Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea
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 phenoty...
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Maximum Academic Press
2024-01-01
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| Series: | Vegetable Research |
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| Online Access: | https://www.maxapress.com/article/doi/10.48130/vegres-0024-0030 |
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| author | Xinyu Zhao Li Chen Kaiwen Yuan Yuxiang Liu Limei Yang Mu Zhuang Yangyong Zhang Yong Wang Jialei Ji Ryo Fujimoto Honghao Lv |
| author_facet | Xinyu Zhao Li Chen Kaiwen Yuan Yuxiang Liu Limei Yang Mu Zhuang Yangyong Zhang Yong Wang Jialei Ji Ryo Fujimoto Honghao Lv |
| author_sort | Xinyu Zhao |
| collection | DOAJ |
| description | 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 operates in chloroplasts. These results revealed that the BoAP1 mutation disrupted the normal development of chloroplasts in 6-219W, leading to a lethal albino phenotype. The present research lays a foundation for the in-depth study of the molecular mechanism regulating chloroplast development. |
| format | Article |
| id | doaj-art-e173ba270b954742bb8bc1cdc2f28d86 |
| institution | OA Journals |
| issn | 2769-0520 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Maximum Academic Press |
| record_format | Article |
| series | Vegetable Research |
| spelling | doaj-art-e173ba270b954742bb8bc1cdc2f28d862025-08-20T02:12:19ZengMaximum Academic PressVegetable Research2769-05202024-01-01411810.48130/vegres-0024-0030vegres-0024-0030Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleraceaXinyu Zhao0Li Chen1Kaiwen Yuan2Yuxiang Liu3Limei Yang4Mu Zhuang5Yangyong Zhang6Yong Wang7Jialei Ji8Ryo Fujimoto9Honghao Lv10State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaCollege of Life Sciences, Zaozhuang University, Zaozhuang 277160, Shandong, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaGraduate School of Agricultural Science, Kobe University, Kobe, JapanState Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaChloroplasts 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 operates in chloroplasts. These results revealed that the BoAP1 mutation disrupted the normal development of chloroplasts in 6-219W, leading to a lethal albino phenotype. The present research lays a foundation for the in-depth study of the molecular mechanism regulating chloroplast development.https://www.maxapress.com/article/doi/10.48130/vegres-0024-0030chloroplast developmentalbino lethal mutantmap-based cloningcrispr/cas9 |
| spellingShingle | Xinyu Zhao Li Chen Kaiwen Yuan Yuxiang Liu Limei Yang Mu Zhuang Yangyong Zhang Yong Wang Jialei Ji Ryo Fujimoto Honghao Lv Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea Vegetable Research chloroplast development albino lethal mutant map-based cloning crispr/cas9 |
| title | Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea |
| title_full | Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea |
| title_fullStr | Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea |
| title_full_unstemmed | Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea |
| title_short | Genetic mapping and gene editing reveal BoAP1 as a crucial factor regulating chloroplast development in Brassica oleracea |
| title_sort | genetic mapping and gene editing reveal boap1 as a crucial factor regulating chloroplast development in brassica oleracea |
| topic | chloroplast development albino lethal mutant map-based cloning crispr/cas9 |
| url | https://www.maxapress.com/article/doi/10.48130/vegres-0024-0030 |
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