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1.
It is crucial to select stable references in gene expression analyses using quantitative real-time PCR (qRT-PCR). In this
work, seven frequently used reference genes, 18S, Actin, EF1α, α-tubulin, β-tubulin, Cyclophilin and Cytoplasmic ribosomal protein L2 (L2), from Bupleurum chinense DC. were evaluated as the internal control in five tissues, roots, stems, leaves, flowers and fruits, before tissue specific
gene expression assays. The results showed that β-tubulin was the most stable and reliable reference gene among the seven candidate genes in the measured tissues. The expression levels
of four genes involved in saikosaponins (the pharmacological active compounds of B. chinense) biosynthesis, HMGR, IPPI, FPS and β-AS, were assayed with β-tubulin as the internal control in the five tissues. All the four genes were expressed in the five tissues with different profiles
and HMGR in the order of roots > flowers, stems and leaves > fruits, IPPI of stems > leaves and fruits > roots and flowers, FPS of flowers > fruits > stems and roots > leaves and β-AS of roots > flowers, stems and fruits > leaves. The genes of FPS and β-AS were expressed predominantly in flowers and roots, respectively. This study may provide a suitable internal control for quantitative
gene expression assays in various tissues and give insight into the tissue expression profiles of four saikosaponins biosynthesis-involved
genes of medicinal B. chinense. 相似文献
2.
Z. -J. Zhao Y. -G. Song Y. -L. Liu M. Qiao X. -L. Zhai F. -N. Xiang 《Biologia Plantarum》2013,57(1):139-143
Gentiana straminea is native to the Qinghai-Tibet plateau, where it is exposed to extremes of cold and strong UV-B radiation. Here we showed that low temperature, but not UV-B radiation, affected the accumulation of the triterpenoid oleanolic acid. Neither of these stresses altered the expression of known triterpenoid synthesis genes. However, the application of methyl jasmonate (MeJA), but not salicylic acid (SA), significantly enhanced the accumulation of oleanolic acid, and up-regulated the triterpenoid synthesis genes, especially the expression of βAS, the gene encoding β-amyrin. 相似文献
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Muhammad Yaqoob Li Ping Wang Tang Fang Cheng-Ping Lu 《World journal of microbiology & biotechnology》2011,27(9):2041-2050
Thirty four avian Escherichia coli isolates were collected from different cities of Punjab province, Pakistan during 2008–2009. Twenty five phenotypic highly
ampicillin-resistant (MICs ≥ 256 μg/ml) avian E. coli strains were selected for the investigation of occurrence and transmission of class 1, 2 and 3 integrons and β-lactamase
genes. Amoxicillin, sulfonamide, trimethoprim, enrofloxacin, pefloxacin and tetracycline were the most common phenotypic resistant
among ampicillin-resistant avian E. coli strains. Integrons and β-lactamase were found 60 and 72% respectively. Class 1 and 2 integrons were found 52 and 8%, while class 3 integrons were
not found in all strains. All class 1 positive strains had variable fragments associated with gene cassettes dfrA7, dfrA1-aadA1, aadA1, aadA22 and dfrA12-orfF-aadA2 respectively, which confer resistance to trimethoprim and streptomycin. Class 2-positive strains had similar gene cassettes
array dfrA1-sat1-aadA1 conferring resistance to trimethoprim, streptothricin and spectinomicin/streptomycin. Integrons are frequently found in β-lactamase positive isolates and widely disseminate multidrug resistance genes but they do not play role in the spreading
of β-lactamase genes. Class 1 integrons gene cassette aadA22 is reported for the first time in avian E. coli. Findings of this study may provide important and useful information reflecting specific antibiotic selective pressure in
Punjab province, Pakistan. 相似文献
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O. B. Kazakova N. I. Medvedeva I. P. Baikova G. A. Tolstikov T. V. Lopatina M. S. Yunusov L. Zaprutko 《Russian Journal of Bioorganic Chemistry》2010,36(6):771-778
The synthesis of a new group of triterpenoid acylates has been conducted on the basis of oleanolic, glycyrrhetic, and ursolic
acids and betulin. 28-ortho-Methoxycynnamoylbetulin has been demonstrated to possess high activity against the influenza type A (H1N1) virus with the
selectivity index SI > 100 while studying the activity of the synthesized compounds in relation to the reproduction of viral
pathogens of respiratory infections. The high activity of 3,28-dinicotinoylbetulin against the papilloma virus (strain HPV-11)
was detected with the selectivity index SI 35. 相似文献
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Capel CS de Souza AC de Carvalho TC de Sousa JP Ambrósio SR Martins CH Cunha WR Galán RH Furtado NA 《Journal of industrial microbiology & biotechnology》2011,38(9):1493-1498
The goal of this study is to produce oleanolic acid derivatives by biotransformation process using Mucor rouxii and evaluate their antimicrobial activity against oral pathogens. The microbial transformation was carried out in shake flasks
at 30°C for 216 h with shaking at 120 rpm. Three new derivatives, 7β-hydroxy-3-oxo-olean-12-en-28-oic acid, 7β,21β-dihydroxy-3-oxo-olean-12-en-28-oic
acid, and 3β,7β,21β-trihydroxyolean-12-en-28-oic acid, and one know compound, 21β-hydroxy-3-oxo-olean-12-en-28-oic acid, were
isolated, and the structures were elucidated on the basis of spectroscopic analyses. The antimicrobial activity of the substrate
and its transformed products was evaluated against five oral pathogens. Among these compounds, the derivative 21β-hydroxy-3-oxo-olean-12-en-28-oic
acid displayed the strongest activity against Porphyromonas gingivalis, which is a primary etiological agent of periodontal disease. In an attempt to improve the antimicrobial activity of the
derivative 21β-hydroxy-3-oxo-olean-12-en-28-oic acid, its sodium salt was prepared, and the minimum inhibitory concentration
against P. gingivalis was reduced by one-half. The biotransformation process using M. rouxii has potential to be applied to the production of oleanolic acid derivatives. Research and antimicrobial activity evaluation
of new oleanolic acid derivatives may provide an important contribution to the discovery of new adjunct agents for treatment
of dental diseases such as dental caries, gingivitis, and periodontitis. 相似文献
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Jing Yin Chunxiao Li Yaguang Zhan Hongran Sun Yuan Gong Mengyan Zhang Jialei Xiao 《Plant Molecular Biology Reporter》2016,34(2):427-439
The pentacyclic triterpenoids from birch (Betula platyphylla suk) have broad pharmacological activities and can be potentially used for the development of anti-cancer and anti-AIDS drugs. In this study, we explored the effects of spraying 3-year-old white birch with different concentration of methyl jasmonate (MeJA) and salicylic acid (SA) on the expression of key genes in triterpenoid biosynthesis pathways and on the accumulation and physiological characteristics of triterpenoids in birch saplings. The results showed that spraying different concentration of MeJA and SA could obviously promote accumulation of total triterpenoids in 3-year-old white birch. The triterpenoid content in the stem bark was increased by 46.11 %, reaching 81.86 mg/g, after 1 day of treatment with 1 mmol·L?1 MeJA (MJ2), and by 45.07 %, reaching 91.4 mg/g, after 14 days of treatment with 5 mmol·L?1 SA (SA1). In addition, MeJA and SA treatment increased the contents of chlorophyll a and b, antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as photosynthetic performance, and affected the content of soluble sugar and soluble protein in birch leaf. Fluorescence quantitative polymerase chain reaction (qPCR) results showed that MeJA and SA treatment deferentially enhanced the key gene expression of cycloartenol synthase (BPX and BPX2), lupeol synthase (BPW) and beta-amyrin synthase (BPY) in triterpenoid synthesis pathway in birch bark and leaves. The results showed that MeJA and SA induced triterpenoid synthesis of birch plant is closely related with not only the expression of key genes of triterpenoid synthesis pathway but also photosynthesis, anti-stress response and physiological indexes, suggesting that regulation of triterpenoid synthesis of birch by MeJA and SA may involve in more complex mechanisms at physiological and molecular levels. 相似文献
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Jean Baptiste Bassene Yann Froelicher Claudie Dhuique-Mayer Waffa Mouhaya Rosa Mar Ferrer Gema Ancillo Raphael Morillon Luis Navarro Patrick Ollitrault 《Plant cell reports》2009,28(11):1689-1697
Allopolyploidy is known to induce novel patterns of gene expression and often gives rise to new phenotypes. Here we report
on the first attempt to relate phenotypic inheritance in an allotetraploid somatic hybrid with gene expression. Carotenoid
compounds in the fruit pulp of the two parental species and the hybrid were evaluated quantitatively by HPLC. Only very low
levels of β-carotene and β-cryptoxanthin were observed in Citrus limon, while β-cryptoxanthin was a major component of C. reticulata, which also displayed high levels of phytoene, phytofluene, β-carotene, lutein, zeaxantin and violaxanthin. Total carotenoid
content in mandarin juice sacs was 60 times greater than that in lemon. The allotetraploid hybrid produced all the same compounds
as mandarin but at very low levels. Transgressive concentration of abscisic acid (ABA) was observed in the somatic hybrid.
Real-time RT-PCR of total RNA from juice sacs was used to study expression of seven genes (CitDxs, CitPsy, CitPds, CitZds, CitLcy-b, CitChx-b, and CitZep) of the carotenoid biosynthetic pathway and two genes (CitNced1 and CitNced2) involved in abscisic acid synthesis from carotenoid. Gene expression was significantly higher for mandarin than lemon for
seven of the nine genes analyzed. Lemon under expression was partially dominant in the somatic hybrid for three upstream steps
of the biosynthetic pathway, particularly for CitDxs. Transgressive over expression was observed for the two CitNced genes. A limitation of the upstream steps of the pathway and a downstream higher consumption of carotenoids may explain the
phenotype of the somatic hybrid. 相似文献
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Maofu Li Kaikai Zhu Songling Bai Zhi Liu Tianzhong Li 《Molecular breeding : new strategies in plant improvement》2011,28(2):171-180
Apple (Malus domestica Borkh.) possesses gametophytic self-incompatibility (GSI) which is controlled by S-RNase in the pistil as well as a pollen S-determinant that has not been well characterized. The identification of S-locus F-box brother (SFBB) genes, which are good candidates for the pollen S-determinant in apple and pear, indicated the presence of multiple S-allelic polymorphic F-box genes at the S-locus. In apple, two SFBB gene groups have been described, while there are at least three groups in pear. In this report, we identified five MdSLFB (S-RNase-linked F-box) genes from four different S-genotypes of apple. These genes showed pollen- and S-allele-specific expression with a high polymorphism among S-alleles. The phylogenetic tree suggested that some of them belong to SFBBα or β groups as described previously, while others
appear to be different from SFBBs. In particular, the presence of MdSLFB3 and MdSLFB9 suggested that there are more S-allelic polymorphic F-box gene groups in the S-locus besides α and β. Based on the sequence polymorphism of MdSLFBs, we developed an S-genotyping system for apple cultivars. In addition, we isolated twelve MdSLFB-like genes, which showed pollen-specific expression without S-allelic polymorphism. 相似文献
14.
Soybean seeds contain substantial amount of diverse triterpenoid saponins that influence the seed quality, although little
is known about the physiologic functions of saponins in plants. We now describe the modification of saponin biosynthesis by
RNA interference (RNAi)-mediated gene silencing targeted to β-amyrin synthase, a key enzyme in the synthesis of a common aglycon
of soybean saponins. We identified two putative β-amyrin synthase genes in soybean that manifested distinct expression patterns
with regard to developmental stage and tissue specificity. Given that one of these genes, GmBAS1, was expressed at a much higher level than the other (GmBAS2) in various tissues including the developing seeds, we constructed two RNAi vectors that encode self-complementary hairpin
RNAs corresponding to the distinct regions of GmBAS1 under the control of a seed-specific promoter derived from the soybean gene for the α′ subunit of the seed storage protein
β-conglycinin. These vectors were introduced independently into soybean. Six independent transgenic lines exhibited a stable
reduction in seed saponin content, with the extent of saponin deficiency correlating with the β-amyrin synthase mRNA depletion.
Although some transgenic lines produced seeds almost devoid of saponins, no abnormality in their growth was apparent and the
antioxidant activity of their seeds was similar to that of control seeds. These results suggest that saponins are not required
for seed development and survival, and that soybean seeds may therefore be amenable to the modification of triterpenoid saponin
content and composition through molecular biologic approaches. 相似文献
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Łukasz Kuźma Zuzanna Skrzypek Halina Wysokińska 《Plant Cell, Tissue and Organ Culture》2006,84(2):100152-100160
Hairy root culture of Salvia sclarea L. was established following infection with Agrobacterium rhizogenes LBA 9402. The culture was grown in growth regulator-free half-strength B5 Gamborg medium with 30 g l−1 sucrose and was investigated with respect to its capability of producing diterpenoids and triterpenoids. Four diterpenoids
(ferruginol, salvipisone, aethiopinone and 1-oxoaethiopinone) and two ursene-type triterpenoids (2α,3α-dihydroxy-urs-12-en-28-oic acid and 2α,3α,24-trihydroxy-urs-12-en-28-oic acid) were isolated from the hairy roots. The presence of three sterols (β-sitosterol, stigmasterol and campesterol), as well as oleanolic and ursolic acids was also shown by GC–MS analysis. The quantitative
and qualitative differences in diterpenoid and triterpenoid production patterns between hairy roots grown in the light and
in the dark were described. 相似文献
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Consensus amino acid sequences of FADH2-dependent bacterial halogenases were used to design PCR primers amplifying a halogenase gene fragment from the chloramphenicol
producer Streptomyces venezuelae ISP5230. The sequence-specific degenerate primers (MPF1 and MPR2) were used with a touchdown PCR procedure in the first PCR-assisted
cloning of a halogenase gene fragment. In the region of the 290-bp PCR product containing the reverse primer, the deduced
amino acid sequence exhibited characteristics of a β–α–β fold present in FAD-binding sites of certain monooxygenases. When
used to probe Southern blots of restriction-enzyme-digested DNA, the [α-32P]dCTP-labeled PCR product hybridized specifically with DNA fragments from genomic DNA of S. venezuelae ISP5230. Primers MPF1 and MPR2 also allowed amplification by PCR of approximately 290-bp DNA fragments from several other
streptomycetes. The fragments from Streptomyces aureofaciens NRRL2209 and Streptomyces coelicolor A3(2) showed sequence identity with halogenase genes from these species. Thus, the PCR primers are of potential value for
amplification and subsequent isolation of actinomycete halogenase genes. Journal of Industrial Microbiology & Biotechnology (2002) 29, 1–5 doi:10.1038/sj.jim.7000263
Received 25 June 2001/ Accepted in revised form 02 April 2002 相似文献
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Wen Xian Liu Hua Liang Liu Zhi Jian Chai Xiu Ping Xu Yan Ru Song Le Qing Qu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2010,121(7):1267-1274
5′ untranslated regions (UTRs) are important sequence elements that modulate the expression of genes. Using the β-glucuronidase
(GUS) reporter gene driven by the GluC promoter for the rice-seed storage-protein glutelin, we evaluated the potential of the 5′-UTRs of six seed storage-protein
genes in enhancing the expression levels of the foreign gene in stable transgenic rice lines. All of the 5′-UTRs significantly
enhanced the expression level of the GluC promoter without altering its expression pattern. The 5′-UTRs of Glb-1 and GluA-1 increased the expression of GUS by about 3.36- and 3.11-fold, respectively. The two 5′-UTRs downstream of the Glb-1, OsAct2 and CMV35S promoters also increased GUS expression level in stable transgenic rice lines or in transient expression protoplasts. Therefore,
the enhancements were independent of the promoter sequence. Real-time quantitative RT-PCR analysis showed that the increase
in protein production was not accompanied by alteration in mRNA levels, which suggests that the enhancements were due to increasing
the translational efficiencies of the mRNA. The 5′-UTRs of Glb-1 and GluA-1, when combined with strong promoters, might be ideal candidates for high production of recombinant proteins in rice seeds. 相似文献
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The structure of the sulfated analogue of cauloside C, a biologically active triterpenoid glycoside, was elucidated to be
3-O-[β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl]-hederagenin 23,4′,4″,6″-tetrasulfate pentasodium salt by the comparison of its13C NMR spectrum with that of cauloside C potassium salt. 相似文献