Effects Of Temperature And Water On The Content Of Saponins In Hedera Helix And FPS Gene Expression Analysis

Hedera helix is the high-quality materials, which with medicinal, ornamental and other multi-functional used. As a traditional medicinal plant, Hedera helix’s main active ingredients are triterpene glycosides (hederacoside C and α-hederin). Study finds that triterpene saponins has a broad application prospects in antitumor, antibacterial, antioxidant and other areas.The experiment used HPLC technology to detect the content of hederacoside C and α-hederin from Hedera helix leaves under the cultivation in different temperature and soil moisture. It analysis the effects of temperature and soil moisture on the yield of saponin compounds in Hedera helix, and in order to know environment factors on the yield of saponin compounds in Hedera helix, which can use related technology, to improve production and guarantee the quality.The experiment used clone and RT-qPCR technology to analysis the information about FPS gene’s, and explored the relation between the key enzymes of FPS gene and the content of spaonin in the metobolic pathway of spaonin compounds.From this, we can understand the Hedera helix’s triterpene glycoside synthesis of mechanism of gene regulation, and it is not only helps to reveal the mechanism of genuine traditional Chinese drugs, but also provide a comprehensive and in-depth approach to understand the relationship between the environment and plants, so as to ultimately achieve the predicted goal that human can control the environmental factors to ensure the quality of Chinese drugs.The main results of this study were as follows:1. This study, combined with HPLC method and with Hedera helix as the materials, was designed four different soil moisture levels and four temperature levels to detect the effects of saponins (hederacoside C and α-hederin) accumulation from Hedera helix under soil moisture and temperature control. The result shows that the total content of saponins in Hedera helix’s leaves was much higher than that in stem, and in the stems the content of hederacoside C was far higher than α-hederin, what’s more, the trend of content of hederacoside C and α-hederin in different soil moisture and temperature levels were not changing much. The accumulation of hederacoside C and α-hederin from Hedera helix in different soil moisture and temperature levels have different or even opposite response trends, and moderate soil moisture and temperature could increase the content of hederacoside C, but drought and high temperature helps the accumulation of a-hederin. The conclusion is that different soil moisture and temperature levels have some effects on the accumulation of hederacoside C and a-hederin, and it’s related to Hedera helix’s intrinsic chemical composition of saponins.2. Taking up with homology RT-PCR method,we used leaves of Hedera helix as material to get FPS of Hedera helix by cloning.The analysis result revealed that the cDNA encoding FPS of Hedera helix, with open reading frame (ORF) for 1 026 bp and encoding 342 a mino acids residues, had 1050 bp base. The FPS sequence of nucleotides and a mino acids,compared with araliaceae plants, has homology; According to the online analysis:the FPS, with the relative molecular of 39 735.7,might be stored in cytoplasm and located in the outer membrane; The FPS contains two aspartic acid conserved domains, and the secondary structure mainly with alpha helix and random coil;The result of dicotyledons plants system evolution shows that the FPS was the closest related to Eleutherococcus senticosus, Panax ginseng, Panax notoginseng, Panax quinquefolius and Aralia elata, which was consistent with the traditional classification rules.3. This experiment, which has extracted Hedera helix’s RNA of different organs of leaves, petiole, stems and root, used RT-qPCR method to detect its expression volume. The result shows that in Hedera helix’s different organ,the expression volume of FPS gene from Hedera helix can be followed by shoot apex, root, petiole, leaves and stems. The results further confirmed that the expression volume of FPS gene were differences in different organ, and also laid the foundation for further research of Hedera helix’s FPS gene.