compared with the unedited plants. These lines also showed either an absence or a drastic reduction in the levels of lutein and α-carotene, suggesting metabolic reprogramming, without any significant effect on the agro-morphological parameters. In addition, differential expression of carotenoid pathway genes was observed in the edited lines in comparison to unedited plants. Overall, this is the first report in banana to improve nutritional trait by using a precise genome editing approach.

香蕉是世界上最重要的经济作物之一。由于单性结实和三倍体基因组，香蕉的遗传改良是一项艰巨的任务。通过CRISPR / Cas9模块对农作物进行遗传修饰已成为发展重要性状的有前途的工具。在目前的工作中，基于CRISPR / Cas9的方法被用于开发富含β-胡萝卜素的Cavendish香蕉栽培种（cv。）Grand Naine（AAA基因组）。靶向番茄红素ε-环化酶（LCYε）基因的第五外显子。还通过其在LCYε中形成插入缺失的能力测试了设计的指导RNA的靶向特异性。简历A基因组的基因。拉斯塔利（AAB基因组）。序列分析揭示了Grand NaineLCYε（GN-LCYε）基因组区域中的多种插入缺失。所选编辑品系的果肉的代谢谱分析显示，与未编辑植物相比，β-胡萝卜素含量的累积积累提高了6倍（〜24μg/ g）。这些品系还显示出叶黄素和α-胡萝卜素水平的缺乏或急剧降低，表明代谢重新编程，而对农业形态参数没有任何显着影响。另外，与未编辑的植物相比，在编辑的品系中观察到类胡萝卜素途径基因的差异表达。总体而言，这是香蕉中使用精确基因组编辑方法改善营养性状的第一份报告。