Sara Zafar1,Erum Dilshad2,Hammad Ismail3,Chahat Batool Rizvi1,Bushra Mirza1*.Rol genes enhance content of artemisinin and other secondary metabolites in Shennong hybrid of Artemisia annua[J].Chinese Herbal Medicines (CHM),2019,11(2):209-215
Rol genes enhance content of artemisinin and other secondary metabolites in Shennong hybrid of Artemisia annua
  
DOI:10.1016/j.chmed.2018.11.002
中文关键词:  
英文关键词:Artemisia annua L.  artemisinin  genetic transformation  Hyb1209r  rol genes
基金项目:
Author NameAffiliation
Sara Zafar1 1.Department of Biochemistry, Quaid-i-Azam University Islamabad, Islamabad 44000, Pakistan 
Erum Dilshad2 2.Department of Bioinformatics and Biosciences, Faculty of Health and Life Sciences, Capital University of Science and Technology (CUST) Islamabad, Islamabad 44000, Pakistan 
Hammad Ismail3 3.Department of Biochemistry and Molecular Biology, University of Gujarat, Gujrat 50700, Pakistan 
Chahat Batool Rizvi1 1.Department of Biochemistry, Quaid-i-Azam University Islamabad, Islamabad 44000, Pakistan 
Bushra Mirza1* 1.Department of Biochemistry, Quaid-i-Azam University Islamabad, Islamabad 44000, Pakistan 
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中文摘要:
      
英文摘要:
      Objective Artemisia annua is the chief source of artemisinin, a potent antimalarial agent, in which other bioactive phytochemicals present. Due to low levels of bioactive compounds including artemisinin and flavonoids, it is necessary to increase the level of the secondary metabolites by regulating rol genes. Methods A hybrid variety of A. annua (Hyb1209r, Shennong) developed by the Centre for Novel Agricultural Products, University of York, UK, was selected to produce transgenics of rolB and rolC genes. Genetic transformation was carried out via Agrobacterium tumefaciens GV3101 harboring rolB and rolC genes of Agrobacterium rhizogenes cloned separately. HPLC was used for the qualitative and quantitative analysis of flavonoids and artemisinin. Furthermore, thin layer chromatography (TLC) was also used to analyse artemisinin content. Results Comparative analysis via HPLC revealed considerable enhancement in the phytochemical content of transgenic A. annua plants as compared to the wild type plant. Transgenics of rolB gene showed an average increase of 321% in rutin, 97.2% in caffeic acid, and 218.4% in myricetin, respectively. In the case of rolC gene transgenics, an average increase of 197.5% in rutin, 76.3 % in caffeic acid, and 209.3% in myricetin was observed. Transgenics of rolB and rolC genes showed a 14.3%?28.6% and 2.8%?12.7% increase in artemisinin content respectively by HPLC analysis. TLC analysis showed that an average 142.2% and 110.2% enhancement in artemisinin for rolB and rolC transgenics respectively, compared with the wild type. An enhanced production of total flavonoids (average 30.2% and 25.5% increase in rolB and rolC transgenics, respectively) and total phenolics (average 34.3% and 25.8% increase in rolB and rolC transgenics, respectively) was observed as a result of transformation. Transformed A. annua plants showed improved free radical scavenging activity (average 46.5% and 29.1% increase in rolB and rolC transgenics, respectively) and total reducing power (average 32.7% and 26.4% increase in rolB and rolC transgenics, respectively) compared with untransformed plant. Conclusion rolB and rolC genes were effective for developing A. annua plants with an enhanced level of phytochemicals.
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