生物技术产业是各大经济体在21世纪优先发展的战略性产业,逐步成为世界经济的主导产业.英国生物技术产业规模在欧洲排名第一,创新创业文化浓厚.通过孵化器运营案例详细阐述英国生物技术产业发展的生态系统构建.针对我国生物技术产业创新环境提出相应建议,以期对我国生物医药产业的创新创业发展提供借鉴.

目的 为了解我国现有葛根产品的现状,利用数据挖掘的方法对其进行网络分析.方法 通过检索原国家食品药品监督管理总局数据库已经公布的葛根保健食品和药品注册信息,统计1997-2018年间批准的产品数量、标志性成分、产品剂型等信息,并结合Bibexcel分析软件对葛根保健食品的主要配伍关系进行抽取和整理,采用Ucinet 6.0软件中NetDraw构建可视化网络;以“葛根”为关键词或“篇名”检索中国知网相关文献.结果 1.1997-2018年间葛根保健品共注册549个,获批葛根药品231个;葛根保健食品的保健功能主要为保护肝损伤(30.14％),降血糖(18.47％),降血脂(18.85％),增强免疫力(11.85％)等;葛根主要药理作用为糖尿病(17.43％),肝损伤(8.25％);3.葛根保健食品剂型以胶囊(63.37％)、片剂(13.97％)颗粒剂(5.51％)为主,药品以注射剂(68.45％),片剂(22.46％)为主;4.已获批含葛根产品最常见配伍为葛根配伍黄芪;5.含葛根产品的主要有效成分为黄酮类、粗多糖等.结论 目前葛根产品市场规模较大,但仍然存在较大的发展空间.葛根保健食品需要着重开发新的保健功能,剂型要向食品化发展且形式上需要创新;葛根药品仍需要注重基础药理研究,为新的药理作用奠定基础.

PYR/PYLs function as ABA receptors and are key regulators during plant drought stress response. Previously we screened drought tolerance of Arabidopsis ABA receptors PYR/PYLs under the control of five different promoters. In this study, we characterized drought stress tolerance of AtPYL5 transgene under the control of one guard cell specific promoter, pGC1. pGC1::AtPYL5 transgenic Arabidopsis exhibited reduced transpiration rate and decreased water loss after drought treatment. Transformation of pGC1::AtPYL5 in Arabidopsis also decreased oxidative stress damage and improved photosynthesis under drought stress condition. These results indicated that pGC1::AtPYL5 construct is effective and might pave new way to develop genetically engineered plants to improve drought stress tolerance.

BACKGROUND:Quantitative real-time reverse transcription PCR (qRT-PCR) requires a stable internal control to avoid misinterpretation of data or errors for gene expression normalization. However, there are still no validated reference genes for stable internal control in Poria cocos (Schw.) Wolf (Fuling). This study aims to validate the reference genes of P. cocos.METHODS:This study firstly collected the 14 candidate reference genes by BLASTP from the genome of P. cocos for qRT-PCR analysis to determine the expression levels of 14 housekeeping genes (GAPDH, MAPK, β-Act, RPB2, RPB1-1, RPB1-2, his3-1, his3-2, APT, SAMDC, RP, β-Tub, EIF, and CYP) under different temperatures and in response to different plant hormones (indole-3-acetic acid, abscisic acid, 6-benzylaminopurine, methyl jasmonate, and gibberellic acid), and the threshold cycle (Ct) values. The results were analyzed by four programs (i.e., geNorm, NormFinder, BestKeeper, and RefFinder) for evaluating the candidate reference genes.RESULTS:SAMDC, his3-2, RP, RPB2, and his3-1 were recommended as reference genes for treating P. cocos with indole-3-acetic acid, abscisic acid, 6-benzylaminopurine, methyl jasmonate, and gibberellic acid, respectively. Under different temperatures RPB2 was the most stable reference gene. CYP was the most stable gene for all 90 samples by RefFinder.CONCLUSION:SAMDC, his3-2, RP, RPB2, and his3-1 were evaluated to be suitable reference genes for P. cocos following different treatments. RPB2 was the most stable reference gene under different temperatures and CYP was the most stable gene in the mycelia under all six evaluated conditions.

Drought stress is an important environmental factor limiting plant productivity. In this study, we screened drought-resistant transgenic plants from 65 promoter-pyrabactin resistance 1-like (PYL) abscisic acid (ABA) receptor gene combinations and discovered that pRD29A::PYL9 transgenic lines showed dramatically increased drought resistance and drought-induced leaf senescence in both Arabidopsis and rice. Previous studies suggested that ABA promotes senescence by causing ethylene production. However, we found that ABA promotes leaf senescence in an ethylene-independent manner by activating sucrose nonfermenting 1-related protein kinase 2s (SnRK2s), which subsequently phosphorylate ABA-responsive element-binding factors (ABFs) and Related to ABA-Insensitive 3/VP1 (RAV1) transcription factors. The phosphorylated ABFs and RAV1 up-regulate the expression of senescence-associated genes, partly by up-regulating the expression of Oresara 1. The pyl9 and ABA-insensitive 1-1 single mutants, pyl8-1pyl9 double mutant, and snrk2.2/3/6 triple mutant showed reduced ABA-induced leaf senescence relative to the WT, whereas pRD29A::PYL9 transgenic plants showed enhanced ABA-induced leaf senescence. We found that leaf senescence may benefit drought resistance by helping to generate an osmotic potential gradient, which is increased in pRD29A::PYL9 transgenic plants and causes water to preferentially flow to developing tissues. Our results uncover the molecular mechanism of ABA-induced leaf senescence and suggest an important role of PYL9 and leaf senescence in promoting resistance to extreme drought stress.

The C-REPEAT-BINDING FACTOR (CBF) pathway has important roles in plant responses to cold stress. How the CBF genes themselves are activated after cold acclimation remains poorly understood.In this study, we characterized cold tolerance of null mutant of RNA-DIRECTED DNA METHYLATION 4 (RDM4), which encodes a protein that associates with RNA polymerases Pol V and Pol II, and is required for RNA-directed DNA methylation (RdDM) in Arabidopsis.The results showed that dysfunction of RDM4 reduced cold tolerance, as evidenced by decreased survival and increased electrolyte leakage. Mutation of RDM4 resulted in extensive transcriptomic reprogramming. CBFs and CBF regulon genes were down-regulated in rdm4 but not nrpe1 (the largest subunit of PolV) mutants, suggesting that the role of RDM4 in cold stress responses is independent of the RdDM pathway. Overexpression of RDM4 constitutively increased the expression of CBFs and regulon genes and decreased cold-induced membrane injury. A great proportion of genes affected by rdm4 overlapped with those affected by CBFs. Chromatin immunoprecipitation results suggested that RDM4 is important for Pol II occupancy at the promoters of CBF2 and CBF3.We present evidence of a considerable role for RDM4 in regulating gene expression at low temperature, including the CBF pathway in Arabidopsis.

目的:采用一年生的铁皮石斛组培苗茎段为外植体,以茎段→原球茎诱导→原球茎增殖分化→幼苗壮苗生根为途径,对铁皮石斛组织培养进行系统研究,以期建立稳定的铁皮石斛再生体系.方法:主要以植物组织培养的方法,研究培养基中不同激素配比对铁皮石斛原球茎诱导、原球茎增殖分化、幼苗壮苗生根等方面的影响.结果:铁皮石斛原球茎诱导最佳培养基为MS+活性炭1.0 g/L+NAA 1.5 mg/L+6-BA 0.5 mg/L,诱导率达31.33％;原球茎增殖最佳培养基为1/2 MS+香蕉100 g/L+NAA 1.0 mg/L+6-BA 1.5 mg/L,此条件下培养的原球茎质地紧密,颜色墨绿,不易分化;原球茎分化最佳培养基为MS+香蕉100 g/L+NAA 1.0 mg/L+6-BA 2.0 mg/L;壮苗生根最佳培养基为MS+香蕉100 g/L+NAA 1.0 mg/L,生根率达75.00％.结论:在西北地区,有效建立铁皮石斛再生体系,对保护濒危植物和扩大铁皮石斛种植区域具有重要意义,同时,为大规模生产提供相关数据参考.

Quantitative real-time polymerase chain reaction (qRT-PCR) is widely used for the accurate analysis of gene expression. However, high homology among gene families might result in unsuitability of reference genes, which leads to the inaccuracy of qRT-PCR analysis. The release of the Ganoderma lucidum genome has triggered numerous studies to be done on the homology among gene families with the purpose of selecting reliable reference genes. Based on the G. lucdum genome and transcriptome database, 38 candidate reference genes including 28 novel genes were systematically selected and evaluated for qRT-PCR normalization. The result indicated that commonly used polyubiquitin (PUB), beta-actin (BAT), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were unsuitable reference genes because of the high sequence similarity and low primer specificity. According to the evaluation of RefFinder, cyclophilin 5 (CYP5) was ranked as the most stable reference gene for 27 tested samples under all experimental conditions and eighteen mycelial samples. Based on sequence analysis and expression analysis, our study suggested that gene characteristic, primer specificity of high homologous genes, allele-specificity expression of candidate genes and under-evaluation of reference genes influenced the accuracy and sensitivity of qRT-PCR analysis. This investigation not only revealed potential factors influencing the unsuitability of reference genes but also selected the superior reference genes from more candidate genes and testing samples than those used in the previous study. Furthermore, our study established a model for reference gene analysis by using the genomic sequence.

从天山雪莲叶片低温诱导的cDNA文库中克隆了一个未知蛋白基因(SikSD-82)cDNA全长序列.生物信息学分析表明SikSD-82基因共编码262个氨基酸,具有小麦铝诱导蛋白Wali7的保守结构域,属Gn_AT_Ⅱ家族的可溶性蛋白;其氨基酸序列与蓖麻的茎部特异性表达蛋白相似性最高为60.89％;进化树分析显示,该蛋白与番茄的茎部特异性表达蛋白关系最近.亚细胞定位结果表明,SikSD-82蛋白主要定位于细胞核.在铝胁迫条件下,转SikSD-82烟草幼苗根的伸长明显优于野生型,根部铝离子和丙二醛含量均显著低于野生型烟草,转SikSD-82烟草显示出较强的耐铝性.

Use of model organisms in biology research enables understanding of mechanisms and properties of biological systems. The investigation of model organisms has also led to the development of experimental techniques for different areas of biology. Species like Ganoderma lucidum and Salvia miltiorrhiza have been nominated for candidate model medicinal organisms. In this review, we discuss the concept of model medicinal organisms and define the aims of its research. Then, we analyze the feasibility of applying model organism research strategies to medicinal species. We also summarize the selection criteria for model medicinal organisms. Further, we introduce strategies for creating model medicinal organisms from different aspects including genome sequencing, genetic transformation platforms, and mutagenesis. Finally, we discuss the application of the model medicinal species system in gene function analysis, metabolic pathways, and biosynthetic metabolite analysis. We conclude that the model medicinal organism system will enhance understanding of the genetic basis of synthetic metabolites and accelerate the improvement of medicinal species research.

Quantitative real-time reverse transcription PCR (qRT-PCR) is a rapid, sensitive, and reliable technique for gene expression studies. The accuracy and reliability of qRT-PCR results depend on the stability of the reference genes used for gene normalization. Therefore, a systematic process of reference gene evaluation is needed. Ganoderma lucidum is a famous medicinal mushroom in East Asia. In the current study, 10 potential reference genes were selected from the G. lucidum genomic data. The sequences of these genes were manually curated, and primers were designed following strict criteria. The experiment was conducted using qRT-PCR, and the stability of each candidate gene was assessed using four commonly used statistical programs—geNorm, NormFinder, BestKeeper, and RefFinder. According to our results, PP2A was expressed at the most stable levels under different fermentation conditions, and RPL4 was the most stably expressed gene in different tissues. RPL4, PP2A, and β-tubulin are the most commonly recommended reference genes for normalizing gene expression in the entire sample set. The current study provides a foundation for the further use of qRT-PCR in G. lucidum gene analysis.

以导入大肠杆菌过氧化氢酶基因KatE的T3代转基因棉花为供试材料,经卡那霉素检测和PCR鉴定,将筛选出的阳性转基因植株与对照棉花进行整个生育期的持续水分胁迫处理直至收获,比较材料间的生理生化指标的差异,鉴定转基因植株的耐旱能力.结果显示:(1)干旱胁迫持续至初蕾期时,转基因棉花与对照植株间各项抗旱生理指标差异均未达到显著水平.(2)水分胁迫持续至盛蕾和盛花期时,转基因棉花叶片相对含水量、光系统Ⅱ最大光化学效率(Fv/Fm)、CAT活性,以及叶片的净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)均显著或极显著高于对照植株,叶绿素含量也都明显高于对照植株.干旱胁迫持续至吐絮期时,转基因棉花的株高、果枝数和铃数均显著或极显著高于对照植株,且转基因棉花和对照的籽棉产量分别比正常灌溉处理降低57.5％和60.1％,全生育期的水分胁迫严重影响了棉花籽棉产量,但转基因棉花的籽棉产量仍显著高于对照.研究表明,在新疆石河子当地自然降水(干旱胁迫)条件下,转KatE基因棉花表现出了较好的生理和生长优势,KatE基因有助于提高棉花的抗旱性.

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.

Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.

A novel germin-like protein gene clone was analyzed from a cDNA library of Sasussured involucrate, named SikGLP.Bioinformatics analysis showed that the SikGLP gene contains a 633 bp open reading frame encoding 210 amino acids with a theoretical molecular mass of 21.6 kDa and an isoelectric point of 6.81. SikGLP was a hydrophobic protein and contained a signal peptide.Multiple sequence alignment of GLP proteins revealed that the GLP gene shares high identities in sequence,and possesses typical characteristics of plant germin-like proteins.Phylogenetic analysis among 17 species indicated that SikGLP was clustered with Vitis vinifera.Real time-PCR analysis showed that the expression of SikGLP gene by low temperature,MV and PEG stresses.It may plays a role in improving resistance capacity of early stress.

Salt stress is one of the major abiotic stresses in agriculture worldwide. Analysis of natural genetic variation in Arabidopsis is an effective approach to characterize candidate salt responsive genes. Differences in salt tolerance of three Arabidopsis ecotypes were compared in this study based on their responses to salt treatments at two developmental stages: seed germination and later growth. The Sha ecotype had higher germination rates, longer roots and less accumulation of superoxide radical and hydrogen peroxide than the Ler and Col ecotypes after short term salt treatment. With long term salt treatment, Sha exhibited higher survival rates and lower electrolyte leakage. Transcriptome analysis revealed that many genes involved in cell wall, photosynthesis, and redox were mainly down-regulated by salinity effects, while transposable element genes, microRNA and biotic stress related genes were significantly changed in comparisons of Sha vs. Ler and Sha vs. Col. Several pathways involved in tricarboxylic acid cycle, hormone metabolism and development, and the Gene Ontology terms involved in response to stress and defense response were enriched after salt treatment, and between Sha and other two ecotypes. Collectively, these results suggest that the Sha ecotype is preconditioned to withstand abiotic stress. Further studies about detailed gene function are needed. These comparative transcriptomic and analytical results also provide insight into the complexity of salt stress tolerance mechanisms.

通过花粉管通道技术,以该实验室自育陆地棉品系TH1和TH2为材料,将诸葛菜(Orychophragmus vidaceus)抗逆转录因子OvDREB2B基因构建到植物表达载体后,导入棉花基因组,经卡那霉素筛选和分子鉴定表明目的基因已整合到棉花基因组中并表达.将T1代转基因植株和受体对照在温室中栽培,待植株生长至四叶一心时,用不同渗透势的PEG-6000水溶液进行渗透胁迫处理,分析探讨转基因植株的抗旱效果及其抗旱机理.结果显示:当渗透势为0和0.5 MPa处理时,转基因植株和对照无明显差异;当渗透势为0.8 MPa和1.1 MPa处理时转基因植株较对照抗旱性明显提高.当渗透势为1.1 MPa处理96h时,对照植株Fv/Fm降至0.2左右,而转基因植株仍正常生长,Fv/Fm值约为0.51,而且初始荧光(F0)值、净光合速率(Pn)、胞间CO2浓度(Ci)、蒸腾速率(Tr)等一系列参数转基因植株都明显优于对照,表明DREB2B基因能够提高棉花对水分胁迫的耐受性.

Model organisms play critical roles in life science. At present, lack of mature model systems is one of the major bottlenecks that hinder the study of the biosynthesis of natural medicines. Ganoderma lucidum is one of the most widely studied medicinal organisms and has the general features of model organisms: short life cycle, ability to produce large number of progenies, small genome size, easy to culture and genetically transform, and no harm for human being and the environment. In addition, G. lucidum involves multiple biosynthetic pathways of secondary metabolites, making it an ideal model to study the biosynthesis of secondary metabolites and their regulations. Recently, the elucidation of the chromosome-level genome has laid the solid foundation for further application of G. lucidum as a medicinal model organism. G. lucidum is supposed to play important roles in the study on the diversity of secondary metabolites, development of medicinal fungi and synthetic biology of natural medicines. In addition, the proposal of G. lucidum as a medicinal model fungus will greatly contribute to introducing state-of-the-art technologies and strategies of modern life sciences into the study of medicinal organisms and uncovering the common mechanisms and principles in the biosynthesis of secondary metabolites, thus paving the way for the construction of effective and controllable biosynthetic platform of natural medicines.

SYMRK is a plant receptor-like kinase with a role in root endosymbiosis. Heterologous expression of SYMRK from non-legumes can complement the loss-of-function effects of the mutant symrk in legumes. However, it is unclear whether the development of arbuscular mycorrhiza (AM) is affected along with the enhanced expression of SYMRK. In the present study, the full-length LsSYMRK gene was cloned from Lathyrus sativus. Overexpression of LsSYMRK in tobacco roots was essential for AM development, and affected the expression of genes which are involved in the potential signaling pathway of AM. In addition, the pattern of hyphal penetration changed from radial to longitudinal when hyphae reached the root cortex. Furthermore, overexpression of LsSYMRK increased tobacco biomass in the presence of AM fungi. These results suggest that increased expression of SYMRK in roots of AM-infected tobacco can increase the colonization and biomass.

An important characteristic of arbuscular mycorrhizal (AM) symbiosis is the transfer of phosphate from AM fungi to plants. This transfer is facilitated by mycorrhiza-inducible phosphate transporter proteins. Lysophosphatidylcholine (LPC) stimulates mycorrhiza-inducible phosphate transporter gene expression. This study aims to detect the pattern of five Petunia hybrida phosphate transporter (PhPT1–5) genes associated with AM development and their response to LPC in petunia roots. Using the method of real-time reverse transcription-polymerase chain reaction (RT-PCR), we explored the transcriptional changes of PhPT1–5 genes. Our results reveal that the PhPT4 transcript level is enhanced by 8,320-fold at 40 days after inoculation in mycorrhizal roots compared with uninoculated roots. However, PhPT4 is depressed by LPC, which acts as a mycorrhiza signal in several mycorrhizal plants. Our results suggest that PhPT4 is a mycorrhiza-specific phosphate transporter and LPC is not always a universal signal in AM fungi–plant symbiosis.