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Editing of ORF138 restores fertility of Ogura cytoplasmic male sterile broccoli via mitoTALENs

文献类型： 外文期刊

作者： Xu, Fengyuan 1 ; Su, Tongbing 3 ; Zhang, Xiaochen 1 ; Qiu, Lei 4 ; Yang, Xiaodong 4 ; Koizuka, Nobuya 5 ; Arimura, Shin-ichi 6 ; Hu, Zhongyuan 1 ; Zhang, Mingfang 1 ; Yang, Jinghua 1 ;

作者机构： 1.Zhejiang Univ, Hainan Inst, Yazhou Bay Sci & Technol City, Sanya, Peoples R China

2.Zhejiang Univ, Inst Vegetable Sci, Lab Germplasm Innovat & Mol Breeding, Hangzhou, Peoples R China

3.Beijing Acad Agr & Forestry Sci, Beijing Vegetable Res Ctr, Beijing, Peoples R China

4.Yangzhou Univ, Coll Hort & Landscape Architecture, Yangzhou 225000, Peoples R China

5.Tamagawa Univ, Coll Agr, Tokyo, Japan

6.Univ Tokyo, Grad Sch Agr & Life Sci, Lab Plant Mol Genet, Tokyo, Japan

7.Minist Agr & Rural Affairs, Key Lab Hort Plant Growth & Dev, Hangzhou, Peoples R China

关键词： cytoplasmic male sterility; fertility restoration; mitoTALENs; cold-sensitive male sterility; broccoli

期刊名称：PLANT BIOTECHNOLOGY JOURNAL （ 影响因子：10.1； 五年影响因子：11.6 ）

ISSN： 1467-7644

年卷期： 2024 年 22 卷 5 期

页码：

收录情况： SCI

摘要： Cytoplasmic male sterility (CMS), encoded by the mitochondrial open reading frames (ORFs), has long been used to economically produce crop hybrids. However, the utilization of CMS also hinders the exploitation of sterility and fertility variation in the absence of a restorer line, which in turn narrows the genetic background and reduces biodiversity. Here, we used a mitochondrial targeted transcription activator-like effector nuclease (mitoTALENs) to knock out ORF138 from the Ogura CMS broccoli hybrid. The knockout was confirmed by the amplification and re-sequencing read mapping to the mitochondrial genome. As a result, knockout of ORF138 restored the fertility of the CMS hybrid, and simultaneously manifested a cold-sensitive male sterility. ORF138 depletion is stably inherited to the next generation, allowing for direct use in the breeding process. In addition, we proposed a highly reliable and cost-effective toolkit to accelerate the life cycle of fertile lines from CMS-derived broccoli hybrids. By applying the k-mean clustering and interaction network analysis, we identified the central gene networks involved in the fertility restoration and cold-sensitive male sterility. Our study enables mitochondrial genome editing via mitoTALENs in Brassicaceae vegetable crops and provides evidence that the sex production machinery and its temperature-responsive ability are regulated by the mitochondria.

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