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DNA barcoding provides distinction between Radix Astragali and its adulterants

Science China(Life Sciences) 2010年第8期53卷 992-999页

作者： GUO HaiYan1,2, WANG WeiWei2, YANG Ning2, GUO BaoLin3, ZHANG Sun2, YANG RuiJing2, YUAN Ye2, YU JunLin4, HU SongNian2, SUN QiShi1 & YU Jun2 1School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China 2Key laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 101300, China 3Chinese Academy of Medical Sciences, Peking Union Medical College Institute of Medicinal plant Development, Beijing 100094, China 4Department of Chemistry, Tonghua Normal College, Tonghua 134002, China 1. School of Traditional Chinese Materia Medica Shenyang Pharmaceutical University Shenyang 110016 China2. Key Laboratory of Genome Sciences and Information Beijing Institute of Genomics Chinese Academy of Sciences Beijing 101300 China3. Peking Union Medical College Institute of Medicinal Plant Development Chinese Academy of Medical Sciences Beijing 100094 China4. Department of Chemistry Tonghua Normal College Tonghua 134002 China

Based on variable nuclear and/or organellar DNA sequences among vastly divergent species as well as morphologically indistinguishable species, DNA barcoding is widely applicable in species identification, biodiversity studies, forensic analyses, and authentication of medicinal plants. The roots of Astragalus membranaceus and A. membranaceus var. mongholica are commonly used as Radix Astragali in several Asian countries, including China, Japan, and Korea. However, in addition to the two species recorded in the Chinese Pharmacopoeia, there are twenty-three species from different genera including Astragalus, Oxytropis, Hedysarum, and Glycyrrhiza, which have been used as adulterants not only in trading markets but also by the herbal medicine industry. Therefore, a simple, reliable, and accurate classification method is important for distinguishing authentic Radix Astragali from its adulterants. In this study, we acquired data for 37 samples from four related genera within the family Fabaceae. Then we compared four candidate DNA barcoding markers using ITS, matK, rbcL, and coxI sequences from nuclear, chloroplast, and mitochondrial genomes, all commonly used for plants to identify genetic variations among genera, intraspecies, and interspecies. We observed higher divergences among genera and interspecies for ITS, which have the average Kimura 2-parameter distances of 4.5% and 14.1%, respectively, whereas matK was found to have sufficient divergence at the intraspecific level. Moreover, two indels detected in the matK sequence are useful for PCR studies in distinguishing Radix Astragali from its adulterants. This study suggests that the combined barcoding regions of ITS and matK are superior barcodes for Radix Astragali and further studies should focus on evaluating the applicability and accuracy of such combined markers for a wide range of traditional Chinese herbs.

关键词： Radix Astragali DNA barcode Astragalus adulterant identification

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New insights into the species problem

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Science China(Life Sciences) 2010年第8期53卷 964-972页

作者： YANG Yong1 & ZHOU ZheKun2 1State Key laboratory of Systematic and Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China 2Key laboratory of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650204, China 1. State Key laboratory of Systematic and Evolutionary Botany Institute of Botany Chinese Academy of Sciences Beijing 100093 China2. Key Laboratory of Biodiversity and Biogeography Kunming Institute of Botany Chinese Academy of Sciences Kunming 650204 China

Historical opinions of the "species problem" are briefly reviewed, and four salient stages are recognized according to origin of species concepts. We propose that species is the unit preserving superior gene assembly and is maintained by specific mechanisms. Based on characteristics of plant evolution, we assume that understanding plant species may include three stages, i.e. morphological recognition stage, multidisciplinary verification stage, and illuminating mechanisms preserving superior gene assembly.

关键词： species concept plant evolution phylogeny

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Cloning,expression and binding specificity analysis of odorant binding protein 3 of the lucerne plant bug,Adelphocoris lineolatus(Goeze)

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Science Bulletin 2010年第34期55卷 3913-3923页

作者： GU ShaoHua1,SUN Yang1,2,REN LiYan1,2,ZHANG XueYing1,ZHANG YongJun1,WU KongMing1 & GUO YuYuan1 1State Key laboratory for Biology of plant Diseases and Insect Pests,Institute of plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China 2College of Agriculture and Biotechnology,China Agricultural University,Beijing 100193,China

Credible evidence shows that odorant binding proteins(OBPs)are required for insect olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum lymph to the olfactory receptors(ORs).In the present study,a novel OBP(AlinOBP3)gene from the lucerne plant bug,Adelphocoris lineolatus,was cloned and *** expression pattern of AlinOBP3 was evaluated by qPCR,which indicated that AlinOBP3 was dominantly expressed in *** binding properties of AlinOBP3 with 9 cotton volatiles and 5 sex pheromone analogs were measured by fluorescence competitive binding assays with the fluorescence probe *** results revealed that of 9 cotton volatiles,Myrcene,β-Ocimene and α-Phellandrene can bind with AlinOBP3.α-Phellandrene especially bound to AlinOBP3 with a high binding affinity,with a dissociation constant of 56.68 μmol/*** the 5 sex pheromone analogs,Hexyl butyrate had the strongest binding affinity with AlinOBP3,with a dissociation constant as 59.53 μmol/*** butyrate,trans-2-Hexenyl butyrate and Ethyl butyrate had medium binding affinities with AlinOBP3,with dissociation constants of 227.39,108.77 and 143.47 μmol/L,*** results suggest that AlinOBP3 might be a pheromone binding protein(PBP)with a dual-function for the perception of sex pheromones and plant volatiles.

关键词： Adelphocoris lineolatus(Goeze) odorant binding protein gene expression Western blot expression pattern fluorescence competitive binding assays

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plant fertility defects induced by the enhanced expression of microRNA167

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Cell Research 2006年第5期16卷 457-465页

作者： Peng Ru~1 Lin Xu~1 Hong Ma~(2,3) Hai Huang~(1,2) ~1National laboratory of plant Molecular Genetics,Shanghai Institute of plant Physiology and Ecology,Shanghai Institute for Biological Sciences,300 Fenglin Road,Shanghai 200032,China ~2SIBS-SJTU-PSU Joint Center for Life Sciences,Shanghai Institute of plant Physiology and Ecologgy,Shanghai Institute for Biological Sciences,300 Fenglin Road,Shanghai 200032,China ~3Department of Biology and the Huck Institute for the Life Sciences,The Pennsylvania State University,University Park,PA 16802,USA NationallaboratoryofplantMolecularGenetics ShanghaiInstituteofPlantPhysiologyandEcologyShanghaiInstituteforBiologicalSciences300FenglinRoadShanghai200032China SIBS-SJTU-PSUJointCenterforLifeSciences

The plant hormone auxin plays a critical role in regulating plant growth and *** advances havebeen made in the understanding of auxin response pathways,primarily by the characterization of auxin response mu-tants in *** addition,microRNAs (miRNAs) have been shown to be critical regulators of genes importantfor normal plant development and ***,little is known about possible interactions between miRNAsand hormonal signaling during norlnal *** we show that an Arabidopsis microRNA,miR167,whichhas a complementary sequence to a portion of the A UXIN RESPONSE FACTOR6 (ARF6) and ARF8 mRNAs,can causetranscript degradation for ARFS,but not for *** report phenotypic characterizations of35S::MIR167b transgeniclines,and show that severe 35S::MIR167b transgenic lines had phenotypes similar to those of an arf6 arf8 double *** transgenic phenotypes suggest that miR167 may repress ARF6 at the level of *** demonstrate that thetransgenic plants are defective in all four whorls of floral *** the transgenic flowers,filaments were abnormallyshort,anthers could not properly release pollen,and pollen grains did not *** results provide an importantlink between the miRNA-mediated regulatory pathway of gene expression and the auxin signaling network promotingplant reproductive development.

关键词： Arabidopsis auxin response factor microRNA167 floral organ pollen germination

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The Inhibitory Effect of Quercetin on IL-6 Production by LPS-Stimulated Neutrophils

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Cellular & Molecular Immunology 2005年第6期2卷 455-460页

作者： Jiajia Liu~(1,2) Xin Li~1 Yang Yue~1 Juan Li~1 Tao He~1 Yanzheng He~1 ~1laboratory of Molecular Biology,Luzhou Medical College,Luzhou 646000,Sichuan,China ~2laboratory of Molecular Biology,Luzhou Medical College,Luzhou 646000,Sichuan,China. laboratory of Molecular Biology Luzhou Medical College Luzhou 646000 Sichuan China.

Quercetin is a herbal flavonoid derived from various foods of plant origin and plays a role in anti-inflammation. Although a number of researches in the field have been done, the mechanism of anti-inflammatory effect of quercetin should be further darified. In the present study, we investigated the effects of quercetin on IL-6 production by LPS-stimulated neutrophils in human. Neutrophils were were pre-treated with quercetin at the final concentrations of ranging from 0-80 ~M for 30 rain, or not treated, and then incubated in the presence or absence of lipopolysaccharide （LPS） at a final concentration of 100 ng/ml for indicated time. The secretion level of IL-6 in the culture supernatants was assayed by ELISA, the intracellular level of IL-6 was detected by flow cytometry and the expression of IL-6 mRNA was analyzed by RT-PCR. The experiment results showed that neutrophils cultured with medium or quercetin alone did not express IL-6, but LPS （100 ng/ml） induced IL-6 expression of neutrophils. However, after pre-treatment of neutrophils with quercetin （40 ~Vl） for 30 rain, the inducible effects of LPS on the increase of IL-6 secretion, intracellular IL-6 level and IL-6 mRNA expression by neutrophils were abrogated. IL-6 is one of the important pro-inflammatory factors, especially in early phage of inflammation. Thus, our data suggested that quercetin might exert its anti-inflammatory effect through negatively modulating pro-inflammatory factors, such as IL-6. The inhibitory effects of quercetin on IL-6 production by neutrophils may provide a theoretical basis on future therapy of inflammation. Cellular ＆ Molecular Immunology. 2005;2（6）：455-460.

关键词： quercetin neutrophil IL-6 LPS inflammation

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Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: silencing of the tomato ferric chelate reductase gene, FRO1, as an example

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Science China(Life Sciences) 2008年第5期51卷 402-409页

作者： HE XiuXia1,2,JIN ChongWei1, LI GuiXin3, YOU GuangYi4, ZHOU XuePing3 & ZHENG ShaoJian4 1 Key laboratory for Environmental and Ecosystem Health of the Ministry of Education, College of Environmental and Resource Science, Zhejiang University, Hangzhou 310029, China 2 College of Life Science, Changchun University of Science and Technology, Changchun 130022, China 3 Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China 4 State Key laboratory of plant Physiology and Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China 1. Key laboratory for Environmental and Ecosystem Health of the Ministry of Education College of Environmental and Resource Science Zhejiang University Hangzhou 310029 China 2. College of Life Science Changchun University of Science and Technology Changchun 130022 China 3. Institute of Biotechnology Zhejiang University Hangzhou 310029 China 4. State Key laboratory of plant Physiology and Biochemistry College of Life Science Zhejiang University Hangzhou 310058 China

Virus-induced gene silencing (VIGS) is potentially an attractive reverse-genetics tool for studies of plant gene function, but whether it is effective in silencing mineral nutritional-related genes in roots has not been demonstrated. Here we report on an efficient VIGS system that functions in tomato roots using a modified viral satellite DNA (DNAmβ) associated with Tomato yellow leaf curl China virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FRO1) from tomato was inserted into the DNAmβ vector. Tomato roots agro-inoculated with DNAmβ carrying both a fragment of FRO1 and TYLCCNV used as a helper virus exhibited a significant reduction at the FRO1 mRNA level. As a consequence, ferric chelate reductase activity, as determined by visualization of the pink FeBPDS3 complex was significantly decreased. Our results clearly demonstrated that VIGS system can be employed to investigate gene function associated with plant nutrient uptake in roots.

关键词： virus-induced gene silencing roots ferric chelate reductase

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Comparing Genetic Characteristics of Retrotransposon TOS17 During Different Tissue Culture Processes in the Rice Cultivars Nipponbare and Shishoubaimao

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Agricultural Sciences in China 2010年第2期9卷 157-162页

作者： LI Kuan, WANG Chuang, YING Shan and SHOU Hui-xia State Key laboratory of plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, P.R.China [a] State Key laboratory of plant Physiology and Biochemistry College of Life Sciences Zhejiang University Hangzhou 310058 P.R. China

As a retrotransposon, TOS17 was a useful tool for rice genetic and functional genomic research. To ascertain the feasibility of constructing a TOS17 insertion mutation library in the rice cultivar Shishoubaimao, the genetic and expression characteristics of TOS17 were analyzed. We made solid and suspension tissue cultures and confirmed the copy numbers of TOS17 at different time points in both tissue culture processes by real-time quantitative PCR （RT-qPCR）. Three primary copies of TOS17 were detected in naturally grown Shishoubaimao. TOS17 was activated by tissue culture, and the copy numbers of TOSI7 increased along with a prolonged tissue culture time in both the Nipponbare and the Shishoubaimao cultivars. Therefore, Shishoubaimao is a potential candidate for constructing a TOS17 insertion mutant library. Compared with Nipponbare, TOS17 was more active in Shishoubaimao during tissue culture. Higher copy numbers of TOS17 were obtained with the suspension tissue culture process than with the solid tissue culture process over the same time courses. We concluded that 3-4 months of suspension tissue culture time is suitable for constructing a TOS17 insertion mutant library in Shishoubaimao.

关键词： Shishoubaimao tissue culture TOS17 copy number insertion mutant library

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Cloning and characterization of two putative seven-transmembrane receptor genes from cotton

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Progress in Natural Science:Materials International 2008年第10期18卷 1225-1231页

作者： Peng Gao Piming Zhao Juan Wang Haiyun Wang Guiling Wang Guixian Xia National Key laboratory of plant Genomics,Institute of Microbiology,Chinese Academy of Sciences,Datun Road,Chaoyang District,Beijing 100101,China National Center for plant Gene Research,Beijing 100101,China National Key laboratory of plant Genomics Institute of Microbiology Chinese Academy of Sciences Datun Road Chaoyang District Beijing 100101 China National Center for plant Gene Research Beijing 100101 China

Using rapid amplification of cDNA ends (RACE)-PCR,two full-length cDNAs encoding putative seven-transmembrane receptors (designated Gh7TMpRI and Gh7TMpR2) were cloned from cotton *** blot and an ApaL1 restriction site polymorphism analyses revealed that Gh7TMpRI was derived from the ancestral A diploid genome,while Gh7TMpR2 was from the D subgenome. Northern blot hybridization indicated that both Gh7TMpR1 and GhTTMpR2 were expressed preferentially in the elongation phase of fiber *** of the Gh7TMpRl proteins were located within the membrane structure and displayed a punctuate pattern of *** of Gh7TMpR1 in fission yeast disrupted the polar growth and caused the formation of rounded cells. These results suggest that GhT7MpRl may play a critical role in cotton fiber development,perhaps as a signaling receptor that is involved in controlling fiber elongation.

关键词： Cotton Seven-transmembrane protein Fiber elongation Alloalleles

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Analysis of Five Differentially Expressed Gene Families in Fast Elongating Cotton Fiber

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 51-56页

作者： Jian-Xun FENG, Sheng-Jian JI, Yong-Hui SHI, Yu XU, Gang WEI, and Yu-Xian ZHU( National laboratory of Protein Engineering and plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China ) National laboratory of Protein Engineering and plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China

Using the suppression subtractive hybridization method, we isolated five gene families, including proline-rich proteins (PRPs), arabinogalactan proteins (AGPs), expansins, tubulins and lipid transfer proteins (LTPs), from fast elongating cotton fiber cells. Expression profile analysis using cDNA array technology showed that most of these gene families were highly expressed during early cotton fiber developmental stages (0-20 days post anthesis, DPA). Many transcripts accumulated over 50-fold in 10 DPA fiber cells than in 0 DPA samples. The entire gene family -AGP, together with 20 individual members in other 4 gene families, are reported in cotton for the first time. Accumulation of cell wall proteins, wall loosening enzymes, microtubules and lipid transfer proteins may contribute directly to the elongation and development of fiber cells.

关键词： cotton fiber cDNA array cell elongation arabinogalactan protein lipid transfer protein

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plantlet regeneration from mature zygotic embryos andembryonic explants of masson pine (Pinus massoniana Lamb.)

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Cell Research 1994年第2期4卷 155-162页

作者： HUANG JIANQIU ZHIMING WEI WEI ZHOU ZHIHONG XU.(National laboratory of plant Molecular Genetics, Shanghai Institute of plant Physiology, Academia Sinica, Shanghai 200032,China) National laboratory of plant Molecular Genetics Shanghai Institute of Plant Physiology Academia Sinica Shanghai China

Excised zygotic embryos, cotyledons and hypocotyls of juvenile seedlings of masson pine were grown on DCR medium supplemented with several concentrations of various plant phytohormones. BA (1.0 mg/ L) in combination with NAA (0.05 mg/L)in DCR medium was found to increase the formation of adventitious buds from mature zygotic embryos, but most of them were formed at the tips of embryonic cotyledons. Adventitious buds were obtained from cotyledons and hypocotyls from juvenile seedlings when they were cultured on DCR medium containing BA 3-5 mg/L and NAA 0.1-0.2 mg/L. Elongation of buds were observed on hormone-free DCR medium with or without activated charcoal (0.5%). Root initiation was achieved with full or half strength DCR inedium supplemented with IBA 1.0 mg/L and NAA 0.25-0.5 mg/L. Approximately 11-20 axillary buds formed on each explant when juvenile seedling explants were treated (3-20h) with BA 50-100 mg/L, followed by transfer to hormone-free DCR medium. The maximum number of shoots obtained per explant within six months was 33.

关键词： Pinus massoniana Lamb. DCR medium mature zygotic embryos embryonic explants plantlet regeneration

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