Feng Xin, Yang Xinru, Zhong Meiqin, Li Xin, Zhu Pengfang
College of Forestry, Shenyang Agricultural University, Shenyang 110866, China.
Key Laboratory of Forest Tree Genetics, Breeding and Cultivation of Liaoning Province, Shenyang 110866, China.
Hortic Res. 2022 Jun 15;9:uhac137. doi: 10.1093/hr/uhac137. eCollection 2022.
The morphological diversity of leaf margin shapes is an identifying characteristic of many plant species. In our previous work, BoALG10 (α-1,2 glycosyltransferase) was predicted to be a key regulator of leaf margin shape in ornamental kale ( var. ). An alanine and a leucine residue in the conserved domain of the smooth-margined S0835 were replaced by an aspartate and a phenylalanine, respectively, in the corresponding positions of the feathered-margined F0819. However, the expression pattern and function of this gene remain unclear. Here, we examined the expression patterns of using quantitative real-time PCR, and found that statistically significant differences in expression existed between F0819 and S0835 in nine developmental stages. The BoALG10 protein localized to the endoplasmic reticulum. The function of was then examined using complementary mutant assays. The overexpression strains phenocopied the smooth leaf margin after introduction of into the feathered-margined inbred line F0819. Simultaneously, irregular dissections appeared in the leaf margins of knockout mutants KO-1 and KO-2, which were generated by CRISPR/Cas9 technology from the smooth-margined inbred line S0835. Microscopic observation showed that the leaf margin cells of the smooth-margined plants S0835 and OE-3 were arranged regularly, while the cells of the feathered-margined plants F0819 and KO-1 were of inconsistent size and distributed in an irregular manner, particularly around the indentations of the leaf. This elucidation of function provides a novel insight into the morphological regulation of leaf margin shape.
叶缘形状的形态多样性是许多植物物种的一个识别特征。在我们之前的工作中，预测BoALG10（α-1,2糖基转移酶）是观赏羽衣甘蓝（变种）叶缘形状的关键调节因子。在光滑叶缘的S0835保守结构域中的一个丙氨酸和一个亮氨酸残基，在羽状叶缘的F0819相应位置分别被一个天冬氨酸和一个苯丙氨酸取代。然而，该基因的表达模式和功能仍不清楚。在这里，我们使用定量实时PCR检测了 的表达模式，发现在九个发育阶段F0819和S0835之间存在统计学上显著的表达差异。BoALG10蛋白定位于内质网。然后使用互补突变体分析检测了 的功能。将 导入羽状叶缘自交系F0819后，过表达菌株表现出光滑叶缘的表型。同时，通过CRISPR/Cas9技术从光滑叶缘自交系S0835产生的敲除突变体KO-1和KO-2的叶缘出现不规则切割。显微镜观察表明，光滑叶缘植株S0835和OE-3的叶缘细胞排列规则，而羽状叶缘植株F0819和KO-1的细胞大小不一致且分布不规则，特别是在叶片的凹陷处周围。对 功能的这一阐释为叶缘形状的形态调控提供了新的见解。
