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| Effect of W-box Mutation on the Activity of TaGAPC5 Gene Promoter from Wheat (Triticum aestivum) Under Abiotic Stress |
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Abstract In wheat (Triticum aestivum), the cytosolic glyceraldehyde-3-phosphate dehydrogenase gene TaGAPC5 was induced under abiotic stresses. Sequence analysis showed that the TaGAPC5 gene promoter region contained three W-box cis-elements. In this study, PCR-mediated mutagenesis method was performed to introduce -640 site mutagenesis, -640 and -700 both sites mutagenesis, and -640 and -700 and -997 three sites mutagenesis in TaGAPC5 gene promoter region. Then, recombinants of the three mutagenesis sequences with pC0390-GUS vector were constructed, respectively, and transformed into Nicotiana tabacum by Agrobacterium-mediated approach. The transformed tobaccos growth under normal conditions and were treated with 100 nmol/L abscisic acid (ABA) and 20% PEG8000, respectively. Histochemical staining results showed that the three mutations of TaGAPC5 promoter all had promoter activity. The results of GUS enzyme activity in tobacco leaves revealed that under normal growth condition, compared with the TaGAPC5 gene promoter without mutagenesis，-640 alone site mutant didn't affect the activity of the promoter, the activity of mutant at -640 and -700 and -997 three sites was decreased significantly(P＜0.01). What's more, under the 100 nmol/L ABA treatment, -640 site mutant, -640 and -700 both sites mutant, and -640 and -700 and -997 three sites mutant all significantly reduced the activity of the TaGAPC5 gene promoter (P＜0.01); under 20% PEG8000 treatment, the activities of -640 alone site mutant and the -640, -700, -997 mutant were both decreased more significantly (P＜0.01). Taken together, the results proved that -640, -700, -997 three W-box sites in TaGAPC5 gene promoter region had a great role in promoter activity, particularlly in 100 nmol/L ABA、20% PEG8000 conditions, and suggested that the W-box cis-elements of TaGAPC5 gene promoter may participate in the regulation of TaGAPC5 gene expression under abiotic stress. Thus, the results of this study could provide important clues for regulatory mechanism of TaGAPC5 gene under abiotic stress in wheat.
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Received: 03 July 2017
Published: 14 February 2018
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