Isolation, Characterization, and Expression Profiling of Nucleoside Diphosphate Kinase Gene from Tall Fescue ( Festuca a
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ORIGINAL PAPER
Isolation, Characterization, and Expression Profiling of Nucleoside Diphosphate Kinase Gene from Tall Fescue (Festuca arundinaceous Schreb.) (FaNDPK) under Salt Stress Zeinab Shahabzadeh 1 & Reza Darvishzadeh 1,2
&
Reza Mohammadi 3 & Morad Jafari 1
# Springer Science+Business Media, LLC, part of Springer Nature 2019
Abstract Fifty tall fescue (Festuca arundinacea Schreb.) half-sib families were screened under salinity stress during germination stage, and five tolerant and five sensitive half-sib families were selected. After studying morphological and biochemical characteristics of 10 selected families, the most salt-tolerant family was used for gene expression analysis. Real-time PCR analysis showed that the NDPK expression in plants treated by 200-mM sodium chloride has increased 36.782-fold greater than that in the control ones. The full length of encoding region of NDPK in tall fescue was obtained using the rapid amplification of cDNA ends technique. The homology comparison of DNA sequences, deduced amino acid sequences, and protein sequences were carried out using CLASTALW program in NCBI. The fragment composed of 822 nucleotides (gene’s ORF was 508 nucleotides), and corresponding protein consist 168 amino acids with a predicted molecular mass of 19.4 kD. Blast search for isolated fragment showed that our sequenced fragment is highly similar to UMP/CMP kinase of Oryza sativa japonica (95%). In Blastp, the amino acid sequence of isolated fragment was similar to UMP/CMP kinase of Sorghum bicolor (95%). The results of study confirm that the NDPK is related to UMP/CMP kinase. Keywords Gene expression . NDPK gene . qPCR . Tall fescue . RACE technique
Key message Some tall fescues presented high morphological and biochemical performance under salt stress. Expression of NDPK in salttolerant genotype plants was 36.782-fold more than control ones under salt stress. Full-length of encoding region of NDPK was obtained in tall fescue using RACE. Bioinformatics studies confirmed FaNDPK related to UMP/CMP kinase. * Reza Darvishzadeh [email protected] Zeinab Shahabzadeh [email protected] Reza Mohammadi [email protected] Morad Jafari [email protected] 1
Department of Plant Production and Genetics, Faculty of Agriculture and Natural Resources, Urmia University, Urmia, Iran
2
Department of Agricultural Biotechnology, Institute of Biotechnology, Urmia University, Urmia, Iran
3
Branch for Northwest & West region, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Tabriz, Iran
Introduction Salinity is one of the most important environmental stresses, which limits growth and production of crop plants worldwide. A soil is considered saline if the electrical conductivity of its saturation extract (ECe) is above 4 ds m−1 (US Salinity Laboratory Staff, 1954). Progress in breeding for salinity tolerance is slow due to several factors such as its multigenic behavior, complex mechanism, inadequate screening techniqu
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