Establishment Of Regeneration System In Brassica Juncea And Genetic Transformation Of HO-1 Gene Mediated By Agrobacterium Tumefaciens

Indian mustard belonging to the Brassica family is an importantly economic crop. Prompting plants to endure stress by genetic engineering has becomed a mainly developed focus. The heme oxygenase catalyses the degradation of heme to biliverdin, CO and Fe2+, and its capability of heavy metal-risistant is well identified in animals. We carried out the research in three aspects:(1) Using the cotyledon petiole of Indian mustard cultivar long-standing as explants, different kinds and concentrations of hormones which affected the regeneration frequency were studied. An efficient system for the production of transgenic Indian mustard plants was established by optimizing the various factors including seeding ages, hygromycin (Hyg) concentrations, antibiotic concentrations, AgN03 concentrations, the duration time of pre-culture, infection and co-culture, and infection concentrations.2. We cloned a novel gene HO-1 in Indian mustard encoding an enzyme which catalyzes heme degrading, with an open reading frame (ORF) of 844 bp, encodes a 281-amino acid protein.3. The HO-1 gene was transferred into the pCAMBIA1304, and then the vector of pCAMBIA1304-HO-1 jwas introduced into Indian mustard via Agrobacterium tumefaciena, and 32 regenerated plants with Hyg resistant were obtained. Experimental validation revealed that the HO-1 gene had been intergrated into the genome of Indian mustard by GUS staining and genomic PCR detceting. In total 15 plants were tested positive, withi a transformation frequency being 46.88%. This work lays a basis for Indian mustard improvement.