ABSTRACT.Finding an efficient and affordable treatment against malaria is still a challenge for medicine. Artemisinin is an effective antimalarial drug isolated from Artemisia annua. However, the artemisinin content of A. annua is very low. We used transgenic technology to increase the artemisinin content of A. annua by overexpressing cytochrome P450 monooxygenase (cyp71av1) and cytochrome P450 reductase (cpr) genes. CYP71AV1 is a key enzyme in the artemisinin biosynthesis pathway, while CPR is a redox partner for CYP71AV1. Eight independent transgenic A. annua plants were obtained through Agrobacterium tumefaciens-mediated transformation, which was confirmed by PCR and Southern blot analyses. The real-time qPCR results showed that the gene cyp71av1 was highly expressed at the transcriptional level in the transgenic A. annua plants. HPLC analysis showed that the artemisinin content was increased in a number of the transgenic plants, in which both cyp71av1 and cpr were overexpressed. In one of the transgenic A. annua plants, the artemisinin content was 38% higher than in the non-transgenic plants. We conclude that overexpressing key enzymes of the biosynthesis pathway is an effective means for increasing artemisinin content in plants.
Rht18, derived from Triticum durum (tetraploid) wheat, is classified as a gibberellic acid (GA)-responsive dwarfing gene. Prior to this study, the responses of Rht18 to exogenous GA on agronomic traits in hexaploid wheat were still unknown. The response of Rht18 to exogenous GA 3 on coleoptile length, plant height, yield components and other agronomic traits were investigated using F 4:5 and F 5:6 hexaploid dwarf lines with Rht18 derived from two crosses between the tetraploid donor Icaro and tall Chinese winter wheat cultivars, Xifeng 20 and Jinmai 47. Applications of exogenous GA 3 significantly increased coleoptile length in both lines and their tall parents. Plant height was significantly increased by 21.3 and 10.7% in the GA 3 -treated dwarf lines of Xifeng 20 and Jinmai 47, respectively. Compared to the untreated dwarf lines, the partitioning of dry matter to ears at anthesis was significantly decreased while the partitioning of dry matter to stems was significantly increased in the GA 3 -treated dwarf lines. There were no obvious changes in plant height and dry matter partitioning in the GA 3 -treated tall parents. Exogenous GA 3 significantly decreased grain number spike -1 while it increased 1000-kernel weight in both the dwarf lines and tall parents. Thus, applications of exogenous GA 3 restored plant height and other agronomic traits of Rht18 dwarf lines to the levels of the tall parents. This study indicated that Rht18 dwarf mutants are GA-deficient lines with impaired GA biosynthesis.
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