Haploidization is invaluable for basic genetic research and crop breeding. The haploid bio-induction principle is an important topic that remains largely unexplored. In this study, both CenH3 RNAi and in vitro inhibition were used to simulate and induce haploids in allopolyploid crop. Notably, in vitro CenH3 inhibition showed that the results were much the same to that of RNAi in phenotype, chromosome behavior, microspore production, and haploid induction. Cytological analyses of RNAi and inhibitor-treated progenies revealed elimination of chromosomes, defective microspores with empty nuclei, thereby giving rise to pseudo male gametes, and haploid parthenogenesis induction. We found distinct defective empty microspores that were positively correlated with the decrease of CenH3 during RNAi manipulation. Investigation through both in vivo and in vitro studies revealed that haploidization was induced through the pseudo male gamete-mediated mock sexual reproduction. The present results provide insights for the haploid parthenogenesis induction process.

单倍体化对于基础遗传研究和农作物育种是无价的。单倍体生物诱导原理是一个重要的话题，至今仍未开发。在这项研究中，CenH3 RNAi和体外抑制都用于模拟和诱导同种多倍体作物中的单倍体。值得注意的是，体外CenH3抑制作用在表型，染色体行为，小孢子产生和单倍体诱导方面与RNAi的结果非常相似。RNAi和抑制剂处理的子代的细胞学分析表明，消除了染色体，空核缺陷的小孢子，从而引起了假雄配子和单倍体孤雌生殖的诱导。我们发现了明显的缺陷空微孢子，与空泡的减少呈正相关。RNAi操纵过程中的CenH3。通过体内和体外研究的调查表明，单倍体化是通过假雄配子介导的模拟性生殖而诱导的。本结果为单倍体孤雌生殖诱导过程提供了见识。