The human insulin gene modified with a C-peptide was synthesized according to the plant-preferred codon,and a fusion gene expression vector of insulin combined with green fluorescent protein(GFP)was constructed.The op...The human insulin gene modified with a C-peptide was synthesized according to the plant-preferred codon,and a fusion gene expression vector of insulin combined with green fluorescent protein(GFP)was constructed.The optimization of the flax callus culturing was undertaken,and a more efficient Agrobacterium-mediated genetic transformation of the flax hypocotyls was achieved.The critical concentration values of hygromycin on the flax hypocotyl development,as well as on its differentiated callus,were explored by the method of antibiotic gradient addition,and the application of antibiotic screening for the verification of positive calluses was assessed.The fusion gene of insulin and GFP was successfully inserted into the flax genome and expressed,as confirmed through polymerase chain reaction and Western blotting.In conclusion,we have established a flax callus culture system suitable for insulin expression.By optimizing the conditions of the flax callus induction,transformation,screening,and verification of a transgenic callus,we have provided an effective way to obtain insulin.Moreover,the herein-employed flax callus culture system could provide a feasible,cheap,and environmentally friendly platform for producing bioactive proteins.展开更多
In main foxtail millet growing regions of China, natural disasters happen frequently, causing losses in production and finance. Therefore, it is urgently needed to breed new superior quality foxtail millet varieties w...In main foxtail millet growing regions of China, natural disasters happen frequently, causing losses in production and finance. Therefore, it is urgently needed to breed new superior quality foxtail millet varieties with stress resistance, stable and high production, and, so as to stabilize millet production and promote millet industry development. Jigu32, a new foxtail millet variety with stable, high-yield and superior qualities, was developed using Target Character Gene Bank breeding method, and its physiological mechanism was studied as well. Results showed that the prominent characteristics of Jigu32 were as follows: 1) strong stress resistance and stable yielding;2) high yielding;3) rich calcium content and superior qualities;4) excellent comprehensive characteristics. In 2010 National Foxtail Millet Regional Trials, the weather was tough. Severe drought occurred in some experimental stations while in some others, continuous rain, low temperature and little sunlight appeared. However, with the outstanding stress resistance, Jigu32 achieved the highest yields, and the yields were very stable under different conditions. Per unit yield of Jigu32 reached to5133.3 kg/hm2, which was the highest in the trials, increasing 9.42% compared with the controls. Calcium content of Jigu32 was 121 mg/kg in the grain, and the taste, nutrition and commodity qualities were optimal. Therefore, Jigu32 was rated as the national secondary superior quality foxtail millet. The study showed that the physiological mechanism of Jigu32’s merits was based on the improved activities of peroxidase (POD), superoxide dismutase (SOD), 6 phosphate dehydrogenase (G6PDH), glutamine synthetase (GS) and glutamic dehydrogenase (GDH), and on its higher absorption ability and conversion efficiency of N, P, K. POD, SOD and G6PDH of Jigu32 were more active in each development phase, leading to higher resistance to adversity and aging;glutamine synthetase (GS) and glutamate dehydrogenase (GDH) of Jigu32 were more active, resulting in higher assimilation and transformation ability of nutrients. It is of great significance to promote the development of Jigu32, and it will be beneficial to sustainable, stable agricultural development, and thus orderly and stably boost the development of the millet industry in our country. The research on its physiological mechanism of stable and high yielding will provide theoretical support while breeding new stable and high-yield foxtail millet varieties later.展开更多
基金funded by Hebei Development and Reform Commission of China,Hebei Engineering Laboratory,grant number(2021)1157 and the Innovation Fund Project of Hebei University of Engineering(114/SJ2401002149).
文摘The human insulin gene modified with a C-peptide was synthesized according to the plant-preferred codon,and a fusion gene expression vector of insulin combined with green fluorescent protein(GFP)was constructed.The optimization of the flax callus culturing was undertaken,and a more efficient Agrobacterium-mediated genetic transformation of the flax hypocotyls was achieved.The critical concentration values of hygromycin on the flax hypocotyl development,as well as on its differentiated callus,were explored by the method of antibiotic gradient addition,and the application of antibiotic screening for the verification of positive calluses was assessed.The fusion gene of insulin and GFP was successfully inserted into the flax genome and expressed,as confirmed through polymerase chain reaction and Western blotting.In conclusion,we have established a flax callus culture system suitable for insulin expression.By optimizing the conditions of the flax callus induction,transformation,screening,and verification of a transgenic callus,we have provided an effective way to obtain insulin.Moreover,the herein-employed flax callus culture system could provide a feasible,cheap,and environmentally friendly platform for producing bioactive proteins.
文摘In main foxtail millet growing regions of China, natural disasters happen frequently, causing losses in production and finance. Therefore, it is urgently needed to breed new superior quality foxtail millet varieties with stress resistance, stable and high production, and, so as to stabilize millet production and promote millet industry development. Jigu32, a new foxtail millet variety with stable, high-yield and superior qualities, was developed using Target Character Gene Bank breeding method, and its physiological mechanism was studied as well. Results showed that the prominent characteristics of Jigu32 were as follows: 1) strong stress resistance and stable yielding;2) high yielding;3) rich calcium content and superior qualities;4) excellent comprehensive characteristics. In 2010 National Foxtail Millet Regional Trials, the weather was tough. Severe drought occurred in some experimental stations while in some others, continuous rain, low temperature and little sunlight appeared. However, with the outstanding stress resistance, Jigu32 achieved the highest yields, and the yields were very stable under different conditions. Per unit yield of Jigu32 reached to5133.3 kg/hm2, which was the highest in the trials, increasing 9.42% compared with the controls. Calcium content of Jigu32 was 121 mg/kg in the grain, and the taste, nutrition and commodity qualities were optimal. Therefore, Jigu32 was rated as the national secondary superior quality foxtail millet. The study showed that the physiological mechanism of Jigu32’s merits was based on the improved activities of peroxidase (POD), superoxide dismutase (SOD), 6 phosphate dehydrogenase (G6PDH), glutamine synthetase (GS) and glutamic dehydrogenase (GDH), and on its higher absorption ability and conversion efficiency of N, P, K. POD, SOD and G6PDH of Jigu32 were more active in each development phase, leading to higher resistance to adversity and aging;glutamine synthetase (GS) and glutamate dehydrogenase (GDH) of Jigu32 were more active, resulting in higher assimilation and transformation ability of nutrients. It is of great significance to promote the development of Jigu32, and it will be beneficial to sustainable, stable agricultural development, and thus orderly and stably boost the development of the millet industry in our country. The research on its physiological mechanism of stable and high yielding will provide theoretical support while breeding new stable and high-yield foxtail millet varieties later.
