Comparison of metabolomics of Dendrobium officinale in different habitats by UPLC-Q-TOF-MS

Dendrobium officinale has been considered over past centuries to be extremely valuable for use as an herbal medicine in South Asian countries. In this work, the chemical profiles of D. officinale from different habitats were systematically characterized using ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and multivariate analysis. The principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA) and orthogonal partial least squares analysis (OPLS-DA) of UPLC-Q-TOF-MS data displayed an obvious separation. Several flavonoids and terpenoids derivatives contribute to the quantitative chemotypic variation within and between the samples as observed. These findings lead to a better understanding of the phytochemical variation of D. officinale which can aid in quality control of raw material.     

Purification and Structural Characterization of Dendrobium officinale Polysaccharides and Its Activities

In this study, Dendrobium officinale polysaccharide (named DOPS-1) was isolated from the stems of Dendrobium officinale by hot-water extraction and purified by using Sephadex G-150 column chromatography. The structural characterization, antioxidant and cytotoxic activity were carried out. Based on the results of HPLC, GC, Congo red experiment, together with periodate oxidation, Smith degradation, SEM, FT-IR, and NMR spectral analysis, it expressed that DOPS-1 was largely composed of mannose, glucose and galacturonic acid in a molar ratio of 3.2 : 1.3 : 1. The molecular weight of DOPS-1 was 1530 kDa and the main chain was composed of (1→4)-β-D-Glcp, (1→4)-β-D-Manp and 2-O-acetyl-(1→4)-β-D-Manp. The measurement results of antioxidant activity showed that DOPS-1 had the strong scavenging activities on hydroxyl radicals, DPPH radicals and superoxide radicals and the high reducing ability in vitro. Moreover, DOPS-1 was cytotoxic to all three human cancer cells of MDA-MB-231, A549 and HepG2.     

微生物共同作用下的铁皮石斛组培苗生长效应研究

植物体和植物根际均是复杂的微生态系统,其内栖息着关系复杂的共生微生物,共同影响植物的生长发育.为探讨混合接种真菌与细菌对兰科植物生长的影响,筛选出真菌与细菌的优势促生组合,本研究选取经分离、筛选获得的4株促生内生真菌(铁皮石斛内生真菌C22、C35,美花石斛内生真菌L12、L28)和3株促生内生细菌(铁皮石斛菌内生细菌TX-7、TX-16、TX-19),以"真菌+细菌"的方式混合接种于铁皮石斛无菌组培苗中,共培养120 d.结果获得了3组优势组合:C22+TX-19、L28+TX-16和L28+TX-19,它们对铁皮石斛组培苗的生长均表现出正效应,其中C22+TX-19和L28+TX-19对促进组培苗生物量的增长具有协同效应,L28+TX-19对提高组培苗的根分枝率具有协同效应,3个组合对增加组培苗的分蘖数和根尖数均表现为累加效应.研究结果表明,内生真菌与内生细菌的共同作用可显著促进铁皮石斛的生长,混合接种有可能更大地发挥微生物的效能.     

美花石斛化学成分及其护肤活性研究

本文研究美花石斛(Dendrobium loddigesii)的化学成分及其护肤活性。采用色谱法从美花石斛粗提物中分离得到12个化合物,利用波谱学方法分别鉴定为拖鞋状石斛素(1)、2,4,7-三羟基-9,10-二氢菲(2)、3,6,9-三羟基-3,4-二氢蒽-1(2H)-酮(3)、3,5′-二甲氧基-3′,4,9′-三羟基-7′,9-环氧-8,8′-木酚素(4)、5-(4-羟基-3-甲氧基苯乙基)-2-(4-羟基-3-甲氧基苯基)苯并二氢呋喃-3,7-二醇(5)、5-(4′′-羟基-3′′-甲氧基苯乙基)-2-(4′-羟基-5′-甲氧基苯基)-7-甲氧基苯并二氢吡喃-3-醇(6)、丁香脂素(7)、异香草醛(8)、松柏醛(9)、2-甲氧基-5-羟甲基苯酚(10)、二氢松柏醇(11)、4-羟基-3-甲氧基苯乙醇(12)。其中化合物3～5、7～12为首次从美花石斛中分离得到;化合物1、2、6(100μg/mL)有一定的自由基清除活性(>80%);化合物2(10μg/mL)有一定促胶原蛋白分泌活性(>20%)。结果显示从美花石斛的茎中分离得到了12个化合物,3个表现出良好的护肤活性。     

磷对霍山石斛类原球茎悬浮培养细胞生长和多糖合成的影响

在确定了最适接种量和外植体细胞生理时间的基础上,研究了在不同起始磷浓度下,霍山石斛类原球茎生长、碳、氮消耗和多糖积累的动力学特性。以生长30d的类原球茎为材料,在接种量为100g/L时,类原球茎生长的最佳起始磷浓度为2.5mmol/L,培养36d时,类原球茎鲜重达496.5g/L。动力学分析表明,磷是霍山石斛类原球茎生长的限制性因素,胞内磷的积累水平与细胞生长具有相关性,2.5mmol/L的磷酸盐有利于碳、氮等营养物质的吸收;而多糖积累的最佳起始磷浓度为0.312mmol/L,培养36d时,其产量为2.22g/L。     

冰冻切片技术在铁皮石斛显微结构上的应用

为了快速高效的观察兰科植物铁皮石斛的显微结构,利用光镜和改进的蔗糖保护--液氮速冻冰冻切片法,观察铁皮石斛根、茎、叶的显微结构。该技术方法是将铁皮石斛器官经过蔗糖磷酸缓冲液保护液处理后抽真空,再经过液氮速冻、包埋、切片、展片观察、染色以及拍照等步骤,制作出铁皮石斛根、茎和叶较完整的显微结构切片。研究结果表明,适合铁皮石斛根的最适条件为:蔗糖质量体积分数为8%、冷凝温度-25℃、切片厚度20μm;茎的最适条件为:蔗糖质量体积分数为16%、冷凝温度-20℃、切片厚度15μm;叶的最适条件为:蔗糖质量体积分数为4%、冷凝温度-20℃、切片厚度10μm。该研究在兰科植物显微结构观察和组织化学研究中将具有广阔的应用前景。     

铁皮石斛GRAS转录因子家族基因SCL3克隆及表达分析

GRAS家族是一类植物特有的转录调控因子,参与调控植物生长发育及抵御逆境胁迫,并且在赤霉素(gibberellins, GAs)信号转导过程中具有重要作用。本研究采用RT-PCR结合RACE方法从铁皮石斛(Dendrobium officinale Kimura et Migo)原球茎中克隆到了一个转录因子GRAS家族基因SCL3 (scarecrow-like 3)并对其进行了表达分析。该基因cDNA序列全长2 278 bp,命名为DoSCL3 (GenBank注册号:MG252261),其编码425个氨基酸,分子量为47.88 kD,等电点(PI)为6.21。DoSCL3基因编码的蛋白不含跨膜域和信号肽,具有GRAS转录因子家族特有的保守结构域(22~422);多序列比对表明,DoSCL3与小兰屿蝴蝶兰的SCL3蛋白高度同源(相似性达到83%),同时进化树分析结果显示其与小兰屿蝴蝶兰亲缘关系非常相近。qRT-PCR实验结果显示,DoSCL3基因在铁皮石斛种子共生萌发中随着萌发进程的变化(1~3级,萌发至原球茎阶段)其表达量逐渐升高,并且在种子萌发至2级时,共生萌发组表达量显著高于无菌萌发组(为无菌组2.2倍),表明DoSCL3在兰科药用植物铁皮石斛种子共生萌发中菌根共生关系的建立过程起到重要的调控作用。     

A novel case of autogamy and cleistogamy in Dendrobium wangliangii: A rare orchid distributed in the dry‐hot valley

Dendrobium wangliangii is an epiphytic orchid distributed in the Jinshajiang dry‐hot valley in Luquan County, Yunnan Province, China. Most Dendrobium spp. typically have a low fruit set, but this orchid shows a higher fruit set under natural conditions despite the lack of effective pollinators. The pollination biology of the critically endangered D. wangliangii was investigated in this study. A fruit set rate of 33.33 ± 4.71% was observed after bagging treatment in 2017 and a high fruit set rate (65.72 ± 4.44% in 2011; 50.79 ± 5.44% in 2017) was observed under natural conditions, indicating that D. wangliangii is characterized by spontaneous self‐pollination. The anther cap blocked the growing pollinium; thus, the pollinium slid down and reached the stigmatic cavity, leading to autogamous self‐pollination. Specifically, 51.50% of 162 unopened flowers (total 257 flowers) of this Dendrobium species under extreme water‐deficit conditions developed into fruits, suggesting the presence of cleistogamy in D. wangliangii. Here, cleistogamy may represent the primary mode of pollination for this orchid. Spontaneous self‐pollination and specific cleistogamous autogamy could represent major adaptions to the drought and pollinator‐scarce habitat in the Jinshajiang dry‐hot valley.