广西植物
廣西植物
엄서식물
GUIHAIA
2015年
4期
591-597
,共7页
京大戟%茉莉酸甲酯%3-羟基-3-甲基戊二酰辅酶 A 还原酶%鲨烯合酶%法尼基焦磷酸合酶%实时定量 PCR%总三萜
京大戟%茉莉痠甲酯%3-羥基-3-甲基戊二酰輔酶 A 還原酶%鯊烯閤酶%法尼基焦燐痠閤酶%實時定量 PCR%總三萜
경대극%말리산갑지%3-간기-3-갑기무이선보매 A 환원매%사희합매%법니기초린산합매%실시정량 PCR%총삼첩
Euphorbia pekinensis%methyl jasmonate%3-hydroxy-3-methylglutaryl coenzyme A reductase%squalene synthase%farnesyl pyrophosphate synthase%quantitative real-time PCR%total triterpenoids
京大戟是多年生草本药用植物,入药部分是其干燥根,但可入药的京大戟资源由于生长缓慢以及环境污染的加剧而越发匮乏,因此解决大戟资源日益紧张的问题是当今药用植物资源开发与利用方向的重要课题。京大戟含有三萜类、二萜类、黄酮类等丰富的活性成分,一些常见药用植物的有效成分是三萜类化合物,其在抗病毒、抗肿瘤、免疫调节等方面具有很好的活性。对植物萜类物质代谢起重要作用的关键酶,如3-羟基,3-甲基戊二酰辅酶 A 还原酶(hmgr)、鲨烯合酶(sqs)、法尼基焦磷酸合酶(fps)的基因克隆及活性研究取得了进展和突破,但通过调控萜类物质代谢途径中关键酶基因的表达来诱导终产物合成的研究鲜有报道。通过研究大戟萜类物质代谢途径进而利用基因工程手段提升目的物质的产量来解决京大戟药源短缺问题具有重要意义。该研究以大戟愈伤组织为材料,使用茉莉酸甲酯分别按时间梯度和浓度梯度进行诱导,将诱导后的愈伤组织分为两部分：一部分提取其总 RNA,以 actin 为内参基因进行反转录,实时定量 RT-PCR 分析大戟三萜类代谢途径中 hmgr、sqs 与 fps 基因的相对表达差异；另一部分用于提取其总三萜并使用分光光度法进行含量测定。实时定量 RT-PCR 分析结果表明,茉莉酸甲酯可诱导3个基因的表达,但其表达模式不一样。相应的京大戟愈伤组织中总三萜的含量明显提高,最高可较未处理样品增加27％。研究结果可为茉莉酸甲酯促进药用植物大戟三萜类物质积累的分子机制研究提供参考。
京大戟是多年生草本藥用植物,入藥部分是其榦燥根,但可入藥的京大戟資源由于生長緩慢以及環境汙染的加劇而越髮匱乏,因此解決大戟資源日益緊張的問題是噹今藥用植物資源開髮與利用方嚮的重要課題。京大戟含有三萜類、二萜類、黃酮類等豐富的活性成分,一些常見藥用植物的有效成分是三萜類化閤物,其在抗病毒、抗腫瘤、免疫調節等方麵具有很好的活性。對植物萜類物質代謝起重要作用的關鍵酶,如3-羥基,3-甲基戊二酰輔酶 A 還原酶(hmgr)、鯊烯閤酶(sqs)、法尼基焦燐痠閤酶(fps)的基因剋隆及活性研究取得瞭進展和突破,但通過調控萜類物質代謝途徑中關鍵酶基因的錶達來誘導終產物閤成的研究鮮有報道。通過研究大戟萜類物質代謝途徑進而利用基因工程手段提升目的物質的產量來解決京大戟藥源短缺問題具有重要意義。該研究以大戟愈傷組織為材料,使用茉莉痠甲酯分彆按時間梯度和濃度梯度進行誘導,將誘導後的愈傷組織分為兩部分：一部分提取其總 RNA,以 actin 為內參基因進行反轉錄,實時定量 RT-PCR 分析大戟三萜類代謝途徑中 hmgr、sqs 與 fps 基因的相對錶達差異；另一部分用于提取其總三萜併使用分光光度法進行含量測定。實時定量 RT-PCR 分析結果錶明,茉莉痠甲酯可誘導3箇基因的錶達,但其錶達模式不一樣。相應的京大戟愈傷組織中總三萜的含量明顯提高,最高可較未處理樣品增加27％。研究結果可為茉莉痠甲酯促進藥用植物大戟三萜類物質積纍的分子機製研究提供參攷。
경대극시다년생초본약용식물,입약부분시기간조근,단가입약적경대극자원유우생장완만이급배경오염적가극이월발궤핍,인차해결대극자원일익긴장적문제시당금약용식물자원개발여이용방향적중요과제。경대극함유삼첩류、이첩류、황동류등봉부적활성성분,일사상견약용식물적유효성분시삼첩류화합물,기재항병독、항종류、면역조절등방면구유흔호적활성。대식물첩류물질대사기중요작용적관건매,여3-간기,3-갑기무이선보매 A 환원매(hmgr)、사희합매(sqs)、법니기초린산합매(fps)적기인극륭급활성연구취득료진전화돌파,단통과조공첩류물질대사도경중관건매기인적표체래유도종산물합성적연구선유보도。통과연구대극첩류물질대사도경진이이용기인공정수단제승목적물질적산량래해결경대극약원단결문제구유중요의의。해연구이대극유상조직위재료,사용말리산갑지분별안시간제도화농도제도진행유도,장유도후적유상조직분위량부분：일부분제취기총 RNA,이 actin 위내삼기인진행반전록,실시정량 RT-PCR 분석대극삼첩류대사도경중 hmgr、sqs 여 fps 기인적상대표체차이；령일부분용우제취기총삼첩병사용분광광도법진행함량측정。실시정량 RT-PCR 분석결과표명,말리산갑지가유도3개기인적표체,단기표체모식불일양。상응적경대극유상조직중총삼첩적함량명현제고,최고가교미처리양품증가27％。연구결과가위말리산갑지촉진약용식물대극삼첩류물질적루적분자궤제연구제공삼고。
Euphorbia pekinensis is a perennial herb.The part of medicine is the dried root and is our traditional Chi-nese medicine.E .pekinensis have distributed in many provinces in China except for Tibet and Xinjiang.In recent years,medicine studies have showed that it can also be used on carbuncle swollen,sore-toxin,etc.So there are an in-creasing number of people who pay attention to it nowadays and later greatly exploit and unearth to it.But E .pekin-ensis with dried roots need many years cultivation and the deterioration of environmental pollution problem leads to the resources become more and more deficient.Therefore,many research workers faced the problem that how to re-lieve the resource tension of E .pekinensis .This is an important topic in development and utilization of the medicinal plant resources and it is also a huge challenge to science research personnel.Modern research shows that E .pekinen-sis mainly contains the active ingredient of three terpenoids,two terpenoids,flavonoids,alkaloids,organic acid,ect.The effective components of some commom medicinal plants are the three terpenoids and they have good activity in antiviral,antitumor,immune regulation and so on.So the terpenoids of Euphorbia pekinensis plays an important role in disease treatment.With the rapid development of molecular biology and the increasingly research in technolo-gy,the plant terpenoid metabolism pathway has been well studied.Now the key enzymes such as hmgr,sqs and fps play an important role in the metabolic flow.Now the researcher has made greatly progress in gene clone and activity research.But there are few reports in controlling the expression of the key enzyme of terpenoid metabolic pathway to induce the synthesis of terpenoids.Therefore,it is significantly important to study the metabolic pathway of the terpe-noid synthesis in E .pekinensis and use the means of gene engineering and technology to greatly increase the output of target product in order to solve the problem of the shortage in medicinal resource of E .pekinensis .In this study,the callus of E .pekinensis was induced by methyl jasmonate with different concentrations in different treatment times.These treated calluses were respectively divided into two parts:one was used to extract total RNA,then re-verse-transcribed to cDNA,transcript levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase (hmgr),farnesyl pyrophosphate synthase (fps)and squalene synthase (sqs)were determined by quantitative real-time PCR using actin gene as internal reference.The other part was used to extract triterpenoids whose content was detected by spectro-photometry.The results of quantitative real-time PCR showed that methyl jasmonate could induce the expression of these three genes,but the expression patterns were different.The result of total triterpenoids detection showed that methyl jasmonate induced the accumulation of total triterpenoids up to 27% compared with untreated sample.These results would afford the reference for the research on the molecular mechanism that methyl jasmonate promotes the accumulation of total triterpenoids.
