Scientia Agricultura Sinica ›› 2018, Vol. 51 ›› Issue (23): 4514-4521.doi: 10.3864/j.issn.0578-1752.2018.23.010

• HORTICULTURE • Previous Articles     Next Articles

Molecular Mechanism of Apple MdWRKY18 and MdWRKY40 Participating in Salt Stress

XU HaiFeng(),YANG GuanXian,ZHANG Jing,ZOU Qi,WANG YiCheng,QU ChangZhi,JIANG ShengHui,WANG Nan,CHEN XueSen   

  1. College of Horticulture Science and Engineering, Shandong Agricultural University/State Key Laboratory of Crop Biology, Tai’an 271018, Shandong
  • Received:2018-05-25 Accepted:2018-07-26 Online:2018-12-01 Published:2018-12-12

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Abstract:

【Objective】 In order to improve the molecular mechanism of salt stress, we studied several aspects of MdWRKY18 and MdWRKY40 in apple WRKY transcription factors, including the protein structure, the expression level and the function in salt stress. 【Method】 We cloned the MdWRKY18 and MdWRKY40 genes in ‘Hongcui No.2’ apple and analysed their protein structure. The expression levels of MdWRKY18 and MdWRKY40 were studied by the qRT-PCR under the salt stress, and their promoter activities were analyzed using the GUS staining. We analyzed the interaction relationship between MdWRKY18 and MdWRKY40 proteins by yeast two-hybrid and verified their function by transgenosis. 【Result】 Analysis of protein structure showed that both MdWRKY18 and MdWRKY40 proteins contained a WRKY, Cx5C and HxH structural domains. The expression levels and promoter activities of MdWRKY18 and MdWRKY40 were induced by the 150 mmol·L -1 NaCl. The yeast two-hybrid experiments showed that MdWRKY18 and MdWRKY40 could respectively interact with itself to form homodimers, and MdWRKY18 could also interact with MdWRKY40 to form heterodimers. When MdWRKY18 and MdWRKY40 was overexpressed respectively in orin callus, they could increase the callus weight under salt stress and promote the expression of MdSOS1 and MdNHX1. When MdWRKY18 and MdWRKY40 were co-overexpressed in orin callus, it could also promote the expression of MdSOS1 and MdNHX1, however, the weight of callus was heavier than the weight of callus overexpressing MdWRKY18 or MdWRKY40. 【Conclusion】 MdWRKY18 and MdWRKY40 were induced by the salt stress, and they could form homodimers or heterodimers, overexpressing MdWRKY18 or MdWRKY40 in orin callus could increase its salt tolerance.

Key words: apple, WRKY transcription factor, salt stress, GUS staining, yeast two-hybrid

Fig. 1

Protein structure analysis of MdWRKY18 and MdWRKY40"

Fig. 2

The expression level of MdWRKY18 and MdWRKY40 treated with 150 mmol·L-1 NaCl"

Fig. 3

The GUS staining analysis of MdWRKY18 and MdWRKY40 promoter treated with 150 mmol·L-1 NaCl"

Fig. 4

Yeast two-hybrid analysis between MdWRKY18 and MdWRKY40"

Fig. 5

The orin callus that overexpressing MdWRKY18 and MdWRKY40 treated with 150 mmol·L-1 NaCl"

Fig. 6

The weight of orin callus that overexpressing MdWRKY18 and MdWRKY40 treated with 150 mmol·L-1 NaCl"

Fig. 7

The expression level analysis of related gene in four types of callus 1：Orin;2：OEWRKY18;3：OEWRKY40;4：OEWRKY18+OEWRKY40"

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