Genetic mapping and gene editing reveal <i>BoAP1</i> as a crucial factor regulating chloroplast development in <i>Brassica oleracea</i>

作者：Xinyu Zhao, Li Chen, Kaiwen Yuan, Yuxiang Liu, Limei Yang, Mu Zhuang, Yangyong Zhang, Yong Wang, Jialei Ji, Ryo Fujimoto, Lv Honghao · 发表于：Vegetable Research · 年份：2024 · DOI：10.48130/vegres-0024-0030 · 被引用次数：3 · 研究领域：Photosynthetic Processes and Mechanisms、Algal biology and biofuel production、Plant Molecular Biology Research

Chloroplasts are crucial cellular components that plants need to carry out photosynthesis. Exploring the relevant underlying molecular regulatory mechanism could help elucidate this complex process. In this study, a spontaneous chloroplast-deficient mutant, 6-219W, exhibiting a lethal albino phenotype in curly kale ( Brassica oleracea var. acephala ) was identified. The number of chloroplasts in the 6-219W mutant were considerably reduced, those chloroplasts appeared crumpled, and the thylakoid membranes could not be observed under transmission electron microscopy. Genetic analysis revealed that boap1 ( Brassica oleracea albino plant1 ), a single recessive gene, is responsible for this lethal albino trait. Fine mapping demonstrated that boap1 is located at a 300 kb interval between the InDel markers PW404 and PW406 on chromosome 8. Based on the B. oleracea genome annotation, a candidate gene, BolC08g019310.2J , was identified within the target interval. Sequence analysis revealed a 3-nucleotide (GAT) deletion in the coding sequence of BolC08g019310.2J in the 6-219W mutant, leading to the absence of the amino acid methionine at position 298 that is conserved in Arabidopsis thaliana , B. rapa , and B. oleracea . CRISPR/Cas9 technology was used to knock out the BolC08g019310.2J gene in NB11, and the generation of boap1 mutants with an albino phenotype confirmed BolC08g019310.2J as the causal gene. Subcellular localization indicated that the BoAP1 protein oper...