Germplasm Screen Of Gardenia Jasminoides From A Base And Study On The High-yield Cultivation Techniques

Gardenia (Gardenia jasminoides Ellis), also known as mountain gardenia, yellow gardenia and red gardenia, is belongs to the Rubiaceae. It is a perennial evergreen shrub, and distributes in southern China such as Jiangxi, Sichuan and Hunan. Gardenia is relatively drought tolerant and grows well in the climate with warm, high humidity and abundant light, but grows worse in soil■with abundant water. Gardenia is the original plant of Fructus Gardeniae which recorded in Chinese pharmacopoeia (2010version). There are many researches concerning on medicinal chemistry and pharmacology about gardenia, but few concerning on germplasm identification and screen by DNA fingerprint.We collected14samples from Kangcheng gardenia base located in Wucheng country, Zhangshu city, Jiangxi province, which looked large difference in configuration. On the basis of former researches, RAPD (random amplification of polymorphic DNA) and HPLC (high performance liquid chromatography) were employed to analyze the genetic relationships among14samples and detect the contents of active compounds in them. Combining other indexes of plant morphological characteristics, leaf and fruit characters, yield per plant,14samples are evaluated comprehensively. To improve the quality and yield of gardenia and control pests and diseases, an experiment was conducted by using gardenia plants of main cultivated variety in base. The study purposes of this paper are:(1) To identify different germplasms of gardenia plants objectively by morphological indicators.(2) To study the genetic relationships among14samples and give further evidences for identification by analyzing fingerprints using the method of RAPD.(3) To investigate the relationship between germplasm and quality by detecting the contents of geniposide, crocin-1and chlorogenic acid using the method of HPLC.(4) To investigate the relationship between germplasm and yield by calculating the yield per plant of different germplams.(5) At last, to establish a method for screening gardenia germplasm and obtain a good germplasm in the base.(6) To provide evidences for formulating rules about high-yield cultivation techniques, relationship between the contents of three active compounds detected by HPLC in gardenia fruits, which are geniposide, crocin-1and chlorogenic acid, and treatments by different fertilizers and pesticides should be investigated, the effect of pesticides for controlling pests should also be evaluated.(7) Overall, to obtain good germplasm of gardenia and high-yield cultivation techniques.The main results of this study are as below.1. The morphological identification of Gardenias in Base.Gardenia germplasm resources were investigated and14samples were collected. The14samples are yellow gardenias according to the description of morphological characters in 《Flora of China》. The morphological parameters of the14samples were analyzed by the software of SPSS19. The result showed that all the samples can be divided into four groups:Group A includes gardenia of No.14, Group B includes gardenias of No.1,4,5,7and12, Group C includes gardenias of No.2,6,8,9,10,11and13and Group D includes gardenia of No.3.The characters of Group A are leaf is the smallest among14samples, the fruit size is relative smaller, and the length of calyx is also relative shorter. The fruits in a plant are more inducing higher yield. The characters of Group B are leaf is relative larger, and the shape of fruit is wide. For the yield per plant, all in this group are low except No.1gardenia. The characters of Group C are the leaf size is between Group A and B. The yield per plant in this group is not similar; the yield of No.8is highest. The weight of100fruits of No.8is heaviest, but that of No.9is lightest. Fruits in three gardenias of No.10,11and13are more than other plants. The characters of Group D are the stipule is small. The remaining leaf indicators in this group are at the highest level, but the weight of100fruits is low.2. Molecular identification of gardenias in base.Three primers screened from12RAPD primers were employed to amplify DNA bands.146out of the total150bands amplified are polymorphic bands; the ratio is93.33%. The map of polymorphic RAPD fingerprints was obtained to distinguish the14samples effectively. The result of cluster analysis showed that14gardenias were clustered into2main clades. Gardenias of No.2and6clustered together, and other gardenias clustered into another main clade. The main clade containing12samples was divided into4branches. Gardenias of No.1and3clustered the first subclade; Gardenias of No.10,11,13and12clustered the second subclade; Gardenias of No.4,7,14and5clustered the third subclade; and gardenias of No.8and9clustered the fourth subclade. The first and second subclades clustered further, and same as happened in subclade of the third and fourth. Molecular identification confirms that there are differences among14gardenias. Comparing to the result of morphological cluster, the results of genetic identification and morphological identification are similar.3. Screening of high-quality strains.The results about contents of three active compounds and yields of seven gardenia groups showed that the active compounds are the highest in gardenias of fourth group, but the yield is lowest; the gardenias in first group are with highest yields, but lower active compounds; the gardenias in sixth group are with higher contents of three active compounds, and higher yields. Comparing the yield and contents of active compounds, we suggest that gardenias in sixth group are good strains. Gardenias in sixth group (i. e., the8th and the9th gardenias) should be recommended as seed source.4. The high-yield cultivation techniques were obtained.The yield can be increased to43.3%by treated with farmyard manure, but no effect by treated with pesticide of60%emulsion of Dual-effect of Phosphorus (DEP). That treated with low concentration of DEP is more effectively for controlling pests.