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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (4): 503-512.doi: 10.3724/SP.J.1006.2020.94082

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Pathogenesis-related protein gene SfPR1a from Salsola ferganica enhances the resistances to drought, salt and leaf spot disease in transgenic tobacco

HENG You-Qiang,YOU Xi-Long,WANG Yan()   

  1. Xinjiang Key Laboratory of Biological resources and Genetic Engineering / College of Life Science and Technology, Xinjiang University, Urumqi 830046, Xinjiang, China
  • Received:2019-06-04 Accepted:2019-12-26 Online:2020-04-12 Published:2020-01-15
  • Contact: Yan WANG E-mail:wangyanxju@126.com
  • Supported by:
    This study was supported by the Xinjiang Key Laboratory of Biological Resources Genetic Engineering(2017D04026);the National Natural Science Foundation of China(31860061)

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Abstract:

In order to investigate whether SfPR1a, a pathogenesis-related protein gene from an annual halophytic species Salsola ferganica Drob., was involved in the response to plant defense, qRT-PCR was employed to detect its expression patterns under abscisic acid (ABA), jasmonic acid (JAs), ethylene synthesis direct precursor (ACC), and NaCl treatments. We also identified resistances to salt, drought and Pseudomonas syringae tomato (PstDC3000) of transgenic tobacco were identified. The expression of SfPR1a gene in roots was significantly higher than that in shoots, and positively induced by ABA, JAs, ACC, and NaCl treatments. The malondialdehyde (MDA) content of transgenic tobacco was significantly lower than that of wild-type tobacco, showing a strong resistance to drought. The ectopic expression of SfPR1a gene improved plant growth under salt stress. After infection of P. syringae, transgenic tobacco leaves showed serious necrosis reaction, but the overall resistance phenotype of the plants was significantly better than that of WT. Subcellular localization analysis showed SfPR1a was localized in the plant cell apoplastic space. The above results indicated that the SfPR1a gene is involved in plant resistance to abiotic and biotic stresses.

Key words: pathogenesis-related protein gene SfPR1a, expression pattern, transgenic tobacco, resistance function, subcellular localization

Fig. 1

Alignment of the amino acid sequences of SfPR1a with other PR1 proteins GenBank accession numbers of different species: Oryza sativa indica Group OsPR1 (AJR16763.1), Glycine max GmPR1 (XP003545775.1), Hordeum vulgare HvPR1 (CAA79703.1), Hevea brasiliensis HbPR1 (ALS87256.1), and Salsola ferganica SfPR1a (AFR90191.1)."

Fig. 2

Expression of SfPR1a gene in different tissues and stress conditions The relative expression of SfPR1a gene in aerial part and root (A) and under the conditions of ABA (B), ACC (C), JAs (D), and NaCl (E), respectively. Three biological replicates were set for each experiment. Bars labelled with different letters are significantly different at P < 0.05."

Fig. 3

Expression of SfPR1a gene in the transgenic tobacco plants Bars labelled with different letters are significantly different at P < 0.05."

Fig. 4

Drought tolerance of transgenic SfPR1a tobacco A: phenotype of tobacco under drought stress treatment for two weeks; B: MDA content of tobacco under for two weeks drought treatment; C: phenotype of tobaccos under drought stress for four weeks and then re-water for one week. D: survival rate (D) of tobaccos under drought stress for four weeks and then re-water for one week. Bars labelled with different letters are significantly different at P < 0.05."

Fig. 5

Salt tolerance of transgenic SfPR1a tobaccos The phynotypic growth of tobaccos after germination (A) and relative growth in 200 mmol L-1 NaCl medium (B); C: tolerance of tobacco at the adult stage under 250 mmol L-1 NaCl for 2 weeks. Bars labelled with different letters are significantly different at P < 0.05."

Fig. 6

Resistance to leaf spot disease of transgenic SfPR1a tobacco A: plaque area after inoculation of Pseudomonas syringae for 10 d; B: tobacco leaves inoculated with Pseudomonas syringae for 45 d; C: plaque area after inoculation of Pseudomonas syringae for 45 d; D: growth phenotype of tobaccos after 60 d of Pseudomonas syringae inoculation. Bars labelled with different letters are significantly different at P < 0.05."

Fig. 7

Subcellular localization of SfPR1a A: control empty plasmid (GFP) viewed under fluorescence filter; B: SfPR1a-mGFP transformed cells viewed under fluorescence filter (B) to show the location of GFP protein; C: SfPR1a-mGFP transformed cells shape under bright field; D: merge of B and C. Bar = 50 µm."

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