title: A novel male sterility-fertility restoration system in plants for hybrid seed production.
abstract: Hybrid seeds are used for stimulated crop production, as they harness heterosis. The achievement of complete male-sterility in the female-parent and the restored-fertility in F1-hybrids are the major bottlenecks in the commercial hybrid seed production. Here, we report a male sterility-fertility restoration system by engineering the in most nutritive anther wall layer tapetum of female and male parents. In the female parent, high-level, and stringent expression of Arabidopsis autophagy-related gene BECLIN1 was achieved in the tapetum, which altered the tapetal degeneration program, leading to male sterility. This works on our previously demonstrated expression cassette based on functional complementation of TATA-box mutant (TGTA) promoter and TATA-binding protein mutant3 (TBPm3), with modification by conjugating Long Hypocotyle in Far-Red1 fragment (HFR1(NT131)) with TBPm3 (HFR1(NT131)-TBPm3) to exercise regulatory control over it. In the male parent, tapetum-specific Constitutive photo-morphogenesis1 (COP1) was expressed. The F1 obtained by crossing these engineered parents showed decreased BECLIN1 expression, which was further completely abolished when COP1-mutant (COP1(L105A)) was used as a male parent, leading to normal tapetal development and restored fertility. The system works on COP1-HFR1 interaction and COP1-mediated degradation of TBPm3 pool (HFR1(NT131)-TBPm3). The system can be deployed for hybrid seed production in agricultural crops.
authors.affiliation: Plant Molecular Biology Laboratory, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, INDIA.
authors.forename: Surendra Pratap
authors.initials: SP
authors.lastname: Singh
authors.affiliation: National Agri-Food Biotechnology Institute, Department of Biotechnology (DBT), C-127, Industrial Area, Phase VIII, Mohali, 160071, INDIA.
authors.forename: Sudhir P
authors.initials: SP
authors.lastname: Singh
authors.affiliation: Plant Molecular Biology Laboratory, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, INDIA.
authors.forename: Tripti
authors.initials: T
authors.lastname: Pandey
authors.affiliation: Department of Botany, University of Lucknow, Lucknow-226007, INDIA.
authors.forename: Ram Rakshpal
authors.initials: RR
authors.lastname: Singh
authors.affiliation: 1] Plant Molecular Biology Laboratory, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, INDIA [2] Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2 Rafi Marg, New Delhi-110 001, India.
authors.forename: Samir V
authors.initials: SV
authors.lastname: Sawant
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D001343:Autophagy / Q000235:genetics; D000071186:Beclin-1; D002678:Chimera / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003433:Crosses, Genetic; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D035264:Flowers / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D040582:Inheritance Patterns; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D000069600:Plant Breeding / Q000379:methods*; D051479:Plant Infertility / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D035181:TATA-Box Binding Protein / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D029681:Arabidopsis Proteins; D000071186:Beclin-1; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D035181:TATA-Box Binding Protein; D044767:Ubiquitin-Protein Ligases; C414438:At1g02340 protein, Arabidopsis; C519561:beclin 1 protein, Arabidopsis; C471983:AT2G32950 protein, Arabidopsis