剑叶龙血树4CL基因的克隆及其功能预测
投稿时间:2022-05-17     点此下载全文
引用本文:孙惠芳,宋美芳,张越,李海涛,王云强,张忠廉.剑叶龙血树4CL基因的克隆及其功能预测[J].中国现代中药,2023,25(3):513-522
DOI:10.13313/j.issn.1673-4890.20220517004
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作者中文名作者英文名单位中文名单位英文名E-Mail
孙惠芳 SUN Hui-fang 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100 Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China  
宋美芳 SONG Mei-fang 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100 Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China  
张越 ZHANG Yue 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100 Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China  
李海涛 LI Hai-tao 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100 Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China  
王云强 WANG Yun-qiang 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100 Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China  
张忠廉* ZHANG Zhong-lian 中国医学科学院 北京协和医学院 药用植物研究所 云南分所/云南省南药可持续利用研究重点实验室, 云南 景洪 666100
道地药材国家重点实验室,北京 100700
Yunnan Key Laboratory of Southern Medicinal Utilization, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Jinghong 666100, China
State Key Laboratory of Dao-di Herbs, Beijing, 100700, China
 
基金项目:中央本级重大增减支项目(2060302);国家重点研发计划重点专项(2019YFC1712305);云南省重大科技专项(202002AA100007);云南省科技人才和平台计划项目(202105AG070011);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-032)
中文摘要:目的 对剑叶龙血树4CL家族基因进行克隆及功能预测,筛选出可能参与黄酮类生物合成相关的4CL基因,为深入阐述龙血竭的形成机制奠定基础。方法 在前期转录组测序数据的基础上,初步筛选4CL家族基因并对其进行基因克隆及功能预测,筛选可能与黄酮类及木质素类化合物合成相关的4CL基因;以剑叶龙血树组培苗为研究对象,对其进行过氧化氢、茉莉酸、水杨酸等胁迫处理,实时荧光定量聚合酶链式反应(qRT-PCR)分析不同胁迫处理条件下4CL基因的相对表达变化规律。结果 在剑叶龙血树转录组数据库中共注释得到6个4CL基因并成功完成其基因克隆;其中3个基因(Dc4CL1Dc4CL2Dc4CL3)在龙血竭形成过程中呈现差异表达,Dc4CL1基因和Dc4CL2基因的表达模式与木质素类化合物生物合成的关键酶基因表达变化趋势一致,而Dc4CL3基因的表达模式与黄酮类化合物生物合成的关键酶基因表达变化趋势一致;系统发育分析结果显示,Dc4CL1Dc4CL2属于Ⅰ类4CL基因,而Dc4CL3基因属于Ⅱ类4CLs基因;胁迫处理结果显示,Dc4CL均有差异性表达变化,其中Dc4CL3基因的相对表达量显著高于其他Dc4CL基因。结论 对龙血竭形成过程中起关键催化作用的4CL基因进行初步的筛选,其中Dc4CL1基因和Dc4CL2基因(Ⅰ类)可能主要参与木质素类化合物的生物合成,Dc4CL3基因(Ⅱ类)可能是龙血竭形成过程中黄酮类化合物生物合成的关键催化酶基因。
中文关键词:剑叶龙血树  4CL基因  木质素  黄酮类
 
Cloning and Function Prediction of 4CL Genes in Dracaena cochinchinensis (Lour.) S. C. Chen
Abstract:Objective To perform cloning and function prediction of 4CL family genes in Dracaena cochinchinensis (Lour.) S.C.Chen and screen for 4CL genes that might be involved in flavonoid biosynthesis,aiming to lay the foundation for in-depth exploration of the mechanism of dragon's blood formation.Methods On the basis of existing transcriptome sequence data, we preliminarily screened for 4CL family genes and then performed gene cloning and function prediction to screen for 4CL genes that might be related to the biosynthesis of flavonoids and lignins. D.cochinchinensis seedlings were subjected to stress treatments of hydrogen peroxide, jasmonic acid, and salicylic acid, and the relative expression patterns of 4CL genes under various stress conditions were analyzed through quantitative real-time polymerase chain reaction (qRT-PCR).Results In this study, six 4CL genes were annotated in the transcriptome database of D. cochinchinensis and successfully cloned. Among them, three genes (Dc4CL1, Dc4CL2, and Dc4CL3) showed differential expression during the dragon's blood formation. The expression patterns of Dc4CL1 and Dc4CL2 were consistent with the expression trends of key enzyme genes related to lignin biosynthesis, while the expression pattern of Dc4CL3 conformed to the trends of key enzyme genes related to flavonoid biosynthesis. Phylogenetic analysis revealed that Dc4CL1 and Dc4CL2 were class Ⅰ 4CL genes while Dc4CL3 belonged to class Ⅱ 4CL gene. By stress treatment, it was found that all Dc4CL genes were differentially expressed under stress, and the relative expression of Dc4CL3 was significantly higher than that of the other Dc4CL genes.Conclusion In this study, we preliminarily screened for the 4CL genes that play key catalytic roles in the dragon's blood formation. Among them, Dc4CL1 and Dc4CL2 (class Ⅰ) might be mainly responsible for lignin biosynthesis, while Dc4CL3 (class Ⅱ) might be involved in flavonoid biosynthesis.
keywords:Dracaena cochinchinensis (Lour.) S.C.Chen  4CL gene  lignins  flavonoids
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