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  • 1.
    Nahar, Noor
    et al.
    Högskolan i Skövde, Forskningscentrum för Systembiologi. Högskolan i Skövde, Institutionen för vård och natur.
    Rahman, Aminur
    Högskolan i Skövde, Forskningscentrum för Systembiologi. Högskolan i Skövde, Institutionen för vård och natur.
    Mós, Maria
    University of Agriculture in Krakow.
    Warzecha, Tomasz
    University of Agriculture in Krakow.
    Algerin, Maria
    Högskolan i Skövde, Forskningscentrum för Systembiologi. Högskolan i Skövde, Institutionen för vård och natur.
    Ghosh, Sibdas
    Dominican University of California.
    Johnson-Brousseau, Sheila
    Dominican University of California.
    Mandal, Abul
    Högskolan i Skövde, Forskningscentrum för Systembiologi. Högskolan i Skövde, Institutionen för vård och natur.
    In silico and in vivo studies of an Arabidopsis thaliana gene, ACR2, putatively involved in arsenic accumulation in plants2012Ingår i: Journal of Molecular Modeling, ISSN 1610-2940, E-ISSN 0948-5023, Vol. 18, nr 9, s. 4249-4262Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Previously, our in silico analyses identified four candidate genes that might be involved in uptake and/or accumulation of arsenics in plants: arsenate reductase 2 (ACR2), phytochelatin synthase 1 (PCS1) and two multi-drug resistant proteins (MRP1 and MRP2) [Lund et al. (2010) J Biol Syst 18:223–224]. We also postulated that one of these four genes, ACR2, seems to play a central role in this process. To investigate further, we have constructed a 3D structure of the Arabidopsis thaliana ACR2 protein using the iterative implementation of the threading assembly refinement (I-TASSER) server. These analyses revealed that, for catalytic metabolism of arsenate, the arsenate binding-loop (AB-loop) and residues Phe-53, Phe-54, Cys-134, Cys-136, Cys-141, Cys-145, and Lys-135 are essential for reducing arsenate to arsenic intermediates (arsenylated enzyme-substrate intermediates) and arsenite in plants. Thus, functional predictions suggest that the ACR2 protein is involved in the conversion of arsenate to arsenite in plant cells. To validate the in silico results, we exposed a transfer-DNA (T-DNA)-tagged mutant of A. thaliana (mutation in the ACR2 gene) to various amounts of arsenic. Reverse transcriptase PCR revealed that the mutant exhibits significantly reduced expression of the ACR2 gene. Spectrophotometric analyses revealed that the amount of accumulated arsenic compounds in this mutant was approximately six times higher than that observed in control plants. The results obtained from in silico analyses are in complete agreement with those obtained in laboratory experiments.

  • 2.
    Svensson, Maria
    Högskolan i Skövde, Institutionen för vård och natur.
    Studies of genes involved in regulating flowering time in Arabidopsis2006Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Transition from a vegetative growth phase to flowering in plants occurs in response to both environmental conditions and endogenous signals. Identification of genes that are involved in regulating the time of flowering is of great importance in agri- and horticulture. Flowering-time genes can be used for crop improvement by, for instance, engineering plants to flower earlier. This shortening of the time to flowering could result in an extended growing season that could enable farmers to grow more than one crop each year. In this work, a gene knockout approach using T-DNA tagging and in vivo gene fusion has been employed to identify and characterise genes that are involved in regulating flowering time in the model plant Arabidopsis thaliana. This approach resulted in the identification of two genes, At4g20010 and its homologue At1g31010. Expression studies and GUS histochemical analysis of a reporter gene revealed that At4g20010 is mainly expressed in rapid growing tissues such as root tips, shoot apex, flowers and stem nodes. T-DNA insertional mutants of At4g20010 and At1g31010 exhibit a late-flowering phenotype that can largely be repressed by application of gibberellin. Plants with an insertional mutation in At4g20010 contain a reduced amount of the bioactive gibberellin GA4 compared to wild-type plants. The decreased level of GA4 is not due to a transcriptional repression of the GA-biosynthetic genes AtGA3ox1 or AtGA20ox1, since their expressions were increased in the mutant plants. In silico analyses revealed that the C-terminal protein sequences encoded by At4g20010 and At1g31010 contain RNA-binding motifs, whereas the N-terminal sequences have three-dimensional structures similar to single stranded nucleic acid-binding proteins. To conclude, At4g20010 and At1g31010 may encode two RNA-binding proteins that are involved in regulating flowering time in A. thaliana by affecting the metabolism of GA. This can be possible either by a positive regulation of GA3ox at the post-transcriptional level or by a negative regulation of GA2ox.

  • 3.
    Svensson, Maria
    et al.
    Högskolan i Skövde, Institutionen för vård och natur.
    Lundh, Dan
    Högskolan i Skövde, Institutionen för kommunikation och information.
    Bergman, Per
    Department of Plant Biology and Forest Genetics, SLU, SE-750 07 Uppsala, Sweden.
    Mandal, Abul
    Högskolan i Skövde, Institutionen för vård och natur.
    Characterisation of a T-DNA-tagged gene of Arabidopsis thaliana that regulates gibberellin metabolism and flowering time2005Ingår i: Functional Plant Biology, ISSN 1445-4408, E-ISSN 1445-4416, Vol. 32, nr 10, s. 923-932Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    A gene (At4g20010) involved in regulating flowering time in Arabidopsis thaliana (L.) Heynh. was identified by promoter trap T-DNA tagging. Plants containing a T-DNA insert in the 3′-UTR of At4g20010 flowered later under both long- and short-day conditions compared with control plants. Histochemical assays of the mutant plants showed that the promoterless gus gene is expressed predominantly in the shoot apex, but it is also expressed in root tips, stem nodes and in the abscission zone of developing siliques. Measurement of endogenous gibberellin (GA) showed that bioactive GA4 levels in mutant plants were reduced compared with wild type (WT) plants. Like other known mutants defective in GA biosynthesis, the late-flowering phenotype observed in our T-DNA-tagged line could be largely repressed by application of exogenous GA3. The T-DNA-tagged gene At4g20010 encodes a previously uncharacterised protein belonging to the DUF731 family. Sequence analysis showed similarity to a single-stranded binding domain and to an RNA-binding protein of Chlamydomonas reinhardtii. Considering the above results (sequence similarity, mutant phenotype and level of endogenous GA), we propose that At4g20010 is an RNA-binding protein involved in regulating GA biosynthesis, possibly at the post-transcriptional level.

  • 4.
    Svensson, Maria
    et al.
    Högskolan i Skövde, Institutionen för vård och natur.
    Lundh, Dan
    Högskolan i Skövde, Institutionen för kommunikation och information.
    Ejdebäck, Mikael
    Högskolan i Skövde, Forskningscentrum för Systembiologi. Högskolan i Skövde, Institutionen för vård och natur.
    Mandal, Abul
    Högskolan i Skövde, Institutionen för vård och natur.
    Functional prediction of a T-DNA tagged gene of Arabidopsis thalianaby in silico analysis2004Ingår i: Journal of Molecular Modeling, ISSN 1610-2940, E-ISSN 0948-5023, Vol. 10, nr 2, s. 130-138Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    We have employed a gene-knockout approach using T-DNA tagging and in vivo gene fusion in Arabidopsis thaliana for identification and isolation of specific plant genes. Screening of about 3,000 T-DNA tagged lines resulted in identification of a mutant line (no. 197) exhibiting a significant delay in flowering. From this line a 600-bp plant DNA fragment downstream of the left T-DNA junction was cloned by inverse PCR. BLAST searching in the A. thaliana genomic database indicated a putative gene, frf (flowering regulating factor), with unknown function downstream of the T-DNA insert. Bioinformatic tools were used to predict possible protein structure and function. The protein structure predicted by fold recognition indicates that frf is a transcriptional regulator, a ligand-binding receptor responsive to steroids and hormones. Analyzing the predicted results and the phenotype of the T-DNA tagged plant we hypothesized that FRF might be involved in hormone response in A. thaliana. For verification of this hypothesis we exposed the plants of line no. 197 to gibberellic acid (GA3), a potential growth regulator in higher plants. This treatment resulted in an earlier onset of flowering, almost similar to that in wild type control plants.

  • 5.
    Wallenhammar, Ann-Charlotte
    et al.
    HS Konsult AB, Örebro, Sweden.
    Algerin, Maria
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    Tilevik, Diana
    Högskolan i Skövde, Institutionen för biovetenskap. Högskolan i Skövde, Forskningscentrum för Systembiologi.
    Ny metod bedömer risk för bomullsmögel2017Ingår i: Arvensis, ISSN 2000-0871, nr 3Artikel i tidskrift (Övrigt vetenskapligt)
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