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Objective: Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation. Both pathogens and antagonistic fungi have been reported to induce plant resistance responses, thereby promoting the accumulation of secondary metabolites. The purpose of this experiment is to compare the advantages of one of the two fungi, in order to screen out more effective elicitors. The mechanism of fungal elicitor-induced plant resistance response is supplemented. Methods: A gradient dilution and the dural culture were carried out to screen strains. The test strain was identified by morphology and 18 s rDNA. The effect of different concentrations (0, 50, 100, 200, 400 mg/L) of Penicillium sp. YJM-2013 and F. oxysporum on fresh weight and ginsenosides accumulation were tested. Signal molecules transduction, expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors. Results: Antagonistic fungi of F. oxysporum was identified as Penicillium sp. YJM-2013, which reduced root biomass. The total ginsenosides content of Panax ginseng adventitious roots reached the maximum (48.95 ± 0.97 mg/g) treated with Penicillium sp. YJM-2013 at 200 mg/L, higher than control by 2.59-fold, in which protopanoxadiol-type ginsenosides (PPD) were increased by 4.57 times. Moreover, Penicillium sp. YJM-2013 activated defense signaling molecules, up-regulated the expression of PgWRKY 1, 2, 3, 5, 7, 9 and functional genes in ginsenosides synthesis. Conclusion: Compared with the pathogenic fungi F. oxysporum, antagonistic fungi Penicillium sp. YJM-2013 was more conducive to the accumulation of ginsenosides in P. ginseng adventitious roots. Penicillium sp. YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules, activating the expression of transcription factors and functional genes.
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Rehmannia glutinosa is one of the most common bulk medicinal materials in China and it has a long history of medicinal use. In recent years, with the development of molecular biology, especially for the emergence of high-throughput sequencing technology, it not only provides new ways to identify R. glutinosa quickly, and reveal the genetic diversity and relationship of R. glutinosa, but also lays the vital foundation for explaining the mechanism on metabolism, root tuber growth, stress response and continuous cropping obstacles of R. glutinosa. The present paper reviews the recent study progress in molecular biology research of R. glutinosa from molecular systematics, molecular identification and functional genes, and puts forward three research prospects in order to provide a reference for further study on molecular biology of R. glutinosa.
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Transcriptome sequencing technology is a newly developed transcriptomics method. In the absence of the genome information of the species, sequencing is carried out to obtain genetic information. Transcriptome sequencing technology is an important molecular biology method with wide application in various research fields. Due to the deficience of the genetic information of medicinal plants, and the transcriptome sequencing research of medicinal plants is an active field in genomics. Researchers can investigate plant gene expression and analyze its function and regulatory mechanisms. Transcriptomics research on medicinal plants can help to solve problems such as genetic breeding, excellent resistance genes screening, and etc. This paper introduces the development of transcriptome sequencing, and reviews the application of transcriptome sequencing in medicinal plants in recent years from functional gene mining and secondary metabolite pathway exploration, which provide more basic data for medicinal plant studies.
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Iridoids glycosides are a class of compounds which have pharmacological functions of anti-inflammatory, antitumor, hepato-protection, cardio-protection, etc. This review summarizes the biosynthetic pathway, related enzymes (GPPS, GES, G10, G10H, 10-HGO, IS, 7-DLS, 7-DLGT, 7-DLH, LAMT, and SLS) and the application of functional genes in iridoids glycosides, in hopes of regulating the production of metabolites and providing the valuable reference for discovering new drugs.
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Objective: To compare the expression of genes in the leaves of Tussilago farfara that involved in biosynthesis of phenylpropanoids in different developmental stages, and infer the accumulation period of biosynthesis of phenylpropanoids and provide a scientific basis for the resource utilization of leaves of T. farfara. Methods: The Illumina HiSeq2500 highthroughput sequencing method was used to analyze the transcriptome of the leaves of T. farfara in different periods. After obtaining transcriptome data, bioinformatics analysis of gene function annotation was performed to compare the expression of genes related to phenylpropanoid biosynthesis in different periods. Results: A total of 46 793 unigenes were obtained by transcriptome sequencing and the average length was 952.144 8 bp. Among them, 4 774 unigenes were annotated in the public databases NR, Swiss-Prot, eggNOG, GO, and KEGG. According to the assignment of KEGG pathway, 144 unigenes were involved in terpenoid biosynthesis, phenylpropanoid biosynthesis and flavonoids, 65 unigenes were involved in terpenoid biosynthesis, 64 unigenes were involved in phenylpropanoid and 15 unigenes were involved in flavonoids biosynthesis. The enzyme genes involved in the phenylpropanoid biosynthesis were also compared in different development stages, and the results indicated that the expression of PAL, 4CL, HCT, and CCoAOMT, which were closely related to biosynthesis of phenylpropanoids, were highest in September, which means that the contents of these compounds might be highest in September. Conclusion: This study lays the foundation for the biosynthetic pathway and regulation analysis of phenylpropanoids, and provides a scientific basis for the development and the resource utilization of leaves of T. farfara.
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Objective Previous studies suggested that overerpression of Slit2 results in abnormal Alzheimer's disease-like behavior and cognition impairment in mice. The aim of this study is to investigate the relationship between overerpression of Slit2 and accumulation and clearance of amyloid-β in aging mice by comparing the differential expression of genes for accumulation and clearance of amyloid-β in aging Tg-Slit2 and Tg-2576 mice. Methods 14-month old male C57BL/6, Tg-Slit2 and Tg-2576 mice were used to detect the expression of Aβ1 - 40 and Aβ1 - 42 in brain by immunohistochemistry. Further, the total RNA in the brain of these mice were extracted, identified and inversely transcripted to cDNA, then the cDNA was detected by PCR array. The expression of genes in the brain of Tg-Slit2 and Tg-2576 mice were analyzed. Results Comparing with the Tg-2576 mice in the same age, accumulation of Aβ was not found in the brain of Tg-Slit2 and C57BL/6 mice. The result from PCR array analysis showed that comparing with the same aged C57BL/6 mice, there were 16 up-regulated genes and 8 down-regulated genes in the brain of Tg-Slit2 mice and 14 up-regulated genes and 17 down-regulated genes in the brain of Tg-Slit2 mice. The expression of amyloid beta precursor protein (APP) in the brain of the three group mice was not changed. The expression of presenilin 2 ( Psen2) related with Aβ production was significantly up-regulated in the Tg-2576 mice. In addition, the expression of low density lipoprotein receptor-related protein ( LRP) 6 and 9 were markedly decreased in the Tg-2576 mice. Notably, these genes were not changed in the brain of the aging Tg-Slit2 mice. Conclusions The accumulation of Aβ in the brain are not found in 14-month Tg-Slit2 mice, In addition, different from Tg-2576 mice, the significant changes of expression of Aβ-related genes is not found in the brain of Tg-Slit2 mice.
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Objective: To study the biosynthesis mechanism and transshipment law of limonin compounds in Citri Reticulatae Semen, traditional medicinal variety Citrus reticulata "Dahongpao". Methods: In the process of seed growth and development, the contents of limonin, nomilin, and obacunone in stem, vane, peel, and seeds were determined by UPLC tangerine; Using grey relational analysis method, the correlation analysis between the contents and the cloning of squalene synthase gene (ss), squalene epoxidase gene (se), and glucose transferase (lgt) expression was performed. Results: Limonin compounds in different organs had the largest accumulation in seeds; The accumulation of limonin compounds in different organs was significantly correlated with ss, se, and lgt gene expression, but in the seeds the accumulation of limonin has the most closely correlation with ss, se, and lgt gene expression; The ss gene expression had the largest contribution on the accumulation of limonin, nomilin, and obacunone. Conclusion: The study provides reliable and scientific envidendce for the synthesis mechanism and transshipment law of limonin compounds.
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Objective To understand the transcriptome data of flowers and leaves of Ocimum basilicum, and analyze the transcriptome sequencing and bioinforamtics of O. basilicum. Methods Selecting fresh flowers and leaves of O. basilicum as samples, the transcriptome libraries of O. basilicum were constructed using Illumina HiSeqTM 2500 sequencing technique and analyzed using the bioinformatics methods subsequently, such as sequencing assess, transcriptome data assembly, and gene function annotation. Results After transcriptome sequencing and removing insignificant reads, 86 331 137 reads of O. basilicum were obtained. All of the reads contained 6 455 365 309 nucleotides. After de novo splicing, 90 341 Unigenes were obtained. The Unigenes were aligned in COG database, and searching result demonstrated that UniGenes were devided into 25 classes according to function. The Unigene GO functions could be broadly divided into biological processes, cellular components and molecular function categories of 43 branches. In KEGG database, the data in transcriptome could be divided into 111 classes according to the metabolic pathway which included the biochemical pathway in plants-Pathogens interaction, terpenoid and steroid compounds synthesis, lipid metabolism, RNA degradation and so on. Totally 15 617 pairs of SSR primers were successful designed by MISA software, and 10 254 SNP loci were detected. Conclusion The results of this study can provide the further development of functional gene excavation, mentabolic pathways and their regulatory mechanism for O. basilicum with theatrical basis.
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ABSTRACT The microbial composition of different types,in ecosystems (including agro-ecosystems), has been investigated in a rapidly growing number of studies in the past few years. The importance of microorganisms, regarding the maintenance and stability of nutrients in agroecosystems, is a key to maintain the sustainability of a crop. Molecular tools to study microbial communities are possible through many methods such as RISA, DGGE, TGGE, clone libraries, T-RFLP, RAPD, SSCP and more recently NGS (Next-Generation Sequencing). DGGE is widely employed to characterize the diversity and the community dynamics of microorganisms in the environment, making possible to find out specific groups through functional genes, allowing access to data that cannot be obtained by cultural methods. The aim of this paper is to review the functional groups related to agroecosystems and to indicate the critical choice of DNA primers pairs and targeted DNA regions that may be used in PCR-based methods such as the DGGE technique in order to evaluate the microbial communities in a variety of environments.
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The average yield of safflower blooming from 1 to 7 day was recorded and calculated, HPLC was used to detect the percentage composition of HYSA,quercetin,naringenin and kaempferol, and the real-time PCR was used to analyze the expression of chs and chi. The average yield,percentage composition of HYSA and naringenin as well as functional genes' expression presented similar trends. The average yield reached the highest peak at the third day, showing highpositive correlation with the contents of HYSA (r=0.756,P<0.05), and significant correlation with the expression of chi (r=0.892,P<0.01). The contents of naringenin showed a high positive correlation with the expression of chs(r=0.766,P<0.05). The study provides a theory basis for the composition and regulation mechanism of the flavonoid constituents and lays foundation for molecular mechanisms which lead to the difference of quality in C. tinctorius.
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Phylogeography is a hot point in the field of Chinese materia medica (CMM) in the study on the genetic diversity of medicinal plants, geo-herbs formation mechanisms, and the construction of core collection. In recent years, the scholars in China studied the functional gene screening and the genomics associated with DNA differences in the field of CMM resources. And they have in-depth studied the functional genes of Glycyrrhiza uralensis, Artemisia annua, Salvia miltiorrhiza, and other medicinal plants. It will be the mainstream for functional gene screening of medicinal plants based on molecular phylogeography and geo-herbs genomics in the future researches and applications. In the paper, the authors review the recent progress in molecular phylogeography in the molecular identification of CMM, the formation mechanism of CMM genuineness, particularly the medicinal plant functional genes screening on which the genomic study is carried, and so on.
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Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to be the marker compound to evaluate the quality of G. uralensis based on Chinese Pharmacopoeia. Many studies showed that glycyrrhizin possesses various pharmacological activities, such as antibacterial, antiviral, antitumor, anti-inflammatory, and immune-stimulating activities. In this paper, we summarized the cloning, characterization, expression, and polymorphism analysis of several functional genes involved in glycyrrhizin biosynthesis in G. uralensis.
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Objective: To analyze the codon usage of functional genes in Eleutherococcus senticosus and their influencing factors. Methods: The multivariate statistical analysis and correspondence analysis were carried out using CodonW and SPSS software with codon of 17 genes selected from the functional genes of E. senticosus. Results: GC contents at the three positions of functional gene codons in E. senticosus was 51.03%, 41.23%, and 40.04%, all of which had a significant correlation coefficient (P < 0.05). The correlation coefficients with GC12 and GC3 were 0.262, and both were insignificant. The relative synonymous codon usage of 27 codons was greater than 1, among which 22 codons ended with A or T base. In the corresponding analysis, the first axis showed the variation of 22.78%, and there were significant correlation coefficients in codon adaptation, codon bias index, and GC3 (P < 0.01). The correlation coefficients were 0.686, 0.617, and 0.786, respectively, but they were insignificantly related with the effective number of codons (ENC). The second axis showed the variation of 19.28% and it was only significantly related with ENC (r = 0.635). Seventeen optimized codons in functional genes of E. senticosus were defined. Conclusion: All codons of functional genes in E. senticosus prefer to ending with A or T base. The codon usage bias is formed under the effects of mutation and selection.
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During the past several decades, researches on parasite genetics have progressed from biochemical and serodiagnostic studies to protein chemistry, molecular biology, and functional gene studies. Nowadays, bioinformatics, genomics, and proteomics approaches are being applied by Korean parasitology researchers. As for Clonorchis sinensis, investigations have been carried out to identify its functional genes using forward and reverse genetic approaches and to characterize the biochemical and biological properties of its gene products. The authors review the proteins of cloned genes, which include antigenic proteins, physiologic and metabolic enzymes, and the gene expression profile of Clonorchis sinensis.
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Animals , Humans , Clonorchiasis/parasitology , Clonorchis sinensis/enzymology , Gene Expression Regulation , Helminth Proteins/geneticsABSTRACT
Unicellular green alga,Dunaliella salina(D.salina),is a biflagellar alga without cell wall,which is a kind of very important eukaryotic microalga.In the previous study,the research of D.salina focus on the morphology,the mechanism of salt tolerance and ?-carotene,however,with the rapid development of microalgal biotechnology,a lot of work about D.salina was reported in recent years.In the area of molecular biology,the studies of D.salina mainly place emphasis on the cloning and analysis of important functional genes,regulatory sequences,and the expression of foreign genes using D.salina as host.The research advance in these aspects were reviewed.
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In this mini review,some research advance on Marinomonas from domestic and overseas was briefly summarized,mainly including of its classification、ecological distribution、functional genes and bioactive molecules.Furthermore,some suggestion and perspectives for further studies on Marinomonas were also proposed.