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Isolation and characterization of gene encoding G protein α subunit protein responsive to plant hormones and abiotic stresses in Brassica napus

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Abstract

G protein plays an important role in signal pathways and involved in various signal transduction systems in plant. A full-length cDNA encoding a putative G protein α subunit (Gα), designated as BnGA1, was isolated from Brassica napus. The expression of BnGA1 in different B. napus tissues and developmental stags was analyzed using real-time PCR. The results showed that BnGA1 expressed was high in root, cotyledon and shoot apex. Stage expression pattern analysis revealed that BnGA1 expressed strongly at the 7th day, the bolting stage and fruiting stage. In addition, the expression of BnGA1 was analyzed under different concentrations of four plant hormones. The expression of BnGA1 was significantly induced by the high concentrations of abscisic acid (ABA) and brassinosteroid (BR). The expression of BnGA1 was also induced by low gibberellins acid 3 (GA3) concentrations and higher GA3 concentrations inhibit the expression of BnGA1. However, the expression of BnGA1 did not significantly regulated by exogenous indole-3-acetic acid (IAA). Moreover, the expression of BnGA1 under different abiotic stresses was analyzed at different time points. The BnGA1 was up-regulated in salt and drought stress and down-regulated in heat and cold stress. These expression results suggested that BnGA1 play an important role in plant hormones signal pathways and BnGA1 may be involved in plant defense system against environmental stresses in B. napus.

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Abbreviations

G protein:

Heterotrimeric GTP-binding proteins

Gα:

G proteins α subunit

ORF:

Open reading frame

ABA:

Abscisic acid

GA:

Gibberellins

BR:

Brassinosteroid

IAA:

Indole-3-acetic acid

PEG:

Polyetheleneglycol

RGS:

Regulator of G-protein signaling proteins

GPCR:

G protein-coupled receptor

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Acknowledgments

This work was supported by the National ‘863’ Programme (SN: 2001AA241104) and the 10th ‘five-year’ key task project in crop breeding of Sichuan Province (SN: 200107001).

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Authors and Affiliations

  1. Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610064, Sichuan, People’s Republic of China

    Yong Gao, Tingting Li, Yang Liu, Caixia Ren, Yun Zhao & Maolin Wang

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  1. Yong Gao
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  2. Tingting Li
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Correspondence to Maolin Wang.

Additional information

Yong Gao and Tingting Li contributed equally to this work.

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11033_2010_54_MOESM1_ESM.jpg

Fig. S1 Comparison of the putative amino acid sequences of BnGA1 with some representative Gα proteins. The identical amino acids were showed in white with black background and the conserved amino acids were showed in white with gray background. The specific sites for NTP binding, GTP hydrolysis, guanine recognition, two RGS-box interaction sites (RIS 1-2) and three binding to Gβγ–subunit complex (βγBS 1-3) were highly conserved in the listed Gα-subunits of plants. Putative sites for N-myristoylation (*) and ADP-ribosylation by cholera toxin (↓) were also presented. The aligned Gα subunit sequences were from Arabidopsis thaliana (GPA1, GenBank accession no. AT2G26300), Nicotiana tabacum (NtGA2, GenBank accession no. BAB84093), Pisum sativum (PGA2, GenBank accession no. AAB57826), Triticum aestivum (TaGA1, GenBank accession no. BAC10501) and Oryza sativa (Os05g0333200, GenBank accession no. AY792541)

11033_2010_54_MOESM2_ESM.jpg

Fig. S2 Phylogenetic tree analysis of BnGA1 from plants by MEGA version 3.1. The neighbor-joining method was used to construct the tree. Numbers on nodes indicate the bootstrap values after 1,000 replicates. The amino acids sequences used in phylogenetic tree analysis were from plants including Arabidopsis thaliana (GenBank accession no. AT2G26300), Daucus carota (GenBank accession no. ABK80761), Glycine max (GenBank accession no. X95582), Hordeum vulgare (GenBank accession no. AAF71788), Lotus japonicus (GenBank accession no. X77250), Lupinus luteus (GenBank accession no. X99485), Nicotiana tabacum (GenBank accession no. BAB84093), Oryza sativa (GenBank accession no. AY792541), Phaseolus lunatus (GenBank accession no. AB234091), Pisum sativum (GenBank accession no. AAB57826), Scoparia dulcis (GenBank accession no. EU489474), Solanum tuberosum (GenBank accession no. CAA61105), Sorghum bicolor (GenBank accession no. ABU48662), Spinacia oleracea (GenBank accession no. CAA76186), Triticum aestivum (GenBank accession no. BAC10501) and Vitis vinifera (GenBank accession no. CAO43897)

Table S1 Gene-specific primer sequences used for the quantitative real-time PCR analysis of B. napus BnGA1 genes

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Gao, Y., Li, T., Liu, Y. et al. Isolation and characterization of gene encoding G protein α subunit protein responsive to plant hormones and abiotic stresses in Brassica napus . Mol Biol Rep 37, 3957–3965 (2010). https://doi.org/10.1007/s11033-010-0054-x

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