Method for improving artemisinin content in Artemisia annua L. by DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene transfer

Abstract

The invention provides a method for improving an artemisinin content in Artemisia annua L. by DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene transfer. The method comprises the following steps of: cloning DXR (1-deoxy-D-xylulose-5-phosphate reductoisomerase) gene in the Artemisia annua L., so as to establish a plant expression carrier containing the DXR gene; transferring the DXR gene into Artemisia annua L. through agrobacterium tumefaciens-mediated transformation to grow a plant; detecting the integration condition of the exogenous target gene DXR by a PCR (Polymerase Chain Reaction); detecting the artemisinin content in the transgenic Artemisia annua L. through high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD); and screening the transgenic Artemisia annua L. plant with improved artemisinin content. The artemisinin content of the transgenic Artemisia annua L. obtained by the invention is obviously improved and high up to 2.4 times that of the non-transferred plant. Therefore, the invention provides the method for improving the artemisinin content in the Artemisia annua L. and lays a foundation for producing artemisinin by using the transgenic Artemisia annua L. in a large scale.

Description

technical field [0001] The invention relates to a method for increasing the content of artemisinin in the field of biotechnology, in particular to a method for increasing the content of artemisinin in Artemisia annua by transducing DXR gene. Background technique [0002] Artemisia annua L. is an annual herb belonging to the family Asteraceae. Artemisinin (artemisinin) is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is currently the most effective drug for treating malaria in the world, especially for cerebral malaria and anti-malaria Chloroquine malaria has the characteristics of quick action and low toxicity. Currently, the most effective treatment for malaria recommended by the World Health Organization is artemisinin combination therapy (ACTs). In addition, with the deepening of pharmacological research on artemisinin, scientists have discovered that artemisinin and its derivatives also have anti-inflammatory,...

AI Technical Summary

Problems solved by technology

[0003] At present, the main source of artemisinin is extracted from the aerial parts of Artemisia annua plants. However, the content of artemisinin in Artemisia annua is very low. The average content of artemisinin in different planting environments and planting varieties is 0.01-1% of the dry weight of leaves of Artemisia annua. , making the large-scale commercial production of this drug limited

Due to the complex structure of artemisinin, the artificial synthesis is difficult, the yield is low, and the cost is high, so it is not feasible

Some people try to produce artemisinin by tissue culture and cell engineering. However, the content of artemisinin in callus is less than 0.1% of dry weight, and the highest in shoot is only 0.16% of dry weight. Artemisinin was not detected in roots

Therefore, the feasibility of using tissue culture and cell engineering to produce artemisinin is not high.