biostudies-literatureLi LNIGMS NIH HHS2039-2046https://www.ebi.ac.uk/biostudies/studies/S-EPMC9968529biostudies-literatureUnknown64(11)In flowering plants (angiosperms), fertilization of the egg cell by one sperm cell produces an embryo, whereas fusion of a second sperm cell with the central cell generates the endosperm. In most angiosperms like Arabidopsis, a pollen grain contains two isomorphic sperm cells required for this double fertilization process. A long-standing unsolved question is whether the two fertilization events have any preference. A tool to address this question is the usage of the cyclin-dependent kinase a1 (cdka;1) mutant pollen, which produces a single sperm-like cell (SLC). Here, we first adopt a complementation-based fluorescence-labeling method to successfully separate and collect cdka;1 mutant pollen containing a single SLC. Single-cell RNA-sequencing analysis revealed that cdka;1 SLCs show a geneJournal of integrative plant biologyThe egg cell is preferentially fertilized in Arabidopsis double fertilization.PMC9968529R35 GM131827Qu LJLi JGu HDresselhaus TZhong SDong JHou SXiang WZhang LWang YLi LSong ZfalseThe egg cell is preferentially fertilized in Arabidopsis double fertilization.In flowering plants (angiosperms), fertilization of the egg cell by one sperm cell produces an embryo, whereas fusion of a second sperm cell with the central cell generates the endosperm. In most angiosperms like Arabidopsis, a pollen grain contains two isomorphic sperm cells required for this double fertilization process. A long-standing unsolved question is whether the two fertilization events have any preference. A tool to address this question is the usage of the cyclin-dependent kinase a1 (cdka;1) mutant pollen, which produces a single sperm-like cell (SLC). Here, we first adopt a complementation-based fluorescence-labeling method to successfully separate and collect cdka;1 mutant pollen containing a single SLC. Single-cell RNA-sequencing analysis revealed that cdka;1 SLCs show a gene2022-01-01T00:00:00Z2022 Nov2025-04-22T09:58:12.649Z2025-02-18T23:57:27.972ZS-EPMC99685293616537310.1111/jipb.13370