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Haploid induction in allotetraploid tobacco using DMPs mutation
Xiaolian Zhang1,2, Lili Zhang2, Jishun Zhang1,2
1Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Guizhou Key Laboratory of Agro-Bioengineering, Institute of Agro-Bioengineering/College of Life Sciences, Guizhou University, Guiyang, 550025, China.
CRISPR/Cas9 technology enabled the creation of dmp1dmp2dmp3 mutants in tobacco, successfully triggering maternal haploids. This breakthrough advances double haploid production in polyploid plants.
Area of Science:
- Plant genetics
- Polyploid breeding
- Haploid induction
Background:
- Double haploid (DH) technology accelerates crop breeding by rapidly producing homozygous lines.
- Haploid induction is crucial for DH production, but its application in polyploid plants remains challenging.
- DMP genes have been utilized for haploid induction in diploid plants.
Purpose of the Study:
- To investigate the potential of haploid induction in the allotetraploid plant Nicotiana tabacum.
- To identify and characterize DMP gene homologues in tobacco.
- To develop a functional haploid inducer system in polyploids.
Main Methods:
- Identification and characterization of three DMP gene homologues (NtDMP1, NtDMP2, NtDMP3) in Nicotiana tabacum.
- CRISPR/Cas9-mediated generation of loss-of-function mutants for all three genes.
- Subcellular localization studies of NtDMP proteins.
- Assessment of haploid induction rates and phenotypic analysis of resulting haploids.
- Development of a visible screening system for haploids using a powdery mildew resistance phenotype.
Main Results:
- Three NtDMP genes were identified, with encoded proteins localized to the endoplasmic reticulum.
- Loss-of-function mutations in NtDMP1, NtDMP2, and NtDMP3 successfully induced maternal haploids at rates of 1.52-1.75%.
- The created haploid inducers showed altered pollen vigor and seed germination compared to wild-type tobacco.
- A visible haploid identification system based on powdery mildew resistance was established.
Conclusions:
- CRISPR/Cas9-induced mutations in NtDMP genes can trigger maternal haploidy in the allotetraploid Nicotiana tabacum.
- This study demonstrates the feasibility of haploid induction in polyploid plants, expanding the utility of DH technology.
- The developed system provides a foundation for applying haploid induction in breeding programs for polyploid crops.
