1.Research progress in biosynthesis of podophyllotoxin and its derivatives.
Zhen MENG ; Tingting YAO ; Wei ZHAO ; Hongmei LI ; Ya-Jie TANG
Chinese Journal of Biotechnology 2021;37(6):2026-2038
Podophyllotoxin (PTOX) is an aryl-tetralin lignan of plant origin found in some species of Podophyllum such as Dysosma versipellis, Diphylleia sinensis, and Sinopodophyllum hexandrum. Etoposide and teniposide are produced semisynthetically from PTOX and used clinically to treat several forms of cancer. As a typical representative of new drug discovery from natural products, the production of PTOX solely depends on extraction from plants, resulting in severe contradiction between supply and demand. With the advantages of unconstrained resources and eco-friendly reaction conditions, biosynthesis method has become a trend in the production of PTOX and its derivatives. In this review, we summarize the research progress of PTOX biosynthesis in plants and expound the functions of the key enzymes as well as their subcellular location. The synthetic biology for production of PTOX intermediates in a tobacco chassis is also introduced. Finally, the heterologous expression and biotransformation of PTOX in microorganisms is summarized, which sets the foundation for the efficient microbial production of PTOX using cell factories.
Genes, Plant
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Podophyllotoxin/biosynthesis*
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Podophyllum/genetics*
2.Genome-wide identification of SUN gene family in Fragaria vesca and stresses-response analysis.
Yao YU ; Ziyao WANG ; Yiling XU ; Bojun MA ; Xifeng CHEN
Chinese Journal of Biotechnology 2023;39(2):724-740
SUN gene is a group of key genes regulating plant growth and development. Here, SUN gene families of strawberry were identified from the genome of the diploid Fragaria vesca, and their physicochemical properties, genes structure, evolution and genes expression were also analyzed. Our results showed that there were thirty-one FvSUN genes in F. vesca and the FvSUNs encoded proteins were classified into seven groups, and the members in the same group showed high similarity in gene structures and conservative motifs. The electronic subcellular localization of FvSUNs was mainly in the nucleus. Collinearity analysis showed that the members of FvSUN gene family were mainly expanded by segmental duplication in F. vesca, and Arabidopsis and F. vesca shared twenty-three pairs of orthologous SUN genes. According to the expression pattern in different tissues shown by the transcriptome data of F. vesca, the FvSUNs gene can be divided into three types: (1) expressed in nearly all tissues, (2) hardly expressed in any tissues, and (3) expressed in special tissues. The gene expression pattern of FvSUNs was further verified by quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, the seedlings of F. vesca were treated by different abiotic stresses, and the expression level of 31 FvSUNs genes were assayed by qRT-PCR. The expression of most of the tested genes was induced by cold, high salt or drought stress. Our studies may facilitate revealing the biological function and molecular mechanism of SUN genes in strawberry.
Fragaria/metabolism*
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Genes, Plant
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Stress, Physiological/genetics*
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Arabidopsis/genetics*
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Plant Development
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Gene Expression Regulation, Plant
;
Plant Proteins/metabolism*
3.Characteristics of the chloroplast genome of Dracaena marginata and phylogenetic analysis.
Zihao WANG ; Jiale GUO ; Qi FAN ; Zeyuan TIAN ; Xueqing WANG ; Wei ZHENG ; Luodong HUANG
Chinese Journal of Biotechnology 2023;39(7):2926-2938
Dracaena marginata is a widely cultivated horticultural plant in the world, which has high ornamental and medicinal value. In this study, the whole genome of leaves from D. marginata was sequenced by Illumina HiSeq 4000 platform. The chloroplast genome were assembled for functional annotation, sequence characteristics and phylogenetic analysis. The results showed that the chloroplast genome of D. marginata composed of four regions with a size of 154 926 bp, which was the smallest chloroplast genome reported for Dracaena species to date. A total of 132 genes were identified, including 86 coding genes, 38 tRNA genes and 8 rRNA genes. Codon bias analysis found that the codon usage bias was weak and there was a bias for using A/U base endings. 46 simple sequence repeat and 54 repeats loci were detected in the chloroplast genome, with the maximum detection rate in the large single copy region and inverted repeat region, respectively. The inverted repeats boundaries of D. marginata and Dracaena were highly conserved, whereas gene location differences occurred. Phylogenetic analysis revealed that D. serrulata and D. cinnabari form a monophyletic clade, which was the closest relationship and conformed to the morphological classification characteristics. The analysis of the chloroplast genome of D. marginata provides important data basis for species identification, genetic diversity and chloroplast genome engineering of Dracaena.
Phylogeny
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Dracaena
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Genome, Chloroplast/genetics*
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Base Sequence
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Genes, Plant
4.Functional characterization of SsNES responsible for nerolidol biosynthesis in Senecio scandens.
Qin-Qin SHEN ; Li-Ping WANG ; Jin LIANG ; Li-Jun LIU ; Qiang WANG
China Journal of Chinese Materia Medica 2019;44(7):1334-1340
A short terpene synthase gene was obtained by screening the transcriptome data of Senecio scandens. The phylogenetic tree and sequence alignment putatively identified this gene as a nerolidol synthase gene, named SsNES(GenBank MH518312). Protein homology modeling indicated that SsNES contained a complete conserved domain and folded correctly. SsNES was cloned and successfully expressed in Escherichia coli as soluble protein. The biochemical function of SsNES was characterized by E. coli metabolic engineering, which showed that SsNES catalyzed formation of trans-nerolidol with(E, E)-farnesyl diphosphate as the substrate. Nerolidol was also detected in stems and leaves of S. scandens, indicating that SsNES might act as the nerolidol synthase in plant. RT-PCR analysis indicated that SsNES was mainly expressed in stem, flowers and leaves, and no expression was observed in roots. After the treatment of SA, MeJA or Ala, SsNES was induced significantly at 6 h, indicating involvement in the defense response of S. scandens. The identification of SsNES not only clarified biosynthesis of nerolidol in S. scandens, but also provided diversity of sesquiterpene synthase, as well as theoretical basis for disease and pest defense mediated by the terpene metabolites.
Escherichia coli
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Genes, Plant
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Phylogeny
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Senecio
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enzymology
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Sesquiterpenes
;
metabolism
5.Pyramiding of 3-resistant-gene to improve rice blast resistance of a restorer line, Fuhui 673.
Zhiwei CHEN ; Huazhong GUAN ; Xiaofang WANG ; Ruixia DONG ; Chenghai ZHUO ; Damei MAO ; Runsen PAN ; Yuanchang ZHOU ; Weiren WU
Chinese Journal of Biotechnology 2019;35(5):837-846
To improve the blast resistance of elite rice restorer line Fuhui 673, 3 blast resistance genes Pi-1, Pi-9 and Pi-kh were introduced into Fuhui 673 from a good-quality restorer line Jinhui 1059 through 3 successive backcrosses followed by one selfing using the technique of marker-assisted selection. Ten near-isogenic lines (NILs) of Fuhui 673 carrying the 3 introduced resistance genes were created. Genotype analysis using 68 SSR markers evenly distributed in the genome indicated that 92.96%-98.59% of the NILs' genetic background had been recovered to Fuhui 673. Both indoor and field resistance tests indicated that the NILs and their hybrids with sterile line Yixiang A were all resistant to rice blast, with resistance levels significantly higher than those of controls Fuhui 673 and hybrid Yiyou 673 (Yixiang A Fuhui 673). In addition, among the 10 hybrids between the NILs and Yixiang A, 2 showed significantly higher yield than and 4 displayed similar yield to that of control Yiyou 673, suggesting that most of the NILs retained the elite characteristics of Fuhui 673. Two new hybrid rice cultivars Liangyou 7283 and Jintaiyou 683 from NIL Line 9 showed high yield, good resistance to blast and moderate growth period in regional trial, suggesting that the NIL Line 9 has a good prospect for application.
Breeding
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Disease Resistance
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genetics
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Genes, Plant
;
genetics
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Oryza
;
genetics
6.Phylogenetic analyses of some important Paris species based on sequences of matK gene.
Jian MA ; Diqiang LI ; Yuguang ZHANG ; Dayuan XUE
China Journal of Chinese Materia Medica 2010;35(1):18-21
The matK genes of 10 samples in Paris from Hunan, Yunnan and Jilin provinces were sequenced and compared. The phylogenetic tree was constructed based on the matK gene sequences and the ten pairs samples were divided into four groups. The results did not support the reality of four taxa named P. polyphylla var. pseudothibetica, P. polyphylla var. yunnanensis, P. polyphylla var. appendiculata and P. polyphylla var. chinenis. They are supposed to be treated as different forms of P. polyphylla var. polyphylla.
Genes, Plant
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genetics
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Liliaceae
;
classification
;
genetics
;
Molecular Sequence Data
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Phylogeny
7.Identification and bioinformatics analysis of genes associated with MVA pathway in Magnolia officinalis.
Liang-ping ZHA ; Yuan YUAN ; Lu-qi HUANG ; Shu-lin YU
China Journal of Chinese Materia Medica 2015;40(11):2077-2083
Methyl valerate (MVA) pathway is one of the important ways for synthesis of terpenoids. This study was based on data of the transcriptome sequencing of Magnolia officinalis, the associated genes MoACOT, MoHMGS, MoHMGR, MoMK in methyl valerate (MVA) pathway, were completed in detail by using bioinformatics methods. The results of analysis showed that MoACOT and MoMK were stable hydrophobic proteins, MoHMGS and MoHMGR were unstable hydrophobic protein. The secondary structures of all proteins were hybrid architecture,and alpha helical were the major motifs. There were no clear transmembrane domains in MoACOT, MoHMGS and MoMK, but two transmembrane domains were founded in MoHMGR which were from 39-61 aa and 82-104 aa resepectively. The results of evolutionary relationship analysis showed that MoACOT, MoHMGS, MoHMGR and MoMK had relative close relationship to angiosperm or dicotyledonous plants, and accorded with genetic evolution rule. From transcriptome data, transcripted level of MoACOT, MoHMGS, MoHMGR, MoMK in M. officinalis and M. officinalis var. biloba was not significantly different. The result provided theoretical reference for study on Methyl valerate (MVA) pathway of terpenoid of M. officinalis.
Computational Biology
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Genes, Plant
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Magnolia
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genetics
;
metabolism
;
Phylogeny
;
Terpenes
;
metabolism
8.Applylication of new type combined fragments: nrDNA ITS+ nad 1-intron 2 for identification of Dendrobium species of Fengdous.
Li-xia GENG ; Rui ZHENG ; Jie REN ; Zhi-tao NIU ; Yu-long SUN ; Qing-yun XUE ; Wei LIU ; Xiao-yu DING
Acta Pharmaceutica Sinica 2015;50(8):1060-1067
In this study, 17 kinds of Dendrobium species of Fengdous including 39 individuals were collected from 4 provinces. Mitochondrial gene sequences co I, nad 5, nad 1-intron 2 and chloroplast gene sequences rbcL, matK amd psbA-trnH were amplified from these materials, as well as nrDNA ITS. Furthermore, suitable sequences for identification of Dendrobium species of Fengdous were screened by K-2-P and P-distance. The results showed that during the mentioned 7 sequences, nrDNA ITS, nad 1-intron 2 and psbA-trnH which had a high degree of variability could be used to identify Dendrobium species of Fengdous. However, single fragment could not be used to distinguish D. moniliforme and D. huoshanense. Moreover, compared to other combined fragments, new type combined fragments nrDNA ITS+nad 1-intron 2 was more effective in identifying the original plants of Dendrobium species and could be used to identify D. huoshanense and D. moniliforme. Besides, according to the UPGMA tree constructed with nrDNA ITS+nad 1-intron 2, 3 inspected Dendrobium plants were identified as D. huoshanense, D. moniliforme and D. officinale, respectively. This study identified Dendrobium species of Fengdous by combined fragments nrDNA ITS+nad 1-intron 2 for the first time, which provided a more effective basis for identification of Dendrobium species. And this study will be helpful for regulating the market of Fengdous.
DNA, Plant
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genetics
;
DNA, Ribosomal Spacer
;
genetics
;
Dendrobium
;
classification
;
genetics
;
Genes, Chloroplast
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Genes, Plant
;
Introns
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Plants, Medicinal
;
classification
;
genetics
9.Identification and expression profiling of R2R3-MYB transcription factors in Erigeron breviscapus.
Wan-Ling SONG ; Gui-Sheng XIANG ; Ying-Chun LU ; Guang-Hui ZHANG ; Sheng-Chao YANG ; Yan ZHAO
China Journal of Chinese Materia Medica 2021;46(23):6149-6162
R2 R3-MYB transcription factors are ubiquitous in plants, playing a role in the regulation of plant growth, development, and secondary metabolism. In this paper, the R2 R3-MYB transcription factors were identified by bioinformatics analysis of the genomic data of Erigeron breviscapus, and their gene sequences, structures, physical and chemical properties were analyzed. The functions of R2 R3-MYB transcription factors were predicted by cluster analysis. Meanwhile, the expression patterns of R2 R3-MYB transcription factors in response to hormone treatments were analyzed. A total of 108 R2 R3-MYB transcription factors, named EbMYB1-EbMYB108, were identified from the genome of E. breviscapus. Most of the R2 R3-MYB genes carried 2-4 exons. The phylogenetic tree of MYBs in E. breviscapus and Arabidopsis thaliala was constructed, which classified 234 MYBs into 30 subfamilies. The MYBs in the five MYB subfamilies of A.thaliala were clustered into independent clades, and those in E. breviscapus were clustered into four clades. The transcriptome data showed that MYB genes were differentially expressed in different tissues of E. breviscapus and in response to the treatments with exogenous hormones such as ABA, SA, and GA for different time. The transcription of 13 R2 R3-MYB genes did not change significantly, and the expression patterns of some genes were up-regulated or down-regulated with the extension of hormone treatment time. This study provides a theoretical basis for revealing the mechanisms of R2 R3-MYB transcription factors in regulating the growth and development, stress(hormone) response, and active ingredient accumulation in E. breviscapus.
Erigeron/genetics*
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Gene Expression Regulation, Plant
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Genes, myb
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Phylogeny
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Plant Proteins/metabolism*
;
Transcription Factors/metabolism*
10.Identification and expression analysis of R2R3-MYB gene family in Andrographis paniculata.
Jing-Yu LI ; Ming-Yang SUN ; Shi-Qiang XU ; Wei SUN ; Yan GU ; Yu MEI ; Ji-Hua WANG
China Journal of Chinese Materia Medica 2022;47(1):72-84
The plant growth, development, and secondary metabolism are regulated by R2 R3-MYB transcription factors. This study identified the R2 R3-MYB genes in the genome of Andrographis paniculata and analyzed the chromosomal localization, gene structure, and conserved domains, phylogenetic relationship, and promoter cis-acting elements of these R2 R3-MYB genes. Moreover, the gene expression profiles of R2 R3-MYB genes under abiotic stress and hormone treatments were generated by RNA-seq and validated by qRT-PCR. The results showed that A. paniculata contained 73 R2 R3-MYB genes on 21 chromosomes. These members belonged to 34 subfamilies, 19 of which could be classified into the known subfamilies in Arabidopsis thaliana. The 73 R2 R3-MYB members included 36 acidic proteins and 37 basic proteins, with the lengths of 148-887 aa. The domains, motifs, and gene structures of R2 R3-MYBs in A. paniculata were conserved. The promoter regions of these genes contains a variety of cis-acting elements related to the responses to environmental factors and plant hormones including light, ABA, MeJA, and drought. Based on the similarity of functions of R2 R3-MYBs in the same subfamily and the transcription profiles, ApMYB13/21/35/67/73(S22) may regulate drought stress through ABA pathway; ApMYB20(S11) and ApMYB55(S2) may play a role in the response of A. paniculata to high temperature and UV-C stress; ApMYB5(S7) and ApMYB33(S20) may affect the accumulation of andrographolide by regulating the expression of key enzymes in the MEP pathway. This study provides theoretical reference for further research on the functions of R2 R3-MYB genes in A. paniculata and breeding of A. paniculata varieties with high andrographolide content.
Andrographis paniculata
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Gene Expression Regulation, Plant
;
Genes, myb
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Multigene Family
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Phylogeny
;
Plant Proteins/metabolism*