Identification and characterization drought tolerance of gene LEA-D11 soybean (glycine max L. Merr) based on PCR-sequencing

Abstract

Drought is one of the most damaging abiotic stress. Different plants response differently to drought stress. Abiotic stresses such as drought induced diverse physicological and molecular responses in plants. These responses include changes in gene expression. One of drought tolerance gene is a gene encoding dehydrin which is belongs to the group II or D-11 LEA protein family. LEA-D11 gene produce dehydrin protein which has a role in stabilization of membrane structures and protection of macromolecules in the presence of drought. The aims of the study was to identify and to characterize the LEA-D11 gene in various soybean varieties. This research used seven varieties of soybean: Tanggamus, Nanti, Seulawah, Tidar (drought tolerant), Wilis and Burangrang (drought moderate) and Detam-1 (drought susceptible). DNA genome of those varieties was isolated using the methods from Doyle & Doyle [1]. DNA amplification was conducted using Polymerase Chain Reaction (PCR) with specific primers designed based on GmLEA-D11 gene sequence database from the NCBI. The DNA targets were sequenced using automatic sequencing machine, ABI 3130xl Genetic Analyzer, in Eijkman Institution. The result of this study showed that the sequences of Gm-LEA-D11 gene possessed by drought tolerance varieties (Tanggamus, Nanti, Seulawah and Tidar) and moderately tolerance (Wilis and Burangrang) were similar. However, the sequence of GmLEA-D11 gene detected in the drought susceptible variety Detam-1 was different from the two groups. Similarity between drought tolerance and moderately tolerance indicate that there is not only LEA-D11 gene responsible to drought tolerance but also others. The primer and sequences GmLEA-D11 gene can be used as molecular marker and capable of differentiating between drought susceptible and drought moderate to drought tolerant.