Quality Variation And Production Regionalization Of Artemisia Annua L.

Artemisia annua L.is an annual herb of Artemisia annua L.,whose medicinal part is the dry part of the ground.It has the effect of clearing heat,cutting off malaria and so on.It is our Chinese Traditional medicine.Artemisinin isolated from Artemisia annua L.has the advantages of high anti-malarial and non-toxicity,so it is regarded as the preferred antimalarial drug by World Health Organization.Modern research shows that Artemisia annua L.has activities of anti-viral,anti-tumor and anti-arrhythmic.The wild and cultivated Artemisia annua L.are currently the main source of artemisinin,but the wild resources are very limited and it can not meet the market demand.Therefore,the introduction and cultivation of Artemisia annua L.is an important way to solve the industrial development of Artemisia annua L..Although Artemisia annua L.is widely distributed in the world,the artemisinin content from different regions varies greatly.In order to improve the production and quality of Artemisia annua L.,it is critical to analyze spatial variation of artemisinin content and climate characteristics of high quality producing areas of Artemisia annua L..Then production regionalization should be performed to select high quality producing areas for its introduction and cultivation.Meanwhile,we should pay attention to the genetic stability of germplasm and select the better quality germplasm for genetic breeding.This is also the key problem for the introduction and cultivation of Artemisia annua L..In this study,ultra-high performance liquid chromatography(UPLC-ELSD)was established to determine the content of artemisinin in 119 samples of Artemisia annua L.from different regions.The chromatographic conditions were as follows:column temperature 25?;ELSD detector gain 100,sprayer heating mode,power level 60%,drift tube temperature 45?,gas pressure 20psi.The mobile phase was A for acetonitrile,B for water,flow rate 0.2 ml-min-1,injection volume:5?L;elution gradient:0 to 1 min,20%to 40%A,1 to 3 min,40%to 60%A;3-4 min,60%-62%A;4?5 min,62%?65%A;5?7 min,65%?68%A;7?10 min,68%A.Under the chromatographic conditions established in this paper,the degree of artemisinin separation was good.The linearity,precision,stability,repeatability,and sample recovery experiments of the method were all in line with the requirements,and can be well applied to the determination of artemisinin content.According to the annual and monthly average climatic factors,growth needs of medicinal plants can be characterized on different time scales.Principal component analysis(PCA),partial least squares(PLS)and other statistical method were used to analyze the spatial variability of artemisinin content and climate characteristics of Artemisia annua L.high quality producing area.The results showed that the artemisinin content was significantly correlated with the monthly mean temperature range and the coldest average annual precipitation.The two factors both had inhibitory effects on artemisinin content.Morever,the effect of monthly average climatic factors on artemisinin content was studied.It was showed that the average monthly precipitation in February and September had a greater influence on artemisinin content.The average monthly precipitation in February was negatively correlated with artemisinin content and the average monthly precipitation in September had the opposite correlation.Monthly average temperature from January to April had a greater impact and correlated with artemisinin content positively.Study on climate characteristics of Artemisia annua L.production area showed that the ratios of monthly-precipitation and monthly-temperature from April to October were very stable(the value of CV<1),signifing that the change trend of precipitation and temperature relatively consistent in the main phenological period of Artemisia annua L..Furthermore,the CV values in the high-quality area from April to October were less than 0.5 and that in the low-quality area were less than 1.The value of CV in the high quality area was less than that of the low quality area in every month.It was inferred that the smaller ratio(less than 0.5)was more conducive to the accumulation of artemisinin.Higher artemisinin content in high quality areas may be related to the time of accumulation.In the high-quality area,the minimum range of monthly average precipitation from April to October was 10-88mm,the maximum range was 176-3 09mm;the minimum range of monthly average temperature was 3.1-13.2?and the maximum range was 24.9-30.2?.The results of ITS2 sequence identification showed that 119 samples were Artemisia annua L..Sequence analysis showed that there were almost no variation in the ITS2 sequence and psbA-trnH sequence of Artemisia annua L.from different producing areas.It was found that the genetic distance of Artemisia annua L.in different regions was small based on the ITS2 sequence and the psbA-trnH sequence.We obtained other ITS2 sequences and psbA-trnH sequences of the related species from NCBI and analyzed genetic distance among different species.It was showed that the distance within the species was much less than that between species.The result of system cluster tree,which was constructed by Neighbor Method(NJ)based on ITS2 sequences and psbA-trnH sequences,showed that Artemisia annua L.differed significantly from related species and assembled separately.The evaluation results of four ecological niche models(BIOCLIM,DOMAIN,GARP,MAXENT)showed that the AUC values were above 0.75 and the Kappa values were above 0.4,indicating good prediction accuracy and consistency of models.So they can well predict the potential suitable area.The prediction of different niche models had slightly difference because of different principles.Among them,the MAXENT model showed better accuracy.Therefore,the MAXENT model was selected as the base map of the production regionalization of Artemisia annua L..The model between artemisinin and ecological factors was constructed using R language and the production regionalization was based on Geographic Information System(GIS).The result of production regionalization showed that Chongqing,eastern Sichuan,northern Guizhou,southern Guangdong,western Hunan,western Hubei and Hainan had not only high content of artemisinin,but also suitable index of Artemisia annua L.,and can be the preferred area for high quality Artemisia annua L..The content of artemisinin in the area of central China and Yunnan was slightly lower,which belonged to the sub-suitable area of Artemisia annua L..It can be used as the expansion area of Artemisia annua L..During the cultivation,ecological factors can be reasonably controlled to obtain high-quality Artemisia annua L..In southeast area such as southern Hunan,Jiangxi and northern Fujian,artemisinin content and the ecological suitability index of Artemisia annua L.were both the lowest.So,these areas were not recommended for large-scale introduction and cultivation.