1.Advances in gene editing and natural product synthesis of Rhodotorula toruloides.
Qidou GAO ; Yaqi DONG ; Ying HUANG ; Yijuan LIU ; Xiaobing YANG
Chinese Journal of Biotechnology 2023;39(6):2313-2333
Rhodotorula toruloides is a non-conventional red yeast that can synthesize various carotenoids and lipids. It can utilize a variety of cost-effective raw materials, tolerate and assimilate toxic inhibitors in lignocellulosic hydrolysate. At present, it is widely investigated for the production of microbial lipids, terpenes, high-value enzymes, sugar alcohols and polyketides. Given its broad industrial application prospects, researchers have carried out multi-dimensional theoretical and technological exploration, including research on genomics, transcriptomics, proteomics and genetic operation platform. Here we review the recent progress in metabolic engineering and natural product synthesis of R. toruloides, and prospect the challenges and possible solutions in the construction of R. toruloides cell factory.
Gene Editing
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Metabolic Engineering
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Rhodotorula/metabolism*
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Lipids
2.Direct Presumptive Identification of Candida species from Blood Cultures Using CHROMagar Candida.
Jong Hee SHIN ; Deok CHO ; Soo Hyun KIM ; Dong Euk BYUN ; F S NOLTE ; Dong Wook RYANG
Korean Journal of Clinical Pathology 1997;17(1):128-136
BACKGROUND: CHROMagar Candida is a new differential medium that allows selective isolation and identification of clinically significant yeasts. We evaluated the use of this medium to identify Candida species directly from positive blood culture bottles. METHODS: A total of 152 positive blood culture bottles (51 Candida albicans, 29 Candida troficalis, 28 Candida parapsilosis, 26 Candida glabrata, 10 Candida krusei, 4 Candida pelliculosa. 1 Candida guilliermonidii, 3 C. albicans plus C. glabrata) were directly subcultures to CHROMagar (Hardy diagnostics. USA) and incubated for 48 h. Colony appearance on CHROMagar was assessed independently by three observers. RESULTS: CHROMagar correctly identified 95.4%, 92 1% and 91.4% of Candida app. from blood cultures by the three observers. respectively. There was 91.4% agree cent between the observers. Expected colony appearance on CHROMagar was 100% for C. albicans. 97.7% for C. tropicalis, 96.7% for C. krusei, 94 9% for C. glabrata but 88.1% for C. parapsilosis. Three mixed candidemias, not detected by conventional methods, were detected by CHROMagar. CONCLUSIONS: CHROMagar permits earlier recognition of major Cardida app. in positive blood cultures and more reliable detection of mixed candidemias.
Candida albicans
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Candida glabrata
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Candida*
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Candidemia
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Yeasts
3.Optimization of enzymatic extraction of effective constituents from fibrous roots of Salvia miltiorrhiza by central composite design and response surface method.
Gang LU ; Guo-dong DU ; Jun-ru WANG ; Zong-suo LIANG
China Journal of Chinese Materia Medica 2008;33(16):1976-1981
The fibrous roots are the residues of production of cut crude drug of Danshen (Salvia miltiorrhiza). Enzymatic pretreatment and ultrasonic extraction are beneficial to extract effective constituents from fibrous roots more effectively. The present research was to optimize the enzymatic parameters by the central composite design and response surface method. Under the best conditions, the yields of total tanshinones and total salvianolic acids in the extracts of enzymatic pretreatment increased by 113.92% and 30.64%, comparing with the non-enzymatic extraction, respectively. TLC analysis also showed that the types of effective constituents in the two samples were not affected by enzymatic hydrolysis. Meanwhile, the complex correlation coefficients of the mathematical models were high, which provided a good prediction.
Diterpenes, Abietane
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Drugs, Chinese Herbal
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chemistry
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Phenanthrenes
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chemistry
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Plant Roots
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chemistry
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Salvia miltiorrhiza
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chemistry
4.Genome-wide identification and expression analysis of CSLA gene family of Dendrobium catenatum.
Ya-Qian GAO ; Xue-Liang CHEN ; Dong-Hong CHEN ; Jing-Jing LIU ; Jin-Ping SI
China Journal of Chinese Materia Medica 2020;45(13):3120-3127
Glucomannan is the key active ingredient of Dendrobium catenatum, and CSLA family is responsible for glucomannan biosynthesis. In order to systematically evaluate the CSLA family members of D. catenatum, the bioinformatics methods were performed for genome-wide identification of DcCSLA gene family members through the genomic data of D. catenatum downloaded from the NCBI database, and further analyses of their phylogenetic relationship, gene structure, protein conserved domains and motifs, promoter cis-elements and gene expression profiles in response to stresses. The results showed that D. catenatum contains 13 CSLA members, all of which contain 9-10 exons. In the evolutionary relationship, CSLA genes were clustered into 5 groups, DcCSLA genes were distributed in all branches. Among which the ancestral genes of groupI existed before the monocot-dicot divergence, and groupⅡ-Ⅴ only existed in the monocot plants, indicating that group Ⅰ represents the earliest origin group. CSLA proteins are characteristic of the signature CESA_CaSu_A2 domain. Their promoter regions contain cis elements related to stresses and hormones. Under different stress treatments, low temperature induces the expression of DcCSLA5 and inhibits the expression of DcCSLA3. Infection of Sclerotium delphinii inhibits DcCSLA3/4/6/8/9/10 expression. Under the treatment of jasmonic acid, DcCSLA11 expression was significantly up-regulated, and DcCSLA2/5/7/12/13 were significantly down-regulated. These results laid a foundation for further study on the function of DcCSLA genes in glucomannan biosynthesis and accumulation.
Basidiomycota
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Cold Temperature
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Dendrobium
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genetics
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Gene Expression Regulation, Plant
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Genome, Plant
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Multigene Family
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Phylogeny
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Plant Proteins
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genetics
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Stress, Physiological
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Transcriptome
5.Porcine skeletal muscle development regulated by MicroRNA: a review.
Yulin HE ; Jianjun JIN ; Dong LI ; Gongshe YANG ; Taiyong YU
Chinese Journal of Biotechnology 2023;39(4):1514-1524
The growth and development of skeletal muscle is an important factor affecting pork production and quality, which is elaborately regulated by many genetic and nutritional factors. MicroRNA (miRNA) is a non-coding RNA with a length of about 22 nt, which binds to the 3'UTR sequence of the mRNA of the target genes, and consequently regulates its post-transcriptional expression level. In recent years, a large number of studies have shown that miRNAs are involved in various life processes such as growth and development, reproduction, and diseases. The role of miRNAs in the regulation of porcine skeletal muscle development was reviewed, with the hope to provide a reference for the genetic improvement of pigs.
Swine
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Animals
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MicroRNAs/metabolism*
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Muscle, Skeletal/metabolism*
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Muscle Development/genetics*
6.Effects of germplasm and host tree trunk on endophytic fungal communities in epiphytic Dendrobium catenatum.
Shu-Yuan WANG ; Ling-Shang WU ; Hong-Xiu DONG ; Jin-Ping SI ; Yu-Qiu ZHU
China Journal of Chinese Materia Medica 2018;43(8):1588-1595
In order to investigate the effects of germplasm and host tree trunk on endophytic fungal communities in epiphytic Dendrobium catenatum, a total of 3 835 isolates were recovered from roots, stems and leaves of four D. catenatum germplasms attached to one kind of host tree trunk and one germplasm attached to four kinds of epiphyte-host tree trunks. A total of 152 taxa were identified and classified based on the fungal cultural characteristics and phylogenetic analyses of ITS sequences. The taxa were assigned to 60 genera, 35 families, 21 orders and 5 classes of 2 phyla. The results indicated that D. catenatum cultivated in stereo cultivation harbor variety of fungi. The dominant fungal groups were different between Lin'an and Yiwu. Moreover, several groups showed geographical specificity, such as Arthrinium, Coniochaeta, Fusarium, Neofusicoccum and Zopfiella only dominating in Panshan of Lin'an, while Alternaria, Bjerkandera, Cercophora, Nigrospora and Trichoderma only dominating in Shangxi of Yiwu. There was no significant difference in diversity or species richness of endophytic fungi neither among germplasm nor host tree trunk. However, the richness and diversity indices exhibited a strong dependence on tissue type (<0.05). The germplasm and host tree trunk impact the distribution patterns of endophytic fungi less than tissue type. Nevertheless, the relative frequencies of the dominant fungal groups were different among germplasms or host tree trunk types. Furthermore, there were some fungal species specific to certain germplasm or host tree trunk. This might be due to the distinctions in growth traits and chemical compositions of D. catenatum owning to the differences in D. catenatumgenetic background and microenvironment of host tree. Most of fungal taxa exhibit tissue specificity or preference. These results provide the basis for the study on the relationship between endophytic fungi and D. catenatum in stereo cultivation mode.
7.Morphometric Studies on the Genus Septoria in Korea (I).
Hyeon Dong SHIN ; Ekaterina F SAMEVA ; Jeong Dong KIM
Mycobiology 2001;29(3):145-153
The mycoflora of Korea was rather poorly studied in the past and the fungi belonging to the genus Septoria are no exception. For this reason, taxonomic studies on Septoria have been initiated, with the eventual aim of producing a monograph of the Septoria species present in Korea. The present study circumscribes 10 species; viz., Septoria artemisiae, S. callistephi, S. chrysanthemella, S. erigerontis, S. lycopersici, S. lysimachiae, S. oenotherae, S. phlogis, S. rohlenae, and S. sonchi. Distinguishing morphological characters are described and illustrated for each species.
Artemisia
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Classification
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Fungi
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Korea*
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Oenothera
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Primulaceae
8.Investigation on reproductive characteristics of Polygonatum cyrtonema.
Li-Ge LI ; Ze-Rui ZHANG ; Yan SHI ; Jing-Jing LIU ; Ying-Xiong QIU ; Dong-Hong CHEN ; Jin-Ping SI
China Journal of Chinese Materia Medica 2021;46(5):1079-1083
The study is aimed to investigate the reproductive biology characteristics of Polygonatum cyrtonema, especially including phenology, flower bud differentiation, flowering timing, floral traits, pollen vigor and stigma receptivity. The results showed that P. cyrtonema forms inflorescence before the leaves spread. In the wild, P. cyrtonema is mainly pollinated by insects such as bumblebees, with a seed setting rate of 65.12%. The seed setting rate of indoor single plant isolation or self-pollination enclosed by parchment paper bag is 0, indicating that it is self-incompatible. In Lin'an city, seedlings begin to emerge from mid-March to early April(the temperature is higher than 7.5 ℃), buds begin to emerge from the end of March to mid-April, and then undergo the full bloom stage from mid-to-late April, and the final flowering stage from the end of April to mid-May. The whole flowering period lasts 36 to 45 days. There are obvious differences in the phenology of different provenances. The flowers come into bloom from the base to the top along the aboveground main axis, which usually contain 4-22 inflorescences with(2-) 4-10(-21) flowers per inflorescence. The flowering pe-riod for a single plant is 26-38 days. The single flower lasts about 20-25 days from budding to opening and withers 2 days after pollination, and then the ovary will gradually expand. If unpollinated, it will continue to bloom for 3-5 days and then wither. Flower development period is significantly related to pollen vigor and stigma remittance. The pollen viability is the highest when the flower is fully opened with anthers gathering on the stigma, and the receptivity is the strongest when the stigma protrudes out of the perianth and secretes mucus. The fruits and seeds ripen in October, and proper shading can ensure the smooth development and maturity of the seeds. This study provides a basis for the hybrid breeding and seed production of P. cyrtonema.
Flowers
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Plant Breeding
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Pollination
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Polygonatum
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Reproduction
9. The Regulation of mTOR Signaling Pathway on Adipogenesis
Ming-Kun CHAO ; Xu-Dong YI ; Wei-Jun PANG
Chinese Journal of Biochemistry and Molecular Biology 2022;38(11):1477-1485
Adipose tissue is a major energy storage and endocrine organ. Adipogenesis is a complex process of cell differentiation, which is regulated by nutrient levels, hormones and metabolites, etc. The mammalian target of rapamycin (mTOR) complex includes two protein complexes, mammalian target of rapamycin complex 1 (mTORC1) and mTORC2. The lipid kinase-like domain contained in the mTOR complex lays the foundation for the mTOR pathway to regulate adipogenesis. Research on some components of mTORC1 and mTORC2 has verified the roles of mTOR in the regulation of adipogenesis. Based on previous studies, we reviewed the research of miR-199a-3p, miR-103, miR-188, Src-associated substrate in mitosis of 68 kD (Sam68), endostatin and other substances in the regulation of adipogenesis through mTORC1 and mTORC2. At the same time, we had further constructed the adipogenesis network regulated by mTOR signaling pathway, including insulin/IGF pathway, PI3K-AKT pathway, amino acid pathway, AMPK pathway, cAMP pathway, cGMP pathway, NOTCH pathway, and the modulation of bta-miR-150, 4-O-methylasochlorin and a variety of proteins. This article mainly reviewed the characteristics of mTOR complex and the latest research progress in the regulation of adipogenesis by mTOR pathway. It was pointed out that mTORC2 can regulate lipid uptake, lipolysis and regulate the function of mTORC1. However, there are fewer studies on mTORC2 compared to mTORC1, so further researches on adipogenesis and lipid metabolism may be more focused on mTORC2.
10. Effects of tRNA and tRNA-derived Fragments on Skeletal Muscle Development
Xu-Dong YI ; Tian-Tian ZHAO ; Wei-Jun PANG
Chinese Journal of Biochemistry and Molecular Biology 2021;37(9):1180-1187
Skeletal muscle is an important tissue of human and livestock. The study of the muscle development is of great significance for treating muscle diseases and improving livestock meat quality. The process of muscle development is controlled by several myogenic transcription factors and signaling pathways. In addition, recent findings established that several noncoding RNAs play a critical role in the regulation of muscle development such as long non-coding RNA (lncRNA), microRNA (miRNA) and circu- lar RNA (circRNA), etc. The detailed mechanism of muscle development is not well understood. Transfer RNAs (tRNAs) are fundamental components in the translation machinery as an adaptor molecule, and tRNA pool could be differentially exploited to modulate expression of mRNAs. In addition, tRNA can be cleaved into tRNA-derived fragments (tRFs) by a variety of ribonucleases (RNases) upon various stress conditions. Unlike the post-transcriptional regulation of lncRNA and miRNA on muscle development, tRNA has been implicated in various aspects of muscle development. Mitochondria play a central role in a plethora of processes related to the maintenance of muscle cellular homeostasis and genomic integrity. Mitochondrial tRNA(mt-tRNA) gene mutations lead to multiple myopathy because human mitochondrial genome is extremely small. The regulation of tRF is similar to miRNAs in regards to the related physiological processes, but are more conservative than miRNA. It is generally believed that tRF has strong tissue specificity, disease specificity and temporal specificity. Some skeletal muscle-specific tRFs could act posttranscriptionally via RNAi or targeting related genes. However, the tRF-sequencing analysis and functional mechanism of tRF are rarely studied in skeletal muscles. The myopathy caused by mitochondrial tRNA gene mutations are particularly complex, which are one of the challenges to diagnose, treat, or prevent diseases. Compared with other noncoding RNAs, the structural complexity of tRF also brings great challenges to data mining and analysis. In this review, we summarize the formation and function of tRNA and tRF especially in muscle development, which will deepen our understandings of related myopathy, and provide new ideas and directions for the investigation of skeletal muscle.