The Arabidopsis R-SNARE protein YKT61 is essential for gametophyte development

• 来源期刊: 植物学报（英文版）
• 年，卷（期）: 2021,（4）
• 所属期刊栏目: Cell and Developmental Biology
• 页码范围: 676-690
• 页数: 15页
• 语种: 英文

摘要

Gametophyte development is a pre-requisite for plant reproduction and seed yield; therefore, studies of gametophyte development help us un-derstand 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 sensi-tive factor attachment protein receptor) protein YKT61 is essential for both male and female gametogenesis. By using clustered regularly in-terspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-based ge-nome editing, we demonstrated that male and female gametophytes carrying YKT61 loss-of-function alleles do not survive. Specifically, loss of YKT61 function resulted in the arrest of male ga-metophytic 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 as-sociation, suggesting that the N-terminal longin domain of YKT61 plays multiple roles in its func-tion. This study demonstrates that Arabidopsis YKT61 is essential for male and female gameto-genesis and sets an example for functional characterization of essential genes with the com-bination of Cas9-mediated editing and expression from a Cas9-resistant transgene.