Effect Of Exogenous Factors On Chalcone Synthase Gene Expression And Polyphenol Content In Korean Pine

Korean pine distributed in Northeast and Inner Mongolia forest zone is a kindof important natural resource. Korean pine has considerable scientific research andpractical application value for its bioactive substances. In recent years, research onKorean pine biological activity has been continuously strengthened,but metabolicprocess of active substance is still in its infancy. Especially metabolic process ofpolyphenols in Korean pine has not been fully carried out.The study selected tissuecultured Korean pine adventitious bud and natural growth of Korean pine needles asmain objects. Folin-phenol method and real-time fluorescent quantitative PCRtechnique were used to determine polyphenols content in Korean pine andexpression levels of CHS gene respectively. Two-dimensional electrophoresis andmass spectrometry were used to research proteomics. The results achieved in thestudy were as follows:In this study, Korean pine polyphenol was extracted by ultrasonic and65%ethanol and measured by using folin-phenol method. Polyphenol contents of Koreanpine adventitious bud without ultraviolet radiation were35.32mg/g, however,polyphenol contents reached a maximum value of39.41mg/g when Korean pineadventitious bud were treated with ultraviolet radiation for4h. Regard totemperature, Polyphenol contents were43.25mg/g and21.37mg/g when Koreanpine adventitious bud was cultured at20℃and10℃respectively. The order ofpolyphenol contents in different seasons: autumn> summer> winter> spring, themaximum and minimum value was36.12mg/g and24.58mg/g.In this study, according to the kit instruction extracting total RNA of Koreanpine，and real-time fluorescent quantitative PCR technique were used to detectexpression of CHS gene. Compared to internal standard group without ultravioletradiation, expression levels of CHS gene were3.87times and0.16times whenultraviolet radiation time was2h and8h. Compared to group cultured at10℃,expression levels of CHS gene in Korean pine needles rose to3.10times. The orderof expression levels of CHS gene in different seasons:summer> autumn> winter>spring, the maximum times was3.03in summer. By analyzing the relationship ofpolyphenol contents and expression levels of CHS gene, the phenomenon wasilluminated that polyphenol contents increased with the increasing expression levelsof CHS gene. This result showed that expression of CHS gene could improvesynthesis and accumulation of polyphenol. In this study，2-DE was used to identify differential protein. The resultsshowed that ultraviolet radiation, temperature and season have great effect onprotein expression，Also some of the extreme lack of protein spots. The mostrepresentative differential protein in each group was selected for MALDI-TOF-MS.The result as follows: ssp5701is ruBisCO ssp6501is S-adenosylmethioninesynthase; ssp3804is Stromal70kDa heat shock-related protein，ssp7710is enolase；ssp4602is. rubisco activase，ssp3305is abscisic acid and water-stress inducedprotein.