Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Dec;36(4):556-65.
doi: 10.1590/S1415-47572013000400015. Epub 2013 Nov 8.

Introduction of the rd29A:AtDREB2A CA gene into soybean (Glycine max L. Merril) and its molecular characterization in leaves and roots during dehydration

Cibelle Engels  1 , Renata Fuganti-Pagliarini  2 , Silvana Regina Rockenbach Marin  2 , Francismar Corrêa Marcelino-Guimarães  2 , Maria Cristina Neves Oliveira  2 , Norihito Kanamori  3 , Junya Mizoi  4 , Kazuo Nakashima  3 , Kazuko Yamaguchi-Shinozaki  5 , Alexandre Lima Nepomuceno  2
Affiliations

Introduction of the rd29A:AtDREB2A CA gene into soybean (Glycine max L. Merril) and its molecular characterization in leaves and roots during dehydration

Cibelle Engels et al. Genet Mol Biol. 2013 Dec.

Abstract

The loss of soybean yield to Brazilian producers because of a water deficit in the 2011-2012 season was 12.9%. To reduce such losses, molecular biology techniques, including plant transformation, can be used to insert genes of interest into conventional soybean cultivars to produce lines that are more tolerant to drought. The abscisic acid (ABA)-independent Dehydration Responsive Element Binding (DREB) gene family has been used to obtain plants with increased tolerance to abiotic stresses. In the present study, the rd29A:AtDREB2A CA gene from Arabidopsis thaliana was inserted into soybean using biolistics. Seventy-eight genetically modified (GM) soybean lines containing 2-17 copies of the AtDREB2A CA gene were produced. Two GM soybean lines (P1397 and P2193) were analyzed to assess the differential expression of the AtDREB2A CA transgene in leaves and roots submitted to various dehydration treatments. Both GM lines exhibited high expression of the transgene, with the roots of P2193 showing the highest expression levels during water deficit. Physiological parameters examined during water deficit confirmed the induction of stress. This analysis of AtDREB2A CA expression in GM soybean indicated that line P2193 had the greatest stability and highest expression in roots during water deficit-induced stress.

Keywords: Arabidopsis; differential expression; genetically modified organism; water deficit.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Plasmid containing the transgene rd29A:AtDREB2A CA and the map of restriction enzymes used in this work. The enzymes used in Southern blotting were HindIII and EcoRV.
Figure 2
Figure 2
Phylogenetic tree generated from the DREB-coding DNA sequence (CDS) from various plant species. The sequences were obtained from the NCBI website and were aligned and clustered using Clustal X and MEGA 4.0, respectively.
Figure 3
Figure 3
Phylogenetic tree generated from CDS sequences of the soybean DREB gene. The sequences were obtained from the NCBI website and were aligned and clustered using Clustal X and MEGA 4.0, respectively.
Figure 4
Figure 4
Southern blot for generations T0, T1 and T2 of genetically modified P2193 containing the transcription factor AtDREB2A. Lanes 1–8 represent T1 generation plants. BR16 - control (normal) soybean cultivar.
Figure 5
Figure 5
Relative gene expression of the AtDREB2A CA transgene in leaves and roots of the T2 generation of P2193 during stress induced by a water deficit. Transgene expression was normalized to the reference genes GmrRNA18S and Gmβ_actin and then calibrated with the corresponding time points and tissue from control plants (cultivar BR 16) (p < 0.001). Statistical analyses were done using RESTmsc software and all treatments of leaves and roots were significantly higher than the control.
Figure 6
Figure 6
Physiological responses in control plants (cultivar BR 16) and transgenic P2193 plants during stress for 30, 60 and 90 min. (A) Photosynthetic rate, (B) Stomatal conductance, (C) Transpiration rate and (D) Air-leaf temperature. The points are the mean ± SE of three determinations. The non-overlapping points at 30 min were significantly different from each other (p ≤ 0.05, Duncan’s test).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Almogueva C, Prieto-Dapena P, Díaz-Martín J, Espinosa JM, Carranco R, Jordano R. The HaDREB2 transcription factor enhances basal thermotolerance and longevity of seeds through functional interaction with HaHSFA9. BMC Plant Biol. 2009;9:1–12. - PMC - PubMed
    1. Aragão FJL, Sarokin L, Vianna GR, Rech EL. Selection of transgenic meristematic cells utilizing a herbicidal molecule results in the recovery of fertile transgenic soybean [Glycine max (L.) Merril] plants at a high frequency. Theor Appl Genet. 2000;101:1–6.
    1. Bhatnagar-Mathur P, Devi MJ, Reddy DS, Lavanya M, Vadez V, Serraj R, Yamaguchi-Shinozaki K, Sharma KK. Stress-inducible expression of At DREB1A in transgenic peanut (Arachis hypogaea L.) increases transpiration efficiency under water-limiting conditions. Plant Cell Rep. 2007;26:2071–2082. - PubMed
    1. Dalal M, Tayal D, Chinnusamy V, Bansal KC. Abiotic stress and ABA-inducible Group 4 LEA from Brassica napus plays a key role in salt and drought tolerance. J Biotechnol. 2009;17:137–145. - PubMed
    1. Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small amounts of fresh leaf tissue. Phytochem Bull. 1987;19:11–15.

Internet Resources

    1. Associação Brasileira das Indústrias de Óleos Vegetais (ABIO-VE) 2013 May 20; http://www.abiove.com.br.
    1. Companhia Nacional de Abastecimento (CONAB) 2012 May 8; http://www.conab.gov.br.
    1. Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) 2012 May 10; http://www.cnpso.embrapa.br.
    1. United States Department of Agriculture (USDA) 2011 Jul 6; http://www.usda.gov/wps/portal/usda/usdahome.

LinkOut - more resources

  NODES
INTERN 5
twitter 2