Editing melon eIF4E associates with virus resistance and male sterility.
Editing melon eIF4E associates with virus resistance and male sterility.
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DOI：
10.1111/pbi.13885
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发表时间：
2022-10
影响因子：
13.8
通讯作者：
Aranda, Miguel A.
中科院分区：
文献类型：
--
作者：
Pechar, Giuliano S.;Donaire, Livia;Gosalvez, Blanca;Garcia-Almodovar, Carlos;Amelia Sanchez-Pina, Maria;Truniger, Veronica;Aranda, Miguel A.
帽结合蛋白eIF4E通过与eIF4G的相互作用，构成eIF4F复合体的核心，在mRNAs的环化及其随后的帽依赖翻译中发挥关键作用。除了eIF4E在mRNA翻译启动中的基本作用外，还描述或建议了eIF4E的其他功能，包括作为前驱因子和参与性发育。我们利用CRISPR/Cas9基因组编辑技术获得了甜瓜eIF4E基因敲除突变系。编辑在甜瓜中有效，因为我们获得了T0世代中第一个eIF4E外显子纯合子中单核苷酸缺失的转化植株。用摩洛哥西瓜花叶病毒(MWMV)接种分离F2代的编辑植株和非转基因植株；纯合子突变植株表现出病毒抗性，而杂合子和非突变植株受到感染，这与我们之前在eIF4E中沉默的植株的结果一致。有趣的是，T0代和F2代的纯合编辑植株都表现出雄性不育表型，而与野生型植株杂交则恢复了育性，表现出雄性不育表型的分离与eIF4E突变的分离之间的良好相关性。对不同发育阶段甜瓜雄花的形态比较分析表明，小孢子母细胞减数分裂后发育异常，绒毡层退化的时间与野生型明显不同。RNA-Seq分析发现，在eIF4E/eIF4E植株的花中，花粉发育过程中的关键基因表达下调，表明eIF4E特异的mRNA翻译启动是甜瓜雄配子形成的限制因素。对MWMV的抗性与eIF4E编辑有关，eIF4E基因敲除植株表现出减数分裂后的孢子体雄性不育表型。
The cap‐binding protein eIF4E, through its interaction with eIF4G, constitutes the core of the eIF4F complex, which plays a key role in the circularization of mRNAs and their subsequent cap‐dependent translation. In addition to its fundamental role in mRNA translation initiation, other functions have been described or suggested for eIF4E, including acting as a proviral factor and participating in sexual development. We used CRISPR/Cas9 genome editing to generate melon eif4e knockout mutant lines. Editing worked efficiently in melon, as we obtained transformed plants with a single‐nucleotide deletion in homozygosis in the first eIF4E exon already in a T0 generation. Edited and non‐transgenic plants of a segregating F2 generation were inoculated with Moroccan watermelon mosaic virus (MWMV); homozygous mutant plants showed virus resistance, while heterozygous and non‐mutant plants were infected, in agreement with our previous results with plants silenced in eIF4E. Interestingly, all homozygous edited plants of the T0 and F2 generations showed a male sterility phenotype, while crossing with wild‐type plants restored fertility, displaying a perfect correlation between the segregation of the male sterility phenotype and the segregation of the eif4e mutation. Morphological comparative analysis of melon male flowers along consecutive developmental stages showed postmeiotic abnormal development for both microsporocytes and tapetum, with clear differences in the timing of tapetum degradation in the mutant versus wild‐type. An RNA‐Seq analysis identified critical genes in pollen development that were down‐regulated in flowers of eif4e/eif4e plants, and suggested that eIF4E‐specific mRNA translation initiation is a limiting factor for male gametes formation in melon. Resistance to MWMV is associated with eIF4E editing and eif4e knock‐out plants show a post‐meiotic, sporophytic male sterility phenotype.
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影响因子：
13.8
作者：
Bastet A;Lederer B;Giovinazzo N;Arnoux X;German-Retana S;Reinbold C;Brault V;Garcia D;Djennane S;Gersch S;Lemaire O;Robaglia C;Gallois JL
通讯作者：
Gallois JL
DOI：
10.1073/pnas.1904752116
发表时间：
2019-11-26
影响因子：
11.1
作者：
de Oliveira, Luciana Coutinho;Volpon, Laurent;Borden, Katherine L. B.
通讯作者：
Borden, Katherine L. B.
影响因子：
3
作者：
Garcia-Almodovar, R. C.;Gosalvez, B.;Burgos, L.
通讯作者：
Burgos, L.
影响因子：
3.9
作者：
Browning, KS
通讯作者：
Browning, KS
影响因子：
4.9
作者：
Chandrasekaran, Jeyabharathy;Brumin, Marina;Gal-On, Amit
通讯作者：
Gal-On, Amit