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Identification and Applification of an Endogenous Reporter Gene Foc4carS in Fusarium oxysporum f. sp. cubense race 4
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Jun PENG1, 2, Fanyun ZENG1, 2, Yanwei WANG1, Yanxiang QI1, Zhaojian DING3, Shaoling WANG1, Yixian XIE1, Xin ZHANG1, *
Chinese Journal of Tropical Crops | 2024, 45(5) : 873 - 885
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Chinese Journal of Tropical Crops | 2024, 45(5): 873-885
Omics & Biotechnology
Identification and Applification of an Endogenous Reporter Gene Foc4carS in Fusarium oxysporum f. sp. cubense race 4
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Jun PENG1, 2, Fanyun ZENG1, 2, Yanwei WANG1, Yanxiang QI1, Zhaojian DING3, Shaoling WANG1, Yixian XIE1, Xin ZHANG1, *
Affiliations
  • 1.Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs / Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Haikou, Hainan 571101, China
  • 2.Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya, Hainan 572000, China
  • 3.Department of Biological Science, Qiongtai Normal University, Haikou, Hainan 571127, China
Published: 2024-05-25 doi: 10.3969/j.issn.1000-2561.2024.05.001
Outline
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Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt (Panama disease), is one of the most devastating diseases of banana (Musa spp.). The Foc race 4 (Foc4) is currently known as a major concern in global banana production. carS is involved in the biosynthesis of Fusarium carotenoid by regulating downstream car structural genes. In this study, Foc4carS (FOIG_05085) was cloned and identified. Foc4carS had a typical RING-finger protein domain. The fusion fragment of Foc4carS was obtained by using Split-marker PCR, and the pUC-fFuCas9-HTBNLS-hph-Foc4carS gene editing vector containing the sequence of sgRNA591 of Foc4carS was constructed, the PEG- mediated protoplast transformation was used to obtain the knockout mutants, the complement mutants and the gene-editing mutants. The biological characteristics and pathogenicity of the knockout mutants and the complement mutants were analyzed. The results showed that the colony diameter, sporulation and pathogenicity of ΔFoc4carS mutants were not significantly different with wild strain Foc4, but the colony color of ΔFoc4carS mutant was dark orange, deletion of the Foc4carS gene affected the biosynthesis of the secondary metabolite carotenoid, and gene-edited ΔFoc4carS (HDR) mutants showed typical dark orange coloration in both regenerative screening plates and subcultured PDA plates. The results indicated that Foc4carS could be used as an endogenous reporter gene to perform gene plasmid CRISPR/Cas9 editing in F. oxysporum f. sp. cubense Foc4.

Fusarium oxysporum f. sp. cubense race 4  /  Foc4carS  /  carotenoid  /  gene knockout  /  CRISPR/Cas9 gene editing system
Jun PENG, Fanyun ZENG, Yanwei WANG, Yanxiang QI, Zhaojian DING, Shaoling WANG, Yixian XIE, Xin ZHANG. Identification and Applification of an Endogenous Reporter Gene Foc4carS in Fusarium oxysporum f. sp. cubense race 4[J]. Chinese Journal of Tropical Crops, 2024 , 45 (5) : 873 -885 . DOI: 10.3969/j.issn.1000-2561.2024.05.001
Year 2024 volume 45 Issue 5
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Article Info
doi: 10.3969/j.issn.1000-2561.2024.05.001
  • Receive Date:2023-09-07
  • Online Date:2026-06-23
  • Published:2024-05-25
Article Data
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History
  • Received:2023-09-07
  • Revised:2023-10-08
Funding
Affiliations
    1.Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs / Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Haikou, Hainan 571101, China
    2.Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya, Hainan 572000, China
    3.Department of Biological Science, Qiongtai Normal University, Haikou, Hainan 571127, China
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表12种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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