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Cloning of Six1d Promoter of Fusarium oxysporum and Its Application in the Construction of Fungal Secretory Expression Vector
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Cunzhi PENG1, Lili CHANG1, Dan WANG1, Chao HUANG1, Bingqiang XU2, *, Zheng TONG1, *
Chinese Journal of Tropical Crops | 2023, 44(6) : 1203 - 1213
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Chinese Journal of Tropical Crops | 2023, 44(6): 1203-1213
• Plant Protection & Bio-safety •
Cloning of Six1d Promoter of Fusarium oxysporum and Its Application in the Construction of Fungal Secretory Expression Vector
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Cunzhi PENG1, Lili CHANG1, Dan WANG1, Chao HUANG1, Bingqiang XU2, *, Zheng TONG1, *
Affiliations
  • 1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
  • 2.Haikou Experimental Station (Institute of Tropical Fruit Tree Research), Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
Published: 2023-06-25 doi: 10.3969/j.issn.1000-2561.2023.06.014
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The fungal secretory expression vector p74HSP-EGFP constructed in a previous study can express the target gene in fungal pathogens and secrete the target protein into plants during fungal infection and colonization, thus verifying the function of the target gene. However, the promoter of toxin A gene (ToxA) driving the target gene in p74HSP-EGFP is not a endogenous promoter of Fusarium oxysporum f.sp. cubense (Foc). For more efficient expression of target genes in Foc, the promoter of secreted in xylem 1d (Six1d), which is highly expressed in Foc, was cloned using the genomic DNA of Foc race 1 (Foc1) as a template, replacing the ToxA promoter in p74HSP-EGFP, and the newly constructed fungal secretory vector was named pSix1d74HSPG. Foc tropical race 4 (Foc TR4) was transformed by p74HSP-EGFP and pSix1d74HSPG. The transformed strains were cultured in potassium salt liquid medium (KK) containing 100 μg/ml hygromycin for 5 days, respectively. The fluorescence intensity and the abundance of EGFP protein secreted outside the cell were detected. The results showed that the expression intensity of EGFP driven by Six1d promoter in pSix1d74HSPG was 1.6-1.7 times of that driven by ToxA promoter in p74HSP-EGFP, which could replace the p74HSP-EGFP vector for more efficient expression and secretion of target proteins in Foc for functional characterization. Our work provides an important experimental tool for the study of the molecular mechanism of resistance and susceptibility to banana fusarium wilt.

banana fusarium wilt  /  endogenous promoter  /  secretory expression vector  /  Six1d protein
Cunzhi PENG, Lili CHANG, Dan WANG, Chao HUANG, Bingqiang XU, Zheng TONG. Cloning of Six1d Promoter of Fusarium oxysporum and Its Application in the Construction of Fungal Secretory Expression Vector[J]. Chinese Journal of Tropical Crops, 2023 , 44 (6) : 1203 -1213 . DOI: 10.3969/j.issn.1000-2561.2023.06.014
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Year 2023 volume 44 Issue 6
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Article Info
doi: 10.3969/j.issn.1000-2561.2023.06.014
  • Receive Date：2022-05-17
  • Online Date：2026-03-05
  • Published：2023-06-25
Article Data
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History
  • Received：2022-05-17
  • Revised：2022-08-09
Funding
Affiliations
    1.Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
    2.Haikou Experimental Station (Institute of Tropical Fruit Tree Research), Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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