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1.
Xiuna Wang Xiaoling Zhang Ling Liu Meichun Xiang Wenzhao Wang Xiang Sun Yongsheng Che Liangdong Guo Gang Liu Liyun Guo Chengshu Wang Wen-Bing Yin Marc Stadler Xinyu Zhang Xingzhong Liu 《BMC genomics》2015,16(1)
Background
In recent years, the genus Pestalotiopsis is receiving increasing attention, not only because of its economic impact as a plant pathogen but also as a commonly isolated endophyte which is an important source of bioactive natural products. Pestalotiopsis fici Steyaert W106-1/CGMCC3.15140 as an endophyte of tea produces numerous novel secondary metabolites, including chloropupukeananin, a derivative of chlorinated pupukeanane that is first discovered in fungi. Some of them might be important as the drug leads for future pharmaceutics.Results
Here, we report the genome sequence of the endophytic fungus of tea Pestalotiopsis fici W106-1/CGMCC3.15140. The abundant carbohydrate-active enzymes especially significantly expanding pectinases allow the fungus to utilize the limited intercellular nutrients within the host plants, suggesting adaptation of the fungus to endophytic lifestyle. The P. fici genome encodes a rich set of secondary metabolite synthesis genes, including 27 polyketide synthases (PKSs), 12 non-ribosomal peptide synthases (NRPSs), five dimethylallyl tryptophan synthases, four putative PKS-like enzymes, 15 putative NRPS-like enzymes, 15 terpenoid synthases, seven terpenoid cyclases, seven fatty-acid synthases, and five hybrids of PKS-NRPS. The majority of these core enzymes distributed into 74 secondary metabolite clusters. The putative Diels-Alderase genes have undergone expansion.Conclusion
The significant expansion of pectinase encoding genes provides essential insight in the life strategy of endophytes, and richness of gene clusters for secondary metabolites reveals high potential of natural products of endophytic fungi.Electronic supplementary material
The online version of this article (doi:10.1186/s12864-014-1190-9) contains supplementary material, which is available to authorized users. 相似文献2.
Qian Zhang Hongwei Liu Guiling Sun Iain W. Wilson Jianqiang Wu Angela Hoffman Junwen Cheng Deyou Qiu 《PloS one》2015,10(3)
Taxol (paclitaxel) a diterpenoid is one of the most effective anticancer drugs identified. Biosynthesis of taxol was considered restricted to the Taxus genera until Stierle et al. discovered that an endophytic fungus isolated from Taxus brevifolia could independently synthesize taxol. Little is known about the mechanism of taxol biosynthesis in microbes, but it has been speculated that its biosynthesis may differ from plants. The microbiome from the roots of Taxus chinensis have been extensively investigated with culture-dependent methods to identify taxol synthesizing microbes, but not using culture independent methods.,Using bar-coded high-throughput sequencing in combination with a metagenomics approach, we surveyed the microbial diversity and gene composition of the root-associated microbiomefrom Taxus chinensis (Pilger) Rehd. High-throughput amplicon sequencing revealed 187 fungal OTUs which is higher than any previously reported fungal number identified with the culture-dependent method, suggesting that T. chinensis roots harbor novel and diverse fungi. Some operational taxonomic units (OTU) identified were identical to reported microbe strains possessing the ability to synthesis taxol and several genes previously associated with taxol biosynthesis were identified through metagenomics analysis. 相似文献
3.
Constantina Nasopoulou Johanna Pohjanen Janne J. Koskimäki Ioannis Zabetakis Anna Maria Pirttilä 《Journal of plant physiology》2014
Strawberry flavor is one of the most popular fruit flavors worldwide, with numerous applications in the food industry. In addition, the biosynthetic origin of the most important strawberry flavor components, such as 2,5-dimethyl-4-hydroxy-2H-furan-3-one (DMHF), is a challenging research area. DMHF's precursor, 2-hydroxy-propanal (or lactaldehyde), is biosynthesized by the endophytic bacterium Methylobacterium extorquens (M. extorquens). In particular, the alcohol dehydrogenase (ADH) enzymes of M. extorquens are involved in the biogenesis of DMHF precursors since they have the capacity to oxidize the strawberry-derived 1,2-propanediol to lactaldehyde. In this study, the expression of the endophytic ADH and the plant DMHF biosynthesis genes was examined in the tissues of raw and ripe strawberry receptacles by in situ hybridization. The presence of endophytic bacteria was studied in the same tissues by probes targeting bacterial 16S ribosomal ribonucleic acid. Hybridization signals of probes specific for endophytic ADH and plant DMHF biosynthesis genes, as well as bacteria-specific probes, were detected in the same locations. The probes were localized near the plasma membranes or intercellular spaces of cortical and vascular tissues of the receptacle, and intracellularly in the tissues of achenes. By localizing the expression of the endophytic methanol ADH and plant DMHF biosynthesis genes to the same tissues, we have reinforced our original hypothesis that an intimate symbiotic relationship between strawberry and endophytic cells exists and leads to the biosynthesis of DMHF. 相似文献
4.
Endophytic fungi represent an under explored resource of novel lead compounds and have the capacity to produce diverse classes of plant secondary metabolites. Here, we investigated the endophytic fungal diversity of taxol-producing endophytes from Taxus baccata L. ssp. wallichiana (Zucc.) Pilger and also tested the antimitogenic effect of fungal taxol using potato disc tumor assay. A total of 60 fungal endophytes were isolated from the inner bark (phloem-cambium) of T. baccata ssp. wallichiana, collected from different locations of the northern Himalayan region. Two key genes, DBAT (10-deacetylbaccatin III-10-O-acetyl transferase) and BAPT (C-13 phenylpropanoid side chain-CoA acyltransferase), involved in taxol biosynthesis were used as molecular markers for the screening of taxol-producing strains. Five representative species gave positive amplification hits by molecular marker screening with the bapt gene. These fungi were characterized and identified based on morphological and molecular identification. The taxol-producing capability of these endophytic fungi was validated by HPLC-MS. Among the five taxol-producing fungi, the highest yield of taxol was found to be 66.25 μg/l by Fusarium redolens compared with those of the other four strains. 相似文献
5.
内生真菌紫杉醇生物合成的研究现状与展望 总被引:1,自引:0,他引:1
紫杉醇是重要的抗癌药物之一,已经证明其对多种癌症具有显著疗效。目前,人们主要是从红豆杉的树皮中提取、分离和纯化紫杉醇,但由于红豆杉为生长缓慢、散生、濒危的珍稀植物,且随着紫杉醇临床用途的不断拓宽,市场需求的稳定增长,单纯依靠从红豆杉树皮中提取紫杉醇已经无法满足日益增长的市场需求。为了解决紫杉醇的药源不足,科学家已把目光从红豆杉树分离提取紫杉醇转向了其他替代方法,如化学全合成、半合成、组织培养与细胞培养、微生物发酵法生产紫杉醇等。因此,了解内生真菌紫杉醇生物合成的分子基础和遗传调控机制,对解析内生真菌紫杉醇生物合成机制、构建高产紫杉醇基因工程菌株和早日实现内生真菌紫杉醇工业化生产具有重要的科学意义和现实意义。结合本课题组多年来的科研工作,概述了红豆杉细胞紫杉醇生物合成途径、内生真菌发酵生产紫杉醇的优势、产紫杉醇内生菌的分离研究现状和生物多样性及紫杉醇生物合成相关基因的研究现状。内生真菌生物发酵合成紫杉醇是可以无限生产、大量获取紫杉醇、解决紫杉醇药源短缺问题的很有前景的方法之一。 相似文献
6.
Enhancing Taxol Biosynthesis by Overexpressing a 9-Cis-Epoxycarotenoid Dioxygenase Gene in Transgenic Cell Lines of Taxus chinensis 总被引:2,自引:0,他引:2
Shu-tao Li Chun-hua Fu Meng Zhang Yu Zhang Sha Xie Long-jiang Yu 《Plant Molecular Biology Reporter》2012,30(5):1125-1130
Plant secondary metabolites constitute are a wide range of compounds whose biosynthesis takes place in response to biotic and abiotic stresses. The phytohormone abscisic acid (ABA) acts as an important signaling molecule that regulates plant response to various stresses. Moreover, 9-cis-epoxycarotenoid dioxygenase (NCED) is one of the key enzymes in the ABA biosynthesis pathway in higher plants. In this study, a new NCED gene from Taxus chinensis, the TcNCED1, was overexpressed in transgenic T. chinensis cells, resulting in a maximum of 48?% more accumulation of ABA and a 2.7-fold increase of taxol production compared to the untransformed cells, respectively. These results indicate that overexpression of TcNCED1 can significantly increase the ABA and taxol level in T. chinensis cells, which probably provides an alternative approach in metabolic engineering to improve the yield of taxol in T. chinensis cells through genetic manipulation of the related genes in the ABA biosynthetic pathway. 相似文献
7.
Nan Zhang Zhentai Han Guiling Sun Angela Hoffman Iain W. Wilson Yanfang Yang Qian Gao Jianqiang Wu Dan Xie Jungui Dai Deyou Qiu 《Biochemical and biophysical research communications》2014
Taxol is a well-known effective anticancer compound. Due to the inability to synthesize sufficient quantities of taxol to satisfy commercial demand, a biotechnological approach for a large-scale cell or cell-free system for its production is highly desirable. Several important genes in taxol biosynthesis are currently still unknown and have been shown to be difficult to isolate directly from Taxus, including the gene encoding taxoid 9α-hydroxylase. Ginkgo biloba suspension cells exhibit taxoid hydroxylation activity and provides an alternate means of identifying genes encoding enzymes with taxoid 9α-hydroxylation activity. Through analysis of high throughput RNA sequencing data from G. biloba, we identified two candidate genes with high similarity to Taxus CYP450s. Using in vitro cell-free protein synthesis assays and LC–MS analysis, we show that one candidate that belongs to the CYP716B, a subfamily whose biochemical functions have not been previously studied, possessed 9α-hydroxylation activity. This work will aid future identification of the taxoid 9α-hydroxylase gene from Taxus sp. 相似文献
8.
Haruo Ikeda Kazuo Shin-ya Satoshi Omura 《Journal of industrial microbiology & biotechnology》2014,41(2):233-250
To date, several actinomycete genomes have been completed and annotated. Among them, Streptomyces microorganisms are of major pharmaceutical interest because they are a rich source of numerous secondary metabolites. S. avermitilis is an industrial microorganism used for the production of an anthelmintic agent, avermectin, which is a commercially important antiparasitic agent in human and veterinary medicine, and agricultural pesticides. Genome analysis of S. avermitilis provides significant information for not only industrial applications but also understanding the features of this genus. On genome mining of S. avermitilis, the microorganism has been found to harbor at least 38 secondary metabolic gene clusters and 46 insertion sequence (IS)-like sequences on the genome, which have not been searched so far. A significant use of the genome data of Streptomyces microorganisms is the construction of a versatile host for heterologous expression of exogenous biosynthetic gene clusters by genetic engineering. Since S. avermitilis is used as an industrial microorganism, the microorganism is already optimized for the efficient supply of primary metabolic precursors and biochemical energy to support multistep biosynthesis. The feasibility of large-deletion mutants of S. avermitilis has been confirmed by heterologous expression of more than 20 exogenous biosynthetic gene clusters. 相似文献
9.
红豆杉内生真菌产紫杉醇相关基因BAPT的鉴定及初步研究 总被引:2,自引:0,他引:2
从几种红豆杉中先后分离了30余种内生真菌,深入研究了三种能够产紫杉醇的内生真菌。形态学观察及18srDNA鉴定它们分属于Fusarium(属)和Pestalotiopsis(属),三个菌株均可以在离体培养的条件下产生紫杉醇,经两周培养产量可分别达到8.5,31.5,31.1μg/L。(其中Pestalotiopsis1分离于南方红豆杉,Fusarium1分离于东北红豆杉,Pestalotiopsis2分离于中国红豆杉)。对这些内生真菌产紫杉醇的初步机理作了研究。BAPT(C-13phenylpropanoidsidechain-CoAacyltransferase)是红豆杉中紫杉醇合成途径里支链合成的关键酶之一,我们根据其保守区序列设计了引物,首次在能产生紫杉醇的上述三种红豆杉内生真菌中克隆得到了BAPT基因片段,而分离的其它真菌并没有得到扩增。序列分析表明,来自内生真菌的BAPT基因片段序列与红豆杉BAPT基因片段序列具有非常高的相似性(98.9%)。推测红豆杉内生真菌之所以能够合成紫杉醇,相关基因可能直接源于其宿主植物,即其遗传学起源是基因转移而不是共进化。这同时也建立了一种快速经济的鉴定产紫杉醇真菌的辅助方法。内生真菌的遗传稳定性及改良在进一步研究中。 相似文献
10.
11.
Mingzi Wang Shaosong Liu Yaoyao Li Ren Xu Chunhua Lu Yuemao Shen 《Current microbiology》2010,61(4):254-260
Tubercularia sp. TF5 is an endophytic fungal strain isolated from the medicinal plant Taxus mairei. Previously, taxol has been detected in the fermentation products of this strain. However, it lost the capability of producing
taxol after long-term laboratory culture. Herein, we tried to reactivate the production of taxol by protoplast mutations and
genome shuffling. The protoplasts of Tub. sp. TF5 were prepared from its mycelia, and mutated by UV and NTG. The mutant strains regenerated from the mutated protoplasts
were selected and classified into four groups on the basis of their phenotypes, the profile of their metabolites analyzed
by TLC, MS, and bioassay data. Then, genome shuffling was subsequently carried out with eight mutant strains, with two representatives
from each protoplast mutant group, and genome shuffling mutant strains were obtained and screened using the same screening
procedure. Although taxol has not been detected in any mutant, two important mutants, M-741 and G-444 were selected for metabolites
isolation and determination due to their phenotypes, and differences in TLC analysis result from TF5 and other mutants. Three
new sesquiterpenoids, namely tuberculariols A–C (1–3), and a known dihydroisocoumarin (4) were obtained from M-741. Eighteen novel compounds were isolated from G-444, including five new sesquiterpenoids (5-9), two new dihydroisocoumarins (10, 11), one new tetralone (12), together with 10 known compounds (13–20, 1, and 2). The compounds isolated from the M-741 and G-444 were different in structure types and substitutions from those of TF5 (15, 21–29). The results showed, for the first time, that protoplast mutations and genome shuffling are efficient approaches to mining
natural products from endophytic fungi. Understanding the mechanisms of unlocking the biosynthesis of new metabolites will
facilitate the manipulation of the secondary metabolism in fungi. 相似文献
12.
《Journal of Molecular Catalysis .B, Enzymatic》2011,68(3-4):184-188
Penicillium griseoroseum, isolated as an endophytic microorganism from Coffeea arabica seeds, was grown in Czapeck's medium containing 5,7,3′,4′,5′-pentamethoxyflavanone. The fungus incorporated a dimethylated tetraketide, clavatol, a typical fungal secondary metabolite, into the flavanone structure at C-6, resulting in a novel benzylated flavonoid. Clavatol was also found free in the fungus extract. The compounds were isolated by chromatographic procedures and identified by extensive spectroscopic studies, including MS/MS and 1D and 2D NMR. The process probably involves enzymes catalyzing C–C bound formation, which is uncommon in fungi. The possibility of fungi participation in the biosynthesis of plant benzylated flavonoids is discussed. 相似文献
13.
Transcript profiling of jasmonate‐elicited Taxus cells reveals a β‐phenylalanine‐CoA ligase
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Karla Ramírez‐Estrada Teresa Altabella Miriam Onrubia Elisabeth Moyano Cedric Notredame Lidia Osuna Robin Vanden Bossche Alain Goossens Rosa M. Cusido Javier Palazon 《Plant biotechnology journal》2016,14(1):85-96
Plant cell cultures constitute eco‐friendly biotechnological platforms for the production of plant secondary metabolites with pharmacological activities, as well as a suitable system for extending our knowledge of secondary metabolism. Despite the high added value of taxol and the importance of taxanes as anticancer compounds, several aspects of their biosynthesis remain unknown. In this work, a genomewide expression analysis of jasmonate‐elicited Taxus baccata cell cultures by complementary DNA‐amplified fragment length polymorphism (cDNA‐AFLP) indicated a correlation between an extensive elicitor‐induced genetic reprogramming and increased taxane production in the targeted cultures. Subsequent in silico analysis allowed us to identify 15 genes with a jasmonate‐induced differential expression as putative candidates for genes encoding enzymes involved in five unknown steps of taxane biosynthesis. Among them, the TB768 gene showed a strong homology, including a very similar predicted 3D structure, with other genes previously reported to encode acyl‐CoA ligases, thus suggesting a role in the formation of the taxol lateral chain. Functional analysis confirmed that the TB768 gene encodes an acyl‐CoA ligase that localizes to the cytoplasm and is able to convert β‐phenylalanine, as well as coumaric acid, into their respective derivative CoA esters. β‐phenylalanyl‐CoA is attached to baccatin III in one of the last steps of the taxol biosynthetic pathway. The identification of this gene will contribute to the establishment of sustainable taxol production systems through metabolic engineering or synthetic biology approaches. 相似文献
14.
15.
X. Zhou Z. Wang K. Jiang Y. Wei J. Lin X. Sun K. Tang 《Applied Biochemistry and Microbiology》2007,43(4):439-443
A total of 38 endophytic fungus strains were isolated from Taxus chinensis var. mairei by the aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction
(PCR) analysis for the presence of the Taxus taxadiene synthase (TS) gene, a rate-limiting enzyme gene in the taxol biosynthetic pathway. Twelve out of 38 isolated endophytic fungus strains
showed PCR positive for the ts gene. Subsequently, taxol and its related compounds were extracted from culture filtrates and mycelia of the PCR positive
strains, separated by column chromatography, and analyzed by High Performance Liquid Chromatography and Mass Spectrum. The
analysis result showed that 3 strains could produce taxol and its related compounds at the detectible level. This study indicates
that molecular detection of the ts gene is an efficient method for primary screening of taxol or its related compound-producing endophytic fungi, which can
improve prominently screening efficiency.
Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2007, Vol. 43, No. 4, pp. 490–494.
The text was submitted by the authors in English. 相似文献
16.
Yan Chi Dong-Li Zhao Dong-Po Zhou 《World journal of microbiology & biotechnology》2008,24(11):2601-2605
The endophytic fungus Nodulisporium
sylviforme produces taxol, a diterpene alkaloid compound that has unique anti-neoplastic activity. In this study, suppression subtractive
hybridization was performed to identify genes in N. sylviforme that were involved in the up-regulation of taxol production. A total of 91 clones represented the differentially expressed
genes were selected based on reverse blotting results. These clones were then sequenced and analyzed using BLAST-X comparisons
that revealed that about 37.8% of the ESTs were homologous to 17 different proteins with known function, 20.7% were homologous
to nine hypothetical proteins and the remaining 41.5% had no identifiable homology; these may be novel genes. Two enzymatic
genes involved in taxol production were found. The data presented in this study provide the first primary overview of a set
of genes that are differentially expressed in N. sylviforme during taxol biosynthesis and form a solid foundation for the construction of high taxol-yielding genetically engineered
strains. 相似文献
17.
Plant-associated actinobacteria are rich sources of bioactive compounds including indole-derived molecules such as phytohormone indole-3-acetic acid (IAA). In view of few investigations concerning the biosynthesis of IAA by endophytic actinobacteria, this study evaluated the potential of IAA production in endophytic streptomycete isolates sourced from medicinal plant species Taxus chinensis and Artemisia annua. By HPLC analysis of IAA combined with molecular screening approach of iaaM, a genetic determinant of streptomycete IAA synthesis via indole-3-acetamide (IAM), our data showed the putative operation of IAM-mediated IAA biosynthesis in Streptomyces sp. En-1 endophytic to Taxus chinensis. Furthermore, using the co-cultivation system of model plant Arabidopsis thaliana and streptomycete, En-1 was found to be colonized intercellularly in the tissues of Arabidopsis, an alternative host, and the effects of endophytic En-1 inoculation on the model plant were also assayed. The phytostimulatory effects of En-1 inoculation suggest that IAA-producing Streptomyces sp. En-1 of endophytic origin could be a promising candidate for utilization in growth improvement of plants of economic and agricultural value. 相似文献
18.
B. Jasim C. Anisha Sabu Rohini Jacob Manoj Kurian Mathew Jyothis E. K. Radhakrishnan 《World journal of microbiology & biotechnology》2014,30(5):1649-1654
Ginger (Zingiber officinale) is cultivated commercially in most parts of the world especially in India for its culinary and medicinal applications. One of the major challenges that limit the yield of ginger is rhizome rot disease caused by organisms including Pythium myriotylum. A feasible ecofriendly method is yet to be devised to prevent the plant from this threatening disease. Recent studies on plant microbiome show the possibility of having endophytic organisms with plant protective characteristics associated with the plants. Because of the uniquely evolved underground nature of the ginger rhizome and its peculiar survival in soil for a long time, many interesting endophytic microbes with plant protective characters can be well expected from it. In the current study, previously isolated endophytic Pseudomonas aeruginosa from ginger was investigated in detail for its effect on Pythium myriotylum. The rhizome protective effect of the organism was also studied by co-inoculation studies, which confirmed that Pseudomonas aeruginosa has very potent inhibitory effect on Pythium myriotylum. On further studies, the active antifungal compound was identified as phenazine 1-carboxylic acid. 相似文献
19.
Gene Duplication in the Diversification of Secondary Metabolism: Tandem 2-Oxoglutarate–Dependent Dioxygenases Control Glucosinolate Biosynthesis in Arabidopsis
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Daniel J. Kliebenstein Virginia M. Lambrix Michael Reichelt Jonathan Gershenzon Thomas Mitchell-Olds 《The Plant cell》2001,13(3):681-694
Secondary metabolites are a diverse set of plant compounds believed to have numerous functions in plant–environment interactions. The large chemical diversity of secondary metabolites undoubtedly arises from an equally diverse set of enzymes responsible for their biosynthesis. However, little is known about the evolution of enzymes involved in secondary metabolism. We are studying the biosynthesis of glucosinolates, a large group of secondary metabolites, in Arabidopsis to investigate the evolution of enzymes involved in secondary metabolism. Arabidopsis contains natural variations in the presence of methylsulfinylalkyl, alkenyl, and hydroxyalkyl glucosinolates. In this article, we report the identification of genes encoding two 2-oxoglutarate–dependent dioxygenases that are responsible for this variation. These genes, AOP2 and AOP3, which map to the same position on chromosome IV, result from an apparent gene duplication and control the conversion of methylsulfinylalkyl glucosinolate to either the alkenyl or the hydroxyalkyl form. By heterologous expression in Escherichia and the correlation of gene expression patterns to the glucosinolate phenotype, we show that AOP2 catalyzes the conversion of methylsulfinylalkyl glucosinolates to alkenyl glucosinolates. Conversely, AOP3 directs the formation of hydroxyalkyl glucosinolates from methylsulfinylalkyl glucosinolates. No ecotype coexpressed both genes. Furthermore, the absence of functional AOP2 and AOP3 leads to the accumulation of the precursor methylsulfinylalkyl glucosinolates. A third member of this gene family, AOP1, is present in at least two forms and found in all ecotypes examined. However, its catalytic role is still uncertain. 相似文献
20.
Melinda L. Micallef Paul M. D’Agostino Deepti Sharma Rajesh Viswanathan Michelle C. Moffitt 《BMC genomics》2015,16(1)