Heterologous Overexpression of Soybean GmNF-YB24 Improves the Resistance of Transgenic Tobacco to Drought

doi:10.13560/j.cnki.biotech.bull.1985.2025-0046

Abstract

Abstract:

Objective Nuclear factor (NF-Y) genes are involved in the regulation of drought resistance in plants. The function of resistance to drought and mechanism of resistance to drought of soybean GmNF-YB24 were analyzed, which may provide theoretical basis for the subsequent application of NF-YB genes in the genetic breeding of soybean with high resistance. Method The expression of GmNF-YB24 under salt, drought and cold stresses was detected by RT-qPCR. GmNF-YB24 was cloned and its plant expression vector was constructed to transformed tobacco. The resistance of GmNF-YB24 transgenic tobacco to drought was evaluated. Result Salt, drought and cold stresses induced the expression of GmNF-YB24, and the expression of GmNF-YB24 was the most obvious in response to drought stress. The open reading frame of GmNF-YB24 was 516 bp, and the encoded protein contained 171 amino acid residues. The GmNF-YB24 protein contained a histone-fold motif (HFM). The plant expression vector pRI101-GmNF-YB24 was constructed and transformed into tobacco. Five GmNF-YB24 transgenic tobacco plants (OE1-OE5) were obtained. After drought and rewater treatment, the performance of GmNF-YB24 transgenic tobacco was superior to that of wild-type tobacco. Under drought stress, compared with wild-type tobacco, GmNF-YB24 transgenic tobacco had more proline and soluble carbohydrate, lower relative electrolyte leakage and malondialdehyde content, and antioxidant enzyme activity increased and accumulation of peroxide reduced. The expressions of stress-related genes NtOsmotin and NtERD10B in GmNF-YB24 transgenic tobacco were higher than those in wild-type tobacco. Conclusion Heterologous overexpression of GmNF-YB24 in tobacco improves the resistance of transgenic tobacco to drought.

Key words: soybean, nuclear factor Y, NF-YB, transgenic tobacco, resistance to drought

ZHAI Ying, JI Jun-jie, CHEN Jiong-xin, YU Hai-wei, LI Shan-shan, ZHAO Yan, MA Tian-yi. Heterologous Overexpression of Soybean GmNF-YB24 Improves the Resistance of Transgenic Tobacco to Drought[J]. Biotechnology Bulletin, 2025, 41(8): 137-145.

Figures/Tables 11

Table 1 Primer sequences for RT-qPCR

基因（GenBank登录号）

Gene （GenBank accession no.）

引物序列

Primer sequences （5′‒3′）

GmNF-YB24 （XM003546151）

F-AGACAGGTTCCTTCCGATAGCG

R-TTATGAAGCTGATGAACTCCGACAC

Gmβ-Tubulin （GMU12286）

F-GGAAGGCT TTCTTGCATTGGTA

R-AGTGGCATCCTGGTACTGC

NtActin （AB158612）

F-TTGCTGGTCGTGATCTTACTGATTG

R-CAGTCTCCAACTCTTGCTCATAGTC

NtOsmotin （M29279）

F-CTCCTTGCCTTGGTGACTT

R-ACCGCCTATGGGTGTCG

NtERD10B （AB049336）

F-TCCCATTCGTCAAACCG

R-CCCACCAAGTATGCCAGT

NtP5CS （HM854026）

F-GACACGGACTGATGGAAGATTAG

R-TTCATAGCCTTGCGAGTTAAGC

Fig. 1 Expression of GmNF-YB24 under abiotic stressDifferent letters indicate significant difference (P<0.05). The same below

Fig. 2 Cloning of GmNF-YB24M: DL2000 marker; 1: PCR amplified products of GmNF-YB24

Fig. 3 Nucleotide and protein amino acid sequences of GmNF-YB24Shadow refers to the HFM domain; * refers to the termination codon

Fig. 4 Construction of GmNF-YB24 plant expression vector and A. tumefaciens transformationM: DL2000 marker. 1-2: Double enzyme digestion of pRI101-GmNF-YB24 plasmid. 3‒4: PCR of Agrobacterium tumefaciens

Fig. 5 Identification of GmNF-YB24 transgenic tobaccoA: Genomic DNA PCR detection of GmNF-YB24 transgenic tobacco; B: RT-qPCR detection of GmNF-YB24 transgenic tobacco; M: DL2000 marker for molecular weight; +: pRI101-GmNF-YB24 plasmid; WT: wild-type tobacco; 1-5: GmNF-YB24 transgenic tobacco; OE1-OE5: GmNF-YB24 transgenic tobacco. The same below

Fig. 6 Phenotype of GmNF-YB24 transgenic tobacco under drought and rewater treatmentA: Before stop watering; B: stop watering for 10 d; C: stop watering for 20 d; D: rewater for 3 d

Fig. 7 Proline content, soluble carbohydrate content, relative electrolyte leakage,and malondialdehyde content of GmNF-YB24 transgenic tobacco under drought stress

Fig. 8 SOD activity and POD activity of GmNF-YB24 transgenic tobacco under drought stress

Fig. 9 Leaf DAB staining of GmNF-YB24 transgenic tobacco under drought stress

Fig. 10 Expressions of stress-related genes in GmNF-YB24 transgenic tobacco

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