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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (9): 1347-1356.doi: 10.3724/SP.J.1006.2018.01347

• RESEARCH PAPERS • Previous Articles     Next Articles

Cloning and Salt Resistance Function Identification of GmHDL57 Gene from Glycine max

Dan-Xia KE(),Kun-Peng PENG,Meng-Ke ZHANG,Yan JIA,Jing-Jing WANG   

  1. College of Life Sciences / Institute for Conservation and Utilization of Agro-bioresources in Dabie Mountains, Xinyang Normal University, Xinyang 464000, Henan, China
  • Received:2017-11-03 Accepted:2018-06-12 Online:2018-09-10 Published:2018-06-30
  • Contact: Dan-Xia KE E-mail:kdx_029@163.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31400213);Science and Technology Research Projects of Henan Province(182102110448);Funding Scheme for Young Core Teachers of Xinyang Normal University(2015);Nanhu Scholars Program for Young Scholars of Xinyang Normal University, and the Scientific Research Foundation of Graduate School of Xinyang Normal University

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Abstract:

The HD-Zip I class transcription factor plays an important role in plant resistance to abiotic stresses. An HD-Zip I class gene GmHDL57 (Glycine max homeodomain-leucine zipper protein 57) was cloned from soybean in this study. Sequence analysis showed that GmHDL57 gene contained a 1038 bp ORF, encoding 345 amino acids, and featured with HD-Zip family proteins’ typical conserved domain. GmHDL57 was expressed in different organs of soybean plants and the highest expression occurred in flowers. The effects of abiotic stresses (abscisic acid, NaCl, PEG, and cold) on GmHDL57 gene expression in soybean seedling stage were analyzed by real-time quantitative PCR. The expression level of GmHDL57 gene was obviously increased under high salinity stress and less affected by ABA and drought stress but decreased by cold stress. The expression level of GmHDL57 gene in roots was significantly higher than that in stems and leaves before and after salt stress, and reached the peak at 48 h, then decreased slowly at 72 h and 96 h after salt stress. The overexpression vector of GmHDL57 was constructed and transformed into Agrobacterium tumefaciens strain EHA105 to obtain the stable transgenic Lotus japonicus plants. After being treated with 200 mmol L -1 NaCl for 14 d, the shoot height, root length, chlorophyll content, root activity as well as K + and Ca 2+ content increased significantly in transgenic plants compared with the wild type. The malondialdehyde content, relative membrane permeability and Na +content were obviously reduced in transgenic plants compared with the wild type. It is hypothesized that the GmHDL57 gene participates in the abiotic stress response of soybean, and over-expression of GmHDL57 gene could enhance resistance to saline in Lotus japonicus.

Key words: Glycine max, GmHDL57 gene, abiotic stress, salt resistance

Fig. 1

Predicted spatial structure of GmHDL57"

Fig. 2

Conserved amino acid sequence alignment analysis of GmHDL57 and its homologous proteins in some other plantsThe sequence marked with solid line demonstrates the homeobox domain, and the sequence marked with dotted line demonstrates the homeobox associated leucine zipper domain."

Fig. 3

Phylogenetic tree of GmHDL57 and its homologsThe scale represents genetic similarity, indicating the proximity relationships among species."

Fig. 4

Relative expression levels of GmHDL57 in different tissuesAbscissa represented young leaf, flower, 1 cm pod, pod shell at 10 d, pod shell at 14 d, seed at 10 d, seed at 14 d, seed at 21 d, seed at 25 d, seed at 28 d, seed at 3 d, seed at 42 d, root and nodule. DAF means days after flower."

Fig. 5

Expression of GmHDL57 treated with various abiotic stresses"

Fig. 6

Expression of GmHDL57 gene in different tissues of soybean under salt stress"

Fig. 7

Positive plants detection and salt stress phenotype analysis of transgenic Lotus japonicusA: detection of GUS gene expression in plants by PCR; B: detection of GmHDL57 gene expression in plants by RT-PCR; C: growth state of Lotus japonicus in different salt concentration treatment for 14 d. Lj 9-5, Lj 1-1: transgenic lines. M: DL2000 DNA marker."

Fig. 8

Changes of shoot height (A) and root length (B) of transgenic Lotus japonicus under salt stressLj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05) and ** means extremely significant difference (P<0.01)."

Fig. 9

Physiological characteristics of transgenic Lotus japonicus under salt stressA: MDA content; B: relative membrane permeability; C: chlorophyll content; D: root activity. Lj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05), ** means extremely significant difference (P<0.01)."

Fig. 10

Cation content in transgenic Lotus japonicus under salt stressA: Na+ content in leaves; B: Na + content in roots; C: K + content in leaves; D: K + content in roots; E: Ca2+ content in leaves; F: Ca2+ content in roots. Lj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05) and ** means extremely significant difference (P<0.01)."

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