Cloning and functional analysis of the gene NtUFGT in Nitraria tangutorum

doi:10.11686/cyxb2019334

Abstract

Abstract: Anthocyanidins are important secondary metabolites of flavonoids. Their significant antioxidant effects can assist plants to alleviate oxidative damage caused by various abiotic stressors. Based on transcriptome data, a flavonoid 3-O-glucosyltransferase gene was cloned from the cDNA of Nitraria tangutorum, and was named NtUFGT. The open reading frame of NtUFGT has 1407 bp, and encodes a protein with 468 amino acids. The relative molecular mass of the protein encoded by this gene is predicted to be 51.37 kDa. Multiple sequence alignment analysis showed that the NtUFGT protein belongs to the UDP-glycosyltransferase protein family. Quantitative real-time PCR (qRT-PCR) was carried out to analyze gene expression patterns. NtUFGT had tissue-specific expression patterns. From high to low expression: flower>fruit>stem>leaf>root. Also, it was rapidly up-regulated by polyethylene glycol and abscisic acid treatments. The plasmid of pPZP221:35S:NtUFGT was constructed and introduced into a wild-type (WT) Arabidopsis thaliana by a floral-dip method. qRT-PCR was carried out to confirm that the NtUFGT gene was expressed in three T₃ generations. The growth status and stress-resistant physiological and biochemical indexes of wild-type and transgenic A. thaliana (OE lines) under drought stress were measured. The OE lines showed a stronger phenotype with longer root length, heavier fresh weight and higher chlorophyll content than the WT. OE lines also accumulated more flavonoids and anthocyanin. Consistent with phenotype, OE lines had increased levels of antioxidant enzyme (superoxide dismutase, peroxidase, and catalase) activity, and accumulated more glutathione, proline and less malondialdehyde and H₂O₂. The results of qRT-PCR analysis showed that the expression levels of the stress-related genes (AtCAT1, AtPOD1, AtRD29A) and the proline synthesis gene AtP5CS in OE lines were significantly higher than those in the WT. Our observations indicate that the NtUFGT can facilitate stronger active oxygen scavenging capacity and osmotic adjustment ability by effective promotion of the synthesis of anthocyanidins and flavonoids in transgenic Arabidopsis, thus enhancing the tolerance of plants to drought stress.

Key words: NtUFGT, Nitraria tangutorum, drought stress, anthocyanins, ROS

GAO Zi-qi, WANG Jia, TANG Yu-chen, WANG Ying-chun. Cloning and functional analysis of the gene NtUFGT in Nitraria tangutorum[J]. Acta Prataculturae Sinica, 2020, 29(5): 159-170.

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