Phosphatidylethanolamine-binding protein (PEBP) genes regulate flowering and architecture in many plant species. Here, we study kiwifruit (Actinidia chinensis, Ac) PEBP genes with homology to BROTHER OF FT AND TFL1 (BFT). CRISPR-Cas9 was used to target AcBFT genes in wild-type and fast-flowering kiwifruit backgrounds. The editing construct was designed to preferentially target AcBFT2, whose expression is elevated in dormant buds. Acbft lines displayed an evergrowing phenotype and increased branching, while control plants established winter dormancy. The evergrowing phenotype, encompassing delayed budset and advanced budbreak after defoliation, was identified in multiple independent lines with edits in both alleles of AcBFT2. RNA-seq analyses conducted using buds from gene-edited and control lines indicated that Acbft evergrowing plants had a transcriptome similar to that of actively growing wild-type plants, rather than dormant controls. Mutations in both alleles of AcBFT2 did not promote flowering in wild-type or affect flowering time, morphology and fertility in fast-flowering transgenic kiwifruit. In summary, editing of AcBFT2 has the potential to reduce plant dormancy with no adverse effect on flowering, giving rise to cultivars better suited for a changing climate.

中文翻译：

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CRISPR-Cas9 介导的猕猴桃 BFT 基因诱变导致持续生长但不早开花的表型

磷脂酰乙醇胺结合蛋白 (PEBP) 基因调节许多植物物种的开花和结构。在这里，我们研究了与BROTHER OF FT AND TFL1 ( BFT ) 同源的猕猴桃 ( Actinidia chinensis , Ac ) PEBP 基因。CRISPR-Cas9 用于靶向野生型和快速开花猕猴桃背景中的AcBFT基因。编辑结构被设计为优先靶向AcBFT2，其在休眠芽中的表达升高。交流电品系表现出不断增长的表型和增加的分枝，而对照植物则建立了冬季休眠。不断增长的表型，包括落叶后的延迟萌芽和高级萌芽，在多个独立的系中被鉴定，并在AcBFT2的两个等位基因中进行了编辑。使用来自基因编辑和对照品系的芽进行的 RNA-seq 分析表明，Acbft 常生植物的转录组与活跃生长的野生型植物的转录组相似，而不是休眠对照。AcBFT2两个等位基因的突变不促进野生型开花，也不影响快开花转基因猕猴桃的开花时间、形态和生育力。总之，AcBFT2的编辑具有减少植物休眠的潜力，而不会对开花产生不利影响，从而培育出更适合气候变化的品种。