Journal of Plant Biochemistry and Biotechnology

, Volume 22, Issue 4, pp 441–452 | Cite as

Molecular cloning and sequence comparison of a cDNA encoding α-glucan, water dikinase (GWD) from potato (Solanum tuberosum L.), and analysis of gene expression

ORIGINAL ARTICLE

Abstract

Starch phosphorylation is an important biochemical aspect of plant starch metabolism as it influences the overall structure of the starch granule, and a prerequisite for its degradation. There is a growing interest on the isolation and characterization of α-glucan/glucan-like, water dikinases (GWDs) from plants, particularly agriculturally important crops, because GWD is known to catalyze starch phosphorylation both in leaves and different plant storage organs. In the present study, a 4,789-bp full-length cDNA encoding a GWD isoform was isolated from a commercially important Indian potato cultivar, Kufri Chipsona-1 by RT-PCR approach using tuber RNA. The predicted protein consisted of 1,463 amino acids having N-terminal 77-amino acid transit peptide, and 1,386-amino acid mature protein shorter by one amino acid as compared to the other mature GWDs from potato and tomato. The mature GWD showed 98 % sequence identity with the GWD isolated earlier from the potato cv. Desiree. Variations were found at 25 locations representing mostly non-conservative substitutions. The GWD represents a distinct isoform from potato, as revealed by sequence and phylogenetic analyses. Amino acid composition, segment-wise hydrophobic characters, predicted secondary structures were also analyzed and documented in this report. Broadly, the level of GWD expression as analyzed by semi-quantitative RT-PCR approach was found to be nearly uniform both in the mature tubers and leaves from most of the potato cultivars. By immunodetection technique, a band corresponding to ~155 kDa protein was detected only in the tuber protein extracts. The tuber starch-bound phosphorus content data showed minor variations between the potato cultivars.

Keywords

GWD cDNA cloning GWD expression Indian potato cultivars RT-PCR Sequence analysis Tuber starch-bound phosphorus 

Abbreviations

CBM

Carbohydrate binding module

cDNA

Complementary deoxyribonucleic acid

EMBL

European Molecular Biology Laboratory

GWD

α-glucan water dikinase

MEGA

Molecular Evolutionary Genetics Analysis

NCBI

National Center for Biotechnology Information

PWD

Phosphoglucan water dikinase

RT-PCR

Reverse transcription-polymerase chain reaction

SBD

Starch binding domain

SEX

Starch excess

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Copyright information

© Society for Plant Biochemistry and Biotechnology 2012

Authors and Affiliations

  1. 1.Department of Biotechnology and Environmental SciencesThapar UniversityPatialaIndia

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