933 resultados para Arabidopsis
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RT-PCRCdCuCdCuCdCu 1 CdCu 2 Cd Cu 3 MMRPCNA MMRPCNACdCu 4 MMRPCNA CdCuCdCu
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M&SMutS 2 homolog (atMSH2)atMSH3atMSH71 2 (atPCNA1atPCNA2)-RT-PCRCd00.1250.251.03.0 mg•L-1 atMSH2Cd 1. 00.1250.251.03.0 mg•L-1 Cd7Cd 2. 00.1250.251.03.0 mg•L-1 Cd7 Cd0.125 mg•L-1 Cd0.251.03.0 mg•L-1 Cd 3. atMSH2atPCNA1atPCNA2CdU0.125mg•L-10.25mg•L-10.125mg•L-1 CdatMSH2atPCNA1atPCNA2Cd 4. 00.1250.251.03.0 mg•L-1 Cd7atMSH2PCR0.25 mg•L-1 CdatMSH2891.0 mg•L-1 Cd10
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205070(Dasypyrum villosum2n14VV)KO20(GA20ox)203-(GA3ox)GA20ox1. 110cm32cm1/3 I2-KI90%20%2. F1F1168F280cm40cm13038313. ELISAGA1336 ng/ml900 ng/ml4. PCR20KO616GA20ox105. GenBankEU142950RT-PCRGA3oxcDNAcDNA1206bp1104bp36840.063KD6.272FeGA3ox98%96%86%cDNA478-715bp879-1019bp238bp140bp6. RT-PCRGA20oxDvGA20oxGenBankEU1429491080359GA20oxGA20ox98%97% 91%pET-32a(+)pET-32a(+)-DvGA20oxBL21pLysSIPTGSDS-PAGEDvGA20ox55kDaPCR(1)(2)1/30(3)PCR(4)GA20ox Plant height is an impotrant agronomic trait of triticeae crops.Semi-dwarf cropcultivars, including those of wheat, maize and rice, have significantly increased grainproduction that has been known as green revolution. The new dwarf varieties couldraise the harvest Index at the expense of straw biomass, and, at the sametime, improvelodging resistance and responsiveness to nitrogen fertilizer. Moreover, dwarf traits ofplant are crucial for elucidating mechanisms for plant growth and development aswell. In many plant species, various dwarf mutants have been isolated and theirmodles of inheritance and physiology also have been widely investigated.The causesfor their dwarf phenotypes were found to be associated with plant hormones,especially, gibberellins GAs.Dasypyrum villosum Candargy (syn.Haynaldia villosa) is a cross-pollinating,diploid (2n = 2x = 14) annual species that belongs to the tribe Triticeae. It is native toSouthern Europe and West Asia, especially the Caucasuses, and grows underconditions unfavorable to most cultivated crops. The genome of D. villosum,designated V by Sears, is considered an important donor of genes to wheat for improving powdery mildew resistance, take-all, eyespot, and plant and seed storageprotein content. A spontaneous dwarf mutant was found in D. villosum populations.The biological character and modles of inheritance of this dwarf mutant are studied.The cell length of stem cell is observed. The influence of extraneous gibberellin tothe dwarf mutant is also examined; the transcript level of key enzyme of gibberellinbiosynthesis pathway in mutant and control plants is compared. GA3ox and GA20oxare cloned and its expression pattern is researched.1. The dwarf mutant showed no difference with control plants at seedlingstage.At mature stage, the average height of control plants were 110cm and the dwarfplants were 33cm. The height of the mutant plant was only one third of the normalplants due to the shortened internodes. Cytology observation showed that theelongation of stem epidermal and the parenchyma cells were reduced. The dwarfmutant also shows partly male sterile. Pollen viability test indicates that more than80% of the pollen of the mutant is not viable.2. The inheritance modle of this dwarf mutant is studied. All The F1 plantsshowed normal phenotype indicating that the dwarfism is controlled by recessivealleles. Among the 168 F2 plants, there are 130 normal plants and 30 dwarf plants, thesegregation proportion accord with Mendels 3:1 segregation. We therefore proposethat this dwarf phenotype is controlled by a single recessive gene.3. Quantitative analyses of endogenous GA1+3 in the young seeds indicated thatthe content of GA1+3 was 36ng/ml in mutant plants and 900ng/ml in normal plants.The endogenous bioactive GA1+3 in mutant plants are only about 1/30 of that innormal plants. In addition, exogenously supplied GA3 could considerably restore themutant plant to normal phenotype. These results showed that this mutant wasdefective in the GA biosynthesis.4. More than ten enzymes are involved in GA biosynthesis. KO catalyzes thefirst cytochrome P450-mediated step in the gibberellin biosynthetic pathway and themutant of KO lead to a gibberellin-responsive dwarf mutant. GA20ox catalyze therate-limited steps so that their transcript level will influence the endogenous GAbiosynthesis and modifies plant architecture. The relative expression levels of genesencoding KO and GA20ox were quantified by real time PCR to assess whether thechanges in GA content correlated with the expression of GA metabolism genes andwhere the mutant occurred during the GA biosynthesis pathway. In mutant plants,the transcript levels of KO increased about 6-fold and 16-fold at the seedling stage and elongating stage respectively comparing with the normal plants. For theseedlings, there was no notable difference in the expression of GA20ox betweenmutant and normal plants. At the elongating stage, GA20ox transcript increased 10times in mutant plants, suggesting that the GA biosynthesis pathway in mutant plantshad changed from the early steps rather than the late steps.5. A full length cDNA of D. villosum gibberellin 3-hydroxylase homology(designated as DvGA3ox) was isolated and consisted of 1206bp containing an openreading frame of 1104bp encoding 368 predicted amino acid residues. Identityanalysis showed that the gibberellin 3-hydroxylase nucleotide sequence shared 98%,96% and 86% homology with that of wheat, barley and rice. The predicted peptidecontained the active-site Fe of known gibberellin 3-hydroxylase and the regionhomologous to wheat, barley and Arabidopsis. The genomic clone of gibberellin3-hydroxylase has two introns.6. The full-length cDNA of D. villosum gibberellin 20 oxidase (designated asDvGA20ox) was isolated and consisted of 1080-bp and encoded 359 amino acidresidues with a calculated mol wt of 42.46 KD. Comparative and bio-informaticsanalyses revealed that DvGA20ox had close similarity with GA20ox from otherspecies and contained a conserved LPWKET and NYYPXCQKP regions. Tissueexpression pattern analysis revealed DvGA20ox expressed in all the tissues that wereexamined and the highest expression of DvGA20ox in expanding leaves followed byroots. Heterologous expression of this cDNA clone in Escherichia coli gave a fusionprotein that about 55KD. Transcript levels of DvGA20ox dramatically reduced twohours after application of biologically active GA3, suggesting that the biosynthesis ofthis enzymes might be under feedback control.
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In order to investigate the biological effects of heavy ion radiation at low closes and the different radiosensitivities of growing and non-growing plants. rice at different lift stages (dry seed, wet seed and seedling) were exposed to carbon ions at closes of 0 02, 0.2, 2 and 20 Gy. Radiobiological effects on survival, root growth and mitotic activity, as well as the induction of chromosome aberrations in root meristem. were observed The results show that radiation exposure induces a stimulatory response at lower close and an inhibitory response at higher dose on the mitotic activity of wet seeds and seedlings Cytogenetic damages are induced in both seeds and seedlings by carbon ion radiation at doses as low as 0.02 Gy Compared with seedlings. seeds are more resistant to the lethal damage and the growth rate damage by high doses of carbon ions, but are more sensitive to cytogenetic damage by low closes of irradiation Different types of radiation induced chromosome aberrations are observed between seeds and seedlings. Based on these results, the relationships between low close heavy ion-induced biological effects and the biological materials are discussed.
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(RT-PCR)Cd(Arabidopsis thaliana),,Cd:(0.250.51.0 mg.L-1)Cd;Cd0.25 mg.L-1,(P<0.05),Cd0.51.0 mg.L-1,(P<0.05);Cd(P>0.05)Cd0.25 mg.L-1,18S rRNA,PCNA1PCNA2MSH2MSH3MSH6MSH7 6,Cd1.0 mg.L-1,MSH6MSH3,,0.5 mg.L-1Cd,Cd
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The dichloromethane dehalogenase gene (dcm) of Methylophilus was cloned into the plasmid pKYLX71-35S2 and fused with the 35S2 promotor,then transformed Arabidopsis by the infiltration mediated with Agrobacterium tumefaciens. Homozygous dcm seeds were obtained after several generations selection on the medium with 50 mg L -1 kanamycin.Northern blotting showed dcm mRNA was high in cytoplasm and the detection of DCM enzyme indicated that the lines containing high dcm mRNA expressed high DCM enzyme activities. Fig 4, Ref 9
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The study on the response of a mutant and a wild type of Arabidopsis to 660 lL -1 CO 2 and ambient CO 2 showed that under elevated CO 2,the stomatal numbers of the mutant increased,while those of the wild type decreased. The chlorophyll content and NR (nitrate reductase) activity of the mutant increased,but those of the wild type had no obvious response. The mutant was not reproductively mature after the continuous exposure to increased CO 2 for five months. The results provided evidence of plant response to the changes of atmospheric CO 2 concentration,and the clues to related studies on other plants.
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RNACTAB[2% (w/v) CTAB,100 mM Tris-HC1 (pH 8.0), 50 mM EDTA, pH 7.5, 2 M NaCl, 50 mM DTT]RNA (1) 50 mM DTT(2) 1/4 1/9 3 M pH 4.8(3) 1/103 mol/LpH 5.0RNA(4) -8030 minRNACTABRNAA260/280 =1.960.05)68 g/gRNART-PCRRNA (Arabidopsis thaliana) (Chlamydomonas reinhardtii) (Adiantum capillus-veneris)(Mougeotia scalaris)(Vaucheria frigida)RT-PCR4783 bp-1-RACE35
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Janet Taylor, Ross D King, Thomas Altmann and Oliver Fiehn (2002). Application of metabolomics to plant genotype discrimination using statistics and machine learning. 1st European Conference on Computational Biology (ECCB). (published as a journal supplement in Bioinformatics 18: S241-S248).
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Enot, D. P., Beckmann, M., Overy, D., Draper, J. (2006). Predicting interpretability of metabolome models based on behavior, putative identity, and biological relevance of explanatory signals. Proceedings of the National Academy of Sciences of the USA, 103(40), 14865-14870. Sponsorship: BBSRC RAE2008
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Igor E. Moshkov, Galina V. Novikova, Luis A.J. Mur, Aileen R. Smith, and Michael A. Hall. (2003). Ethylene rapidly up-regulates the activities of both monomeric GTP-binding proteins and protein kinase(s) in epicotyls of pea. Plant Physiology, 131(4), 1718-1726 RAE2008
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Armstead, I. P., Donnison, I. S., Aubry, S., Harper, J. A., H?rtensteiner, S., James, C. L., Mani, J., Moffet, M., Ougham, H. J., Roberts, L. A., Thomas, A. M., Weeden, N., Thomas, S., King, I. P. (2007). Cross-species identification of Mendel's/locus. Science, 315 (5808), 73. Sponsorship: BBSRC RAE2008
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John Draper, Luis A.J. Mur, Glyn Jenkins, Gadab C. Ghosh-Biswas, Pauline Bablak, Robert Hasterok,and Andrew P.M. Routledge (2001). Brachypodium distachyon. A new model system for functional genomics in grasses. Plant Physiology, 127 (4), 1539-1555. Sponsorship: BBSRC / Gatsby Foundation RAE2008
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Wydzia Biologii
