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  • 1.
    Karim, Sazzad
    et al.
    University of Skövde, School of Life Sciences.
    Lundh, Dan
    University of Skövde, School of Humanities and Informatics.
    Holmström, Kjell-Ove
    University of Skövde, School of Life Sciences.
    Mandal, Abul
    University of Skövde, School of Life Sciences.
    Pirhonen, Minna
    Department of Applied Biology, University of Helsinki, Box 27, 00014 Helsinki, Finland.
    Structural and functional characterization of atPTR3, a stress-induced peptide transporter of Arabidopsis2005In: Journal of Molecular Modeling, ISSN 1610-2940, E-ISSN 0948-5023, Vol. 11, no 3, p. 226-236Article in journal (Refereed)
    Abstract [en]

    A T-DNA tagged mutant line of Arabidopsis thaliana, produced with a promoter trap vector carrying a promoterless gus (uidA) as a reporter gene, showed GUS induction in response to mechanical wounding. Cloning of the chromosomal DNA flanking the T-DNA revealed that the insert had caused a knockout mutation in a PTR-type peptide transporter gene named At5g46050 in GenBank, here renamed AtPTR3. The gene and the deduced protein were characterized by molecular modelling and bioinformatics. Molecular modelling of the protein with fold recognition identified 12 transmembrane spanning regions and a large loop between the sixth and seventh helices. The structure of AtPTR3 resembled the other PTR-type transporters of plants and transporters in the major facilitator superfamily. Computer analysis of the AtPTR3 promoter suggested its expression in roots, leaves and seeds, complex hormonal regulation and induction by abiotic and biotic stresses. The computer-based hypotheses were tested experimentally by exposing the mutant plants to amino acids and several stress treatments. The AtPTR3 gene was induced by the amino acids histidine, leucine and phenylalanine in cotyledons and lower leaves, whereas a strong induction was obtained in the whole plant upon exposure to salt. Furthermore, the germination frequency of the mutant line was reduced on salt-containing media, suggesting that the AtPTR3 protein is involved in stress tolerance in seeds during germination.

  • 2.
    Nahar, Noor
    et al.
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Rahman, Aminur
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Nawani, Neelu N.
    Microbial Diversity Research Centre, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Tathawade, Pune, India.
    Ghosh, Sibdas
    School of Arts and Science, Iona College, New Rochelle, NY, USA.
    Mandal, Abul
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Phytoremediation of arsenic from the contaminated soil using transgenic tobacco plants expressing ACR2 gene of Arabidopsis thaliana2017In: Journal of plant physiology (Print), ISSN 0176-1617, E-ISSN 1618-1328, Vol. 218, p. 121-126Article in journal (Refereed)
    Abstract [en]

    We have cloned, characterized and transformed the AtACR2 gene (arsenic reductase 2) of Arabidopsis thaliana into the genome of tobacco (Nicotiana tabacum, var Sumsun). Our results revealed that the transgenic tobacco plants are more tolerant to arsenic than the wild type ones. These plants can grow on culture medium containing 200μM arsenate, whereas the wild type can barely survive under this condition. Furthermore, when exposed to 100μM arsenate for 35days the amount of arsenic accumulated in the shoots of transgenic plants was significantly lower (28μg/g d wt.) than that found in the shoots of non-transgenic controls (40μg/g d wt.). However, the arsenic content in the roots of transgenic plants was significantly higher (2400μg/g d. wt.) than that (2100μg/g d. wt.) observed in roots of wild type plants. We have demonstrated that Arabidopsis thaliana AtACR2 gene is a potential candidate for genetic engineering of plants to develop new crop cultivars that can be grown on arsenic contaminated fields to reduce arsenic content of the soil and can become a source of food containing no arsenic or exhibiting substantially reduced amount of this metalloid.

  • 3.
    Nahar, Nour
    et al.
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Rahman, Aminur
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Ghosh, Sibdas
    School of Arts and Science, Iona College, New Rochelle, NY, USA.
    Nawani, Neelu
    Microbial Diversity Research Centre, Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Tathawade, Pune, India.
    Mandal, Abul
    University of Skövde, School of Bioscience. University of Skövde, The Systems Biology Research Centre.
    Functional studies of AtACR2 gene putatively involved in accumulation, reduction and/or sequestration of arsenic species in plants2017In: Biologia (Bratislava), ISSN 0006-3088, E-ISSN 1336-9563, Vol. 72, no 5, p. 520-526Article in journal (Refereed)
    Abstract [en]

    Food-based exposure to arsenic is a human carcinogen and can severely impact human health resulting in many cancerous diseases and various neurological and vascular disorders. This project is a part of our attempts to develop new varieties of crops for avoiding arsenic contaminated foods. For this purpose, we have previously identified four key genes, and molecular functions of two of these, AtACR2 and AtPCSl, have been studied based on both in silico and in vivo experiments. In the present study, a T-DNA tagged mutant, (SALK-143282C with mutation in AtACR2 gene) of Arabidopsis thaliana was studied for further verification of the function of AtACR2 gene. Semi-quantitative RT-PCR analyses revealed that this mutant exhibits a significantly reduced expression of the AtACR2 gene. When exposed to 100 μM of arsenate (AsV) for three weeks, the mutant plants accumulated arsenic approximately three times higher (778 μg/g d. wt.) than that observed in the control plants (235 μg/g d. wt.). In contrast, when the plants were exposed to 100 μM of arsenite (AsIII), no significant difference in arsenic accumulation was observed between the control and the mutant plants (535 μg/g d. wt. and 498 μg/g d. wt., respectively). Also, when arsenate and arsenite was measured separately either in shoots or roots, significant differences in accumulation of these substances were observed between the mutant and the control plants. These results suggest that AtACR2 gene is involved not only in accumulation of arsenic in plants, but also in conversion of arsenate to arsenite inside the plant cells. © 2017 Institute of Molecular Biology, Slovak Academy of Sciences.

  • 4.
    Svensson, Maria
    et al.
    University of Skövde, School of Life Sciences.
    Lundh, Dan
    University of Skövde, School of Humanities and Informatics.
    Bergman, Per
    Department of Plant Biology and Forest Genetics, SLU, SE-750 07 Uppsala, Sweden.
    Mandal, Abul
    University of Skövde, School of Life Sciences.
    Characterisation of a T-DNA-tagged gene of Arabidopsis thaliana that regulates gibberellin metabolism and flowering time2005In: Functional Plant Biology, ISSN 1445-4408, E-ISSN 1445-4416, Vol. 32, no 10, p. 923-932Article in journal (Refereed)
    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.

  • 5.
    Svensson, Maria
    et al.
    University of Skövde, School of Life Sciences.
    Lundh, Dan
    University of Skövde, School of Humanities and Informatics.
    Ejdebäck, Mikael
    University of Skövde, The Systems Biology Research Centre. University of Skövde, School of Life Sciences.
    Mandal, Abul
    University of Skövde, School of Life Sciences.
    Functional prediction of a T-DNA tagged gene of Arabidopsis thalianaby in silico analysis2004In: Journal of Molecular Modeling, ISSN 1610-2940, E-ISSN 0948-5023, Vol. 10, no 2, p. 130-138Article in journal (Refereed)
    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.

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