桑黄Sanghuangporus Sheng H. Wu, L.W. Zhou & Y.C. Dai是一类多年生大型药用真菌的统称(戴玉成2003;Dai et al. 2009;Zhou et al. 2016;吴声华和戴玉成 2020)。现代药理研究表明，桑黄子实体中富含多糖(Yang et al. 2020;Yi et al. 2020;王豪等 2021)、黄酮(Yeo et al. 2007)、三萜(Jiang et al. 2018)及多酚(吕国英等 2021)等多种活性成分，在抗肿瘤(Wang et al. 2018;刘谟浩和曾建红 2023)、抗氧化(宋吉玲等 2022, 2023)、抗炎(Lee et al. 2017)、保肝(Huang et al. 2018;Shan et al. 2019)、降血糖(Cheng et al. 2020)和调节免疫(Wen et al. 2019)等方面具有良好的功效。桑黄作为天然的免疫调节剂和生物抗癌中最有效的一种食药兼用真菌，在医药和保健领域具有重要的应用价值(Wu et al. 2019)。
桑黄作为一类药用真菌，因其在抑制肿瘤细胞生长和提高免疫方面具有显著疗效(高雯雯等 2014;史帧婷和包海鹰 2016)，桑黄及其提取物在世界范围内获得广泛认可。桑黄类真菌在世界范围内分布18个种，而在我国就有11个种(吴声华和戴玉成 2020)，其中多个种类仅限于野生状态分布，仅有瓦尼桑黄Sanghuangporus vaninii (Ljub.) L.W. Zhou & Y.C. Dai和鲍姆桑黄S. baumii (Pilát) L.W. Zhou & Y.C. Dai可以实现人工栽培(宋吉玲等 2020)。其中瓦尼桑黄因其抗性强、产量高和生长周期短等优势，在全国范围内均有栽培，已成为桑黄类真菌开发与研究的主导品种之一。而在人工栽培过程中，会因栽培品种和栽培模式的不同，使得桑黄在活性成分和生物活性上存在显著的差异(楚文琪等 2023)。侯玉浩等(2022)研究发现，人工代料栽培的瓦尼桑黄子实体的总黄酮和总酚含量较高，体外抗肿瘤活性表现最好;而李小欢等(2021)则认为人工代料栽培的子实体在活性成分和抗氧化活性方面表现相对较弱，出现这一情况可能是与人工代料栽培的培养基质有所差异有关。结合本团队的研究(宋吉玲等 2023)发现，在瓦尼桑黄栽培的培养基质中添加一定量的桑枝屑，可有效促进多糖、黄酮和多酚含量积累，并对其抗氧化活性具有显著的促进作用，且随着桑枝屑添加量的增多而逐渐增强。
1 材料与方法
1.1 供试材料
1.1.1 供试菌株
瓦尼桑黄Sanghuangporus vaninii菌株S1，GenBank中序列登录号为MN153566，保存于杭州市农业科学研究院。
1.1.2 栽培配方
配方①：杂木屑79%，麸皮20%，石灰1%;配方②：桑枝屑79%，麸皮20%，石灰1%。按照常规方法进行管理，于原基形成后培养75 d采收子实体。其中以配方①栽培的瓦尼桑黄子实体编号为P10，配方②栽培的瓦尼桑黄子实体编号为P14。
1.1.3 主要试剂和仪器
试剂：甲醇(LC-MS级，Merck)、乙腈(LC-MS级，Merck)、l-2-氯苯丙氨酸(2-chloro-l- phenylalanine，纯度≥98%，上海阿拉丁)、甲酸(formic acid，LC-MS级，TCI)。仪器：超高效液相(Waters UPLC Acquity I-Class PLUS，Waters);高分辨质谱(Waters UPLC Xevo G2-XS QTOF，Waters);色谱柱(Acquity UPLC HSS T3 1.8 μm，2.1 mm×100 mm，Waters)。
1.2 样品制备
选取样品各3份于真空冷冻干燥，研磨(30 Hz，1.5 min)成粉末。100 mg样品粉末溶解于甲醇:乙腈[内标浓度20 mg/L]提取液中;溶解后于冰水浴中超声10 min，-20 ℃静置1 h，离心15 min (4 ℃、12 000 r/min)后，吸取上清液，在真空浓缩器中进行干燥。再加入160 μL提取液[乙腈:水=1:1 (体积比)]，涡旋30 s进行复溶，将混合样品于冰水浴中超声10 min，在4 ℃、12 000 r/min离心15 min，取10 μL样品进行LC-MS分析(Want et al. 2010)。
1.3 液相条件
色谱柱：Acquity UPLC HSS T3 (1.8 µm，2.1 mm×100 mm);流动相A：0.1%甲酸水溶液;流动相B：0.1%甲酸乙腈;洗脱梯度：B相比例为2% (0 min)，增加到98% (10 min)，并在98%比例维持3 min，13.00-13.10 min比例降至2%，并以2%维持至15 min;流速400 µL/min，进样量4 μL。
1.4 质谱条件
采用Xevo G2-XS Qtof高分辨质谱仪(Waters)对UHPLC分离后的样品进行分析。电喷雾离子源(electrospray ionization, ESI)技术参数参照陈灿等(2021)方法，并做相应的修改。具体参数：毛细管电压2 500 V (正离子模式)或-2 000 V (负离子模式);锥孔电压30 V;离子源温度100 ℃;脱溶剂气流速800 L/h;脱溶剂气温度500 ℃;反吹气流速50 L/h;质核比50-1 200 m/z。低碰撞能量2 V，高碰撞能量区间为10-40 V，扫描频率为0.2 s/谱。
1.5 数据处理和分析
依据检测数据，计算差异代谢产物的差异倍数，并使用T检验计算每个代谢产物的差异显著性值。使用R语言包Ropls进行OPLA-DA建模，并进行200次置换测试验证模型的可靠性。依据OPLS-DA模型的差异倍数和P值相结合的方法筛选显著差异代谢产物。筛选标准为上调代谢物fold change≥1、下调代谢物fold change<0.05，P value≤0.01，且VIP>1。运用clusterProfiler超几何检验的方法对差异代谢物KEGG的注释结果进行富集分析(Yu et al. 2012)，并进行绘图。
2 结果与分析
2.1 主成分分析
图1
图1
主成分分析图
P10：杂木屑栽培的瓦尼桑黄子实体;P14：桑枝屑栽培的瓦尼桑黄子实体. 下同
Fig. 1
Principal component analysis diagram.
P10: Sanghuangporus vaninii basidiomata cultivated with mixed sawdust; P14: S. vaninii basidiomata cultivated with mulberry sawdust. The same below.
图2
2.2 正交偏最小二乘法判别分析
为了能够更为准确地鉴别出具有差异的代谢产物，依据偏最小二乘判别分析(OPLS-DA)对代谢组数据进一步开展相关分析，从模型评价指标R2X、R2Y和Q2Y来看，R2X=0.669，R2Y=0.992，Q2Y=0.889 (图3)，R2Y和Q2Y均接近于1，说明模型构建良好。
图3
图3
代谢产物的正交偏最小二乘法判别分析
Fig. 3
Metabolite orthogonal partial least squares discriminant analysis.
图4
2.3 差异代谢物筛选
基于最小二乘判别分析结果，依据上调代谢物fold change≥1、下调代谢物fold change<0.5和VIP≥1、P value≤0.05标准，筛选出差异显著的代谢物。在瓦尼桑黄子实体中共检测出2 996种代谢物，其中差异显著代谢物有628种，P14样品中上调435种，下调193种，上调表达远大于下调表达(图5)，进一步表明桑枝屑的添加对瓦尼桑黄子实体中次级代谢产物的合成和积累具有显著的促进作用，这与本团队前期的研究结果相符(宋吉玲等 2023)。差异代谢物共可分为22种类型，主要以脂质和类脂分子，氨基酸、多肽和蛋白质，有机杂环化合物，有机含氧化合物，苯丙素类和聚酮类，有机酸及其衍生物和核苷、核苷酸及其类似物等为主(表1)。对所检测到的代谢物的差异倍数(fold change)进行log转换处理后，变化最为显著的有20个代谢物(图6)，P14样品中1种核苷酸及其衍生物，1种含氮有机化合物、萜类、糖苷类、苯环型化合物、杂环化合物和其他，2种有机羟基化合物和1种有机酸及其衍生物的含量显著增加;P10样品中的2种其他，3种氨基酸类，3种脂质，1种有机酸及其衍生物和1种苯丙素类和聚酮类含量显著增加。
表1 差异代谢物定性结果
Table 1
| 序号 No. | 代谢物类别 Metabolite category | 代谢物数量 Number of metabolites | 上调数量 Increased number of metabolites | 下调数量 Reduced number of metabolites |
|---|---|---|---|---|
| 1 | 脂质和类脂分子 Lipids and lipid-like molecules | 93 | 53 | 40 |
| 2 | 氨基酸、多肽和蛋白质 Amino acids, peptides, and proteins | 87 | 58 | 29 |
| 3 | 有机杂环化合物 Organoheterocyclic compounds | 81 | 64 | 17 |
| 4 | 有机含氧化合物 Organic oxygen compounds | 68 | 51 | 17 |
| 5 | 苯环型化合物 Benzenoids | 51 | 39 | 12 |
| 6 | 苯丙素类和聚酮类 Phenylpropanoids and polyketides | 50 | 37 | 13 |
| 7 | 有机酸及其衍生物 Organic acids and derivatives | 49 | 38 | 11 |
| 8 | 核苷、核苷酸和类似物 Nucleosides, nucleotides, and analogues | 36 | 24 | 12 |
| 9 | 糖苷类 Glycosides | 18 | 17 | 1 |
| 10 | 有机羟基化合物 Organic hydroxy compounds | 10 | 7 | 3 |
| 11 | 萜类 Terpenoid | 8 | 3 | 5 |
| 12 | 含氮有机化合物 Organic nitrogen compounds | 7 | 2 | 5 |
| 13 | 有机磷化合物 Organophosphorus compounds | 4 | 3 | 1 |
| 14 | 烃类Hydrocarbons | 3 | 0 | 3 |
| 15 | 生物碱和衍生物 Alkaloids and derivatives | 2 | 0 | 2 |
| 16 | 维生素 Vitamins | 2 | 1 | 1 |
| 17 | 胺类 Amines | 2 | 2 | 0 |
| 18 | 木脂素、新木脂素及相关化合物 Lignans, neolignans and related compounds | 1 | 0 | 1 |
| 19 | 混合金属/非金属化合物 Mixed metal/Non-metal compounds | 1 | 1 | 0 |
| 20 | 色素Pigments | 1 | 1 | 0 |
| 21 | 醌类Quinones | 1 | 0 | 1 |
| 22 | 其他Others | 53 | 34 | 19 |
图5
图6
2.4 代谢通路分析
为进一步了解差异代谢产物所涉及的代谢通路，对筛选出的差异代谢物进行了KEGG显著性富集分析(宋吉玲等 2021)。本研究中，鉴定的差异代谢物共628个，在KEGG代谢通路中注释的有151个，涉及79条代谢通路。依据P value值大小(图7)，筛选出显著性富集的20条代谢通路，其中最为显著的5条代谢通路，分别为硫胺素代谢(ko00730，thiamine metabolism)、果糖和甘露糖代谢(ko00051，fructose and mannose metabolism)、糖基磷脂酰肌醇(GPI)锚生物合成[ko00563，glycosylphosphatidylinositol (GPI)-anchor biosynthesis]、磷酸戊糖途径(ko00030, pentose phosphate pathway)和嘧啶代谢(ko00240, pyrimidine metabolism)。
图7
结合DA Score来看(图8)，对每条通路途径中富集到的差异代谢物表达进一步分析，在木屑和桑枝屑栽培的瓦尼桑黄子实体中硫胺素代谢、果糖和甘露糖代谢2个代谢通路富集到的差异代谢物的含量上调和下调的数目一致，参与的差异代谢物多与有机含氧化合物、有机杂环化合物和氨基酸及其衍生物有关(图9);在桑枝屑栽培的瓦尼桑黄子实体中糖基磷脂酰肌醇(GPI)锚生物合成、磷酸戊糖途径和嘧啶代谢3个代谢通路富集到的差异代谢物上调数目均高于下调，参与的代谢物多与有机酸及其衍生物、核苷酸及其类似物和有机含氧化物有关(图9)。磷酸戊糖途径作为葡萄糖氧化除三羧酸循环、糖酵解和柠檬酸循环外的另一主要途径，在糖代谢中占据重要的作用，为生物体合成提供还原力NADPH、ATP和核酸合成所需的戊糖的同时，其代谢途径中的中间产物d-核糖-5磷酸和1,5-磷酸核糖，为合成更多的嘌呤核苷酸、嘧啶核苷酸和生糖或生酮氨基酸及其衍生物提供了重要的前体物质。而嘧啶类化合物作为核酸的重要组成部分，也是以糖代谢途径中的中间产物为合成前体，这类化合物在抗菌和抗病毒等方面具有很好的生物活性，临床上常常被用于治疗肿瘤和艾滋病等疾病(张海彬 2021)。硫胺素的主要生物活性形式为硫胺素焦磷脂(thiamine pyrophosphate, TPP)，其在添加桑枝屑处理中的含量呈极显著上升，其作为α-酮戊二酸脱氢酶复合体(α-ketoglutarate dehydrogenase complex，KGDHC)、磷酸戊糖途径的转酮醇酶(tran-sketolase, TK)和丙酮酸脱氢酶复合体(pyruvate dehydrogenase complex, PDHC)反应中的重要辅助因子(李文霞和柯尊记 2013)，在利用葡萄糖产生ATP和参与糖异生代谢方面发挥着重要的作用。因此，通过本研究可以清楚地揭示，桑枝屑的添加对促进桑黄子实体中次级代谢产物合成和积累方面具有较好的促进作用。
图8
图8
差异代谢物差异丰度得分图
Fig. 8
Differential metabolite differential abundance score plot.
图9
图9
差异代谢物KEGG富集网络图
注释到代谢通路的具体代谢物，硫胺素代谢：pos_1247：一磷酸硫胺素;neg_385：l-酪氨酸;pos_1414：磷酸吡哆醛;neg_995：甘氨酸;neg_986：5-(2-羟乙基)-4-甲基噻唑;pos_1143：NAD. 果糖和甘露糖代谢：neg_281：山梨糖;neg_993：2-(α-d-甘露糖)-3-磷酸甘油酸酯;neg_1366：d-甘露醇-1-磷酸;neg_311：l-岩藻糖. 糖基磷脂酰肌醇(GPI)锚生物合成：neg_870：1-磷脂酰-d-肌醇;neg_436：O-磷酸乙醇胺. 磷酸戊糖途径：neg_878：1,5-二磷酸核糖;neg_107：糠醛;pos_944：葡糖酸内酯;pos_1602：2-脱氢-d-葡萄糖酸;neg_1327：d-葡萄糖酸-1,5-内酯;neg_825：β-d-葡萄糖-6-磷酸;pos_232：2-脱氧核糖. 嘧啶代谢：pos_2042：l-氢化乳清酸;pos_1443：尿嘧啶;pos_1084：尿苷;neg_994：dUDP;neg_977：l-谷氨酰胺;neg_280：甲基丙二酸;neg_1013：尿嘧啶核苷酸;neg_222：dUMP;neg_534：假尿苷三磷酸;neg_331：脱氧尿苷;pos_945：乳清酸核苷酸
Fig. 9
KEGG enrichment network diagram of differential metabolites.
Specific metabolites annotated to metabolic pathways. Thiamine metabolism: pos_1247: Thiamine monophosphate; neg_385: l-tyrosine; pos_1414: Pyridoxal 5′-phosphate; neg_995: Glycine; neg_986: 5-(2-hydroxyethyl)-4-methylthiazole; pos_1143: NAD. Fructose and mannose metabolism: neg_281: l-sorbose; neg_993: 2-(alpha-d-mannosyl)-3-phosphoglycerate; neg_1366: d-mannitol 1-phosphate; neg_311: l-fucose. Glycosylphosphatidylinositol (GPI)-anchor biosynthesis: neg_870: Phosphatidyl-d-inositol; neg_436: O-phosphoethanolamine. Pentose phosphate pathway: neg_878: Ribose 1,5-bisphosphate; neg_107: Furfural; pos_944: Gluconolactone; pos_1602: 2-dehydro-d-gluconate; neg_1327: d-glucono-1,5-lactone; neg_825: Beta-d-glucose 6-phosphate; pos_232: 2-deoxy-d-Ribose. Pyrimidine metabolism: pos_2042: l-dihydroorotate; pos_1443: Uracil; pos_1084: Uridine; neg_994: dUDP; neg_977: l-glutamine; neg_280: Methylmalonic acid; neg_1013: Uridine 5′-monophosphate; neg_222: dUMP; neg_534: Pseudouridine 5′-phosphate; neg_331: Deoxyuridine; pos_945: Orotidylic acid.
3 讨论
本研究采用广泛非靶向代谢组学技术，在以桑枝为主要基质栽培的瓦尼桑黄子实体中共检测出2 996种代谢产物，其中差异代谢物有628种，主要包含苯环型化合物、有机杂环化合物、有机酸及其衍生物、脂质和类脂分子、核苷酸及其衍生物、苯丙素类和聚酮类等次级代谢产物。与传统木屑配方相比，桑枝屑的添加在代谢途径调节方面起着至关重要的作用，主要涉及硫胺素(维生素B1)代谢、果糖和甘露糖代谢、糖基磷脂酰肌醇(GPI)锚生物合成、磷酸戊糖途径、嘧啶代谢和嘌呤代谢等，其中以糖代谢较为集中，这可能是与桑枝屑的添加可有效促进子实体中多糖含量的累积有很大关联(徐建俊等 2016;胡桂萍等 2022;宋吉玲等 2023)。其中有7个代谢产物与磷酸戊糖途径相关，为生物体内多种代谢提供能量来源的同时，也是其他代谢产物的前体，具有多种多样的生理生化和药理功能。嘧啶类化合物作为核酸的重要组成部分，也是以糖代谢途径中的中间产物为合成前体，在抗菌和抗病毒方面具有很好的生物活性，在临床上常常被用于治疗肿瘤和艾滋病等疾病(张海彬 2021)。而硫胺素作为参与生物体内糖及能量代谢的重要维生素之一，在脂质过氧化产物和谷胱甘肽还原酶活性方面也发挥重要作用，也是维持神经、心脏及消化系统正常机能的一类重要生物活性物质(向仁伸等 2020)。本研究清楚地揭示了桑枝屑的添加能够显著促进次级代谢产物的合成，这为瓦尼桑黄质量评价、药用价值开发提供了重要信息，同时为瓦尼桑黄的高效栽培技术奠定了良好的基础。
参考文献
UPLC/Q-TOF MS-based metabolomics analysis of chemical differences in different Polygala tenuifolia varieties
PMID:26930975
The chemical differences of Polygala tenuifolia varieties-JinYuan 1 (JY1), FenYuan 2 (FY2) and traditional FenYang (FY) were studied, in order to provide reference for the breeding of Polygala tenuifolia.The samples of JY1, FY2 and FY were subjected to ultra-high performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry (Q-TOF MS) analysis. The obtained data were analyzed using Principal Component Analysis (PCA) and other statistical analysis methods, and differential metabolites were further figured out.Compared with FY,sucrose esters (such as sibiricoses A5 and tenuifoliside B) and oligosaccharides (such as tenuifoliose K) in JY1 and FY2 contributed more to the separation of Polygala tenuifolia varieties in the PCA score plot. Compared with JYl, The sugar esters (such as tenuifoliside B and tenuifoliside A) and oligosaccharides( such as tenuifoliose A) in the FY2 also contributed more to the separation of Polygala tenuifolia varieties in the PCA score plot. In addition, the relative contents of sibiricaxanthone A,3,6'-disinapoly sucrose and senegin III showed significant differences among FY, JY1 and FY2.As new Polygala tenuifolia varieties, JY1 and FY2 had certain differences and respective advantages on the chemical composition compared with FY,which could provide data support for the directional breeding of Polygala tenuifolia based on the contents of some active compounds.
Metabolomics reveals potential biomarkers of rice gall midge (Orseolia oryzae) interactions
Differential analysis of secondary metabolites of medicinal and edible polygonati rhizoma and study on potential active components and mechanism for improving insulin resistance
Study on anthocyanin pathway of tea tree flower based on metabolome and transcriptome
Structural characterization and hypoglycemic activity of an intracellular polysaccharide from Sanghuangporus sanghuang mycelia
DOI:10.1016/j.ijbiomac.2020.08.202 URL [本文引用: 1]
Correlation between composition characteristics and activities of graded ethanol precipitation derived crude polysaccharides of Sanghuangporus vaninii basidiomata cultivated on artificial substrate prepared from mulberry sawdust
The new knowledge on the medicinal fungus-Phellinus baumii
Species diversity and utilization of medicinal mushrooms and fungi in China
DOI:10.1615/IntJMedMushr.v11.i3 URL [本文引用: 1]
Metabonomics analysis on the difference of drought resistance in seedling of winter wheat variety
Research progress on antitumor effects and mechanisms of Phellinus
Research advance in efficacy evaluation, active substances and action mechanism of traditional Chinese medicine based on metabonomics
Effect of sRNA SaaS on adhesion response and extracellular metabolites of Salmonella
Effects of different origins, cultivation pattern, substrates on content of main components and in vitro antitumor activity of Sanghuang mushroom
Effect of substitute cultivation with mulberry sawdust on the growth and nutrient composition of Stropharia rugosoannulata
Hepatoprotective principles and other chemical constituents from the mycelium of Phellinus linteus
DOI:10.3390/molecules23071705
URL
[本文引用: 1]
In the dimethylnitrosamine (DMN)-induced hepatic fibrosis Wistar rat model, the mycelium extract of Phellinus linteus (PLE) (20 mg/Kg) displayed significant protection against hepatic fibrosis. The present investigation characterized eleven new ionone derivatives, phellinulins D–N (4–14), from the P. linteus mycelium extract and the relative stereochemical structures were constructed according to the spectroscopic and spectrometric analytical results. Some purified compounds were examined for their inhibitory effects on activated rat hepatic stellate cells (HSCs) and several isolates did exhibit significant protection. The results indicated that the mycelium of P. linteus could be explored as a hepatoprotective drug or healthy food candidate in the near future.
Kinetic models for the effect of temperature on flavonoid production in liquid submerged fermentation by Phellinus baumii
DOI:10.1002/bab.2018.65.issue-5 URL [本文引用: 1]
Anti-inflammatory phenolic metabolites from the edible fungus Phellinus baumii in LPS-Stimulated RAW264.7 cells
DOI:10.3390/molecules22101583 URL [本文引用: 1]
Thiamine deficiency and alzheimer’s disease
Comparison of chemical constituents and antioxidant activities of Sanghuangporus vaninii in different substrates
Chemical components of Phellinus and antitum or mechanism: a review
Comparative study of chemical components and antioxidant activities of wild Sanghuangporus sanghuang and Sanghuangporus vaninii
Research progress on the anti-hepatic fibrosis action and mechanism of natural products
Aluminum stress on root metabolism of peanut based on metabolomics of UPLC-MS/MS
Research progress on effective components and efficacy of Phellinus igniarius
Effects of mulberry sawdust on the content of main active components and antioxidant activities in Sanghuangporus vainii
Antioxidant substances and activity of medicinal fungus Sanghuangporus
Activities of extracellular enzymes and polysaccharides in liquid culture of Sanghuangporus
Study of effects of bran on the metabolism of Sanghuangporus vaninii based on metabolomics of UPLC-MS/MS
Decolorization of disperse red 3B by consortium of the Chlorella sorokiniana XJK and the fungus Aspergillus sp
GC-MS metabolomics analysis of Acer rubrum during autumn leaf coloration
Exploration on metabolic characteritics of pengbo semi-wool sheep under the condition of grazing and barn feeding based on non-targeted metabonomics
Medicinal mushroom Phellinus igniarius induced cell apoptosis in gastric cancer SGC-7901 through a mitochondria-dependent pathway
DOI:10.1016/j.biopha.2018.03.038 URL [本文引用: 1]
Research advances on polysaccharides from sanghuang
Global metabolic profiling procedures for urine using UPLC-MS
DOI:10.1038/nprot.2010.50
PMID:20448546
[本文引用: 1]
The production of 'global' metabolite profiles involves measuring low molecular-weight metabolites (<1 kDa) in complex biofluids/tissues to study perturbations in response to physiological challenges, toxic insults or disease processes. Information-rich analytical platforms, such as mass spectrometry (MS), are needed. Here we describe the application of ultra-performance liquid chromatography-MS (UPLC-MS) to urinary metabolite profiling, including sample preparation, stability/storage and the selection of chromatographic conditions that balance metabolome coverage, chromatographic resolution and throughput. We discuss quality control and metabolite identification, as well as provide details of multivariate data analysis approaches for analyzing such MS data. Using this protocol, the analysis of a sample set in 96-well plate format, would take ca. 30 h, including 1 h for system setup, 1-2 h for sample preparation, 24 h for UPLC-MS analysis and 1-2 h for initial data processing. The use of UPLC-MS for metabolic profiling in this way is not faster than the conventional HPLC-based methods but, because of improved chromatographic performance, provides superior metabolome coverage.
Applications of metabolomics in develpoment of discovering biomarkers of breast cancer
Immunoregenerative effects of the bionically cultured sanghuang mushrooms (Inonotus sanghuang) on the immunodeficient mice
DOI:10.1016/j.jep.2019.112047 URL [本文引用: 1]
Resource diversity of Chinese macrofungi: edible, medicinal and poisonous species
DOI:10.1007/s13225-019-00432-7 [本文引用: 1]
Species clarification of the medicinal fungus sanghuang
Research progress and prospects of thiamine deficiency and various systemic diseases of human body
Comparison of mulberry sawdust and wood sawdust on quality of Lentinula edodes
Resource, chemical structure and activity of natural polysaccharides against alcoholic liver damages
DOI:10.1016/j.carbpol.2020.116355 URL [本文引用: 1]
Phellinone, a new furanone derivative from the Phellinus linteus KT&G PL-2
DOI:10.1007/BF02993957 URL [本文引用: 1]
Natural polysaccharides experience physiochemical and functional changes during preparation: a review
DOI:10.1016/j.carbpol.2020.115896 URL [本文引用: 1]
ClusterProfiler: an R package for comparing biological themes among gene clusters
DOI:10.1089/omi.2011.0118 URL [本文引用: 1]
Research progress of pyrimidine metabolism in microbes
Analysis of differential metabolites in cabernet sauvignon skins from different rootstock-scion combinations by non-targeted metabolomics
DOI:10.7506/spkx1002-6630-20190617-172
This study aimed to analyze the differences in the metabolite profiles and related metabolic pathways among different rootstock-scion combinations of Cabernet Sauvignon (CS). Liquid chromatography-mass spectrometry (LC-MS)-based non-targeted metabolomics was used to analyze the metabolites in grape skins from CS grafted to 1103P, 5C, SO4, 3309C, 140R and CS rootstocks. Multivariate statistical analyses including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal-partial least squares discriminant analysis (OPLS-DA) showed that a total of 32 metabolites significantly differed (P < 0.05, VIP ≥ 1) among the six rootstock-scion combinations, and 22 related metabolic pathways were found. The results showed that the rootstocks could affect the contents of metabolites such as organic acids and polyphenols in grape skins by controlling metabolic pathways involved in the biosynthesis of isoquinoline alkaloids, flavonoids (flavonols), pantothenic acid and coenzyme A, and the citric acid cycle, thereby altering the quality of grapes. This study can provide a theoretical basis for the selection of grape rootstock-scion combinations, and also provide important inspiration for improving the quality of grape-based products.
Global diversity and taxonomy of the Inonotus linteus complex (Hymenochaetales, Basidiomycota): Sanghuangporus gen. nov., Tropicoporus excentrodendri and T. guanacastensis gen. et spp. nov., and 17 new combinations
DOI:10.1007/s13225-015-0335-8 URL [本文引用: 1]
基于UPLC/Q-TOF MS代谢组学技术研究远志不同品种间的化学差异性
代谢组学揭示水稻——稻瘿蚊互作的潜在生化标识物
药用与食用黄精次生代谢产物差异分析及改善胰岛素抵抗的活性成分与作用机制研究
sRNA SaaS对沙门氏菌粘附响应规律和胞外代谢物的影响效应
DOI:10.11869/j.issn.100-8551.2022.01.0121
[本文引用: 1]
sRNA SaaS (Salmonella adhesion associated sRNA)是近期在肉品源肠炎沙门氏菌(S. Entertidis NCM 61)中筛选出的一种新型调控因子。本研究通过对比野生株与SaaS缺失突变株的粘附响应规律,分析二者胞外代谢物的差异以揭示sRNA SaaS的具体功能及可能的作用机制。结果显示,sRNA SaaS对肠炎沙门氏菌粘附能力的影响具有温度依赖性。在37℃下,SaaS 能够抑制该菌在不锈钢、玻璃和聚丙烯表面形成生物菌膜,且在聚丙烯表面的抑制作用尤为明显。进一步通过广泛靶向代谢组学分析发现,sRNA SaaS可能分别从菌体细胞数量和胞外多聚物产量两方面抑制肠炎沙门氏菌生物菌膜的形成。本研究结果有助于完善生物菌膜的理论体系,为研发生物菌膜新型控制技术提供新思路。
基于UPLC-MS/MS技术的代谢组学方法研究铝胁迫下花生的根系代谢
DOI:10.19802/j.issn.1007-9084.2021178
[本文引用: 1]
为有效降低铝毒害，探讨铝胁迫下花生幼苗根系代谢变化，以花育23（H，铝敏感型）和粤油7号（Y，耐铝型）两个花生品种为材料，基于超高效液相色谱-串联质谱法（UPLC-MS/MS）的广泛靶向代谢组学，研究铝胁迫对花生根系的影响。结果发现，从两个花生品种的幼苗根系中检测出416种代谢产物，与对照（HC）相比，花育23（HA）共筛选出155个差异代谢物（即HC vs HA），其中上调代谢物为27种，下调为128种；粤油7号（即YC vs YA）共筛选出109个差异代谢物，其中上调代谢物为28种，下调为81种。铝胁迫下两个花生品种幼苗根系之间（即HA与YA之间）筛选出145个差异代谢物，其中上调代谢物为76种，下调为69种。代谢物主要集中在酚酸类、黄酮、有机酸、木脂素和香豆素、氨基酸及其衍生物和核苷酸及其衍生物等。KEGG代谢通路富集分析显示，这些差异代谢物主要富集在异黄酮生物合成代谢通路上。铝胁迫下，花生幼苗根系代谢物发生的明显改变，可为花生生产有效降低铝毒害提供依据。
桑枝屑对瓦尼桑黄主要活性成分含量和体外抗氧化活性的影响
基于UPLC-MS/MS技术的代谢组学方法研究麸皮对杨树桑黄代谢的影响
小球藻Chlorella sorokiniana XJK与曲霉Aspergillus sp
