The Arabidopsis R-SNARE protein YKT61 is essential for gametophyte development
The Arabidopsis R-SNARE protein YKT61 is essential for gametophyte development
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拟南芥 R-SNARE 蛋白 YKT61 对于配子体发育至关重要
DOI：
10.1111/jipb.13017
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发表时间：
2021
影响因子：
11.4
通讯作者：
Zhang Yan
中科院分区：
文献类型：
--
作者：
Ma Ting;Li En;Li Lushen;Li Sha;Zhang Yan
配子体发育是植物繁殖和种子产量的先决条件；因此，配子体发育的研究有助于我们了解基本的生物学问题，并在农业中具有潜在的应用。内膜区室的生物发生和动力学对于细胞生存至关重要，其调节机制才刚刚开始被揭示。在这里，我们报道了拟南芥SNARE（可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体）蛋白YKT61对于雄性和雌性都至关重要通过使用基于成簇规则间隔短回文重复序列 (CRISPR)/CRISPR 相关蛋白 9 (Cas9) 的基因组编辑，我们证明携带 YKT61 功能丧失等位基因的雄性和雌性配子体无法存活。具体来说，YKT61 功能的丧失导致雄配子体在花粉有丝分裂 I 时发育停滞，雌性配子体退化。 ykt61-3突变体中的YKT61导致YKT61的longin结构域中单个氨基酸缺失；所得突变蛋白不与多个SNARE相互作用，并且膜关联显着减少，表明YKT61的N端longin结构域在其功能中发挥多种作用。本研究表明拟南芥YKT61对于雄性和雌性配子发生至关重要，并为结合Cas9 介导的 Cas9 抗性转基因的编辑和表达。
Gametophyte development is a pre-requisite for plant reproduction and seed yield;therefore,studies of gametophyte development help us understand fundamental biological questions and have potential applications in agriculture.The biogenesis and dynamics of endomembrane compartments are critical for cell survival,and their regulatory mechanisms are just beginning to be revealed.Here,we report that the Arabidopsis thaliana SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) protein YKT61 is essential for both male and female gametogenesis.By using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-based genome editing,we demonstrated that male and female gametophytes carrying YKT61 loss-offunction alleles do not survive.Specifically,loss of YKT61 function resulted in the arrest of male gametophytic development at pollen mitosis I and the degeneration of female gametophytes.A three-base-pair deletion in YKT61 in the ykt61-3 mutant resulted in a single-amino acid deletion in the longin domain of YKT61;the resulting mutant protein does not interact with multiple SNAREs and showed substantially reduced membrane association,suggesting that the N-terminal longin domain of YKT61 plays multiple roles in its function.This study demonstrates that Arabidopsis YKT61 is essential for male and female gametogenesis and sets an example for functional characterization of essential genes with the combination of Cas9-mediated editing and expression from a Cas9-resistant transgene.