Transient Transformation of Citrus Plants

Agrobacterium tumefaciens cultures were prepared using the same method detailed for subcellular localization. For citrus leaves, the suspensions expressing target genes and empty pBI121 vector (as control) were injected in different sides of the same leaf on opposite sides of the main vein (Li et al., 2017). For citrus fruit, the suspensions carrying control and target genes were injected into the peel on opposite sides of the equatorial plane of the same fruit (Yin et al., 2016). The fruit of Ougan used for infiltration was harvested from an orchard in Hangzhou, China, at 170 DAFB, and the Ougan seedlings used for transient transformation in leaves were maintained under a 16‐h light/8‐h dark cycle at 25 °C. After infiltration, the seedlings and fruits were kept in the dark for 1 day and maintained under 16‐h light/8‐h dark condition. The metabolites were analysed at 5 days after infiltration and measured by HPLC.