Xiaolian Zhang1,2, Lili Zhang2, Jishun Zhang1,2, Mengao Jia2, Linggai Cao2, Jing Yu1,2, Degang Zhao3,4. 1. Key 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. 2. Molecular Genetics Key Laboratory of China Tobacco, Guizhou Academy of Tobacco Science, Guiyang, 550081, China. 3. Key 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. dgzhao@gzu.edu.cn. 4. Guizhou Plant Conservation Technology Center, Guizhou Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, Guiyang, 550006, China. dgzhao@gzu.edu.cn.
Abstract
MAIN CONCLUSION: dmp1dmp2dmp3 mutants created by CRISPR/Cas9 could trigger maternal haploids in the allotetraploid model plant Nicotiana tabacum L. 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.
MAIN CONCLUSION: dmp1dmp2dmp3 mutants created by CRISPR/Cas9 could trigger maternal haploids in the allotetraploid model plant Nicotiana tabacum L. 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.
Authors: Sangam L Dwivedi; Anne B Britt; Leena Tripathi; Shivali Sharma; Hari D Upadhyaya; Rodomiro Ortiz Journal: Biotechnol Adv Date: 2015-07-09 Impact factor: 14.227
Authors: Raheleh Karimi-Ashtiyani; Takayoshi Ishii; Markus Niessen; Nils Stein; Stefan Heckmann; Maia Gurushidze; Ali Mohammad Banaei-Moghaddam; Jörg Fuchs; Veit Schubert; Kerstin Koch; Oda Weiss; Dmitri Demidov; Klaus Schmidt; Jochen Kumlehn; Andreas Houben Journal: Proc Natl Acad Sci U S A Date: 2015-08-20 Impact factor: 11.205