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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (12): 1802-1809.doi: 10.3724/SP.J.1006.2015.01802

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Expression Analysis of Tolerance to Aluminum-toxicity Candidate Gene GmSTOP1 in Soybean

CONG Ya-Hui,WANG Ting-Ting,LIU Ju-Ge,WANG Ning,GAO Meng-Meng,LI Yan*,GAI Jun-Yi*   

  1. National Key Laboratory of Crop Genetics and Germplasm Enhancement / National Center for Soybean Improvement / Key Laboratory for Biology and Genetic Improvement of Soybean (General), Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2015-04-01 Revised:2015-07-20 Online:2015-12-12 Published:2015-08-28
  • Contact: 李艳, E-mail: yanli1@njau.edu.cn; 盖钧镒, E-mail: sir@njau.edu.cn E-mail:cyahui616@163.com
  • Supported by:

    This research was supported by the National Natural Science Foundation of China (31371645), the National Key Basic Research Project (2011CB1093), Program for Changjiang Scholars and Innovative Research Team in University of Ministry of Education of China (PCSIRT13073), Program for New Century Excellent Talents in University of Ministry of Education of China (NCET-12-0891), Program for Soybean Biology and Genetic Breeding Innovative Research Team of Ministry of Agriculture of China, and Program for High-level Innovative and Entrepreneurial Talents in Jiangsu Province.

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Abstract:

Aluminum toxicity is one of the major factors that limits the growth and production of crops in acid soils. AtSTOP1 transcription factors can regulate the expression of genes related to aluminum-toxicity tolerance mechanisms, which plays an important role in aluminum-toxicity tolerance in Arabidopsis. To study the expression features of the STOP1-like gene in soybean, we cloned a STOP1 gene located on chromosome 16 from the aluminum-toxicity tolerant soybean cultivar (Kefeng-1) using RT-PCR, and designated as GmSTOP1. The length of GmSTOP1 coding DNA sequence was 1566 bp, which encoded 521 amino acid residues. Diverse cis-acting promoter elements involved in hormone, heat and stress responses were discovered in the 1500 bp upstream region of GmSTOP1, such as ABRE, HSE, TC-rich repeats, and other elements. Protein structure prediction showed that it did not have any signal-peptide or transmembrane region, but contained four conservative Cys-2-His-2 zinc-finger domains. Phylogenetic analysis demonstrated that GmSTOP1 was similar to the putative STOP1-like protein from Phaseolus vulgaris. Results of subcellular localization showed that GmSTOP1 protein is located in the cell nucleus. The transcripts of GmSTOP1 were detected in all organs tested including root, shoot apical meristem, stem, leaf, flower, pod and seed, with the highest level in seed. GmSTOP1 was up-regulated in soybean roots by 25 μmol L–1 AlCl3 treatment, and reached the highest relative expression level at 24 hours, which was about 9.2 times of the level in control (0 μmol L–1 AlCl3). In addition, Real-time PCR analysis showed that the expression of GmSTOP1 in soybean leaf and root was also up-regulated by ABA, NaCl, and PEG, respectively. These results indicated that GmSTOP1 might participate in soybean response to abiotic stresses including aluminum-toxicity, high salinity and osmosis stress, which provides the basis for further studying the functions of GmSTOP1.

Key words: Acid soil, Aluminum toxicity, Soybean, STOP1, Subcellular location, Real-time PCR

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