Study On Seed Germination Characteristics And Quality Evaluation Of Gentiana Crasicaulis Duthie Ex Burk

Gentiana crasicaulis Duthie ex Burk.,which belongs to gentiana of gentianaceae,a traditional Chinese herbal medicines,mainly distributes in southeast of Tibet,Yunnan province,Sichuan province,the northwest of Guizhou province,the southeast of Qinghai province,the south Gansu province and other places at 2100⁴500 meters above sea level.Mainly cultivated products are in Sichuan province and Yunnan province.The stem of wild Gentiana crasicaulis Duthie ex Burk.has highly medicinal value.Modern research shows that Gentiana crasicaulis Duthie ex Burk.contains a variety of different components,including iridoid glycosides,flavonoids and triterpenoids and other bioactive components,and various pharmacological effects,for instance,the alcohol extraction has anti-inflammatory,analgesic,antibacterial activities,the water extraction has liver,blood pressure,cardiovascular protection activities.Gentiana crasicaulis Duthie ex Burk.is based on sexual reproduction.However,in recent years,with the significantly increasing usage of clincal medicine,the wild resources have been over-excavating and depleting.Gentiana crasicaulis Duthie ex Burk.has been listed as Beijing II protected wild variety,and it is urgent to carry out artificial cultivation to alleviate the resources shortage.The previous sampling measurement results shows that the content of malicotinic acid and gentiopicroside of Gentiana crasicaulis Duthie ex Burk.growed in the high-altitude producing area is 3 to 6 times higher than the content in the "Chinese Pharmacopoeia"?2015 edition?.At present,the primary problem of the cultivation in high-altitude habitat is that the seed has dormancy period and the growth of Gentiana crasicaulis Duthie ex Burk.in high altitude area is slow,which has affected resources cultivation,clinical safety and effective usage of Gentiana crasicaulis Duthie ex Burk..Therefore,this study was aimed to study the effects of seed dormancy,optimum growth conditions,seed germination RNA-seq,dynamic temperature,dynamic content and high-resolution mass spectrometry,in order to lay a foundation in evaluation,quality standards and further development and application for Gentiana crasicaulis Duthie ex Burk..1.The dormancy experiment was carried out by using the germination rate as the evaluation index.The results showed that the seeds were immersed in water at 30 ?for 12 h,1.5 % KMnO₄ was soaked for 10 min,washed with running water and washed with 500 mg/L GA3 for 12 h,and incubated at 25 ?.The seed germination rate was highest of the Gentiana crasicaulis Duthie ex Burk..2.The growth condition at different temperature,light intensity and light time was tested by single factor test method,through the growth rate of sprouts from germination to 1cm.The results showed that the sprouts had the fastest growth at 25 ?,3000 lux light intensity and 12 h/d light time.3.A preliminary experiment on the mechanism of hormone regulation in the process of seed germination of Gentiana crasicaulis Duthie ex Burk.by RNA-seq sequencing.The results showed that there were 386 different genes in the three stages of the germination process of Gentiana crasicaulis Duthie ex Burk.,including 71 differentially different genes in the photosynthetic pathway,71 in the phytohormone transcriptional pathway,88 in the starch and sucrose metabolic pathways,and 66 differentially different genes in the biosynthesis pathway.4.Using DS1921 G temperature automatic recorder,conducted the dynamic temperature test of root of Gentiana crasicaulis Duthie ex Burk.,which were collected in five different elevation areas in Heishui County.The results show that the annual average temperature is negatively correlated with the elevation.The daily maximum temperature,daily minimum temperature and daily average temperature increase first,and then decrease after August;the lowest daily temperature is 9:00,the highest temperature is at 18:00;in May and September,the average daily temperature and the average annual temperature ar equivalent,in May to August,the average temperature is higher than the annual average temperature,in Sepetember to November,the average temperature is higher than the daily average temperature.5.The method of rapid identification and identification of chemical constituents in root of Gentiana crasicaulis Duthie ex Burk.was established by UPLC-Q-Exactive four-pole-electrostatic field orbital well high-resolution mass spectrometry.The results showed that 34 compounds were identified,including 13 iridoid glycosides,5 flavonoids,5 ester components,3 triterpenes,2 organic acid,1 phenylpropanoid and 5 other components,particulary 15 chemical composition for the first time found from the stem root of Gentiana crasicaulis Duthie ex Burk.in the study.6.HPLC method to measure the dynamic changes of secondary metabolites in the root of Gentiana crasicaulis Duthie ex Burk..The results showed that the content of the medicine in Gentiana crasicaulis Duthie ex Burk.was 3-6 times higher than that in the Chinese Pharmacopoeia?2015 edition?,and the content of the secondary metabolites loganic acid was increased monthly,and the content of the Gentiopicroside,Swertiamarin and Swertanin were decrease at first,and then slowly to rise after September.