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| Isolation and Functional Analysis of TaZTP29 in Wheat |
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Abstract High salt is one of the major constraint that adversely affects wheat yield in China. The zinc transporter (ZTP) belongs to the ZRT, IRT-like protein (ZIP) family, and ZTP is involved in the response to salt stress through regulating induced the unfold protein response pathway, zinc levels to induced the unfold protein response pathway. In this study, the TaZTP29, which is the orthologous gene of AtZTP29, was isolated from wheat (Triticum aestivum) by RT-PCR and the expression pattern was analyzed through qRT-PCR. The subcellular localization of TaZTP29 was also investigated through green fluorescent protein (GFP) method. Meanwhile, the salt resistance of TaZTP29 was confirmed by over-expressing of the TaZTP29 in Arabidopsis thaliana. The sequencing analysis showed that TaZTP29 gene (GenBank accession: KY610283) consisted of 834 bp CDS, 81 bp 5'UTR and 117 bp 3'UTR, and encoded 277 amino acids. The conserved domain analysis demonstrated that TaZTP29 had the typical ZIP domain, which was consisted of 8 transmerabrane domains and the V transmerabrane domain had the completed conserved amino acid histine (His). TaZTP29 was localized to the endoplasmic reticulum with proyien predicted. The subcellular location were validated by means of fusing TaZTP29 with N-terminal GFP and then expressed in wheat protoplast using polyethylene glycol (PEG) transfection method. The green fluorescence was observed in endoplasmic reticulume. The information of conserved domain proved that TaZTP29 belonged to the ZIP protein family. The 16 homologues genes from 16 different species of TaZTP29 were searched and found in NCBI database, and the identity of amino acid between 16 homologous genes and TaZTP29 ranged from 64.03% to 94.5%. The highest identity of amino acid was between Aegilops tauschii (EMT27056.1) and TaZTP29, while the lowest identity of amino acid was between Arabidopsis thaliana (Q940Q3.1) and TaZTP29. Then phylogenetic tree of TaZTP29 was constructed using these 16 genes and TaZTP29. The expression pattern disclosed the expression level of TaZTP29 was increased immediately under ZnCl2 treatment and NaCl stress. However, TaZTP29 in response to heat stress was passive. When wheat suffered heat stress and drought stress, expression of TaZTP29 dropped rapidly. Tissue expression pattern analysis showed that TaZTP29 expressed in multiple tissues. Root accumulated higher TaZTP29 transcription when comparing to the other tissues of ground. The seed germination rate and total root length of wild type was similar with that of TaZTP29 transgenic Arabidopsis lines in normal growth condition. While, the seed germination rate and total root length of TaZTP29 transgenic lines were significantly higher and longer than those of wild- type plants under salt treatments (100 and 150 mmol/L NaCl), respectively. These results indicated that TaZTP29 transgenic lines had higher salt tolerance than wild type, and TaZTP29 could increase the salt tolerance of Arabidopsis thaliana, and could be used as a candidate gene for salt tolerance. This study provides a new foundation for further understanding the molecular mechanism of wheat stress resistance.
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Received: 22 March 2017
Published: 30 October 2017
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