Cloning of GmDFR gene from Soybean (Glycine max) and Identification of Its Function on Resistance to Iron Deficiency
SHI Zhuo1, LI Hong1, GAO Ming1, GUO Chang-Hong1, GUO Dong-Lin1,*, BI Ying-Dong2,*
1 College of Life Science and Technology/Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province, Harbin Normal University, Harbin 150025, China; 2 Tillage and Cultivation Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China
Abstract Dihydroflavonol-4-reductase (DFR) plays a key catalytic role in anthocyanin synthesis. Anthocyanin can protect tissues from oxidative damage induced by abiotic stress, and can also be used as chelating agents to promote plant iron absorption. In this study, GmDFR gene (GenBank No. MN547961) and its promoter pGmDFR fragment (GenBank No. MW455109) were cloned from soybean (Glycine max). The CDS length of GmDFR gene was 1 020 bp and encoded 339 amino acids. GmDFR had typical PLN02650 and WcaG superfamily domains, belongs to dihydroflavonol 4-reductase and nucleoside diphosphate sugar isomerase family, GmDFR protein was not a transmembrane protein. Phylogenetic analysis showed that soybean GmDFR was most closely related to wild soybean (G. soja) GsDFR, with a similarity of 99.12%. Semi-quantitative RT-PCR detection showed that GmDFR was expressed in soybean tissues and the highest expression level was in roots. The pGmDFR promoter fragment was obtained by cloning, which contained multiple hormone and stress response elements. Under iron deficiency, the anthocyanin content and chlorophyll content of GmDFR transgenic tobacco (Nicotiana tabacum) were significantly higher than that of WT (P<0.05); the contents of malondialdehyde, hydrogen peroxide and superoxide anion were significantly lower than that of WT (P<0.05); and the activities of antioxidant enzymes (SOD, POD and CAT) were significantly higher than that of WT (P<0.05). These results indicated that transgenic GmDFR tobacco had certain ability to resist iron deficiency. This study provides scientific basis for the application of GmDFR gene in soybean resistance to iron deficiency, and also provides new gene resources for soybean molecular breeding.
SHI Zhuo,LI Hong,GAO Ming, et al. Cloning of GmDFR gene from Soybean (Glycine max) and Identification of Its Function on Resistance to Iron Deficiency[J]. 农业生物技术学报, 2023, 31(2): 259-272.
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