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
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Plant Proteins/metabolism*
3.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
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metabolism
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Phylogeny
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Terpenes
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metabolism
4.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
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classification
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genetics
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Molecular Sequence Data
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Phylogeny
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
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genetics
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Oryza
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genetics
6.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
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metabolism
7.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
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
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DNA, Ribosomal Spacer
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genetics
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Dendrobium
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classification
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genetics
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Genes, Chloroplast
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Genes, Plant
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Introns
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Plants, Medicinal
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classification
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genetics
9.Marker-assisted selection and pyramiding for three blast resistance genes, Pi-d(t)1, Pi-b, Pi-ta2, in rice.
Xue-Wei CHEN ; Shi-Gui LI ; Yu-Qing MA ; Han-Yun LI ; Kai-Da ZHOU ; Li-Huang ZHU
Chinese Journal of Biotechnology 2004;20(5):708-714
G46B is a promising holding line used for three-lines breeding strategy in hybrid rice, but it is susceptible to blast disease caused by Pyricularia grisea. To improve its blast resistance, three rice varieties, Digu, BL-1, and Pi-4, with blast resistance genes, Pi-d(t), Pi-b, and Pi-ta2, respectively, were used to be crossed with G46B, and 15 plants with these three blast resistance genes, Pi-d(t)1, Pi-b, and Pi-ta2, were selected from their F2 and B1C1 populations via a marker-aided crossing procedure. Among them, four plants were heterozygotes in the three resistance genes, with the genotype of Pi-d(t)1 pi-d(t)/Pi-b pi-b/ Pi-ta2 pi-ta2; ten plants were heterozygotes in two of the three resistance genes, of which six with the genotype of Pi-d(t)1 Pi-d(t)1/Pi-b pi-b/Pi-ta2 pi-ta2, three with the genotype of Pi-d(t)1 pi-d(t)1/Pi-b pi-b/Pi-ta2 Pi-ta2, and one with the genotype of Pi-d(t)1pi-d(t)1/Pi-b Pi-b/Pi-ta2 pi-ta2; and only one plant was homozygote in two of the three resistance genes with the genotype of Pi-d(t)1 Pi-d(t)/Pi-b pi-b/Pi-ta2 Pi-ta2. These results demonstrate the capacity of maker-assisted selection (MAS) in gene pyramiding for rice blast resistance and its enhancement for the efficiency in rice resistance breeding.
Crosses, Genetic
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Genes, Plant
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Genetic Markers
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Genotype
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Oryza
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genetics
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Plant Diseases
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genetics
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Selection, Genetic
10.Genetic relationships of Atractylodes plants.
Xiao-Xing ZOU ; Lu-Qi HUANG ; Guang-Hong CUI ; Qing-Jun YUAN ; Yong PENG ; Yong LIU ; Pei-Gen XIAO
Acta Pharmaceutica Sinica 2009;44(6):680-686
The phylogenetic relationships of the genus Atractylodes DC. was analyzed using the nuclear ribosomal ITS and three chloroplast fragments, including atpB-rbcL, psbB-psbF and trnL-trnF intergenic spacer (IGS) sequences. Phylogenetic analyses revealed that A. lancea subsp. luotianensis and A. lancea, A. chinensis var. liaotungensis and A. coreana form monophyletic terminal clade, separately. The trees, within each the pairwise genetic distances, did not support Hu's classification. Authors suggested that A. lancea Subsp. luotianensis should be included in A. lancea rather than be treated as a separate subspecies. A. carlinoides was placed in the basal position of Atractylodes, which had a distant relationship with the others of the genus. The results lead us to suggest that A. chinensis var. liaotungensis be put into A. coreana, A. chinensis as a subspecies of A. lancea.
Atractylodes
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classification
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genetics
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DNA, Intergenic
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genetics
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DNA, Plant
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genetics
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Genes, Plant
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Phylogeny