We use cookies on our website. Some are necessary for the operation of the website. You can also allow cookies for statistical purposes. You can adjust the data protection settings or agree to all cookies directly.
Data Protection Declaration
Imprint
Non-transgenic approaches for the development of haploid inducer lines of Apiaceae vegetable species
Project
Project code: JKI-ZGO-08-4175
Contract period: 01.01.2016
- 31.12.2019
Purpose of research: Applied research
In recent years, there has been an increasing usage of F1 hybrid seed in commercial carrot production systems worldwide. Current hybrid breeding programs in carrots are based on inbred lines produced by repeated self fertilizations or in vitro-techniques such as microspore culture. Manipulating centromeres of the chromosomes has been proposed as universal novel method for the production of haploid plants. By this way, haploids can be generated through manual cross-fertilizations after manipulating a single centromere protein, the centromere-specific histone H3 variant CENH3, in one of the parents designated as 'haploid inducer'. Crosses with haploid inducer genotypes result in karyotypically unstable embryo cells, which have lost one of the parent-specific chromosome sets. Initially, the new haploid technique was based on transgenic haploid inducer plants. Therefore, the major goal of the project is the elaboration of non-transgenic methods to generate partially functional CENH3 alleles through targeted mutagenesis in transient expression systems of protoplasts or alternatively other single cell cultures in vitro. In addition, a very promising new DNA-free genome editing technique will be investigated, which uses preassembled complexes of purified Cas9 protein and the specific guide RNA only, with the aim to develop non-GMO haploidy inducers. For latter approach, existing carrot protoplast regeneration protocols are tested and optimized. Plants regenerated from protoplasts (or other single cells) will be transferred into the greenhouse and analyzed by molecular and cytogenetic methods as for instance the localization of CENH3 signals by immunostaining with CENH3 antibodies. Crosses will be done to create seeds and plants of the next generation, which again have to be studied for the behavior (i.e. centromere loading) of the modified CENH3 and for the occurrence and frequency of putative haploids.
Section overview
Subjects
- Plant Breeding
- Vegetable Gardening
Framework programme
Funding programme
Excutive institution
Institute for Breeding Research on Horticultural Crops (JKI-ZG)
