Main conclusion

dmp1dmp2dmp3 mutants created by CRISPR/Cas9 could trigger maternal haploids in the allotetraploid model plant Nicotiana tabacum L.

Abstract

Double haploid (DH) technology is becoming increasingly important because it can significantly accelerate the breeding process. Haploid induction plays a fundamental role in the production of DH lines. Haploid induction has been realized and applied in diploid plants using DMP genes. However, it has yet to be elucidated whether haploid induction could be established in polyploid plants. In the current study, three homologues of the DMP genes (NtDMP1, 2, and 3) were identified in the allotetraploid plant Nicotiana tabacum, and the encoded proteins localized in the endoplasmic reticulum. Loss-of-function mutations in all three genes triggered maternal haploids with an induction rate of 1.52–1.75%. Compared with wild-type tobacco, the created haploid inducer exhibited differences in pollen vigor and seed germination rate. Furthermore, to rapidly and easily screen haploids, a visible haploid identification system was established based on a powdery mildew resistance phenotype. Findings from this study lay the foundation for the potential application of haploid inducers in allotetraploid plants such as tobacco.

中文翻译：

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利用 DMPs 突变在异源四倍体烟草中诱导单倍体

主要结论

由 CRISPR/Cas9 产生的 dmp1dmp2dmp3突变体可以触发异源四倍体模型植物烟草 中的母体单倍体。

抽象的

双单倍体 (DH) 技术变得越来越重要，因为它可以显着加速育种过程。单倍体诱导在 DH 系的产生中起重要作用。使用DMP基因已实现单倍体诱导并在二倍体植物中应用。然而，尚未阐明单倍体诱导是否可以在多倍体植物中建立。在目前的研究中，在异源四倍体植物普通烟草中鉴定了DMP基因的三个同源物（NtDMP1、2和3 ）。，并且编码的蛋白质位于内质网中。所有三个基因的功能丧失突变均引发母体单倍体，诱导率为 1.52-1.75%。与野生型烟草相比，产生的单倍体诱导剂在花粉活力和种子发芽率方面表现出差异。此外，为了快速轻松地筛选单倍体，建立了基于白粉病抗性表型的可见单倍体识别系统。这项研究的结果为单倍体诱导剂在烟草等异源四倍体植物中的潜在应用奠定了基础。