The generation of haploid plants accelerates the crop breeding process. One of the haploidization strategies is based on the genetic manipulation of endogenous centromere-specific histone 3 (CENH3). To extend the haploidization toolbox, we tested whether targeted in vivo degradation of CENH3 protein can be harnessed to generate haploids in Arabidopsis thaliana. We show that a recombinant anti-GFP nanobody fused to either heterologous F-box (NSlmb) or SPOP/BTB ligase proteins can recognize maternally derived enhanced yellow fluorescent protein (EYFP)-tagged CENH3 in planta and make it accessible for the ubiquitin-proteasome pathway. Outcrossing of the genomic CENH3-EYFP-complemented cenh3.1 mother with plants expressing the GFP-nanobody-targeted E3 ubiquitin ligase resulted in a haploid frequency of up to 7.6% in pooled F1 seeds. EYFP-CENH3 degradation occurred independently in embryo and endosperm cells. In reciprocal crosses, no haploid induction occurred. We propose that the uniparental degradation of EYFP-fused genomic CENH3 during early embryogenesis leads to a decrease in its level at centromeres and subsequently weakens the centromeres. The male-derived wild type CENH3 containing centromere outcompetes the CENH3-EYFP depleted centromere. Consequently, maternal chromosomes undergo elimination, resulting in haploids.
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http://dx.doi.org/10.1093/jxb/erac359 | DOI Listing |
Front Genet
January 2025
National Rapeseed Genetic Improvement Center, Chengdu Academy of Agriculture and Forestry Sciences, Chengdu Research Branch, Chengdu, China.
The Ogura cytoplasmic male sterility (CMS) line of has gained significant attention for its use in harnessing heterosis. It remains unaffected by temperature and environment and is thorough and stable. The Ogura cytoplasmic restorer line of is derived from the distant hybridization of and , but it carried a large number of radish fragments into , because there is no homologous allele of the restorer gene in , transferring it becomes challenging.
View Article and Find Full Text PDFPlant Methods
January 2025
Key Laboratory of Grassland Ecosystem of Ministry of Education, Pratacultural Engineering Laboratory of Gansu Province, Sino-U.S. Centers for Grazingland Ecosystem Sustainability, Collage of Pratacultural Science, Gansu Agricultural University, Lanzhou, Gansu, 730070, China.
Background: The rapid production of doubled haploids by anther culture technology is an important breeding method for awnless triticale. The aim of this study was to explore the effects of triticale genotype and the types and ratios of exogenous hormones in the medium on the efficiency of triticale anther culture.
Results: Anthers of five triticale genotypes were cultured on four different callus induction media and the calli were induced to differentiate into green plants by culture on three different differentiation media.
Plant Biotechnol J
November 2024
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Plant Sci
February 2025
Pollen Biotechnology of Crop Plants Group, Margarita Salas Center of Biological Research, CIB-CSIC, Ramiro de Maeztu 9, Madrid 28040, Spain. Electronic address:
In vivo, microspores in the anthers follow the gametophytic development pathway, culminating in the formation of pollen grains. Conversely, in vitro, under stress treatments, microspores can be reprogrammed into totipotent cells, initiating an embryogenic pathway that produces haploid and double-haploid embryos, which are important biotechnological tools in plant breeding. There is growing evidence that epigenetic reprogramming occurs during microspore embryogenesis through DNA methylation, but less is known about the role of histone modifications.
View Article and Find Full Text PDFFor Res (Fayettev)
May 2024
State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China.
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