| Shikimic acid is a small molecular organic acid.It can be easily dissolved in water but hardly dissolved in nonpolar solvent like benzene.It exists in many plants and microorganisms due to the shikimate pathway.Shikimic acid has lots of physiological activities,such as anti-inflammatory and anti-microbial activity.Furthermore,it is an important starting material for the synthesis of a neuraminidase inhibitor oseltamivir(Tamiflu).Applications in industry,shikimic acid mainly comes from three ways including chemical synthesis,metabolism synthesis and plant extraction.In actual production,shikimic acid is mainly extracted from star anise.However,due to the limited resource of anise and it could not meet the needs of industry.Therefore,the development of new resources of shikimic acid is a potential effective method to alleviate the supply shortage of shikimic acid.According to the reported work,Liquidambar formosana Hance contained high shikimic acid content.Hence,the L.formosana Hance leaves were used as the experimental material in this study.A method for the determination of shikimic acid content by Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was established.The extraction conditions of shikimic acid were optimized by using a central composite design in response surface methodology with four factors(extraction temperature,extraction time,the ethanol concentration and solid-liquid ratio)and five levels.The genetic variation of shikimic acid content and other important traits including growth traits were analyzed in the 14 provinces and 19 provenances.The superior provenances and families with high shikimic acid content and excellent growth performance were screened by genetic parameters including breeding value and selection index.The results were as follows:1.The extraction conditions of shikimic acid were optimized by response surface methodology.A single factor experimental design was used to analyze the effects of extraction time,extraction temperature,extraction solvent and solid-liquid ratio on the yield of shikimic acid,and the optimal level of each factor was determined.Then,taking the yield of shikimic acid as the response value,a response surface design with 4 factors and 5 levels was adopted to further optimize the extraction conditions of shikimic acid in the L.formosana Hance leaves.Finally,the optimal extraction conditions were confirmed as follows: the extraction temperature was 96 ?,the extraction solvent was 50% ethanol aqueous solution,the solid-liquid ratio was0.1 g/100 m L and the extraction time was 3 h.2.The genetic variation of important characters in provenances and families of L.formosana were evaluated.The provenance GKX had the highest content of shikimic acid(4.09%),while the provenance WY had the lowest(0.35%).There were significant differences in shikimic acid content,tree height,diameter,volume,under branch height,leave color and stem form among provenances,families and provinces.The variation range of provenance heritability,family heritability and individual heritability were 0.16-0.53,0.14-0.29,0.17-0.36,respectively.The 14 provinces and 19 provenances both could be divided into 3 clusters based on the 6 traits.The geographical distribution of the shikimic acid content was the highest in the west,followed by the eastern and the lowest in the south.Shikimic acid content was significantly negatively correlated with height,diameter,under branch height and leave color.3.The superior provenances/families of L.formosana with high shikimic acid content and good growth performance were screened.5 provenances and 51 families with good growth were obtained in terms of growth traits according to the selection rate of 20% and 1%,respectively.For the combination selection of shikimic acid content and good growth performance were obtained according to the selection rate of 20% for provenances and 1% for families,respectively. |
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