<scp>TALEN</scp> ‐mediated depletion of the mitochondrial gene <i>orf312</i> proves that it is a Tadukan‐type cytoplasmic male sterility‐causative gene in rice
<scp>TALEN</scp> ‐mediated depletion of the mitochondrial gene <i>orf312</i> proves that it is a Tadukan‐type cytoplasmic male sterility‐causative gene in rice
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<scp>TALEN</scp>介导的线粒体基因<i>orf312</i>的耗竭证明它是水稻中Tadukan型细胞质雄性不育致病基因
DOI：
10.1111/tpj.15715
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Toriyama Kinya
中科院分区：
文献类型：
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作者：
Takatsuka Ayumu;Kazama Tomohiko;Arimura Shin‐ichi;Toriyama Kinya
细胞质雄性不育（CMS）是一种导致花粉或花药功能障碍，导致缺乏结实率的性状。CMS被认为是由一种称为CMS相关基因的独特线粒体开放阅读框的表达引起的。orf312已被报道为水稻Tadukan型CMS （TAA）的CMS相关基因，该基因表现为花药开裂受损；然而，证据尚未被报道。在这里，我们采用了功能缺失的方法，使用线粒体靶向转录激活因子样效应核酸酶（mitoTALEN）来敲除TAA中的orf312，以证明orf312确实是一个CMS致病基因。在携带mitoTALEN表达载体的28株转基因TAA植物中，通过PCR、Southern blot和测序分析，在24株植物中检测到forf312的缺失。根据缺失区域将24株植物分为3组。Allorf312‐枯竭的TAA植株花药开裂和结实率恢复。即使在没有mitoTALEN表达盒的分离中，f312的消耗和育性恢复在下一代中也保持不变。相比之下，orf312‐保留植株是不育的。这些结果为orf312是Tadukan型CMS致病基因提供了强有力的证据。
Cytoplasmic male sterility (CMS) is a trait that causes pollen or anther dysfunctions, resulting in the lack of seed setting. CMS is considered to be caused by the expression of a unique mitochondrial open reading frame referred to as CMS‐associated gene.orf312has been reported as a CMS‐associated gene of Tadukan‐type CMS (TAA) in rice (Oryza sativaL.), which exhibits impaired anther dehiscence; however, evidence thereof has not yet been reported. Here, we took a loss‐of‐function approach, using a mitochondria‐targeted transcription activator‐like effector nuclease (mitoTALEN) designed to knock outorf312in TAA, to prove thatorf312indeed is a CMS‐causative gene. Out of 28 transgenic TAA plants harboring the mitoTALEN expression vector, deletion oforf312was detected in 24 plants by PCR, Southern blot, and sequencing analyses. The 24 plants were grouped into three groups based on the deleted regions. Allorf312‐depleted TAA plants exhibited recovery of anther dehiscence and seed setting. The depletion oforf312and fertility restoration was maintained in the next generation, even in mitoTALEN expression cassette null segregants. In contrast,orf312‐retaining plants were sterile. These results provide robust evidence thatorf312is a Tadukan‐type CMS‐causative gene.