Wei Dan, Yan Yu-Hang, Liu Yu-Ping, Wu Qing-Hua, Chen Jiang, Pei Jin
Pharmacy College, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources Chengdu 611137, China.
Zhongguo Zhong Yao Za Zhi. 2020 Aug;45(16):3826-3836. doi: 10.19540/j.cnki.cjcmm.20200527.104.
Magnolia officinalis is a traditional Chinese medicine,with many years of cultivating process, M. officinalis leaves show more differentiation types due to the exchange of seeds from different provenances. "Da Ao"(DA), "Xiao Ao"(XA), "Chuan Hou"(CH),and "Liu Ye"(LY)are the main types of M. officinalis in Sichuan province of China,and there were obvious differences in growth rate,chemical composition,leaf shape and leaf colour. This study selected different types of M. officinalis leaves(DA,XA,LY and CH)from Sichuan to determine their chlorophyll content. Transcriptomic level sequencing of different types of M. officinalis leaf tissues was by high-throughput sequencing analysis and proteomics used an integrated approach involving TMT labelling and LC-MS/MS to quantify the dynamic changes of the whole proteome of M. officinalis. The results showed that CH had the lowest chlorophyll content while DA had the highest chlorophyll content. Furthermore,transcriptome and proteomics results showed that chlorophyll synthesis pathway in DA glutamine-tRNA reductase,urinary porphyrins decarboxylase(UROD),oxygen-dependent protoporphyrin(ODCO),the original-Ⅲ oxidase protoporphyrin oxidase(PPO),magnesium chelating enzyme subunit ChlD,protoporphyrin magnesium Ⅸ monomethyl ester [oxidative] cyclase(MPPMC)were significantly higher than CH,XA and LY,consistent in the results of determination of chlorophyll content(chlorophyll content was highest of 37.56 mg·g~(-1) FW). Some rate-limiting enzymes related to the chlorophyll synthesis,such as ODCO,PPO and MPPMC were tested by Parallel Reaction Monitoring(PRM),and the results showed that the rate-limiting enzyme content in DA was higher than that in other three types. Therefore,based on the differences in leaf color of four types of M. officinalis,the research conducted a preliminary study on the chlorophyll metabolism pathway in leaves of different types of M. officinalis,and explored relevant genes and proteins causing leaf color differences from the molecular level,so as to lay a foundation for studying the differences in growth and development of different types of M. officinalis.
厚朴是一种传统中药，经过多年的栽培过程，由于不同种源种子的交换，厚朴叶片呈现出更多的分化类型。“大凹”（DA）、“小凹”（XA）、“川厚”（CH）和“柳叶”（LY）是中国四川省厚朴的主要类型，它们在生长速率、化学成分、叶片形状和叶色方面存在明显差异。本研究选取了四川省不同类型的厚朴叶片（DA、XA、LY和CH）来测定其叶绿素含量。不同类型厚朴叶片组织的转录组水平测序采用高通量测序分析，蛋白质组学采用TMT标记和LC-MS/MS的综合方法来定量厚朴全蛋白质组的动态变化。结果表明，CH的叶绿素含量最低，而DA的叶绿素含量最高。此外，转录组和蛋白质组结果显示，DA中叶绿素合成途径中的谷氨酰胺 - tRNA还原酶、尿卟啉脱羧酶（UROD）、氧依赖原卟啉（ODCO）、原卟啉原 - Ⅲ氧化酶（PPO）、镁螯合酶亚基ChlD、原卟啉镁Ⅸ单甲酯[氧化]环化酶（MPPMC）显著高于CH、XA和LY，与叶绿素含量测定结果一致（叶绿素含量最高为37.56 mg·g⁻¹ FW）。通过平行反应监测（PRM）对一些与叶绿素合成相关的限速酶，如ODCO、PPO和MPPMC进行检测，结果表明DA中的限速酶含量高于其他三种类型。因此，基于四种厚朴叶色的差异，本研究对不同类型厚朴叶片中的叶绿素代谢途径进行了初步研究，并从分子水平探索了导致叶色差异的相关基因和蛋白质，为研究不同类型厚朴生长发育差异奠定基础。
