目录
目 录
前 言
第一章 基于基因组学的珍稀植物种质资源可持续利用
Genomic characterization provides new insights into the biosynthesis of the secondary metabolite Huperzine A in the endophyte Colletotrichum gloeosporioides Cg01
1 Introduction
2 Materials and Methods
2.1 Fungal strains
2.2 Extraction of genomic DNA and total RNA for sequencing
2.3 Sequencing and assembly
2.4 Gene annotation and phylogenetic analysis
2.5 Differentially expressed gene analysis
2.6 Gene-knockout and retro-complementation
2.7 High performance liquid chromatography(HPLC)analysis of HupA
3 Results
3.1 Genome sequencing，assembly，and characterization
3.2 RNA sequencing，identification and functional analysis of DEGs
3.3 Identification and expression of the genes in the proposed Hup A biosynthesis pathway
3.4 Identification and expression analysis of epigenetics related genes
3.5 Functional analysis of candidate genes in HupA biosynthesis
4 Discussion
5 Conclusion
References
SMRT sequencing revealed mitogenome characteristics and mitogenome-wide DNA modification pattern in
Ophiocordyceps sinensis
1 Introduction
2 Materials and methods
2.1 Sample collection
2.2 DNA extraction and genome sequencing
2.3 Assembly of the mitochondrial genome
2.4 Mitochondrial genome annotation
2.5 Analysis of repetitive sequences
2.6 Methylation modification
2.7 Phylogenetic analysis
3 Results
3.1 Characteristics of the O.sinensis mt genome
3.2 Phylogenetic analysis of O.sinensis
3.3 Introns
3.4 Open reading frames(ORFs)
3.5 Molecular evolution of rps3
3.6 Repetitive sequences in the mt genome of O.sinensis
3.7 DNA modification analysis
4 Discussion
5 Conclusions
References
The genome of Ganoderma lucidum provides insights into triterpenes biosynthesis and wood degradation
1 Background
2 Results
2.1 The genome characteristics of G.lucidum
2.2 Repeat sequences in the genome
2.3 Predicted Gene models
2.4 Gene annotation
2.5 Comparative genomics analysis of KEGG annotation
2.6 The pathway of the synthesis of triterpenes
2.7 Phylogeny of G.lucidum and multigene families
2.8 Function annotation of putative CAZymes.
2.9 Function annotation of putative FOLymes.
3 Discussion
3.1 Ganoderic acids biosynthesis pathway
3.2 Genes related to biodegradation in G.lucidum
4 Conclusion
5 Materials and Methods
5.1 Strains and culture conditions
5.2 DNA isolation，genome sequencing and assembly
5.3 Annotation methods
5.4 KEGG pathway analysis
5.5 Phylogenetic tree construction and gene families
5.6 Carbohydrate-related enzymes(CAZymes)and Lignin oxidative enzymes(FOLymes)annotation
References
The mitochondrial genome of the lepidopteran host cadaver(Thitarodes sp.)of Ophiocordyceps sinensis and related phylogenetic analysis
1 Introduction
2 Materials and Methods
2.1 Sample Collection
2.2 DNA Extraction and PCR Amplification
2.3 Sequence Assembly and Annotation
2.4 Evolutionary Analysis
3 Results and Discussion
3.1 Genome Organization，Base Composition，and Gene Arrangement
3.2 Protein-Coding Genes(PCGs)
3.3 Ribosomal and Transfer RNA genes
3.4 Intergenic Spacers and Overlapping Sequences
3.5 The AT-rich Region
3.6 Evolutionary Relationships Based on Phylogenetic Analysis
3.7 Evolutionary Relationships Based on Genetic Distances
3.8 Evolutionary Relationships with Intergenic Spacers
4 Conclusions
References
第二章 基于代谢组学的物质组基础研究
GC-MS-based metabolic profiling reveals metabolic changes in anaphylaxis animal models
1 Introduction
2 Materials and methods
2.1 Chemicals
2.2 Animals and sensitization
2.3 Assessment of systemic anaphylactic symptoms
2.4 Measurement of IgE
2.5 Sample treatment for GC-MS
2.6 GC-MSmethod
2.7 Data analysis
3 Results
3.1 Anaphylactic symptoms emerging in sensitized animals challenged by ovalbumin or cattle albumin
3.2 IgE levels markedly increased in animals after challenging
3.3 Metabolomic analysis revealed that nine variables were different between control and model groups
3.4 Function analysis suggested that anaphylaxis might be linked to energy metabolism and inositol signal transduction
4 Discussion
References
组蛋白甲基化与去乙酰化对蛇足石杉内生真菌盘长孢状刺盘孢合成石杉碱甲的影响
1 材料与方法
1.1 材料、试剂与仪器
1.2 方法
2 结果与分析
2.1 CgClr4、CgClr3、CgSir2基因的敲除
2.2 CgClr4、CgClr3、CgSir2回补株的构建
2.3 RT-qPCR检测回补菌株基因表达量
2.4 基因敲除和回补对胶胞炭疽生物量和HupA产量的影响
3 讨论
References
蛹虫草液态发酵过程中有效成分的动态积累变化
1 材料和方法
1.1 菌种来源、仪器和试剂
1.2 试验方法
2 结果与分析
2.1 蛹虫草在发酵过程中生物量的变化情况
2.2 虫草多糖的动态积累变化情况
2.3 虫草酸的动态积累变化情况
2.4 虫草素的动态积累变化情况
2.5 发酵罐培养情况
3 结论
参考文献
虫草素高产菌株的筛选及不同添加物对虫草素产量的影响研究
1 材料与方法
1.1 菌种来源、仪器和试剂
1.2 方法
1.3 虫草素测定
2 结果与分析
2.1 高产菌株的筛选
2.2 蛹虫草发酵过程中生物量的变化情况
2.3 虫草素的动态积累变化规律
2.4 不同添加物对虫草素积累影响情况
3 讨论
References
裸仁南瓜籽油活性成分分析及抗氧化能力评价
1 材料与方法
1.1 材料与仪器
1.2 方法
2 结果与分析
2.1 裸仁南瓜籽油质量检测
2.2 脂肪酸组成分析
2.3 植物甾醇组成分析
2.4 角鲨烯含量测定
2.5 抗氧化活性评价
3 结论
参考文献
第三章 植物活性成分高倍富集与提制
Preparation and adsorption characteristics of cordycepin molecularly imprinted polymers
1 Introduction
2 Materials and methods
2.1 Instrumentation
2.2 Materials
2.3 Preparation of the Cordycepin Molecularly Imprinted Polymer
2.4 Static Adsorption Experiments(https://www.daowen.com)
2.5 Solid-Phase Extraction of Cordyceps militaris Extracts
3 Results and discussion
3.1 Interaction Between Cordycepin and the Functional Monomer
3.2 Polymer Characterization
3.3 Adsorption Properties and Kinetics of Molecularly Imprinted Polymers
3.4 Adsorption Isotherm and Scatchard Analysis
3.5 Adsorption Selectivity of the Molecularly Imprinted Polymer for Cordycepin
3.6 Recovery and Regeneration of the Molecularly Imprinted Polymer
3.7 Molecularly Imprinted Solid Phase Extraction of Cordyceps militaris
4 Conclusions
References
超临界及亚临界萃取澳洲薄荷挥发性成分的对比
1 材料与方法
1.1 材料与试剂
1.2 主要仪器设备
1.3 方法
2 结果与讨论
2.1 不同萃取方法萃取得率比较
2.2 澳洲薄荷挥发性成分的GC-MS分析结果
3 结论
参考文献
高速逆流色谱法分离制备蛹虫草发酵液中虫草素
1 仪器与材料
2 方法
2.1 虫草素粗提物及样品溶液的制备
2.2 虫草素的测定［12］
2.3 HSCCC溶剂体系的选择
2.4 HSCCC分离
2.5 样品结构鉴定及纯度测定
3 结果与分析
3.1 溶剂体系的筛选
3.2 HSCCC分离纯化结果
3.3 样品结构鉴定及纯度分析
4 讨论
参考文献
大孔吸附树脂分离纯化蛹虫草固体培养基中虫草素工艺
1 材料与方法
1.1 材料与试剂
1.2 仪器与设备
1.3 方法
2 结果与分析
2.1 树脂的筛选
2.2 虫草素质量浓度对树脂吸附的影响
2.3 HPD-100树脂静态吸附动力学曲线
2.4 不同体积分数乙醇溶液对解吸效果的影响
2.5 动态吸附曲线
2.6 上样流速对吸附的影响
2.7 解吸流速对解吸效果的影响
2.8 解吸溶剂体积的确定
2.9 HPD 100树脂对蛹虫草固体培养基中虫草素的纯化效果
3 结论
参考文献
酶法提取蛹虫草中虫草素的研究
1 材料与仪器
1.1 仪器
1.2 试剂
2 方法
2.1 UPLC测定虫草素含量
2.2 酶法提取虫草素
2.3 虫草素含量测定
3 结果与讨论
3.1 分析条件的选择
3.2 分析方法线性范围、检测限、重现性
3.3 样品测定及回收实验
3.4 酶种类的选择
3.5 酶用量的影响
3.6 酶解温度的影响
3.7 p H的影响
3.8 酶解时间的影响
3.9 不同提取方法效果的比较
4 结论
参考文献
第四章 植物活性成分功能评价
Xiaokeping-induced autophagy protects pancreaticβ-cells against apoptosis under high glucose stress
1 Introduction
2 Material and methods
2.1 Cell culture
2.2 Preparation of XKP
2.3 Flow cytometry
2.4 Immunofluorescence staining
2.5 Western blotting
2.6 Statistical analysis
3 Results
3.1 XKP decreasedβ-cell apoptotic rate under high glucose stress
3.2 XKP likely induced autophagy inβ-cells via mTOR pathway
3.3 Autophagy may contribute to the effect of XKP protectingβ-cells from apoptosis under high glucose stress
4 Discussion
5 Conclusion
References
Organosulfur compounds induce cytoprotective autophagy against apoptosis by inhibiting mTOR phosphorylation activity in macrophages
1 Introduction
2 Materials and Methods
2.1 Reagents and antibodies
2.2 Cell culture and treatment
2.3 Immunofluorescence staining
2.4 Western blot analysis
2.5 Cell apoptosis assay
2.6 Lentivirus transfection
2.7 Isolation of peritoneal macrophages
2.8 Statistical analysis
3 Results
3.1 Screen for garlic organosulfur compounds that induce autophagy
3.2 SAC，DADSand DTS treatment induces autophagic flux and degradation of autophagy substrate
3.3 Knockdown of ATG5 expression inhibits autophagosome formation
3.4 Autophagy protects DADS-，SACand DTS-induced apoptosis
4 Discussion
References
Gymnemic acid I triggers mechanistic target of rapamycin-mediatedβ-cells cytoprotection through the promotion of autophagy under high glucose stress
1 Introduction
2 Materials and methods
2.1 Reagents and antibodies
2.2 Cell culture
2.3 Immunofluorescence staining
2.4 Western blot analysis
2.5 Cell apoptosis assay
3 Results
3.1 GA I protects MIN-6 cell's apoptosis under high glucose stress
3.2 GA I induces autophagy via inhibition of phosphorylation activity of mTOR
3.3 GA induces autophagy under the high glucose stress
3.4 GA protects MIN-6 cells from apoptosis under high glucose stress
4 Discussion
References
The protective effects and genetic pathways of thorn grape seedsoil against high glucose-induced apoptosis in pancreaticβ-cells
1 Background
2 Methods
2.1 TGSO extraction and fatty acids analysis
2.2 Cells culture and treatments
2.3 Detection of cell apoptosis by fluorescence activated cell sorting(FACS)
2.4 Insulin secretion assay
2.5 Isolation of RNA，cDNA synthesis and qRT-PCR
2.6 Statistical analysis
3 Results
3.1 TGSO contained 87.02% UFA which was 5.7 folds higher than SFA(saturated fatty acid)
3.2 TGSO reduced the percentages of apoptotic cells from 43.80%to 15.44%
3.3 TGSO protectedβ-cells from insulin impairment induced by high glucose
3.4 TGSO down-regulated pro-apoptotic gene expression and up-regulated anti-apoptotic gene expression involved in signal transduction pathways
4 Discussion
5 Conclusions
References
Anti-apoptotic function and mechanism of ginseng saponins in rattus pancreaticβ-cells
1 Introduction
2 Material and methods
2.1 Cells Culture and Drug Treatment
2.2 Apoptosis Assay by Fluorescence Activated Cell Sorting(FACS)
2.3 Insulin Secretion Assay
2.4 Nitric Oxide(NO)Measurement
2.5 Isolation of RNA，cDNA Synthesis and Real-Time Reverse Transcription Polymerase Chain Reaction(RT-PCR)
2.6 Statistical Analysis
3 Results
3.1 Ginsenosides Reduced the Percentages of Apoptotic Cells Induced by High Glucose/Cytokines
3.2 Ginsenosides Protected High Glucose/Cytokines-Impaired Insulin Secretion
3.3 Ginsenosides Inhibited High Glucose/Cytokines-Induced iNOS Expression and Consequent NO Production
3.4 Ginsenosides Affected Bcl-2，Bax，Fas and Caspase-3 Genes Expression Levels ofβ-Cells Induced by High Glucose
3.5 Ginsenosides Affected Bcl-2，Bax，Fas and Caspase-3 Genes Expression Levels in Cytokine-Induced Cells
4 Discussion
References