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Hedayati P.,Institute of Systems Biology INBIOSIS | Monfard H.H.,Institute of Systems Biology INBIOSIS | Hwang D.J.,National Academy of Agricultural Science | Uddin M.I.,Institute of Systems Biology INBIOSIS | And 2 more authors.
Acta Physiologiae Plantarum | Year: 2015

To identify salt-responsive genes, we constructed a cDNA subtractive library from a salt-tolerant rice cultivar, MR219. Treatments of the whole plant were carried out with 150 mM sodium chloride for 0, 2, 4, 8, 12 and 24 h. The extraction of total RNA from 14-day-old seedlings was conducted on the shoots and roots of both treated and untreated plants. The treated plants were utilized as ‘drivers’ while the untreated plants were utilized as ‘testers’. Equal amounts of the extracted total RNA from the 12 samples were pooled together and subjected to mRNA isolation. A reverse transcription of tester and driver cDNAs from the mRNA of two pools of the samples was conducted. In total, 300 clones were sequenced and analysed using bioinformatics tools. The 260 high-quality sequences were assembled and 83 contigs and 42 singletons were generated, producing a total of 125 UTs. The majority of the UTs showed significant homology with sequences in the NCBI database. A total of 330 gene ontology terms were grouped into three categories, of which 144 UTs fell under biological process, 108 fell under cellular component and 78 fell under molecular function. Also, RT-PCR was utilized to substantiate the elevation in expression of six candidate genes ((MR219SAP8 (JZ532324) and salT (JZ532363), low molecular weight heat shock (JZ532334), phospholipid hydroperoxide glutathione (JZ532371), Acetamidase (JZ532408), Ferredoxin-1 and chloroplastic (JZ532374)) subjected to salinity stress at varying timepoints. © 2015, Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.


Hedayati P.,Institute of Systems Biology INBIOSIS | Monfard H.H.,Institute of Systems Biology INBIOSIS | Hwang D.J.,National Academy of Agricultural Science | Uddin M.I.,Institute of Systems Biology INBIOSIS | And 2 more authors.
Plant OMICS | Year: 2015

The stress-associated protein (SAP) family provides salinity stress tolerance in plants. We performed functional studies of SAP MR219, which is a member of the SAP family that is induced after salinity stresses. Computational analysis of the SAP MR219 cDNA clone that was isolated from rice root and shoot revealed significant homology with the SAP gene families from rice (89% to 36% ) and Arabidopsis (44% to 32%). This clone has a 516-bp coding region encoding a 171 amino acid protein with a predicted molecular mass of 18.31 kDa. In silico analysis demonstrated that the SAP MR219 gene product encoded a cytoplasmic zinc finger protein that might perform its functions via protein-protein interactions aided by its AN1 and A20 zinc finger domains. The SAP MR219 gene was isolated, cloned and introduced into Arabidopsis thaliana under the control of the CaMV35S promoter. Five transgenic Arabidopsis lines were obtained by the floral-dip transformation method using Agrobacterium tumefaciens strain GV3101. The survivability of the transgenic lines under salinity stress was evaluated at 100, 150, 200 and 250 mM NaCl. At 250 mM NaCl, the germination rates of transgenic lines were approximately 50%, whereas the wild-type plants did not grow. Our results indicate that SAP MR219 may play a significant role in the response to salt stress tolerance in plants.

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