Zhou Yongmei, Fang Wenqin, Pang Ziqin, Chen Li-Yu, Cai Hanyang, Ain Noor-Ul-, Chang Men-Chi, Ming Ray
FAFU and UIUC Joint Center for Genomics and Biotechnology, Key Laboratory of Sugarcane Biology and Genetic Breeding Ministry of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, China.
Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian Agriculture and Forestry University, Fuzhou, China.
Front Plant Sci. 2022 Jul 7;13:924417. doi: 10.3389/fpls.2022.924417. eCollection 2022.
During sexual reproduction in flowering plants, haploid spores are formed from meiosis of spore mother cells. The spores then undergo mitosis, develop into female and male gametophytes, and give rise to seeds after fertilization. We identified a female sterile mutant from EMS mutagenesis, and analyses of two T-DNA insertion mutants, and , and detected a partial female and male sterility. The mutant gametophyte development was arrested at one nuclear stage. A complementation test using a genomic sequence of with its native promoter restored the function in the three mutant lines. Transcriptome profiling of ovules revealed that four genes encoding clathrin assembly proteins PICALM5A/B and PICALM9A/B, which were involved in endocytosis, were downregulated, which were confirmed to interact with AP1G2 through yeast two-hybrid assays and BIFC analysis. Our result also demonstrated that and several calcium-dependent protein kinases, including , were all downregulated in the ovules of . Moreover, Ca concentration was low in impaired gametophytes. Therefore, we proposed that through interaction with PICALM5A/B and PICALM9A/B, AP1G2 may mediate gametogenesis accompanied by Ca signaling in Arabidopsis. Our findings revealed a crucial role of AP1G2 in female and male gametogenesis in Arabidopsis and enhanced our understanding of the molecular mechanisms underpinning sexual reproduction in flowering plants.
在开花植物的有性生殖过程中,单倍体孢子由孢子母细胞减数分裂形成。然后,孢子进行有丝分裂,发育成雌、雄配子体,并在受精后产生种子。我们从EMS诱变中鉴定出一个雌性不育突变体,并对两个T-DNA插入突变体进行分析,检测到部分雌雄不育现象。该突变体的配子体发育在单核期停滞。使用带有其天然启动子的基因组序列进行的互补试验恢复了三个突变系的功能。对该突变体胚珠进行转录组分析发现,四个编码参与内吞作用的网格蛋白组装蛋白PICALM5A/B和PICALM9A/B的基因表达下调,通过酵母双杂交试验和双分子荧光互补分析证实它们与AP1G2相互作用。我们的结果还表明,该突变体和几个钙依赖蛋白激酶,包括某激酶,在胚珠中均表达下调。此外,受损配子体中的钙浓度较低。因此,我们提出,通过与PICALM5A/B和PICALM9A/B相互作用,AP1G2可能在拟南芥中伴随着钙信号传导介导配子发生。我们的研究结果揭示了AP1G2在拟南芥雌雄配子发生中的关键作用,并增进了我们对开花植物有性生殖分子机制的理解。